The development of British pharmacopoeia monographs for idoxuridine and idoxuridine eye drops using high-pressure liquid chromatography for assay and for controlling related impurities. The monograph published in the 1973 edition of the British Pharmacopoeia (BP) for idoxuridine required revision because it contained a non-specific assay and no tests for related impurities. It was also necessary to prepare a new monograph for idoxuridine eye drops. The Japanese Pharmacopoeia contains a thin-layer chromatographic test for impurities but this was not considered ideal. Improved thin-layer chromatographic tests were sought but when this was unsuccessful, methods using high-pressure liquid chromatography were examined. A system using reversed-phase chromatography was selected for inclusion in BP Addendum 1977 since it provided a specific assay method and limit test for related impurities which could be applied to both the drug substance and the eye drops.


Gas chromatographic determination of nalidixic acid in tablets. A gas chromatographic method has been established for the quantitative analysis of nalidixic acid. The method is based on the derivatization of nalidixic acid with diazomethane, and 5-alpha-cholestane is used as an internal standard. The sample is chromatographed on a glass column packed with 1% OV-1 on Chromosorb W. Quantitation is achieved by measuring peak-height ratios. The improved simplicity, specificity and accuracy of the method has been demonstrated for the quantitation of nalidixic acid in tablets.


Rapid micro-method for the measurement of phenobarbitone, primidone and phenytoin in blood plasma or serum by gas-liquid chromatography. A rapid gas-liquid chromatographic method has been developed for the analysis of phenobarbitone, primidone and phenytoin in small (50 microliter) volumes of either blood plasma or serum. Neither solvent transfer nor evaporation are required in the extraction, which takes less than 3 min to complete, and a quantitative analysis may be performed, in duplicate, within 20 min. Sources of interference in the assay are minimal, and prior treatment of the column with gamma-glycidoxypropyltrimethoxysilane facilitates the measurement of as little as 10 ng of underivatized drug "on-column" using a flame-ionisation detector. The method has proved valuable when used for detection and measurement of these three compounds at concentrations of 2 mg/1 or greater.


Quantitative determination of 1,3-bis(tetrahydro-2- furanyl)-5-fluoro-2,4-pyrimidinedione and its metabolites in plasma by high-pressure liquid chromatography and gas chromatography-mass fragmentography. 1,3-Bis(tetrahydro-2-furanyl)-5-fluoro-2,4-pyrimidinedione has been developed clinically as an antitumor agent. A high-pressure liquid chromatographic method was developed with which it could be measured in plasma with a sensitivity of 0.050 microgram/ml. Two of its metabolites, 1-(tetrahydro-2-furanyl)-5-fluoro-2,4-pyrimidinedione and 3-(tetrahydro-2-furanyl)-5-fluoro-2,4-pyrimidinedione, could be determined at the same time with a sensitivity of 0.025 microgram/ml. A gas chromatographic-mass fragmentographic method was developed for the specific determination of the third metabolite, 5-fluoro-2,4-pyrimidinedione, as its silylated derivative with a sensitivity of 0.001 microgram/ml. The precision and sensitivity of the assay appear to be satisfactory for determination of the plasma level of the drug.


High-pressure liquid chromatography of amino acids, peptides and proteins. V. Separation of thyroidal iodo-amino acids by hydrophilic ion-paired reversed-phase high-performance liquid chromatography. The separation of thyroidal iodoamino acids has been carried out by high-performance liquid chromatography in phase systems consisting of chemically bonded C18-hydrophobic supports as the stationary phase and water-organic solvent mixtures containing phosphoric acid or other ion-pairing reagents as the mobile phase. Under conditions of hydrophilic ion-pair formation, excellent resolution of the iodoamino acids is observed. This method permits the rapid separation and, hence, analysis of mixtures containing thyroxine, 3,3',5-triiodothyronine and 3,3',5'-triiodothyronine and related compounds in ca. 30 min with sensitivity, using a UV monitor at 210 nm, at the 1-10-pmole level.


Ion-pair liquid chromatography of amitriptyline and metabolites in plasma. A method is described for the determination of amitriptyline and nortriptyline in the plasma of patients during treatment. The method is based on liquid chromatography of the amines as ion-pairs with perchlorate. The separation column is packed with silica gel, and a mixture of diisopropyl ether, dichloromethane, methanol and water containing the counter ion is used as eluent. High efficacy and stability of the system is achieved. The separation of metabolites is demonstrated. The cis- and trans-10-hydroxy isomers of amitriptyline and nortriptyline are easily resolved with this chromatographic system.


Immunoglobulin M antibody response against Mycoplasma pneumoniae lipid antigen in patients with acute pancreatitis. Serial serum samples from patients with acute pancreatitis showed a significant increase in antibodies against methanol-chloroform-extracted lipid antigen from Mycoplasma pneumoniae when tested by complement fixation. The antibodies did not react with antigens prepared from other human mycoplasmas or from pancreatic tissue by lipid extraction. The antibodies were predominantly immunoglobulin M (IgM). No correlation with cold agglutinins or cardiolipid complement-fixing antibodies was found. The IgM antibody response seemed to be prolonged: after 3 to 4 weeks the antibodies were still in many cases exclusively IgM. Similar IgM responses were also found in certain cases of acute meningoencephalitis. We postulate that during the disease antigenic components identical or very similar to major determinants in the M. pneumoniae lipid antigen are revealed and elicit the IgM antibody response. Their resemblance to natural antibodies and their possible biological role is discussed.


Determination of human immunoglobulin M rheumatoid factor by a solid-phase radioimmunoassay which uses human immunoglobulin G in antigen-antibody complexes. A solid-phase radioimmunoassay for the rapid determination of human immunoglobulin M (IgM) rheumatoid factor (RF) has been developed. Preparation of the solid phase for the assay involved the formation of complexes between respiratory syncytial virus-specific human IgG antibodies and virus antigen on the surface of polystyrene balls. Binding of serum RF to IgG in the immune complex was subsequently detected by 125I-labeled mu-chain-specific antibodies to human IgM. The amount of radioactive indicator antibody bound was converted to units of RF by comparison to the standard curve for an RF reference-serum pool. This assay should prove useful in studies of the physiological role of RF, since it can effectively measure low levels of circulating RF.


Human infections caused by thiamine- or menadione-requiring Staphylococcus aureus. Stable dwarf forms of Staphylococcus aureus have been identified in clinical specimens as the sole or predominant isolate in eight cases. These organisms have been shown to be menadione or thiamine dependent, i.e., cultivation in the presence of one of these agents has permitted growth of colonies which appear typical of S. aureus. In vitro resistance to aminoglycosides was overcome by cultivation in the presence of menadione or thiamine. Menadione- or thiamine-requiring S. aureus can be considered as causative agents in severe human infections. Special care must be taken if they are to be identified in pathological specimens. Their antibiotic sensitivity testing should be done comparatively on supplemented and nonsupplemented media.


Abortion in mice associated with Pasteurella pneumotropica. Pasteurella pneumotropica was isolated from the uteri, fetuses, lungs, and spleens of aborting Swiss Carworth mice. Male mice in the colony carried P. pneumotropica in pharynges, testes, and seminal vesicles. Normal pregnant and nongravid females carried P. pneumotropica in the eye of 1 and in the uteri of 4 of 11. Pregnant mice from another colony did not abort when injected with P. pneumotropica. Necrotizing and suppurative metritis was found among aborting females with P. pneumotropica infections. Occurrence of malignant lymphoma and mammary adenocarcinoma among animals in this colony likely resulted in immunosuppression which could have predisposed animals to the diseases seen.


Mycoplasmas in human pyelonephritis: demonstration of antibodies in serum and urine. This study was carried out to elucidate by serological examination the etiological significance of mycoplasmas isolated from the upper urinary tract of patients with pyelonephritis. The occurrence of antibodies in patients with acute pyelonephritis, chronic pyelonephritis with or without exacerbation, or noninfectious urinary tract disease was compared by the indirect hemagglutination method. Antibody response was demonstrated significantly more often in patients yielding growth of Mycoplasma hominis from the upper urinary tract than in patients not yielding growth. Antibodies against M. hominis were demonstrated in ureteric and bladder urine from three patients with acute pyelonephritis and from one patient with exacerbation of chronic pyelonephritis. M. hominis was isolated from the upper urinary tract of all four patients. Urine antibodies could not be demonstrated in any other cases. Thus, it seems highly possible that M. hominis may play a role in pyelonephritis of humans. The investigations did not disclose a similar role for Ureaplasma urealyticum.


Possible use of frequency-pulse-modulated electron capture gas-liquid chromatography to identify septic and aseptic causes of pleural effusions. Frequency-pulse-modulated electron capture gas-liquid chromatography was used in conjunction with appropriate derivatization procedures to obtain chromatograms from extracts of pleural effusions. These chromatograms were used to rapidly classify the various types of pleural effusions. With this method we have been able to distinguish among a limited number of effusions caused by congestive heart failure, Mycobacterium tuberculosis, and some other types of bacterial empyemas and pleural effusions.


Evaluation of twenty-three blood culture media. Several investigators have evaluated clinically a variety of commercially available blood culture media. No agreement has been reached as to which of these media is optimal for detection of bacteremia. The purpose of this study was to determine the rate of recovery of microorganisms from various blood culture media. A total of 23 blood culture media were inoculated with 7 to 15 microorganisms per bottle in the presence or absence of an erythrocyte-serum mixture. The results demonstrated that blood culture media differed in their ability to support the growth of microorganisms. At 4 days after inoculation, only 10 of the 23 blood culture media supported the growth of 91% (10 of the 11) or more of the test microorganisms. The recovery rate of microorganisms depended not only upon the type of medium but also upon the manufacturer of the type of blood culture medium. The addition of an erythrocyte-serum mixture to the blood culture media did not influence the difference in the recovery rate of microorganisms among media and the same type of medium prepared by different manufacturers. The majority (15 of the 23) of the blood culture media supplemented with the erythrocyte-serum mixture failed to support the growth of 91% or more of the test microorganisms at 4 days after inoculation. These results have demonstrated that blood culture media need to be improved. Better quality control measures should also be implemented to evaluate commercial blood culture media.


Microtitration agglutination for detection of Treponema hyodysenteriae antibody. A microtitration agglutination test for the detection of Treponema hyodysenteriae antibody in swine and rabbit sera is described. The following methods provided the best test results: antigen produced from the spirochete after a culturing period of 36 to 44 h at 38 degrees C, washed antigen inactivated with 0.01% Merthiolate at 4 degrees C for 24 to 36 h, sera heated at 56 degrees C for 30 min, a diluent of phosphate-buffered saline (0.01 M, pH 7.2), and test results read macroscopically after 18 to 24 h of incubation at 38 degrees C. The test enabled detection of antibody against pathogenic T. hyodysenteriae with a high level of consistency and sensitivity. Sera against nonpathogenic T. hyodysenteriae produced low agglutinating titers (less than or equal to 1:8) when reacted against antigen from pathogenic isolates. Inactivated antigen remained stable for 7 to 10 days. Specificity of the reaction in the agglutination test was shown by absorption studies.


Comparison of three blood culture media with tryptic soy broth. Four consecutive studies comparing commercially prepared bottles that contained brain heart infusion, brucella (vented and unvented), and Trypticase soy broths with Tryptic soy broth failed to demonstrate significant differences in rates of isolation of organisms from blood. Statistically significant differences between bottles were limited to the average times required to detect growth of viridans streptococci in one study and of all organisms in another.


Dental care and spread of hepatitis B virus infection. Sera from 576 healthy adults were tested for the hepatitis B surface antigen (HBsAg) and antibody (anti-HBs) to evaluate the role of routine dental care as a factor in the spread of hepatitis B virus (HBV) infection. Serological evidence of prior HBV infection, manifested by acquisition of anti-HBs, was detected in 97 (16.8%) individuals, and 6 (1.0%) were identified to be asymptomatic HBsAg carriers. The anticipated correlations of HBsAg and anti-HBs with age, country of birth, and socioeconomic status were observed in the study population. However, prevalences of both HBsAg and anti-HBs were inversely related to the lifetime total of dental care visits. These findings indicated that, in a region in which the HBsAg carrier state and hepatitis B are prevalent, routine dental care is not identified as an important factor in the spread of HBV infection. While the results do not exclude the obvious possibility that cross-infections with HBV may occur during dental care in specific situations, they indicate that this mode of infection is exceptional.


Carbon substrate utilization studies of some cultures of Alcaligenes denitrificans, Alcaligenes faecalis, and Alcaligenes odorans isolated from clinical specimens. One hundred and sixty-two cultures of Alcaligenes species (A. denitrificans, A. faecalis, and A. odorans) of clinical origin were characterized by routine diagnostic and carbon substrate utilization techniques. The microorganisms were tested for their ability to utilize a total of 188 substrates. Substrate utilization was assayed by (i) growth stimulation and (ii) substrate alkalinization. The A. denitrificans and A. odorans cultures had unique substrate utilization profiles for each species. The A. faecalis isolates were redefined by colonial morphology into two biotypes: (i) biotype I, morphologically and biochemically similar to the A. denitrificans cultures and (ii) biotype II, morphologically similar to the A. odorans cultures.


Search for Acinetobacter calcoaceticus subsp. anitratus: enrichment of fecal samples. None of 50 healthy individuals and only 2 of 50 hospitalized patients (with Acinetobacter anitratus elsewhere) yielded A. anitratus on enrichment from fecal samples.


Double-label fluorescence immunoassay of bacteria. Fluorescence of sensitized bacterial suspensions, reactive with either fluorescein-labeled or rhodamine-labeled antiglobulins, could be quantitatively distinguished in dual-labeled preparations by fluorescence immunoassay.


Renal net glucose release in vivo and its contribution to blood glucose in rats. This study describes the contribution of de novo glucose synthesis by the kidney to blood glucose homeostasis in rats. The net glucose release by the kidney in vivo was measured by an isotope-dilution method, which calculated the extent of dilution of injected [(14)C]glucose by glucose newly synthesized in the kidney. The extent of dilution was determined from the difference between the decrease of the actual blood glucose concentration and that of the radioactivity of [(14)C]glucose, after injecting [(14)C]glucose into functionally hepatectomized rats. The results indicate that the net glucose release by the kidney in vivo in normal fed rats was 0.75+/-0.13 mg/dl per min, and that its contribution to blood glucose was 25.9+/-5.0%. When unilateral nephrectomy was performed, under the same conditions, renal net glucose release was one-half of that in rats with two intact kidneys, which indicates the quantitative accuracy of the isotope-dilution method employed in this study. In rats starved for 24 h, the renal net glucose release increased to 0.99+/-0.08 mg/dl per min. Diabetic rats showed a remarkably higher renal net glucose of 2.28+/-0.33 mg/dl per min, which was 360% of the normal level. Treatment of diabetic rats with insulin, restored the renal net glucose release to the normal level. In acidotic rats, renal net glucose release was as great as 1.03+/-0.15 mg/dl per min, which suggests that the acid-base balance participates in control of renal glucose output. Measurements every 6 h throughout the day showed that glucose was supplied from the kidney at a constant rate without any circadian rhythm. These data suggest that renal gluconeogenesis is of physiological importance in the maintenance of homeostasis of blood glucose.


Role of plasma vasopressin in impaired water excretion of glucocorticoid deficiency. In the present study, the effect of selective glucocorticoid deficiency on renal water excretion was investigated in conscious, trained, adrenalectomized dogs. The animals were studied before and after a water load while on replacement therapy of desoxycorticosterone acetate, 5 mg/day, and dexamethasone, 0.8 mg/day (group I), and while off dexamethasone for 5-9 days (group II). Before the water load the weight, inulin space, cardiac output, blood pressure, glomerular filtration rate, renal blood flow, plasma osmolality, and plasma antidiuretic hormone measured by radioimmunoassay were similar in both groups I and II. However, after a 40 ml/kg water load a marked impairment in renal water excretion in the glucocorticoid deficient dogs became apparent. Maximal free water clearance was -0.046+/-0.16 vs. 6.51+/-0.72 ml/min (P < 0.001) and minimal urinary osmolality was 425+/-56 vs. 82+/-3.5 mosmol/kg H(2)O (P < 0.001) in group II as compared to group I. Plasma antidiuretic hormone was maximally suppressed during the water load in group I to 0.34+/-0.08 pg/ml but remained elevated at 9.18+/-1.79 pg/ml (P < 0.005) in group II. This nonsuppressibility of plasma antidiuretic hormone during water loading in group II was associated with a significant tachycardia of 145+/-6 vs. 87+/-6 beats/min (P < 0.001) in group I and a significantly lower stroke volume of 27+/-0 vs. 59+/-0.5 ml/beat (P < 0.001). In conclusion, our results implicate a persistent secretion of antidiuretic hormone as an important factor in the impaired water excretion of glucocorticoid deficiency. A deleterious effect of glucocorticoid deficiency on cardiac function was observed and this hemodynamic alteration could be involved in initiating a nonosmolar, baroreceptor-mediated release of vasopressin.


Cold-insoluble globulin (fibronectin) in connective tissues of adult human lung and in trophoblast basement membrane. Cold-insoluble globulin (CIG), which is immunochemically indistinguishable from the fibroblast surface protein known as large external transformation-sensitive glycoprotein and fibronectin, was detected immunologically in connective tissue fractions from adult human lung. The fractions tested were (a) intact parenchyma, (b) acidic structural glycoproteins (ASG) extracted from lung parenchyma with 0.3 M acetic acid, and (c) isolated alveolar basement membrane (ABM). For comparison with ABM, preparations of human glomerular basement membrane and human trophoblast basement membrane (TBM) were tested. CIG was not detected in glomerular basement membrane but was present in large amounts in TBM. The CIG antigen could be solubilized from the parenchyma and from ABM by collagenase digestion which indicates that CIG occurs in lung connective tissue in association with collagen. Fibrinogen antigenic determinants were present in the ASG fraction, but the question of whether CIG and fibrin(ogen) are associated in lung connective tissue requires further study. When CIG was quantified by electroimmunoassay, intact lung parenchyma contained  approximately equal to 0.4% CIG, ASG contained 3-4.5% CIG, ABM contained 0.1-0.9% CIG and TBM contained 1.5%-7.2% Cg. the evidence suggests that CIG is a chemical constituent of lung connective tissue matrix where it may influence the function of alveoli.


Prostaglandin suppression of mitogen-stimulated lymphocytes in vitro. Changes with mitogen dose and preincubation. In this study we further characterize the properties of the prostaglandin-producing suppressor cell. Overnight preincubation of peripheral blood mononuclear cells results in an increased response of the cells to phytohemagglutinin or Concanavalin A compared to the response of fresh cells. This increase in mitogen response with preincubation was similar in magnitude to the increase in mitogen response of fresh cells after the addition of indomethacin. The two manipulations were not additive; that is, after preincubation, indomethacin caused much less enhancement of mitogen stimulation of peripheral blood mononuclear cells (100 +/- 12% increase before preincubation vs. 12 +/- 6% after preincubation; mean+/-SEM, P < 0.001). Preincubated cells also lose sensitivity to inhibition by exogenous prostaglandin E(2). It requires the addition of 100- to > 1,000-fold more exogenous PGE(2) to produce comparable inhibition of phytohemagglutinin-stimulated preincubated cells than is required for inhibition of phytohemagglutinin-stimulated fresh cells. The enhancing effect of indomethacin increases with decreasing doses of phytohemagglutinin. Indomethacin causes a 1,059+/-134% increase in [(3)H]thymidine incorporation at the lowest dose of phytohemagglutinin (0.2 mug/ml), and a 4+/-3% increase at the highest dose (20 mug/ml). This increase in response to indomethacin with a lower dose of phytohemagglutinin is due to increased sensitivity to inhibition by PGE(2) at lower mitogen doses. The prostaglandin-producing suppressor cell assay and the short-lived suppressor cell assay measure over-lapping phenomena. The increased suppressive effect of the prostaglandin-producing suppressor at suboptimal mitogen dose must be taken into account in the interpretation of any study where the response to a range of mitogen doses is studied.


Role of large arteries in regulation of cerebral blood flow in dogs. Previous studies have demonstrated a significant pressure gradient from carotid artery to pial or middle cerebral arteries. This pressure gradient suggests that large cerebral arteries contribute to cerebral resistance. We have tested the hypothesis that large cerebral arteries contribute to regulation of cerebral blood flow during changes in blood gases and arterial pressure. Microspheres were used to measure brain blood flow in anesthetized dogs. Resistance of large cerebral arteries was estimated by determining the pressure gradient between common carotid and wedged vertebral artery catheters. Systemic hypercapnia and hypoxia dilated large cerebral arteries, and hypocapnia constricted large cerebral arteries. Resistance of large arteries was 0.6+/-0.1 (mean +/- SE) mm Hg per ml/min per 100 g during normocapnia. During hypercapnia and hypoxia, large artery resistance decreased significantly to 0.2 +/- 0.03 and 0.3 +/- 0.05, respectively. During hypocapnia large artery resistance increased significantly to 1.0 +/- 0.1. In other experiments, we found that large cerebral arteries participate in auto-regulatory responses to hemorrhagic hypotension. When arterial pressure was reduced from 110 to 58 mm Hg, autoregulation maintained cerebral blood flow constant, and resistance of large cerebral arteries decreased significantly from 1.0 +/- 0.2 to 0.6 +/- 0.1 mm Hg per ml/min per 100 g. In absolute terms, we calculated that 20-45% of the change in total cerebral resistance during these interventions was accounted for by changes in large artery resistance. These studies indicate that large cerebral arteries, as well as arterioles, participate actively in regulation of cerebral blood flow during changes in arterial blood gases and during autoregulatory responses to hemorrhagic hypotension.


Mechanism of effect of hypoxia on renal water excretion. The effect of lowering the pressure of oxygen from 80 to 34 mm Hg was examined in anesthetized dogs that were undergoing a water diuresis. This degree of hypoxia was associated with an antidiuresis as urine osmolality (Uosm) increased from 107 to 316 mosmol/kg H(2)O (P < 0.001) and plasma arginine vasopressin increased from 0.06 to 7.5 muU/ml, (P < 0.05). However, hypoxia was not associated with significant changes in cardiac output (CO, from 4.2 to 4.7 liters/ min), mean arterial pressure (MAP, from 143 to 149 mm Hg), glomerular filtration rate (GFR, from 46 to 42 ml/min), solute excretion rate (SV, from 302 to 297 mosmol/min), or filtration fraction (from 0.26 to 0.27, NS). Hypoxia was associated with an increase in renal vascular resistance (from 0.49 to 0.58 mm Hg/ml per min, P < 0.01). The magnitude of hypoxia-induced antidiuresis was the same in innervated kidneys and denervated kidneys. To further examine the role of vasopressin in this antidiuresis, hypoxia was induced in hypophysectomized animals. The effect of hypoxia on CO, MAP, GFR, SV, and renal blood flow in hypophysectomized animals was the same as in intact animals. In contrast to intact animals, however, hypoxia did not induce a significant antidiuresis in hypophysectomized animals (Uosm from 72 to 82 mosmol/kg H(2)O). To delineate the afferent pathway for hypoxia-stimulated vasopressin release, hypoxia was induced in dogs with either chemo- or baroreceptor denervation. The effect of hypoxia on CO, MAP, GFR, SV, and renal blood flow in the denervated animals was the same as in nondenervated animals. Hypoxia resulted in an antidiuresis in chemoreceptor (Uosm from 113 to 357 mosmol/kg H(2)O, P < 0.001) but not in baroreceptor (Uosm from 116 to 138 mosmol/kg H(2)O, NS) denervated animals. To determine if hypoxia alters renal response to vasopressin, exogenous vasopressin was administered to normoxic and hypoxic groups of dogs. The antidiuretic effect of vasopressin was no different in these two groups. These results demonstrate that hypoxia induces an antidiuresis which is independent of alterations in CO, MAP, SV, filtration fraction, renal nerves, or renal response to vasopressin and occurs through baroreceptor-mediated vasopressin release. The nature of the baroreceptor stimulation remains to be elucidated.


Pemphigus antibody interaction with human epidermal cells in culture. The mechanism of pemphigus acantholysis has been studied with an in vitro system. Freshly prepared human skin epidermal cells were incubated in F-10 medium which contained the immunoglobulin G fraction from either pemphigus serum or normal human serum. During 18-h incubation periods, the pemphigus antibody became bound to the surface of the epidermal cells, caused the destruction of 75% of the viable cells as compared to only 14% in the normal immunoglobulin G controls (trypan blue exclusion), prevented the accumulation of newly synthesized proteins by nearly 60% as determined by radioactive tracer studies, and caused a dramatic shift in distribution of the newly synthesized proteins from an insoluble cell-associated fraction to an extracellular soluble fraction. These effects on the accumulation and partitioning of newly synthesized proteins were antibody concentration-dependent. Kinetic studies showed that at a fixed pemphigus antibody concentration the inhibition of protein accumulation preceded solubilization by about 1 h, at which time rapid solubilization of up to 70% of the insoluble cellular material occurred. Several lines of evidence suggested that this phenomenon was caused by enzymatic activity. Epidermal extracts solubilized a prepared substrate of radioactivity labeled insoluble epidermal cell material. This activity was heat labile and pH dependent, with pH optima ranging from 4.5 to 6.5. Enzymes with pH optima between 6 and 6.5 were recovered in the culture medium after a 2-day incubation of pure, intact epidermis with the pemphigus antibody. We proposed the following hypothesis to account for pemphigus acantholysis. The pemphigus antibody reacts with the epidermal cell surface and produces such a severe disturbance that the integrity of the cell surface is lost. As a result of these primary perturbations, the cell is killed and during the process, responds by release or activiation of soluble hydrolytic enzymes. This autolytic process results in the characteristic acantholysis of pemphigus.


Influence of methylprednisolone of the sequential redistribution of cathepsin D and other lysosomal enzymes during myocardial ischemia in rabbits. Occlusion of the circumflex coronary artery induced a profound redistribution in ischemic rabbit myocardium of several lysosomal acid hydrolases, including cathepsin D, B-acetylglycosaminidase, and acid phosphatase. 30-45 min after ligation non-sedimentable cathepsin D activity rose from 36% of the total activity to 42-48%, and in immunohistochemical preparations cathepsin D appeared to have diffused from lysosomes into the cytosol of injured cells. A pharmacologic dose of methylprednisolone (50mg/kg) significantly delayed the subcellular redistribution of cathepsin D and the other hydrolases in ischemic heart. Thus, in treated hearts the nonsedimentable activity of cathepsin D rose to only 38% after 30 min of ischemia and 42% after 45 min (P is less than 0.05 compared to untreated ischemia at each time). Similarly, unlike untreated hearts, noevidence of enzyme diffusion from lysosomes could be demonstrated immunohistochemically in corticosteroid-treated ischemic hearts for over 45 min. After 1-2 h of ischemia, however, steroid-protected myocytes deteriorated and the biochemical activity and anatomical distribution of cathepsin D were indistinguishable from untreated ischemic hearts. This study demonstrates that corticosteroid pretreatment does not prevent alterations in cardiac lysosomes during severe ischemia indefinitely, but does delay their development significantly.


Adherence of glucan-positive and glucan-negative streptococcal strains to normal and damaged heart valves. The adherence of 18 strains of streptococci to sections of normal canine and human aortic, mitral, and tricuspid valves and to canine interatrial septum was compared in an in vitro system. Quantitative measurements of adherence ratios were performed by two independent methods. Adherence ratios for Streptococcus mutans, S. sanguis, S. bovis, and Group D streptococci were higher (0.0058-0.0101) than for the other streptococcal strains studied (0.0025-0.0041). With the exception of Group D streptococci, adherence ratios for each bacterial strain were similar with the aortic, mitral, and tricuspid valve sections. Adherence ratios with normal human and canine valve leaflets were similar, but adherence ratios with interatrial septum were lower than with normal valve sections. Adherence ratios for glucan-positive and glucan-negative strains of streptococci with normal and with damaged aortic valve leaflets were also compared. The adherence ratios of the glucan-positive streptococci (S. mutans, S. sanguis, and S. bovis) and one glucan-negative enterococcal strain (KG-3) were approximately five times higher with damaged aortic valves (0.039-0.051) than with normal aortic valves (0.009-0.010). For glucan-positive strains, adherence ratios with normal aortic leaflets were similar when bacteria were grown in media which contains or lacks sucrose. In striking contrast, growth of the glucan-positive strains in medium which lacks sucrose, with resultant deficiency of glucan production, decreased the adherence ratios with damaged aortic valve leaflets to those found with normal aortic leaflets. Treatment of glucan-positive strains with dextranase resulted in a decrease in their adherence ratios to levels seen with bacteria grown in medium lacking sucrose, but the higher adherence ratios could be restored in the presence of exogenous dextran.It is concluded that glucan production is one quantitatively important factor that contributes to the greater adherence of glucan-positive streptococci to damaged rather than to normal aortic heart valve leaflets. However, glucan production is not the only factor that determines preferential adherence of streptococci to damaged heart valves, because glucan-negative strains may also show some degree of increased adherence to damaged valves. Thus, bacterial glucan production is one of the factors that could contribute to the pathogenesis of bacterial endocarditis.


Patients with congenital factor V deficiency have decreased factor Xa binding sites on their platelets. Human platelets have binding sites for plasma coagulation Factor X(a) that are available only after the platelet release reaction. Platelets from 15 normal donors bound 216+/-52 (SD) molecules of Factor X(a) per platelet. The association of Factor X(a) with its platelet surface receptor results in a 300,000-fold increase in the catalytic activity of Factor X(a) in forming thrombin from prothrombin. The turnover number for platelet-bound Factor X(a) was 1,850+/-460 mol thrombin/ml per min per mol Factor X(a) in experiments with platelets from 15 normal donors. Platelets from five patients with varying degrees of Factor V deficiency were investigated to determine whether or not coagulation Factor V participates in either aspect of the Factor X(a)-platelet interaction. The binding of Factor X(a) to platelets and the accompanying increase in rate of thrombin formation were either reduced in parallel or absent in each case with values ranging from 0 to 45% of control values. The apparent affinity of Factor X(a) from Factor V-deficient patients was normal when platelet binding was detected. The supernate from thrombin-treated control platelets, which contains Factor V activity, corrected the Factor X(a) binding deficiency of the platelets from three patients tested. Immunoreactive Factor V determined with an homologous antibody corresponded to the functional Factor V activity of platelets from one patient with Factor V deficiency, suggesting that the patient's platelets have a decreased amount of normal Factor V. The ability of platelets from the patients to bind Factor X(a) and increase the rate of thrombin formation correlated with the severity of each patient's bleeding disorder better than the plasma level of Factor V. The results indicate that Factor V is required for the Factor X(a)-platelet interaction and that thrombin formation at the platelet surface is important in normal hemostasis.


Immunoglobulin secreting cells in normal human bronchial lavage fluids. Immunoglobulin secreting cells were quantitated in the bronchial lavage fluids of 12 normal volunteers and compared with immunoglobulin secreting cells in peripheral blood, by a reverse hemolytic plaque assay. The mean number of cells secreting immunoglobulin (Ig)G in bronchial lavage fluids was 489 per million lymphocytes vs. a mean of 175 IgG secreting cells per million lymphocytes in peripheral blood (P < 0.02). The mean number of IgA secreting cells in bronchial lavage fluids was 633 per million lymphocytes as compared to 100 per million lymphocytes in peripheral blood (P < 0.005). Thus, compared to peripheral blood, cells from the lavage fluids were relatively enriched for both IgG and IgA secreting cells. However, IgA secreting cells were the major class of immunoglobulin secreting cells in bronchial lavage fluids, whereas IgG secreting cells predominated in peripheral blood. The prominence of IgA secreting cells in bronchial lavage fluids was further demonstrated by a mean ratio of IgA/IgG secreting cells in bronchial lavage fluids of 1.26 compared to a ratio in peripheral blood of 0.57 (P < 0.02). Cells secreting IgM were identified in only four of seven bronchial lavage fluid samples studied but in all peripheral blood samples. IgE secreting cells were not present in normal peripheral blood but could be demonstrated in 5 of 11 lavage fluid specimens. Thus, cells actively secreting immunoglobulins can be identified in the lower bronchial-alveolar tree of normal human subjects. Cells secreting IgG, IgA, or IgM may function in local lung defenses against infection; cells secreting IgE may contribute to hypersensitivity reactions in the lung.


Interaction of collagen with the lipids of tendon xanthomata. To determine the physical state of lipids in tendon xanthomata, six specimens surgically removed from three patients with familial hypercholesterolemia were studied by microscopy, calorimetry, and x-ray diffraction. The major constituents of the xanthomata were lipid (33% of dry weight) and collagen (24% of dry weight). The principal lipids were cholesterol ester and cholesterol. Light microscopy and thin-section electron microscopy showed occasional clusters of foam cells separated by masses of extracellular collagen. Polarized light microscopy of fresh, minced tissue showed rare droplets of free cholesterol ester. When heated, the tissue shrank abruptly at approximately equal to 70 degrees C and, consequently, a large amount of cholesterol ester was released. Scanning calorimetry of fresh pieces of xanthoma showed a single, broad, reversible liquid crystalline transition of cholesterol ester with peak temperature from 32 to 38 degrees C. The enthalpy (0971 +/- 0.07 cal/g) was reduced compared with the isolated cholesterol ester from each xanthoma (1.1+/-0.01 cal/g). There was a large irreversible collagen denaturation endotherm (peak temperature = 67 degrees C; enthalpy 9.9 cal/g collagen) that corresponded to the tissue shrinkage noted by microscopy. After the collagen denaturation, the sample displayed double-peaked reversible liquid crystalline transitions of cholesterol ester, of enthalpy 1.18 +/- 0.1 cal/g, that were identical to transitions of isolated cholesterol ester. Fibers dissected fron xanthomata were examined by X-ray diffraction at temperatures below and above the cholesterol ester transition. At 20 degrees C there was a weakly oriented equatorial reflection of Bragg spacing 36A, which corresponded to the smectic phase of cholesterol ester, and a series of oriented collagen reflections. At 42 degrees C the cholesterol ester reflection disappeared. Stretched fibers examined at 10 degrees C showed good orientation of collagen and cholesterol ester reflections, and in addition, meridional spacings which indicated oriented crystallization of cholesterol ester. These studies suggest that a major component of tendon xanthomata is extracellular cholesterol ester which displays altered melting and molecular orientation as a result of an interaction with collagen. At xanthoma temperatures, the cholesterol ester is in a smectic liquid crystalline state, probably layered between collagen fibrils, with the long axis of the cholesterolester molecules perpendicular to the axis of the collagen fiber. Such collagen-cholesterol ester interactions may favor the extracellular deposition of cholesterol ester derived either from intracellular sources or directly from plasma lipoproteins.


Effect of aspirin on thrombin-induced adherence of platelets to cultured cells from the blood vessel wall. An in vitro method was used to detect adherence of (51)Cr-labeled platelets to monolayers of cultured human endothelial, fibroblast, and smooth muscle cells. Washed platelets did not adhere to untreated or aspirin-treated endothelial monolayers in the absence of thrombin. In contrast, thrombin-induced platelet aggregates adhered to all of the monolayers but adherence to endothelium was significantly less than to the other cells. Additional evidence for adherence of platelets to the endothelium was provided by scanning and transmission electron microscopy. Thrombin-induced platelet adherence to endothelium was inhibited by hirudin. Platelet adherence induced by thrombin was enhanced significantly by treatment of the endothelial monolayer with 1-2 mM aspirin. This increase in adherence was seen even when aspirin-treated platelets were used; adherence values approached those seen with fibroblasts and smooth muscle cells. An aspirin concentration of 0.1 mM was sufficient to block thrombin-induced malonaldehyde production in platelets but it did not interfere with the inhibitory effect of the endothelium against platelet adherence. The effect of aspirin on the endothelium was temporary and inhibitory activity of the endothelium was restored 1 h after aspirin had been removed from the incubation system. The ability of thrombin to cause adherence of platelets to undamaged endothelium, and the potential for aspirin to enhance this adherence have implications for mechanisms which operate in platelet interaction with the blood vessel wall.


Decreased insulin sensitivity of forearm muscle in myotonic dystrophy. Previous studies of patients with myotonic dystrophy have demonstrated hyperinsulinism after glucose loading. This hyperinsulinism has been attributed by some investigators to tissue insulin resistance. We have directly studied insulin sensitivity of forearm muscle in patients having such hyperinsulinism. The effect of an intrabrachial arterial insulin infusion (100 mu U/kg per min) on glucose uptake was determined in six cases of myotonic dystrophy, six normal subjects, and in seven disease control subjects with myotonia or wasting from other disorders. There was no significant difference in insulin tolerance comparing myotonic dystrophy patients to the normal and disease control groups. Glucose tolerance and basal insulin levels were normal in the myotonic dystrophy patients, but hyperinsulinism occurred after glucose ingestion. After 25 min of intra-arterial insulin, the mean peak muscle glucose uptake in myotonic dystrophy was 2.54 +/- 0.54 mu mol/min per 100 ml forearm compared to 5.24 +/- 0.86 mu mol/min per 100 ml for disease controls (P is less than 0.05). Myotonic dystrophy patients showed a peak glucose uptake increment of only 2.6 +/- 0.2-fold over basal contrasted with the disease control value of 6.5 +/- 1.0-fold (P is less than 0.02) and the normal control value of 8.8 +/- 1.1-fold (P is less than 0.01). Thus, there was an absolute as well as a relative decrease in muscle insulin sensitivity in myotonic dystrophy patients compared to both control groups. The peak increments in arterio-superficial venous glucose concentration differences after insulin infusion were not significantly different comparing myotonic dystrophy and control groups. These data suggest that in myotonic dystrophy, there is insulin insensitivity of skeletal muscle.


Inhibition of erythrocyte sickling in vitro by pyridoxal. To test the antisickling activity of pyridoxal, we compared the oxygen affinity and the percent sickling at low PO2 of untreated erythrocytes with values for cells from the same blood sample incubated with pyridoxal, glyceraldehyde, or pyridoxine. Pyridoxal increased oxygen affinity much more than glyceraldehyde. 20 mM pyridoxal and glyceraldehyde had equivalent antisickling activity. At PO2 levels above 20 mm Hg, both agents reduced sickling to less than 2%. In samples examined by electron microscopy, pyridoxal reduced the percent sickled cells and the percent cells that contain hemoglobin S fibers by the same amount (from 74 to 3%). Pyridoxine had no effect on oxygen affinity or sockling. Pyridoxal reacts with intracellular hemoglobin to increase oxygen affinity, which inhibits hemoglobin S polymerization and sickling.


Cell bodies of origin of reticular projections from the superior colliculus in the cat: an experimental study with the use of horseradish peroxidase as a tracer. By use of the retrograde axonal transport of horseradish peroxidase (HRP), the projection from the superior colliculus (SC) to the brain stem reticular formation (RF) was investigated in the cat. A 0.2-0.5 microliter of a 50% suspension of Sigma VI HRP was injected stereotactically in various portions of the pontomedullary RF, and, as a control to the injection to the RF, in the inferior olive or in the spinal cord. Labeled cells were found within and deep to the intermediate gray layer of the SC in the cats which survived for two or three days after HRP injection. The number of the labeled cells varied, according to the difference in the site of injection and the amount of injected HRP. About 400 labeled cells in twenty 50-micron sections, taken every fifth of the SC, occurred throughout its rostrocaudal extent, particularly in the case where the medial portion of the border zone of the nucleus reticularis pontis oralis and caudalis (R.p.o.-R.p.c. zone) or the border zone of the nucleus reticularis pontis caudalis and the gigantocellularis (R.p.c.-R.gc. zone) was heavily stained after three days of survival period. From 10 to 15% of these labeled cells were large in size (more than 40 micron in diameter), 20-30% were medium sized and the rest (60-70%) were small (10-25 micron). On the other hand, when HRP was placed in the inferior olive only eight cells were labeled in the SC, seven of which were small and medium-sized. When HRP was injected in the gray matter of C1-C3 level of the spinal cord, a total of 70 tectal cells (14, 42, and 14 were large, medium, and small cells, respectively) were observed to be labeled. The findings of the tectoreticular neurons are discussed and compared with those of the tectoolivary and the tectospinal neurons. Thus the three kinds of tectal neurons are located within and deep to the intermediate gray layer. The number of the labeled cells and the percentages of the collicular neurons of different sizes are obviously different among the three different projections. Topographic correlations between the SC and the RF could not be discerned in the present materials. These results were discussed in relation to possible influences of the tectoreticular neurons upon the extraocular and the spinal motoneurons.


Laminar origins of spinothalamic projections in the cat as determined by the retrograde transport of horseradish peroxidase. The method of retrograde axonal transport of horseradish peroxidase (HRP) was used to identify the locations of cells of origin of the spinothalamic tract in the cat. Injections of from 0.2-3.0 microliter of 30% HRP were made unilaterally in various regions of the somatosensory thalamus. Massive injections of the caudal thalamus in several cats showed the spinothalamic cells of origin to be located mainly in laminae I, VII and VIII in the lumbar enlargement, and in laminae I, V and VII-VIII in the cervical enlargement. Small injections of HRP were made into the three major spinothalamic terminal zones in the thalamus, to determine the laminar origins(s) of the spinal projections to each zone. Neurons in lamina I in both cervical and lumbar enlargements were found to project almost exclusively to the rostral VB-caudal VL border zone. A small number of neurons in laminae VII and VIII also project there but a larger number project to the intralaminar region. Neurons projecting to the PO regions were located mainly in laminae IV and V. This anatomical segregation of thalamic afferents probably reflects a functional segregation of input, since the functional properties of spinal neurons vary according to their laminar location. Comparison of these data with the differential projection of spinothalamic neurons in the rat and monkey indicate that it is unlikely that the proposed "paleo-" and "neospinothalamic" systems would arise from anatomically separate groups of spinal neurons.


Anatomical and physiological properties of ipsilaterally projecting spinothalamic neurons in the second cervical segment of the cat's spinal cord. Anatomical and electrophysiological methods were used to investigate the projections and response properties of neurons in the second cervical (C2) spinal segment of the cat giving origin to a previously undescribed projection to the ipsilateral thalamus. The method of retrograde axonal transport of horseradish peroxidase (HRP) was used to identify neurons in C2 giving rise to thalamic projections. Following large (3.0 microliter) thalamic HRP injections, a large number of labeled neurons was observed in lateral laminae VII-VIII of C2 ipsilateral to the injections. They occurred as small clusters of cells along the longitudinal axis of C2. Labeled neurons were also observed contralaterally in the lateral cervical nucleus, dorsal horn (especially medial lamina VI), and loosely distributed in the ventral horn. The ipsilaterally projecting neurons were also labeled following small (0.2--0.5 microliter) HRP injections restricted to individual spinothalamic terminal zones (intralaminar nuclei, ventrobasal complex-nucleus ventralis lateralis border zone, medial division of the posterior nuclei), indicating that as a group they project widely throughout the thalamus. Single unit recording methods were used to obtain complementary information on the functional properties of these neurons. The antidromic stimulation method was applied to identify units in C2 projecting to the ipsilateral thalamus in anesthetized, paralyzed cats. Three categories of ipsilaterally projecting C2 units were identified: (1) units not driven by any type of natural stimulation; (2) units having large cutaneous receptive fields (RFs) and wide dynamic response ranges ("widefield"), and (3) units with smaller RFs and varied properties ("other"). Widefield units with bilaterally symmetrical and asymmetrical RFs were observed. Co-stimulation of different portions of an excitatory RF produced summation of the unit response. Inhibitory RF components were identified in one-third of the widefield units. Unit recordings after spinal tract lesions revealed that the afferent input passed via the ipsilateral lateral and/or ventral funiculi. Widefield unit responses to somatosensory stimuli could be inhibited by dorsal column conditioning stimulation. Several "other" units resembled widefield units, while a second group had small RFs restricted to the C2 dermatome. Possible functional roles of the projecting C2 neurons in somatosensory and non-specific systems are discussed.


Distribution of the crossed olivocochlear bundle in the chinchilla's cochlea. The efferent crossed olivocochlear bundle (COCB) was transected in the brain stem of the chinchilla, and the animals sacrificed 7 to 96 days later. Electron microscopy revealed that all the large efferent nerve endings on outer hair cells in the basalmost 2 mm (round window region) of the cochlea had degenerated, 87.5% in the remainder of the first turn, 70% in the second turn and 43% in the third turn. Only a few degenerating nerve fibers were seen in the medial spiral tract (inner spiral and tunnel bundles) of the experimental animals. Nerve fibers were counted in the medial spiral tracts of the cochleas of control animals as well as in those animals whose COCB had been transected. There were considerable individual variations in the fiber numbers, and statistical analysis showed no significant difference between the numbers of nerve fibers in normal and experimental animals. The cochlear microphonics (CM) and nerve action potentials (AP) of acute animals were assessed before and after COCB section. The CM and AP of the chronic experimental animals were compared with responses from normals. Overall, no changes in a physiological response of the anesthetized chinchilla could be attributed to complete section of the COCB.


Differential projections of cat medullary raphe neurons demonstrated by retrograde labelling following spinal cord lesions. Neurons of the medullary raphe nuclei in cats were retrogradely labelled following injection of horseradish peroxidase (HRP) into the L6 spinal cord segment. Brainstems were cut in sagittal section to facilitate examination of the rostral-caudal extent of raphe neurons projecting to the spinal cord. Large numbers of HRP-labelled neurons were found in nucleus raphe magnus, nucleus raphe pallidus, and nucleus raphe obscurus (as well as a few neurons in nucleus raphe pontis). Dorsal or ventral hemisections at the T12-L1 level restricted HRP retrograde transport to those pathways within the intact portion of spinal cord, allowing a determination of the part of the cord through which raphe neurons project to the lumbar enlargement. Neurons of nucleus raphe magnus were found to project primarily in dorsolateral fasciculus. A significant number of neurons of nucleus reticularis gigantocellularis also project in dorsolateral fasciculus. Nucleus raphe obscurus neurons were found to project primarily in ventral funiculus, while nucleus raphe pallidus neurons project in the ventrolateral fasciculi and ventral funiculus. The serotonergic (5HT) fibers described by Dahlström and Fuxe ('65) to terminate in the dorsal horn, intermediolateral cell column, and ventral horn are likely to coincide with the raphe-spinal projections documented in this work.


The synaptic terminations of certain midbrain-olivary fibers in the opossum. The nuclear origin and distribution of midbrain-olivary fibers has been described in a previous study utilizing axonal transport techniques (Linauts and Martin, '78a). The present report extends their results to the electron microscopic level and details the postsynaptic distribution of such fibers. Lesions within the ventral periaqueductal grey and adjacent tegmentum, the red nucleus or the nucleus subparafascicularis result in electron dense axon terminals within the olive at survival times of 48, 72 and 96 hours. At 72 hours, many degenerating presynaptic profiles shrink, become irregular in shape and are totally or partially surrounded by glial processes. The principal olivary nucleus contains the majority of these profiles. However, the subparafascicular terminals are more abundant in the rostral and intermediate parts of the medial accessory nucleus and the rubral terminals are concentrated within the dorsal lamella of the principal nucleus. The nuclear location of the degenerating terminals was determined by examination of 1 micrometer plastic sections cut in the transverse plane from each block face prior to thin sectioning. Degenerating terminals were counted in three cases, one from each of the three lesion sites described above. When taken together these cases show that just over 50% of the degenerating terminals are presynaptic to spiny appendages and are located within the synaptic clusters (glomeruli) described previously (King, '76). The percentage of degenerating terminals in the glomeruli increases to 70% when the lesion is in the ventral periaqueductal grey and adjacent tegmentum. The remaining degenerating terminals contact dendritic shafts outside the astrocytic boundaries of the synaptic clusters. The synpatic vesicle populations within the degenerating terminals vary with the location of the lesion. Lesions in the ventral periaqueductal grey and the adjacent tegmentum result in the degeneration of terminals with either clear spherical vesicles or endings with both clear spherical vesicles and a variable number of large dense core vesicles. In contrast, the primary degenerative changes that occur after destruction of the red nucleus or the nucleus subparafascicularis are in terminals with clear spherical vesicles. When the synaptic complex was present in the plane of section, regardless of the site of the lesion, the degenerating terminals could be classified as Gray's type I. Thus, we have demonstrated that afferents from the mesencephalon terminate within synpatic clusters located in the principal and medial accessory (part A) subnuclei of the inferior olive. Although the mesencephalic afferents have multiple origins (Linauts and Martin, '78a), many of their synaptic terminals contact spiny appendages within the synaptic clusters. This postsynaptic site also receives cerebellar terminals (King et al., '76). The origin of presynaptic profiles within the synaptic clusters that contain clear pleomorphlic vesicles is yet to be determined.


Experimental studies on the ultrastructural localization of acetylcholinesterase in the mediobasal hypothalamus of the rat. Acetylcholinesterase (AChE) activity has been demonstrated ultrastructurally in neurons of the arcuate nucleus and associated with fibers in the arcuate nucleus neuropil and the median eminence (ME) of the rat. In addition, the effects of neonatal monosodium glutamate (MSG) treatment and Halász deafferentation on the AChE staining and localization have been studied. Neonatal MSG-treatment resulted in loss of the majority of AChE-positive neurons in the arcuate nucleus while leaving neuropil staining intact. Halász deafferentation caused a loss of arcuate neuropil activity while leaving the neuronal staining unaltered. These observations are consistent with previous biochemical results suggesting the existence of a cholinergic tuberoinfundibular system with nerve cells in the arcuate nucleus and terminals in the median eminence. In addition, the deafferentation experiments indicated that extra-hypothalamic cholinergic fibers may innervate the arcuate nucleus. Supporting evidence from other biochemical studies and the curious paucity of histochemical and biochemical AChE activity in the ME are also discussed.


The organization of the hippocampus of the fence lizard: a light microscopic study. The hippocampus of the fence lizard (Sceloporus undulatus) is composed of two laminated regions that are readily distinguishable on the basis of position and perikaryal size in Nissl preparations: the small-celled (Hsc) and the large-celled (Hlc) hippocampal divisions. Based upon cellular and fibrillar characteristics, the layers of the Hsc are: (1) the superficial plexiform layer; (2) the cellular layer; (3) the deep plexiform layer; (4) the layer of deep fibers, or alveus; and, (5) the ventricular ependyma. The layers of the Hlc are: (1) the tangential layer; (2) the molecular layer; (3) the pyramidal layer; (4) the deep plexiform layer; (5) the alveus; and, (6) the ventricular ependyma. The neurons of each hippocampal layer were grouped into classes based upon somal position and appearance, and dentritic and axonal arborization patterns in Golgi sections. Neurons of the Hsc are more numerous and pleomorphic than those of the Hlc. Data do not suggest an intrinsic organization that closely parallels that found in the mammalian hippocampus.


Development of the dorsal lateral geniculate nucleus in the cat. The development of the lateral geniculate nucleus has been studied systematically in Nissl preparations from a series of cats that ranged in age from newborn to adult. In addition, preliminary observations are reported at two stages of fetal development. It was found that laminae develop in the lateral geniculate nucleus near the time of birth and continue to differentiate during the first postnatal week. During development the major axis of the lateral geniculate rotates approximately 180 degrees in the sagittal plane. The rotation begins prenatally and is not completed until after the twentieth postnatal week. The volume of the lateral geniculate was computed at different ages and it was determined that during the first postnatal month the nucleus attains two-thirds of its adult size. However, the rate of growth declines markedly thereafter, and final volume, like final position, is not achieved until late in development. The cross-sectional areas of lateral geniculate neurons were measured at four locations in the nucleus in each animal. The locations represented the following parts of the visual field: the paracentral and inferior peripheral fields in the binocular segment of lamina A; the monocular segment of lamina A; and the paracentral field in lamina A1. Neurons in each of these locations grow at approximately the same rate and are essentially fully grown by 56 days. Cell size histograms show that more large cells are found in lamina A1 and more small cells in the monocular segment than elsewhere in the dorsal laminae. Unlike the retina, there appears not to be a gradient of development in the lateral geniculate nucleus from center to periphery, at least in terms of cell body size at the ages studied. On the contrary, that part of the lateral geniculate nucleus which represents the paracentral visual field is the last segment in the dorsal laminae to achieve a mature cell size distribution. Finally, a discrete class of small spindle-shaped neurons was observed in the lateral geniculate nucleus ventral and caudal to the C laminae during the first two postnatal weeks. These cells possess a leading and trailing cytoplasmic process and are distinctly different from cells in the main laminae. It is suggested that these spindle-shaped cells may be neurons that are still in the process of migration or differentiation in the postnatal animal.


The olivocerebellar projection in the cat studied with the method of retrograde axonal transport of horseradish peroxidase. VII. The projection to lobulus simplex, crus I and II. The olivocerebellar projection to lobulus simplex, crus I and II in the cat was investigated by means of retrograde axonal transport of horseradish peroxidase (HRP). The distribution of labeled cells in the inferior olive following HRP injections in lobulus simplex, crus I and II confirmed the findings by Brodal ('40b) that the rostral half of the principal olive projects to these areas of the cerebellar hemisphere. However, concerning details there are some differences in so far as the heaviest contribution to crus I comes from the medial parts of the ventral and dorsal lamella, that to crus II from its lateral part, especially the ventral bend. The present findings show that in addition the rostral part of the medial and the rostromedial part of the dorsal accessory olive project to these areas of the cerebellar cortex. Further details in the projection are shown in figure 8B. The findings agree fairly well with the electrophysiological results of Armstrong et al. ('74) and the experimental anatomical data of Groenewegen and Voogd ('77a,b). An attempt is made to correlate the findings with the pattern of longitudinal zonal subdivision of the cerebellum. There is evidence for a topical organization within the projection to crus I and II and parts of their projection areas in the principal olive. The distribution of the labeled cells which project to lobulus simplex, crus I and II is discussed in relation to afferent pathways to the inferior olive.


Computer assisted tomography in primary malignant lymphomas of the brain. Computer assisted tomography (CAT) data from 10 patients with histologically verified primary malignant lymphomas of the brain are reported. Studies both prior to and after contrast medium administration were done on nine patients. In all these patients, tumor nodules presented with slightly increased density in the precontrast scan and considerable contrast enhancement. The appearance of malignant lymphomas in the CAT scan may be similar to that of meningiomas. Pituitary adenoma, medulloblastoma, and metastasis of malignant melanoma may not be ruled out in a particular case from the CAT picture alone. Blurred margin of the tumor after contrast enhancement was found in half the patients. Diffusely growing malignant lymphomas produce low density areas in the CAT scan without contrast medium uptake.


Computed tomography in primary malignant lymphoma of the brain. Primary malignant lymphoma of the brain is a rare lesion characterized clinically by varied nonspecific neurologic deficits and usually a rapid fulminant deterioration and death. This course may be altered by radiotherapy, and early diagnosis assumes major importance. Laboratory tests, EEG, radionuclide scans, angiography, and pneumoencephalography may not show definite abnormality in the early stages. This report concerns four cases in which good correlation between the location of the brain lesion and the abnormality on computed tomography scan was demonstrated.


Computed tomography of the optic nerve: part I. Normal results. In vivo visualization of the optic nerve (ON) by computed tomography (CT) has important clinical applications, both realized and potential. An accurate CT analysis of the ON is, however, hampered by a number of technical limitations. Sinuosity and gaze-shift-related motility of the ON tend to increase the difficulties of demonstrating in a consistently satisfactory fashion this anatomical structure. Nevertheless, if rigid imaging criteria are followed, adequate visualization of the ON may be attained in the majority of patients. Simultaneous visualization of the ON and the eyeball permits establishment of a neuroocular index for clinical use.


Computed tomography of the optic nerve: part II. Size and shape modifications in papilledema. Increase in the caliber and course tortuosity of the optic nerve (ON) can be demonstrated by computed tomography (CT) in cases of papilledema due to intracranial tumors, hydrocephalus, pseudotumor cerebri, and other conditions causing increased intracranial pressure. The enlargement of the ON in these conditions is generally bilateral, although one of the nerves, usually of the side of the tumoral pathology, may be thicker. Enlargement on the ON may also be demonstrated by CT in papilledema due to optic neuritis. Occasionally, the thickened ON also display increased attenuation coefficient values.


White matter of the cerebellum demonstrated by computed tomography: normal anatomy and physical principles. Although computed tomography (CT) delineation of normal white matter of the cerebral hemispheres has been well documented, there has been no description of white matter within the cerebellum. Through the use of phantom studies, CT number correlations between cerebellum and cerebral hemispheres, and anatomic correlation with in vitro specimens, the ability to visualize cerebellar white matter is demonstrated. Thin sections decrease volume averaging and enable consistent imaging of these structures. Size and shape of the corpus medullaris on CT scan may vary with the scan angle and level of section. Representative examples of various normal appearances are illustrated.


Computed tomography in surgical pancreatic emergencies. The findings observed with computed tomography (CT) in such pancreatic emergencies as necrotic-hemorrhagic pancreatitis, pancreatic abscess, broken pseudocyst, and pancreatic ascites with mediastinitis are presented. The value of CT in these conditions, which often require surgical intervention, is discussed. Computed tomography appears to be the ideal diagnostic procedure, especially for surgical treatment planning in pancreatic abscess. No deaths occurred in a group of pancreatic abscesses treated surgically with CT assistance.


Localization of the head of the pancreas using the junction of the left renal vein and the inferior vena cava. The junction of the left renal vein with the inferior vena cava is visualized on most computed tomography studies of the abdomen. This junction is a useful guide in localizing the pancreatic head, just as the superior mesenteric artery is used to localize the body of the pancreas.


Absence of thoracic osteophytosis in the area adjacent to the aorta: computed tomography demonstration. Computed tomography of the chest shows that right-sided unilateral thoracic osteophytosis is a common phenomenon. The cross-sectional images obtained appear to support the thesis that aortic pulsations have a preventive effect on the formation of left-sided osteophytes.


Internal structures of the knee visualized by computed tomography. The internal structures of specimens of the knee can be visualized by computed tomography, although visualization is directly dependent on a thin (3 mm) scan in the coronal or sagittal plane with air used as a contrast agent.


Standardization of computed tomography images by means of a material-selective beam hardening correction. Polychromaticity of the X-rays used in computed tomography (CT) has made it difficult to establish an absolute scale for CT values and has made quantitative comparisons between patients unreliable. The spectral shift of the X-rays depends on the material distribution within the structure measured and is significant if substantial amounts of bone, fat, or injected contrast material are present. A material-selective beam hardening correction procedure has been developed that allows the reconstruction of good approximations of linear attenuation coefficients with respect to a reference energy. With the aid of mathematical simulations and measurements on a physical phantom, the feasibility of the procedure and its insensitivity with regard to energy settings and other machine parameters are documented.


Computer reconstructed sagittal and coronal computed tomography head scans: clinical applications. Recent technical advances to computed tomography (CT) have enabled evaluation of the head in multiple planes. A technique that utilizes computer reconstruction of data from 3 mm contiguous transverse scans to display images in sagittal and coronal planes is described. The clinical usefulness of this technique is discussed and illustrative cases are presented. Multiple plane CT evaluation has proven a valuable adjunct to axial plane CT scanning, which can provide better definition of the extent of lesions, facilitate treatment, and improve diagnostic accuracy in many cases.


Effective atomic number and electron density as measured with a computed tomography scanner: computation and correlation with brain tumor histology. The mathematical basis and methodology for determining the effective atomic number and electron density of a tissue are presented. Cranial computed tomography (CT) scanning is performed at two different energy levels, both with and without contrast enhancement. The histology of 15 brain tumors has been correlated with the effective atomic number and electron density of the tumor, with parameters of the linear attenuation coefficient, and with statistical values of the CT numbers. The most useful value in separating gliomas from meningiomas from metastases appears to be the percentage change of effective atomic number following contrast enhancement.


Case report: computed tomography in the diagnosis of primary lymphoma of the central nervous system. A case of primary central nervous system lymphoma is presented with characteristics shown by computed tomography (CT) closely resembling those of the only other case reported in the literature. The combination of the arteriographic and CT appearances suggests the diagnosis of this radiosensitive group of tumors.


Case report: computed tomography of cystadenoma of the pancreas. Cystadenoma is an unusual pancreatic tumor characterized by large-sized polycystic components. It is located totally within the pancreas and may cause retroperitoneal collateral venous return. These characteristics can be easily visualized by computed tomography, thus making it a very effective modality in the differential diagnosis of pancreatic lesions.


Case report: echinococcal cyst of the liver seen on computed tomography. A case of a single, large noncalcified echinococcal cyst of the liver demonstrated by computed tomography (CT) and verified at surgery is presented. To our knowledge, no previous CT pictorial display of such a lesion has been reported in the literature. The cyst appears as a radiolucent, sharply demarcated lesion of water density.


A simple method of localizing the level of computed tomography cross sectioning. Before computed tomography (CT) scanning, a preliminary radiograph of the area of interest is obtained with a Bell-Thompson ruler on the table top parallel to the spine. This allows precise localization of the level of CT sectioning in relation to the vertebral column.


Computed tomography in the evaluation of mediastinal lipomatosis. Mediastinal abnormalities can present a challenging differential diagnosis. One of the many causes of superior mediastinal widening is excess deposition of fat or mediastinal lipomatosis. Mediastinal widening was evaluated by routine chest radiography, conventional tomography, and computed tomography in four patients with steroid induced Cushing's syndrome or simple obesity. Computed tomography was the only modality that definitively diagnosed mediastinal lipomatosis in each case.


Evaluation of abdominal abscess with computed tomography. Computed tomography (CT) is valuable in providing a specific diagnosis of abdominal abscess and in determining its site and extent. Computed tomography is also capable of excluding an abscess with a high degree of certainty. In 31 of 34 patients with proven abdominal abscess, CT suggested the correct diagnosis and accurately defined the extent of involvement. In nine patients, the CT findings were specific for abscess. The CT appearance of abscess is diverse, largely depending on its location. The most common appearance is a low density mass often with higher density at the periphery. The presence of gas distributed within the mass or an extraalimentary air--fluid level allows an extremely confident diagnosis of abscess. Differential diagnostic possibilities are reviewed with emphasis on interpretive problems.


Computed tomography in the evaluation of the solitary or unilateral nonfunctioning kidney. Computed tomography (CT) using the EMI CT5000/5005 scanner was performed on 14 patients with a solitary functioning kidney demonstrated by other radiological investigations. Examinations before and after intravenous injection of contrast material have been undertaken. In the majority of cases, the patients were referred as a "nonfunctioning kidney" following excretion urography. Other referrals included previous nephrectomy or solitary kidneys. In all cases, the CT scan made possible the definitive diagnosis. The diagnosis included unsuspected hydronephrosis, vestigial kidney, contracted kidney secondary to chronic pyelonephritis and calculus, recurrence of a hypernephroma, and lymphomatous infiltration. Valuable information was obtained about the vascular pedicle, the renal collecting systems, the perinephric space, and the retroperitoneal space including the paraaortic lymph nodes. Furthermore, the increased sensitivity of the technique in detecting lower concentrations of iodine provided information regarding renal function. It is concluded that the noninvasive nature of CT makes it a valuable adjunct in the further investigation of such patients.


Computed tomographic patterns of pulmonary thromboembolism and infarction. A wedge shaped pulmonary density with its broad base against a peripheral pleural surface and with its tip pointing to the parahilar area is suggestive of pulmonary infarction. This pattern can be shown more often with computed tomography (CT) scanning than in plain chest X-ray films. In 16 patients with clinical evidence of pulmonary thromboembolism, often corroborated by diagnostic procedures other than CT, a variety of CT patterns of increased attenuation were noted. In seven (44%) of these cases, a distinct wedge shaped appearance was observed.


Lack of visualization of the portal venous tree in cirrhosis of the liver: a computed tomography finding with possible diagnostic significance. During computed tomography (CT) scanning of the liver, the inferior vena cava can be identified as a separate structure of lesser density adjacent to the caudate lobe in two-thirds of patients without known hepatocellular disease. In patients with alcoholic (portal) cirrhosis of moderate to severe degree, intrahepatic portal veins may not be identified on CT scans, even though their inferior vena cavas can be definitely distinguished from the caudate lobe. Portal cirrhosis causes distortion and obliteration of the portal triads, which is presumably reflected by lack of visualization of intrahepatic portal veins during CT scanning.


Computer tomography of the larynx.

Computer tomography of the larynx. Evaluation of computed tomography of the normal larynx has been performed utilizing corresponding tomographic and anatomic planes. The transaxial, coronal, and sagittal planes have been evaluated. While all three planes have distinct potential diagnostic value, the transaxial plane offers a unique view of laryngeal structures not afforded by any other technique. It is hoped that diagnostic criteria developed in this study will prove applicable to the evaluation of clinical laryngeal disease.


Computed tomography in olfactory neuroblastoma: one case of esthesioneuroepithelioma and four cases of esthesioneuroblastoma. Five cases of histologically confirmed olfactory neuroblastoma (esthesioneuroblastomas) have been studied by computed tomography (CT). Both the clinical symptoms (in particular, unilateral nasal obstruction and recurring epistaxis) and radiological findings (opacity of the paranasal sinuses) are nonspecific. Computed tomography shows a contrast enhancing mass-lesion and is more useful than conventional tomography in estimating its extension and the associated bone destruction, secondary reaction of sinusitis, and tumoral calcification. In the two cases in which the tumor was associated with exophthalmos, CT demonstrated that the muscular cone acts as a barrier preventing tumoral invasion. In cases wtih intracranial involvement, it appears that the attenuation values can be useful for differentiating between invasion by contiguity (two cases) versus metastasis (one case). Computed tomography is valuable in the follow-up and during and after radiotherapy of the tumor.


Intracranial pearly tumors: the roles of computed tomography, angiography, and pneumoencephalography. The findings is computed tomography (CT) and other radiological examinations in nine histologically proven cases of intracranial pearly tumors are presented. The CT findings include a low density lesion with no enhancement after contrast medium injection and in some cases a lesser degree of mass effect than would be expected from the size and site of such lesions. Pneumoencephalography is superior to CT in detecting suprasellar pearly tumors when they are isodense with the cerebrospinal fluid.


Computer assisted tomography in experimentally induced orbital pseudotumor. Induction of an entity that is comparable to pseudotumor of the orbit in humans has been performed successfully in the rabbit. Injection of a retrobulbar antigen in a previously sensitized rabbit produced a profound inflammatory mass. Proptosis, soft tissue swelling, and an orbital mass effect were grossly visible after a short interval. Computer assisted tomography disclosed dense orbital mass and uveoscleral thickening. Exenteration specimens confirmed as inflammatory cell mass similar to the histology associated with pseudotumor of the human orbit.


Bismuth encephalopathy and cerebral computed tomography. Five cases of severe encephalopathy due to chronic ingestion of bismuth have been studied by cerebral computed tomography (CT). In every case, areas of increased attenuation coefficient in the basal ganglia and the cortex of the cerebral hemispheres were demonstrated. These regions of increased attenuation were diffuse with irregular, patchy denser areas. The increased attenuation in the gray matter territories contrasted with decreased attenuation of the white matter, particularly evident in the centrum semiovale of each hemisphere and probably related to edema. Some ventricular dilatation was also present in three cases. In one patient, areas of increased attenuation were demonstrated in the cerebellum. Follow-up CT studies of one patient revealed regression of the pathological findings. This regression accompanied the clinical improvement, which, in every case, followed the termination of the bismuth intake.


Computed tomography in leptomeningeal spread of tumor. Computed tomography proved insensitive to leptomeningeal spread of hematologic malignancies including leukemia, lymphoma, and malignant histiocytosis. In only 3% of patients did it directly demonstrate leptomeningeal tumor. In comparison, the detection rate of leptomeningeal tumor secondary to carcinoma was 44% and for melanoma, 100%. Intracranial subarachnoid seeding from primary brain gliomas was detected in each instance. The simultaneous presence of parenchymal metastases with leptomeningeal carcinomatosis occurred in 18% of patients with nonhematologic malignancies. Computed tomography evidence of communicating hydrocephalus, previously thought to be a major factor in clinical symptomatology, occurred in only 11% of patients.


Cranial computed tomography findings in xeroderma pigmentosum with neurologic manifestations (De Sanctis-Cacchione syndrome). Computed tomography (CT) scans in two young patients with xeroderma pigmentosum with neurologic manifestations (De Sanctis-Cacchione syndrome) showed ventricular dilatation and cerebral cortical atrophy. The brainstem appeared small. In addition, an abnormal thickening of the calvarial bones was noted in both patients. These CT findings of the brain were compared with the neuropathologic features of this syndrome reported in the literature.


Computer assisted tomography of unoperated subdural hematoma: short- and long-term follow-up. Six recent and six long-term follow-up patients with subdural hematoma studied by computer assisted tomography are reported. This method was used to demonstrate progressive resolution of the hematoma in five of the recent patients as well as enlargement of the hematoma in one of the patients who subsequently was sent to surgery. Computer assisted tomography was also used to demonstrate absence of subdural hematoma in long-term follow-up of unoperated patients. The possible difficulty in visualizing an isodense subdural hematoma is discussed, as is the use of additional coronal views to improve subdural detection.


Noise limitations in X-ray computed tomography. A lower bound for the statistical accuracy in X-ray computed tomography, which, under certain conditions, is independent of the reconstruction algorithm, is derived. An evaluation of this bound indicates that little improvement is possible over the performance of the convolutional algorithm.


Polychromatic streak artifacts in computed tomography images. A series of artifacts in computed tomography (CT) images, consisting of streaks coming off edges and flares surrounding the object being scanned, is shown to be due to polychromaticity of the X-ray beam. These artifacts are seldom seen on conventional CT brain scans. They are helpful in understanding artifacts such as those seen around the petrous bone of the skull. Following this, a comparison is made between "single energy" and "dual energy" correction schemes for poly-artifacts. The theoretical assumptions underlying either method are analyzed, and the importance of these concepts in dual energy reconstructions is stressed. A new test phantom is suggested to evaluate different poly-correction schemes. All the simulated projection data in this study are noise free and are reconstructed using an industrial convolution-back projection technique. In addition, reconstructions are shown of the phantom using real data taken with and without a water bag path length compensator.


Nuclear scattering radiography of the spine and sphenoid bone. Nuclear scattering radiographs of a portion of a spine and a sphenoid bone have been obtained using a 1 GeV proton beam. The ability of the method to yield three-dimensional representations is illustrated by three series of adjacent pictures corresponding to the three orthogonal planes (elementary volume: 5.2 mm3 and 0.9 mm3). The sensitivity of the method is discussed. Nuclear scattering radiographs are compared with ones obtained by conventional X-ray tomography and computed tomography. Nuclear scattering radiography also may be used to analyze the partition of hydrogen within the tissures. Hydrogen radiographs obtained in this way are shown.


Ruptured teratoid tumor in the pineal region. Distinctive plain X-ray and computer assisted tomography (CAT) findings led to the diagnosis in a case of teratoma in the pineal region and helped avoid invasive investigations. The demonstration by CAT of the location and size of the tumor as well as the presence of marked hydrocephalus directed the patient's therapy to ventriculoperitoneal shunting and radiation.


Computed tomography of anatomic specimens. To obtain high quality computed tomographic (CT) images of specimens, the specimens must be submerged in water and immobilized. We describe a simple, effective technique for imaging brain specimens in various CT scanners. Anatomic sections that correspond exactly to the plane of the CT image may be obtained


Chromomycosis.

Chromomycosis. Chromomycosis is a general term to express human and animal fungal infections caused by members of the family Dematiaceae (dark brown or black fungi). Unique host-parasite relationships produce a wide spectrum of clinical disease, sometimes difficult to treat. Four major categories are recognized: 1) Superficial Chromomycosis--A pigmented macule produced by fungus inhabiting exclusively the dead horny layer (stratum corneum) of the skin, as in tinea nigra palmaris. 2) Chromoblastomycosis--A verrucous, slow growing nodule or plaque produced by fungi, characteristically and exclusively present as large brown spherical bodies (chromo bodies) in the dermis. 3) Chromohyphomycosis--A dermal nodule produced by fungi displaying typical brown septated hyphae. Rarely, the fungus may appear as a yeast. It may occur in deeper tissues, i.e., muscles, bone and brain or it may also involve the eye (Keratomycosis). 4) Chromomycetoma--Tumefaction, draining sinuses and presence of colony of organisms as "grain". Dematiaceae are found more frequently in veterinary than in human medicine.


Surface architecture of X-irradiated hair follicles. The combined use of scanning electron-microscopy and chemical separation of epidermis permitted the study of the irradiated and normal hair follicle surface characteristics. Irradiation reduced the length and density of the hair follicles, producing at the same time marked alterations and deformities of the individual follicles. As most of these changes were dose- and time-dependent, these structures, studied by these techniques, provide an interesting model as biological indicators of radiation dose.


Subcutaneous fat necrosis of the newborn. Subcutaneous fat necrosis of the newborn (SFNN) developed in a 1-week-old black boy. His mother had received numerous medications for eclampsia. Birth was by Caesarean section and complicated by meconium aspiration. There were numerous nodules over the back, buttocks and extremities that yielded a caseous-like material. Microscopically, these nodules showed crystallization and necrosis of the fat. Hypoglycemia, pneumonia, oliguria, thrombocytopenia, seizures and urinary infection were associated with the cutaneous problem and led to a fatal outcome 2 weeks after birth.


Reiter's syndrome, immunodepression and strongyloidiasis. Report of a fatal case. We present the case of a 52-year-old white male who showed arthritis, conjunctivitis and non-gonococcal urethritis. Besides those manifestations, the patient also showed mucocutaneous lesions that were both clinically and histologically indistinguishable from those of pustular psoriasis. Due to the severity of the disease it was necessary to treat this patient with corticosteroids and immunosuppressors. With this therapy the clinical manifestations of Reiter's Syndrome disappeared but sudden abdominal and bronchopulmonary symptoms complicated the picture and the patient died in a septical shock. The autopsy findings revealed massive Strongyloidiasis with the presence of larvae in several organs, particularly in the intestinal wall, the lungs and the liver.


Isolation and partial characterization of rate casein proteins. Casein was isolated from rat milk by high speed centrifugation. Polyacrylamide disc gel electrophoresis of the whole casein yielded three major protein zones designated C.1, C.2, and C.3 in order of their decreasing electrophoretic mobility in the alkaline system. Zone 3 subsequently contained two possibly related bands, C.3.1 and C.3.2. The presence of phosphate in all four zones was indicated by staining and conformed by phosphorus-32 labeling studies. A glycoprotein character was indicated by all zones. Separation of the constituents of rat casein by diethylaminoethyl-cellulose ion exchange chromatography yielded the same four major protein entities. Three milk-specific phosphoproteins unique to rat whey cluted from such columns in the same general region as the casein constituents but appear to be otherwise unrelated to the four major components of micellar casein. Gel electrophoresis in sodium dodecyl sulfate systems yielded apparent molecular weight estimates of approximately 24,000 for C.1, 38,000 for C.2, and 28,000 for c.3.1 and c.3.2.


Comparison between tail vein and jugular vein cannulation in cattle. Blood samples obtained simultaneously from the jugular and tail (coccygeal) veins during induction of bovine mastitis infection and after administration of 250 IU of adrenocorticotropin did not differ in percentage of hematocrit, concentration of total corticosteroid in plasma, or concentrations of total circulating erythrocytes and leukocytes. Patency of the tail vein cannula was maintained for 1 mo. Tail vein cannulation is a practical alternative to jugular cannulation in cattle.


The nasolabial flap for reconstruction of the alae nasi. Cicatrizing basal-cell carcinomas, especially in the area of the alae nasi, require radical therapeutic treatment. Surgical removal, therefore, often results in penetrating defects. The nasolabial flap is recommended in such cases as a relatively simple operative method of reconstructing the lateral part of the nose. This technique offers favorable results in a curative and esthetic respect.


Treatment of an epithelioma cuniculatum plantare by local excision and a plantar skin flap. Differentiation between benign and malignant lesions on the plantar surfaces is sometimes clinically impossible and requires biopsy. A case of epithelioma cuniculatum plantare is reported that was futilely treated for years as a plantar wart and then upon proper diagnosis was successfully treated by complete excision and rotation of a flap of plantar skin.


Management of alopecia of the scalp by a combination of excisions and transplantations. Since the number of grafts that are available in any given patient is a fixed quantity, significant benefits may be derived by surgically reducing the size of the area of alopecia in combination with prior and later punch hair transplantation. This paper outlines and describes techniques of excisions that achieve such reductions.


Beads of a dextran polymer for the local treatment of cutaneous ulcers. A new polymer of dextran in the form of beads, 0.1 to 0.3 mm in diameter, was applied topically to 39 cutaneous ulcers of various causes. The material, which is powerfully hydrophilic, promptly absorbed serous and purulent exudate from the surfaces of the ulcers. This action decreased the bacterial load and prevented crust formation. Healthy granulation tissue then developed quickly in many ulcers and healing time seemed to be shorter in some cases. The exudate-absorbing capability of the beads of dextran was particularly beneficial in ulcers that had uneven surfaces, undercut margins, and sinus tracts. Complete healing was achieved in some of these cases that had previously resisted many kinds of treatment. The most important undesirable effect of the beads of dextran is their tendency to dehydrate the bases of ulcers too much, which slows or stops healing in some cases. This effect may be overcome by alternating applications of the material with hydrating forms of treatment or changing entirely to such treatment when applications of the beads has removed all exudate from the ulcers. No allergic, irritant, or toxic reaction from the dextran was encountered.


Differential diagnosis of ulcers on legs of vascular cause. This article reviews the clinical features of the different types of leg ulcers due to vascular insufficiency. Diagnostic tests available to confirm or deny clinical impressions are discussed.


Multiple vascular anomalies: report of a case. A patient with multiple congenital vascular anomalies is described. The lesions are a nevus flammeus on the face, a nevus anemicus on the trunk, and asymmetric reticulated mottling of a lower extremity that had undergone spontaneous ulceration. The characteristics of these lesions are reviewed and the difficulty of differential diagnosis of the reticulated lesion is discussed. The ulcerations healed after administration of reserpine intraarterially followed by oral administration of phenoxybenzamine hydrochloride.


Treatment with benzoyl peroxide of ulcers on legs within lesions of necrobiosis lipoidica diabeticorum. A lotion of 20% benzoyl peroxide was applied to ulcers on legs from necrobiosis lipoidica diabeticorum. The ulcers healed rapidly and uneventfully, leaving firm, yellow scars. The efficacy and simplicity of use of topical benzoyl peroxide therapy is discussed.


The healing of wounds in the skin of piglets treated with benzoyl peroxide. Clinical and microscopic changes in the healing of dermal wounds in the skin of piglets that were treated with a 20% benzoyl peroxide lotion were compared with untreated wounds in the same animal. The wounds treated with benzoyl peroxide showed more rapid healing clinically, and a very pronounced foreign-body giant-cell infiltrate and very vascular granulation tissue microscopically. It may be that benzoyl peroxide promotes wound healing by attracting macrophages and histiocytes into the wound.


On the use of comfortable listening levels in speech experiments. In order to investigate the effect of presentation subjects' labeling of speechlike sound patterns, synthetic stimuli were constructed, varying systematically FO contours, VOT, and F2 transitions. These stimuli were presented in random sequences at levels between 15 and 105 dB SPL to subjects with normal hearing. No significant response variation was observed in the range 40--100 dB SPL. Subjects' labeling behavior suddenly breaks down below levels of around 35 dB SPL. The secondary findings of the study are also discussed in terms of different specific processing strategies for different specific speech features.


Temporal summation of constant and gliding tones at masked auditory threshold. Masked thresholds for constant and gliding tones were determined by the method of adjustment for durations between 0.5 and 5000 ms in three overlapping frequency regions between 0.25 and 3.3 kHz. The masker was a continuous white noise at 70-dB SPL. Listening was monaural; subjects had normal hearing. Below 10 ms the thresholds for upward glides were lower and those for downward glides higher than the thresholds for constant tones. In the 10--300 ms duration range, which encompasses formant transitions of speech, the highest thresholds are for downward glides and the lowest ones for constant tones. These differences could result from different time courses of neural decay and inhibition for constant tones, upward and downward glides. The differences between upward and downward glides indicate that the phase spectra influence sound detectability. The thresholds for constant tones reach minimum around 1 s. The thresholds for glides continue to decrease at least up to 5 s. The "critical" duration for constant tone integration can result from the overriding of integration effects by adaptation effects, the latter ones being eliminated by changing frequency. The curves for constant-tone threshold between 10- and 1000-ms duration were fitted by a product of exponential and hyperbolic functions.


On the role of spatial and temporal cues in the perception of the pitch of complex tones. This paper describes a new approach to pitch perception. It focuses attention on the slight difference between the pitch of complex tone and the pitch of a pure tone with the same (fundamental) frequency. This approach is based on the assumption that pitch perception is based on both spatial and temporal cue analysis. In this study, the values provided by the temporal cue are calculated from physiological data from the auditory nerve fibers. The possible ratios of the pitch of complex tones relative to the pitch of pure tones at various frequencies are predicted. Psychophysical experimental results strongly support this prediction. In addition, another experiment suggests that the above psychophysical effect is not based upon a mutual masking effect in the spatial domain.


Adaptation of the acoustic reflex.

Adaptation of the acoustic reflex. The characteristics of acoustic reflex adaptation were studied in human subjects. Contralateral stapedial reflex measurements were made by monitoring changes in acoustic conductance and susceptance with a 220-Hz probe. The reflex activators included four tonal stimuli (500, 1000, 2000, and 4000 Hz) and broadband noise presented for 180 s at 96, 104, and 112 dB SPL. The data are presented as change in acoustic admittance and in percent of maximum admittance change. The static admittance of the ear was found to drift systematically toward lower admittance values in the absence of contralateral stimulation. All stimulus conditions demonstrated some reflex adaptation. The half-life and time of onset of adaptation were found to vary directly with intensity and inversely with frequency. The effects of stimulus parameters on the rates of reflex adaptation are discussed. A previously described exponential model of reflex adaptation provided a reasonable first approximation to the data.