Image formation in fundus cameras.

Image formation in fundus cameras. Imaging in a fundus camera depends more on design of the system than on correction of the first fundus image as formed by the ophthalmoscopic lens. We show here that the designer may use the free parameters of the ophthalmoscopic lens (contact or noncontact) to correct the latter for observation and illumination of the fundus. In both contact and noncontact systems the fundus is illuminated by forming a ring of light on the patient's cornea around a central area (the corneal window) reserved for observation. On the first surface of the crystalline lens, the light also forms a ring which must accomodate the total entrance pupil (TEP) of the observation system in its middle and which is limited on the outside by the patient's iris. The restrictions that result from this situation define the entrance pupil of the bundle of rays that image the marginal point of the retina. The limits of this bundle are imposed by the choice of the angular field of view and by the size of the patient's pupil.


Light-triggered rod disc shedding in Xenopus retina in vitro. The present study was designed to determine whether light-induced rod outer segment disc shedding occurs when the eye is isolated from the rest of the organism. Whole eye explants of Xenopus laevis were maintained in vitro and either exposed to 5 min of light or kept in darkness. In both the in vitro eyes and in vivo controls exposed to the same conditions, a significant degree of disc shedding occurred within 1 hr of the light stimulus. The disc shedding response was larger in the explanted eyes than in the intact animals. In explants exposed to [3H]leucine for 24 hr, a radioactive band formed at the base of rod outer segments. Thus both light-stimulated disc shedding and disc synthesis occur in the eye kept in vitro. The results of this study are compatible with other recently reported results suggesting that rod disc shedding is initiated within the eye.


Effect of dipyridamole on prostaglandin-induced ocular hypertension in rabbits. The effect of intraperitoneal injections of dipyridamole on the elevations of intraocular pressure and anterior chamber aqueous humor protein produced by topical application of prostaglandin E2 was studied in rabbits. Pretreatment with 100 mg/kg dipyridamole inhibited the prostaglandin E2--induced ocular hypertension and rise of aqueous humor protein. Systemic administration of dipyridamole did not alter the responses of the eye to instilled nitrogen mustard. Topical instillation of dipyridamole was ineffective. Dipyridamole, a clinically available drug, may be useful in the treatment of ocular inflammatory disease.


Subject cooperation and the visual evoked response. The effects on the visual evoked response (VER) of subject behavior, such as accuracy of fixation, stability of eye position, and degree of concentration, are poorly documented. Experiments were performed on 20 normal subjects to investigate the effects on the VER of eye movements, off-center stimulation, concentration on another task, and voluntary defocusing. It was found that there are some conditions under which a normal subject can produce an abnormal VER which could lead to misdiagnosis.


Vitamin E deficiency and the retina: photoreceptor and pigment epithelial changes. To investigate the role of normal vitamin E levels and the interrelationships between vitamin E and A in maintaining the visual cells of the retina, weanling female Sprague-Dawley rats were fed vitamin E-free diets differing tenfold in their vitamin A content (0.8 and 8.0 mg of retinol per kilogram of diet). Rats on vitamin E-free diets with the higher vitamin A level exhibited marked disruption of photoreceptor outer segment membranes and a fivefold increase in the number of lipofuscin granules in the pigment epithelial cells which ingest these membranes. Rats on vitamin E-free diets with the lower vitamin A level showed the same retinal damages plus significant loss of photoreceptor cells compared to age-matched rats on control diets. Rods and cones were involved equally, and their pattern of loss was not like that found in vitamin A deficiency. Normal levels of vitamin E probably protect photoreceptor membranes from oxidative damage and retard the accumulation of their remnants and other products of lipid breakdown in the pigment epithelium. The vitamin A status of rats has a significant influence on the extent of damage induced by vitamin E deficiency.


Alterations of lens protein synthesis in galactosemic rats. A differential effect on protein synthesis has been demonstrated in the lenses of galactosemic rats. During galactose cataract development the synthesis of lens crystallins is depressed, whereas that of noncrystallin proteins is unaffected. This effect correlates with the influx of Na+ and loss of K+ from the lens. Removal of the galactose diet results in a gradual recovering of crystallin synthesis to normal levels. In vitro the nuclear cataractous lenses leak crystallins into the media; however, upon 5 day's recovery no leak-out of crystallins could be detected. Both decreased synthesis and leak-out probably account for the marked loss of dry weight of cataractous lenses. These results support the hypothesis that crystallin synthesis may be affected by cation imbalance or changes concomitant with such an imbalance.


Rapid assessment of visual function: an electronic sweep technique for the pattern visual evoked potential. We have developed an electronic spatial frequency sweep technique for electrophysiological assessment of visual acuity and pattern vision. The technique allows an accurate and reliable measurement of VEPs to a full range of spatial frequencies in just 10 sec. Because the measurements are so rapid, the technique suggests several new improvements in the assessment of visual function. Sweeping spatial frequency linearly and extrapolating the high-frequency region of the VEP spatial-tuning function to zero voltage allows an estimate of acuity which correlates highly with psychophysical estimates of acuity. Variants of the procedure are appropriate for the assessment of refracture error, determination of equality of visual function for the two eyes and of binocular interactions, and for sequential assessment of therapeutic conditions.


Suprathreshold spatial frequency detection and binocular interaction in strabismic and anisometropic amblyopia. We have investigated suprathreshold contrast sensitivity and binocular interactions in strabismic and anisometropic amblyopes using a reaction time paradigm. For every spatial frequency, reaction time increased as the grating contrast decreased. At all spatial frequencies and contrast values the reaction times using the amblyopic eye were prolonged compared to the nonamblyopic eye, but most markedly at high spatial frequencies. In the middle range of spatial frequencies, the contrast vs. reaction time function for the nonamblyopic eyes was biphasic, suggesting that two separate mechanisms detect gratings at high and low contrast levels. These functions in deep amblyopia were monotonic, and in shallow amblyopia the break in the functions was present but shifted to lower contrast levels. Binocular interaction experiments showed that binocular summation was absent at all contrast levels, but binocular occlusion was evident at high contrast levels for amblyopic observers.


Interocular alignment following visual deprivation in the cat. Kittens were placed in the dark just after birth and then removed at various ages for the study of interocular alignment. It was found that kittens dark-reared for 4 months or longer were characteristically incyclotorted with respect to normal animals. Deprivation periods of less than 2 months were ineffective in producing these changes. Divergence of the visual axes was also observed in some dark-reared cats. Pupillary constriction in response to light was much more pronounced in dark-reared cats than in normal cats. This enhanced pupillary reaction persisted for at least 3 weeks after the deprived animals were brought into an illuninated environment. When dark-reared cats were allowed a recovery period in a normally lit visual environment, their ocular alignment changed markedly. The incyclotorsion and divergence of the visual axes disappeared, and instead cats allowed recovery from deprivation could display excyclotorsion and/or convergence of the visual axes. These anomalies of ocular alignment associated with the recovery from visual deprivation could occur following periods of initial deprivation as short as 30 days or as long as 2 years. The mechanisms and possible significance of such anomalies are considered.


Role of visual cortex in interocular alignment. The role which the visual cortex plays in the development of interocular alignment in the cat was examined by removing this structure bilaterally in 4 groups of subjects. These included (1) kittens 10 to 14 days of age, (2) 10- to 14-day-old kittens in which one eyelid was sutured shut at the same time, (3) normally reared adult cats, and (4) cats dark-reared until 4 months of age. If the cortex is removed in young kittens, interocular alignment appears to develop normally until the kittens are 60 to 80 days of age. At this time, an abrupt change in alignment resulting in incyclotorsion of the optic axes is observed. If binocular vision is prevented in kittens with neonatal visual cortex lesions by suturing one eyelid shut, convergent strabismus and/or incyclotorsion are frequently observed. This characteristic incyclotorsion does not develop if similar lesions are made in adult cats; no significant alterations of eye alignment occur in these animals even after postoperative survival times of more than 6 months. Incyclotorsion characterizes dark-reared cats when they are first brought into the light, but this diminishes with time and may even be replaced by excyclotorsion after the animals spend a few weeks in the light. If dark-reared cats are decorticated on being brought into the light, these changes are largely prevented. Such animals remain permanently incyclotorted relative to normal cats. The results indicate that the visual cortex plays an important role in the development of torsional alignment of the eyes.


The effects of prolonged dark exposure on visual thresholds in young and adult rats. A comparison of electroretinogram (ERG) thresholds measured on rats reared in continuous darkness and under cyclic lighting conditions shows that by 30 days of age the dark-reared animals have achieved significantly lower thresholds than the animals reared under cyclic illumination. Ten days of continuous dark exposure produces this same increase in sensitivity in adult rats reared in cyclic lighting. These changes in sensitivity appear to reflect structural changes occurring within the rod outer segments.


Retinomotor activity and the c-wave of the hypoxic trout retina. The teleost retina exhibits retinomotor activity in response to changing light intensity. We have shown that hypoxia interfers with normal retinomotor activity in the dark-adapted rainbow trout, so that the retina assumes an essentially light-adapted configuration with the cones contracted, the rods extended toward the pigmented epithelium, and epithelial pigment expanded. These cell movements appear to be correlated with the marked increase in ERG c-wave amplitude which we consistently observe during hypoxia in trout. Since the a-wave is not immediately affected by hypoxia, this increase in c-wave amplitude may be related to the movement of the rods toward the pigmented epithelium, which would cause a greater than normal change in extracellular [K+] near the apical membrane in response to a light stimulus, leading to an increase in c-wave amplitude.


Topical induction of delayed hypersensitivity in the bladder. Delayed hypersensitivity reactions have been elicited by topical application of dinitrofluorobenzene to the bladder mucosa of sensitized dogs and rats. The resultant animal models may be of value in assessing the role of topically induced delayed hypersensitivity in the attempted immunotherapy of bladder cancer.


Ureteral reimplantation with proximal urinary diversion: laboratory investigation. In an effort to determine whether or not urine flow is necessary for proper healing after ureteral reimplantation, a laboratory study was carried out using mongrel dogs. Twelve animals underwent three separate surgical procedures in order to show whether ureteroneocystostomy in the presence of proximal urinary diversion would lead to subsequent stenosis of the reimplantation sites. Our findings indicate that stenosis does not occur regardless of whether or not dry ureteral implants are stented. These studies were carried out in ureters of normal caliber and animals otherwise free of disease. It is our contention that under these circumstances, dry ureteral reimplantation will result in satisfactory healing without stricture or stenosis


Static testicular imaging utilizing 201Tl. Successful testicular imaging of rat and rabbit testes was achieved utilizing the radiopharmaceutical Thallium-201. These observations indicate the usefulness of Thallium-201 as an agent for testicular imaging and suggest a clinical application in localizing the impalpable testis in humans.


Testicular steroid secretion and peripheral serum steroid concentrations in patients with prostatic carcinoma after short-term estrogen treatment. The short-term effects of the administration of estradiol on testicular steroid production were assessed by measuring the serum concentrations of testosterone, and a number of its precursors and metabolites, in the spermatic and peripheral veins of five inguinal hernia patients and one untreated prostatic carcinoma patient, as well as in 18 patients with advanced prostatic carcinoma, treated for 1 to 9 days with a single injection of 80 mg of polyestradiol phosphate (Estradurin). With the exception of progesterone, the treatment resulted in a significantly decreased testicular production of all steroids measured in spermatic vein serum; the same finding holds true for concentrations of testosterone, 5 alpha-dihydrotestosterone, and 17 alpha-hydroxyprogesterone in peripheral vein serum before and after treatment. By comparing the steroid concentrations in the spermatic and peripheral veins it is evident that the testicular production of all steroids measured persisted, although at a decreased level, even 7 to 9 days after Estradurin administration; this was most pronounced in the case of testosterone and 17 alpha-hydroxyprogesterone. It is apparent that the effect of estradiol on testicular steroid production is a gradual process which takes several days before it is clearly evident.


Citrate in daily and fasting urine: results of controls, patients with recurrent idiopathic calcium urolithiasis, and primary hyperparathyroidism. In three groups--patients with recurrent calcium urolithiasis (RCU), patients with primary hyperparathyroidism (pHPT), and healthy controls--citrate was measured enzymatically in 24 and in 2-hr urine after an overnight fast. Citrate excretion per 24 hr was significantly lower in RCU than in age and sex matched controls, whereas there was no significant difference in citrate excretion in urines from the 2-hr morning collection. In pHPT citrate was also lower than in controls and fell within the range of RCU of comparable age. Both categories of urines (24 and 2hr) have in common the characteristic that the actual citrate concentration is lower by 50 per cent in RCU and pHPT than in controls, mainly as a result of the higher urine volume. Correction of citrate for creatinine does not disclose further differences among the populations studied but conversely hampers exact interpretation of urinary citrate in the absence of strict separation of individuals according to sex and age. From these data we conclude that (i) a low excretion and concentration of urinary citrate is detectable in calcium lithiasis and may contribute to a deficiency in inhibitory activity against nucleating processes in stone-forming urine; and (ii) the differences in urinary citrate elicited in samples of 24 and 2-hr morning urine are of unknown origin and merit further investigations.


Effect of renal vein ligation with or without suprarenal inferior vena cava ligation on sodium and phosphate excretions during acute extracellular volume expansion in the rat. Right nephrectomy with or without inferior vena cava (IVC) ligation was performed in rats 1 week after left renal vein ligation. Left kidney function and sodium and phosphate excretions by volume expansion were investigated. Inulin (CIN) and para-aminohippuric acid (CPAH) clearances and fractional excretions of sodium (FENa) and phosphate (FEP) were examined before and after volume expansion was induced by normal saline. Kidney functions were well preserved with development of collateral venous channels; biochemical parameters returned to the normal ranges. However, CIN and CPAH were reduced to 33 to 25 per cent of control values and the filtration fraction was elevated. After volume expansion, a marked increment of FENa was observed in cases of IVC ligation but the increment in animals without IVC ligation was less than in the control group. FEP was also increased in experimental groups but the magnitude of phosphaturia was not so prominent compared to the control group. The increment of FEP was not comparable to that of FENa. Thus, from the aspect of kidney function, this experiment showed the rationality of left renal vein ligation with contralateral nephrectomy. From the pathophysiologic aspect, the magnitude of natriuresis and phosphaturia by volume expansion was not prominent and a reduced reabsorptive capacity for sodium and phosphate may exist in a solitary kidney with a ligated renal vein. When suprarenal IVC ligation was also done, prominent natriuresis was observed with saline infusion.


The effect of vitamin A on the migration and DNA synthesis of rat bladder tumor cell line NBT II in culture. In the presence of vitamin A, NBT II cells, derived from a carcinoma of rat bladder, grew as a monolayer with diminished piling up. Keratinization, which normally appeared within stratified cells in postconfluent cultures, was inhibited. A "wounding" technique suitable for quantitative analysis of cell migration was developed for confluent cultures grown on glass coverslips. Vitamin A treatment enhanced the migration of cells from the wound edge. In dense postconfluent monolayer cultures, vitamin A treatment maintained a higher percentage of cells in DNA synthesis than in the control cultures, as determined by 3H-TdR uptake and autoradiography. In contrast, in sparse cultures vitamin A did not stimulate DNA synthesis or increase the mitotic index. This stimulatory effect, limited to dense cultures, may be attributable to vitamin A causing viable cells to be shed into the medium, thereby maintaining the monolayer just at confluence. Thus vitamin A inhibits squamous cell differentiation, enhances migration, and maintains the culture in the proliferative phase. In a different system of high cell density, NBT II aggregates cultured in a combined matrix of chick plasma clot and collagen-coated sponge, vitamin A also enhanced the migration of cells. These results may explain, in part, the failure of vitamin A to inhibit completely the growth of some established tumors.


Establishment and characterization of a human prostatic carcinoma cell line (PC-3). The establishment, characterization, and tumorigenicity of a new epithelial cell line (PC-3) from a human prostatic adenocarcinoma metastatic to bone is reported. The cultured cells show anchorage-independent growth in both monolayers and in soft agar suspension and produce subcutaneous tumors in nude mice. Culture of the transplanted tumor yielded a human cell line with characteristics identical to those used initially to produce the tumor. PC-3 has a greatly reduced dependence upon serum for growth when compared to normal prostatic epithelial cells and does not respond to androgens, glucocorticoids, or epidermal or fibroblast gowth factors. Karyotypic analysis by quinacrine banding revealed the cells to be completely aneuploid with a modal chromosome number in the hypotriploid range. At least 10 distinctive marker chromosomes were identified. The overall karyotype as well as the marker chromosomes are distinct from those of the HeLa cell. Electron microscopic studies revealed many features common to neoplastic cells of epithelial origin including numerous microvilli, junctional complexes, abnormal nuclei and nucleoli, abnormal mitochondria, annulate lamellae, and lipoidal bodies. Overall, the functional and morphologic characteristics of PC-3 are those of a poorly-differentiated adenocarcinoma. These cells should be useful in investigating the biochemical changes in advanced prostatic cancer cells and in assessing their response to chemotherapeutic agents.


Effects of neonatal steroids on male sex tissues. The effects of neonatally administered sex hormones on the growth and hormone responsiveness of the sex accessory tissues of the adult male rat were investigated. If intact neonatal male rats are treated with estradiol diproprionate, the size of the ventral prostate in the adult animal at day 70 of age is less than 10 per cent of the untreated control value. If androgens are administered to intact neonates, the size of the ventral prostate at day 70 is one-third to one-half of the untreated control values. The androgenic response of these abnormally small prostates was tested by treating these adult animals with testosterone propionate daily from day 70 to day 83. Toward ventral prostates of the animals treated with estrogens in the neonatal period show a very limited response to exogenous androgens at adulthood. In contrast, the ventral prostates of the animals receiving testosterone propionate or dihydrotestosterone propionate in the neonatal period were fully responsive to exogenous androgens. In addition, the ability of the adult prostate to respond fully to androgens requires the presence of the testes in the neonatal period. Comparing neonatal castrates with intact neonates, it was shown that the administration of androgen in the neonatal period was only partially effective in restoring the androgenic responsiveness of the adult ventral prostate. This suggests that other testicular factors, or the presence of testosterone at other time periods before the onset of puberty, may be necessary for the normal androgenic induced response of the adult sex accessory tissues.


Experimental penetrating renal trauma: a comparison of bench and in situ repair. Penetrating renal trauma often requires total nephrectomy. To minimize this high rate of renal loss, extracorporeal (bench) surgery, already an accepted method for treating renovascular and renal tumor, might be used in patients with parenchymal injuries to provide a more meticulous repair and improved renal function. To compare the effectiveness of this technique with that of in situ repair, 32 dogs were divided into two equal groups. They all received a standardized injury to the renal parenchyma. Contralateral nephrectomy was performed. Renal function was evaluated for 21 days. Serum creatinine values were comparable in the two groups. Bench repair offered excellent results but not better than in situ repair for parenchymal and collecting system injuries.


A rat prostatic adenocarcinoma as a model for the human disease. A transplantable, metastasizing prostatic adenocarcinoma (Tumor I) in Lobund Wistar rats was examined for activity and distribution of five hydrolytic enzymes and for ability to accumulate radioactive zinc. The results suggest that the tumor had arisen in the ventral lobe of the prostate and that its growth was not affected by orchiectomy, adrenalectomy, or replacement treatment with exogenous androgen or corticosteroids. The androgen independency of the tumor was further shown by the low uptake of 3H-testosterone, in contrast to the high uptake in the ventral prostate. Tumor growth was retarded by Cytoxan but not by 5-fluorouracil, Estracyt, or streptozotocin, three agents clinically effective in the treatment of some patients with prostatic cancer resistant to endocrine therapy. It is concluded that this tumor in Lobund Wistar rats may be an adequate model for human prostatic cancers resistant to the agents mentioned above.


Renal lactate dehydrogenase (LDH) isoenzyme pattern in short-term experimental obstructive nephropathy. The lactate dehydrogenase isoenzyme pattern of different zones of rat kidney has been investigated at different stages after unilateral ureteral obstruction. In the obstructed kidney, the cortical tissue showed an early (3 days) progressive shift toward a cathodic zymogram; at 7 days LDH-5 isoenzyme was the predominant fraction (44.7 +/- 2.5). In the outer medullary tissue a similar change was observed even though middle fractions were always quite evident. No significant changes were found in the inner medullary tissue. The observed enzymatic abnormalities are related to renal hypoxia and to the occurrence of a less differentiated nephronic cell population. They represent a metabolic marker of the morphologic and functional alterations which occur in obstructive nephropathy.


Muscular development of the bladder neck in the human fetus. We conducted a histologic study of the muscular development of the bladder neck in human fetuses of 5 to 23 cm crown-rump length (CRL) and in neonates in special reference to the middle circular layer of the posterior wall of the bladder neck (posterior circular muscle of the bladder neck) and the deep trigonal muscle. Our attempt was to determine the anatomic relationship between and the origins of the two musculatures. The posterior circular muscle had begun to differentiate in the fetus of 8.3 cm CRL in which the trigonal musculature was indistinguishable. Differentiation of the deep trigonal musculature was evident in the fetus of 17 cm CRL; a complete growth in which the muscle was connected to the posterior circular muscle of the bladder neck was evident only in the largest fetus (23 cm CRL). We conclude that the deep trigonal muscle and the posterior circular muscle of the bladder neck are of different origins.


The state of urethral musculature during the detrusor areflexia after spinal cord transection. The state of the urethral musculature was studied using the urethral pressure profile and electromyographic recording in periurethral striated muscle. In dogs under chloralose anesthesia a micturition reflex could be elicited by bladder distension and somatic reflexes could be elicited by various stimuli. Both the urethral profile and electromyographic activity could be recorded immediately after surgical transection of the spinal cord between T2 and T8 but the bladder remained areflexic for more than 12 hr. Pharmacologic analysis of the urethral pressure profile revealed a substantial contribution from both sympathetic and somatic components. The periurethral striated musculature usually responded to bladder filling in a similar manner both before and after transection, although there was no bladder contraction in the latter circumstance. It is concluded that the urethra does not experience the same depression of reflex activity as does the bladder in the acute stage after spinal cord transection.


Patterns of recovery of renal function after surgical relief of chronic bilateral partial ureteral obstruction. A canine model of bilateral chronic partial ureteral obstruction with one side severely obstructed and the other moderately obstructed was used to determine which kidney should be initially released to facilitate maximum recovery of renal function. The data show a significantly greater return of total renal function (20 per cent) if the more severely injured kidney is released first.


The embryologic origin of the caput epididymidis in the rat. The ACI rat with its known incidence of ipsilateral agenesis of the kidney and ductus deferens provides an excellent mammalian model for the study of the embryologic origin or the caput epididymidis. Because a portion of the caput epididymidis remains in these affected animals, it seems to be derived from the mesonephric tubules rather than the mesonephric duct in the rat. Histochemical assays also support the common origin of both the ductuli efferentes and caput epididymidis from the mesonephric tubules.


Effects of estrogen upon the fine structure of epithelium and stroma in the rat ventral prostate gland. We studied the effects of estradiol upon the fine structure of the rat ventral prostate, with special emphasis on the interstitial tissue. Intact and castrated rats were injected with estradiol or testosterone in combination, or with peanut oil (control). Estrogen exerted an antiandrogenic effect on the epithelium of intact animals resulting in lower epithelial height and reduction in the number of cell organelles and secretory bodies; however, the prostate gland did not atrophy to the same degree as that in castrated controls. In castrates, estradiol had no discernible effect upon the glandular epithelium. Although testosterone alone was able to restore glandular morphology to normal in these animals, estrogen in combination prevented regeneration of rough endoplasmic reticulum and elicited the formation of large lipid-like inclusions and the accumulation of secretory bodies in the apical zone of many cells. Intact animals treated with these hormones in combination exhibited many of the same effects except that a more normal configuration of rough endoplasmic reticulum was present. In castrates, the prostatic stroma became thickened, with a large increase in fibrous material between and surrounding each acinus, although smooth muscle cells retained their normal cytology. With estradiol treatment, singly or in combination with testosterone, smooth muscle cells increased in size and number and organelles decreased in number, cytoplasm became more electron dense, and nuclei became more heterochromatic. Surface vesicles were profuse in smooth muscle cells in animals treated with estradiol alone; larger phagocytic-like vacuoles were characteristic of the glands of animals treated with estrogen and testosterone in combination. In these latter treatment groups, testosterone prevented the overall hypertrophy and hyperplasia of the interstitial tissue; to a degree, endogenous testosterone levels in intact animals treated with estradiol alone prevented these effects as well.


Bladder and urethral responses to sympathetic stimulation. Bladder and urethral pressure responses were recorded in 47 cats subjected to electrical stimulation of sympathetic fibers isolated at the caudal mesenteric ganglion before and after complete sacral denervation. Recorded responses were complex and seemed to be at least partially dependent on afferent and efferent pelvic nerve activity. Sacral denervation alone did not influence urethral closing pressure but the combination of sacral and hypogastric denervation resulted in a marked loss of urethral sphincter function.


Patterns of gastrointestinal neoplasms in Israel. Review of the incidence of gastrointestinal neoplasms in Israel reveals that gastric and colorectal cancers present a major problem in the country. Diet has been postulated as a possible etiologic factor on the basis of a) an indirect correlation between disease incidence and the consumption of various food constituents, b) animal experimentation, and c) case-control dietary studies. Studies conducted in Israel point to high starch consumption as a risk factor in gastric cancer and low fiber intake as a risk factor in colon cancer. Mechanisms by which fiber may exert its protective effect include shortening of intestinal transit time, bulkier stool, lowered absorption of potential carcinogens, reduction of the conversion of bile salts to potentially carcinogenic sterols, interference with the cholesterol pool and a lower ratio of anaerobic:aerobic bacteria in the intestinal flora. The potential effect of excessive starch consumption is unclear, although it may facilitate the formation of nitrosamines.


Epidemiology of Crohn's disease and ulcerative colitis: etiologic implications. Good epidemiologic studies of Crohn's disease and ulcerative colitis are available from only a limited number of geographic locations and populations. A review of the data reveals some interesting facts. There is considerable geographic variation in the incidence of both diseases, which is more marked for Crohn's disease. When various populations living in one geographic location were compared, great differences became apparent with respect to both diseases. Studies performed in nine different populations during the last three decades showed a marked rise, up to 400%, in the incidence of Crohn's disease in each population. No such rise was documented for ulcerative colitis. Studies on Ashkenazic Jews in different parts of the world showed very marked variations in the incidence of both diseases, better documented for Crohn's disease. Both diseases are very rare in several nonindustrialized populations and their incidence rises with migration to an urban environment. All these data point to the preponderance of environmental factors in the causation of these two diseases of unknown etiology.


Epidemiology of noninfective intestinal diseases in various ethnic groups in South Africa. The prevalences of bowel diseases (hemorrhoids, appendicitis, polyps, ulcerative colitis, irritable bowel syndrome, diverticular disease, and colon cancer) are similar in South African whites and in populations of prosperous western countries. Among rural South African blacks with a traditional life style, these diseases are very uncommon or almost unknown. Among the urban South African blacks with a partially westernized life style, the diseases remain uncommon. Frequencies of appendectomies and colon cancer in urban blacks have increased little during the last three decades, although an increase undoubtedly has occurred in the frequency of diverticular disease. Frequencies of bowel diseases in South African Indian and colored (Eurafrican, Malay) populations are intermediate. Because the diseases are almost entirely of environmental causation (due to principally to changes in diet), prevalences almost certainly will increase in blacks, Indians and coloreds, as their way of life becomes further westernized.


Epidemiology of chronic intestinal disease in middle Africa. Gastrointestinal disease in middle Africa is changing. The traditional patterns were high frequencies of parasitic and infectious diseases and sigmoid volvulus, and low incidences of colonic polyps, carcinoma of the colon, appendicitis, diverticulosis, ulcerative colitis and Crohn's disease. The current disease patterns are compared with those for gastrointestinal disease in developed countries and etiological factors are discussed.


Epidemiology of colon cancer in Scandinavia. The occurrence of colon cancer in four Scandinavian (Nordic) countries--Finland, Denmark, Norway and Sweden--is described. In addition, a number of correlations relevant to the etiology of colon cancer are presented. The observable differences in the risk for colon cancer among the Nordic countries can largely be accounted for by socioeconomic factors.


Dietary factors and colorectal cancer in Scandinavia. The food consumption, including type and quantity of dietary fiber, of male population samples in Copenhagen, Denmark, and Kuopio, Finland, with a fourfold difference in incidence of colon cancer was studied. A cohort of male brewery workers in Copenhagen, who consumed approximately four times more beer than did the population at large, was investigated for the development of colorectal cancer. The results suggest that there is a protective effect of dietary fiber, which is not related to mouth to anus transit time, against the development of colon cancer. Beer consumption is unrelated to the development of colorectal cancer.


Fecal bacteria in South African rural blacks and other population groups. Quantitative studies were performed on fecal flora of three population groups consuming different diets. Twenty rural black South Africans and 22 Japanese, representing groups at low risk for carcinoma of the colon, were compared with 41 North Americans from a high-risk population. Specimens taken immediately after defecation were mixed and processed under anaerobic conditions. After the initial incubation, roll tubes were shipped to the United States for final identification. Bacterioides and bifidobacteria were present in lower numbers in South African subjects, as were Bacteroides uniformis (thought to be increased by conditions of fear and anger stress), compared with the other two groups. The number of B. vulgatus and B. distasonis and the "Peptostreptococcus productus species complex," showed a positive correlation with the risk of colon cancer, while an inverse relationship was found with Eubacterium aerofaciens II, B. fragilis and Escherichia coli. The percentage of fecal isolates stimulated by bile was slightly higher in populations with a high fat intake and a high risk of cancer of the colon.


Idiopathic subclinical enteropathy in southern Iran. A study of 100 normal subjects from villages in southern Iran revealed that almost one third of this population had one (or more) absorptive or morphological abnormality of the small-bowel mucosa. The finding of morphologically normal bowel mucosa in stillborn fetuses and in newborns who died within two weeks of birth was consistent with the possibility that the abnormalities noted in the village population were postnatally acquired. Finally, speculations are offered regarding the relationship of idiopathic enteropathy and "immunoproliferative small intestinal disease," which is often seen in the region.


Immune disturbances after severe enteritis during the first six months of life. Infants with severe enteritis and associated prolonged thymic atrophy during the first six months of life showed persistent cell-mediated immune deficiency between the ages of three and five years. Experimental investigations show that early intrauterine and extrauterine stress may lead to permanent cell-mediated immune deficiency and an imbalanced immune response. Trends in disease frequency among different population groups may be influenced in part by the immunity of the host.


Disseminated endocrine cells of the gastrointestinal tract and their possible influence on gastrointestinal disease. The role of the disseminated endocrine cells of the gastrointestinal tract is reviewed. Some of these cells are scarce or absent in pernicious anemia, ulcerative colitis and Sheehan's syndrome, but are increased in number in peptic ulceration, chronic gastritis and celiac disease. A subgroup of the diffuse endocrine cells with both endocrine and exocrine functions, the so-called amphicrine cells, suggests that these cells originate from the entoderm. A study of the diffuse endocrine cells in the chronic intestinal diseases of the developing countries is suggested.


Gastrointestinal lymphomas in Hong Kong Chinese. Forty-six Chinese patients with primary gastrointestinal lymphoma were analyzed. The most common presentations were with abdominal pain, bleeding or a palpable mass. Only one case resembled the pattern of the so-called "Mediterranean lymphoma." The most common primary sites were the stomach and the ileum. One case of colonic lymphoma was associated with schistosomiasis but was probably not etiologically related. The pattern of primary gastrointestinal lymphoma in Chinese corresponds in general to the pattern observed in western countries.


Primary malignant lymphoma of the small intestine in Israel. Changing incidence with time. One hundred and eighty-one cases of primary small intestinal lymphoma were diagnosed in residents of Israel--145 Jews and 36 Arabs--during a 16-year period, 1960--75. The male:female ratio was 1.8:1. Incidence rates were higher in children, the middle-aged and the elderly than in teenagers or young adults. Crude and age-adjusted incidence rates were higher in Arabs than in Jews. The pattern among the subgroups of the Jewish population varied with age. In young adults of both sexes the rates were much higher among Jews born in North Africa or Asia than among those born in Europe or Israel. In elderly males, however, the rates were almost twice as high in European-born Jews than among those born in North Africa or Asia. The mean annual incidence of primary small intestinal lymphoma fell from 4.8 per million during 1960--67 to 3.6 per million during 1968--75. This was due to a marked fall in the rates in children and young adults, whereas the rates in those aged 40 years or more rose with time. The pattern of the change with time suggests that environmental conditions are an important factor in the causation of primary intestinal lymphoma in young adults.


Premalignant lesions of the stomach.

Premalignant lesions of the stomach. The histopathogenesis of the three major types of gastric carcinoma--intestinal, diffuse and indeterminate--is reviewed, considering normal and abnormal cell kinetics of the gastric mucosa. Intestinalization is a precursor of intestinal-type carcinoma and is related to environmental factors. Diffuse gastric carcinoma originates from dysplasia of the neck of the gland and is of nonenvironmental, probably genetic, origin.


Epidemiologic studies of gastric carcinoma: comparison between Iranians and two racial groups in the USA. Gastric carcinoma in 226 cases from the two major racial groups--black and white--in Richmond, VA, USA, and in 75 cases from Shiraz, Iran, were studied. The age at onset and the ratio of intestinal to diffuse gastric carcinoma were compared with those for the high- and low-risk groups from Colombia. Poor socioeconomic conditions are directly related to intestinalization of the gastric mucosa and onset of carcinoma at a relatively young age. In Shiraz, the peak incidence of lymphoma, including intestinal lymphoma, is in the third decade of life; for esophageal carcinoma, it is in the fourth decade, and for gastric carcinoma, in the fifth. Comparison of this age distribution with induction times of neoplasia in immunosuppressed patients supports the suggestion of a common cocarcinogenic factor acting early in infancy. In Richmond, the ratio of intestinal to diffuse carcinoma for the black population is consistently higher than for the white. This difference has increased recently due to a more rapid decline in incidence of intestinal-type gastric carcinoma in the white population than in the black.


Anemia and secondary hyperparathyroidism in chronically hemodialyzed patients. A role in the pathogenesis of the anemia of chronic renal failure has been attributed to secondary hyperparathyroidism. An amelioration of anemia after subtotal parathyroidectomy has been described. In the present study, no correlation was found between the severity of anemia and the level of serum parathormone in dialyzed patients. Furthermore, in seven patients, no improvement of the anemia was observed after parathyroidectomy.


Delivery at 40 years of age and over.

Delivery at 40 years of age and over. This is a retrospective review of 494 deliveries during a 10-year period in women aged 40 or more years. A decline in the incidence of deliveries in this age group was noted, from 2.1% in 1965 to 1.2% in 1974. An increased frequency of complications during pregnancy and delivery, a high rate of cesarean sections, and increased perinatal mortality rate, and more congenital malformations were recorded as compared with both a control group of randomly selected grand multiparas and a general obstetric population group of patients under the age of 40 years. The fetal outcome improved during the last years of the study; the perinatal mortality rate dropped from 10.4% in 1965 to 1.6% in 1973 to 0% in 1974. This achievement reflects the significant progress in obstetrics that has been made during these years.


Diagnostic value of cross-sectional echocardiography in left atrial myxoma. Atrial myxoma is a rare condition that may have unusual clinical manifestations. Two patients with left atrial myxoma are described. One presented with paroxysmal nocturnal dyspnea and no abnormal physical signs on clinical examination; the second presented with palpitations and physical findings suggestive of mild mixed mitral valve disease. Echocardiography and cross-sectional echocardiography confirmed the diagnosis of atrial myxoma in both patients, and the tumors were successfully removed by operation. Cross-sectional echocardiography may obviate the need for cardiac catheterization in patients with left atrial tumors.


Sheehan's syndrome with diabetes insipidus. A case study. A case of a woman in whom Sheehan's syndrome and diabetes insipidus were found is presented. This association is very uncommon, but well-documented cases have been reported. Controversial opinions about the primary lesion still exist. To find out whether the pituitary gland responds to hypothalamic releasing factors, three kinds of test were performed. The failure of the pituitary to respond suggested pituitary damage. Because diabetes insipidus was present, presumably the hypothalamus was also damaged. A review of the literature and different opinions about the primary anatomic lesions in this syndrome are discussed.


Effect of dibromochloropropane on human testicular function. Severe impairment of spermatogenesis was found in 18 (78%) of 23 workers after exposure to 1,2-dibromo-3-chloropropane (DBCP). Azoospermia associated with elevated plasma follicle-stimulating hormone was diagnosed in 12 workers whose exposure time to DBCP varied from 100 to greater than 6,000 h. Oligospermia and normal plasma follicle-stimulating hormone were found in six employees with exposure times between 34 and 95 h. Plasma luteinizing hormone, testosterone and thyroxine were within the normal range. Testicular biopsy showed selective atrophy of the germinal epithelium, intact Sertoli cells and normal appearance of Leydig cells. The mechanism and potential reversibility of these testicular changes are still to be evaluated.


Serum concentrations of isosorbide dinitrate produced by a sustained-release capsule. The bioavailability of isosorbide dinitrate (ISDN) and the time course of its serum concentration produced by a standard 20-mg tablet and a sustained-release capsule were compared in six normal volunteers. Following administration of the tablet, serum ISDN concentrations reached a peak of 29 ng/ml at 0.5 h, declined monoexponentially with a t1/2 of 25.5 min, and disappeared after 2 to 3 h. Following the administration of the sustained-release capsule, serum ISDN concentrations reached a plateau of 4 ng/ml between 2 and 5 h, and declined slowly and nonlinearly up to and including the 10-h point. The total amount of ISDN absorbed was 1.32 times greater with the standard tablet; following the absorption of the capsule, there was truncation of the serum concentration curve due to ongoing absorption at and beyond 10 h.


Asplenia syndrome. Report of two cases.

Asplenia syndrome. Report of two cases. Two babies with asplenia syndrome are described. Both presented with the severe cardiovascular anomalies found in this syndrome and had Howell-Jolly bodies in peripheral blood smears. The chest X-rays showed dextrocardia, decreased pulmonary circulation and a horizontal symmetrical liver, and were helpful in the clinical diagnosis. Additional findings at autopsy included a mobile cecum and bilateral trilobed lungs with bilateral eparterial bronchi.


Successful treatment of Cushing's disease with o,p'-DDD followed by pituitary irradiation in a 19-year-old male patient. A 19-year-old male patient with Cushing's disease was treated for 15 months with a gastric-insoluble preparation of o,p'-DDD. The daily o,p'-DDD dose (range, 2 to 12 g) was adjusted periodically according to the urinary excretion of 11-hydroxycorticosteroids. Because of a rise in the plasma ACTH level from 135 to 300 pg/ml 12 months after the start of the o,p'-DDD therapy, the dose was reduced from 6 to 2 g/day and external pituitary irradiation (4,480 rads) was initiated. Insulin-induced hypoglycemia and stimulation tests with luteinizing-hormone-releasing factor and thyrotropin-releasing hormone, performed before initiation of o,p'-DDD and six months after pituitary irradiation, did not reveal any disturbance in the secretion of human growth hormone, thyroid-stimulating hormone, luteinizing hormone, follicle-stimulating hormone or prolactin. The clinical and laboratory signs of Cushing's disease disappeared gradually, and the patient tolerated the drug well, even at a dose of 12 g/day. The only abnormalities found during o,p'-DDD treatment were low serum thyroxine levels, which returned to normal after discontinuation of the drug, and a transient drop in thrombocyte count. At present, two years after the discontinuation of o,p'-DDD therapy and pituitary irradiation, the patient is symptom free and receives no medication.


[Histology of a plexiform neuroma with Vater-Pacini-lamellar-corpuscle-like structures]. From the finger of a 11 year old boy a tumor that had been present since birth was removed. Histological examination showed a great number of immature nerve bundles, and structures resembling Pacinian corpuscles. It could be shown that the organoid structures are a result of a proliferation of the so-called perineurial cells. These formations do not correspond with real tactile corpuscles, rather they can be considered as neoplastic structures of the perineurium.


[Studies on monilethrix].

[Studies on monilethrix]. Monilethrix is a disorder of the hair manifested by fragile, beaded hair and baldness. The four children described below showed no regulary rhythm of node and internode formation. Light microscopy supported the hypothesis that periodic inhibition of keratin synthesis explains the production of the constriction. Scanning electron microscopy shows longitudinal ridging of the internodes in monilethrix and distinctive surface irregularity. Chromatography of urine for argininosuccinic acid do not support the theory that this hair abnormality is a metabolic defect.


[Clinical picture and x-ray therapy of actinic cheilitis]. 97 patients suffering from cheilitis actinica treated with x-rays between 1959 and 1977 were followed up. The cure rate was 98%. Early diagnosis, the necessity of treatment and the problems of prophylaxis are emphasized. Good results can be obtained only by a total dosis of 4,000 R.


[Pathological-anatomical teaching models in moulages. Manufacturing technic and history]. Wax moulages were used as pathological-anatomical teaching objects in medicine well into this century. On the occasion of the restauration of the moulage collection of the Department of Dermatology, University of Munich, the history of Munich wax artists is reviewed. The historical development of reproductions in wax in previous centuries is described. Several moulages showing various dermatological diseases are pictured. The technical details of moulage manufacture are mentioned.


[Overall progeroid aspect of the Louis-Bar syndrome: pigmentary and keratotic phenomena]. A patient with ataxia-teleangiectasia is presented who developed numerous actinic keratoses and pigmented spots predominating the clinical picture with only few teleangiectasis. Premature ageing of the skin is perhaps the reason for the marginal dermatologic symptoms of this disease.


Evaluation of diagnostic procedures for detection of mycoplasmal pneumonia of swine. The complement-fixation test (CFT), a latex-agglutination test (LAT), and Mycoplasma hyopneumoniae isolation procedures were compared with gross and microscopic lung evaluations for the routine diagnosis of mycoplasmal pneumonia (MP) of swine. The difficulty and variability of M hyopneumoniae isolation rendered this technique unacceptable. In market-weight swine, the finding of typical gross lesions was correlated with CFT results (P less than 0.005). The MP status of a herd could not be determined by use of these techniques in 4- to 9-week-old pigs. No technique could be used alone to establish a diagnosis. Gross and microscopic examination of all slaughter specimens, CFT, and clinical evaluation of the herd, used in combination, were the most useful means of determining the MP status of the herd.


Electromyographic and esophagomanometric findings in clinically normal dogs and in dogs with idiopathic megaesophagus. Electromyography of 12 clinically normal dogs and 7 dogs with idiopathic megaesophagus revealed trains of positive sharp waves in the muscles of facial expression and in the lingual muscles of both groups. Positive waves are usually indicative of motor-unit disease; however, they are clinically insignificant in these muscles. Positive sharp waves were detected in the esophageal muscle of one dog with congenital megaesophagus. Esophageal electromyograms obtained in a dog with congenital megaesophagus and in 2 clinically normal dogs were normal. Resting caudal esophageal sphincter pressure was similar in both clinically normal dogs (mean, 22.3 mm of Hg; range, 15--37 mm of Hg) and in dogs with congenital or acquired idiopathic megaesophagus  (mean, 29.6 mm of Hg; range, 20--50 mm of Hg).


Trigonal-colonic anastomosis: a urinary diversion procedure in dogs. Trigonal-colonic anastomosis for diversion of urine into the colon was performed in 12 clinically normal dogs and in 10 incontinent dogs with diseases of the urinary bladder or urethra. Dogs were studied from 1 to 30 months after surgery. The surgical procedure was technically satisfactory. Fifteen of 22 dogs were studied with intravenous urography, and only 1 case of hydronephrosis was found. Pyelitis was a common histopathologic finding in both groups of dogs. Pyelonephritis developed in 30% of dogs, regardless of duration of anastomosis. Glomerular filtration rate was reduced in all dogs studied, but renal failure was infrequent. Values for blood urea nitrogen and serum inorganic phosphorus were elevated due to intestinal recycling of nitrogenous products and phosphate. Electrolyte imbalances were not a problem, but gastrointestinal disturbances developed in 3 of the 10 diseased dogs. Six of 10 diseased dogs survived from 9 months to more than 3 years. Trigonal-colonic anastomosis appears to be a satisfactory salvage procedure for incontinent dogs with diseases of the urinary bladder or urethra that do not respond to other forms of therapy.


Cranial thoracic diskospondylitis in two dogs. Cranial thoracic diskospondylitis was diagnosed in 2 dogs. Staphylococcus aureus was isolated from the vertebral lesion of both dogs, from the urine of one dog, and from the blood of the other dog. In each case, curettage of the affected disk and contiguous vertebrae was accomplished via thoracotomy. Both dogs recovered following curettage and antibiotic therapy.


Immunotherapy in ocular equine sarcoid.

Immunotherapy in ocular equine sarcoid. A modified Bacillus Calmette-Guerin (BCG) preparation was used successfully in the treatment of 7 cases of equine sarcoid. The BCG preparation was injected into the lesions. The longest period of remission has been 24 months, and the shortest period of remission has been 9 months, with no signs of recurrence of the tumor in any of the presented cases.


Roles of predisposing factors and gonadal hormones in the buller syndrome of feedlot steers. In a study of the role of predisposing factors and gonadal hormones in the buller syndrome, it was found that weather, amount of space available to each steer, and entry weights of the steers were not associated with increased occurrence of bulling; however, the type and timing of administration of the hormonal implant, the number of steers in a pen, and the manner in which the steers were grouped after arrival at the feedlot were found to influence the incidence of the syndrome. Serum concentrations of estradiol and testosterone in the buller steers were found to be lower during bulling than after a short period of isolation and apparent recovery from the syndrome. It was concluded that the occurrence of bulling is related to the use of hormonal implants and certain feedlot management procedures but that it is apparently not related to increased serum estrogen or testosterone concentrations.


Availability of oral carbohydrates to neonatal calves. Four experiments were conducted to determine the availability of 3 carbohydrate sources given orally to 1- to 3-week-old clinically normal calves. In terms of effect on plasma glucose concentration, commercial fruit pectin was equivalent to glucose, corn syrup was less effective (P less than 0.05) than glucose, and sucrose was totally unavailable. The addition of 80 mM NaCl to a 10% glucose solution enhanced glucose absorption (P less than 0.01). Increasing the glucose concentration of a commercial product from 1% to 3% resulted in a significant (P less than 0.01) increase in plasma glucose concentration. Lactose availability from suckled fluids was greater (P less than 0.01) than that from fluids given by stomach tube.


Water deprivation test in the dog: maximal normal values. Studies were undertaken to determine maximal urine osmolality and urine specific gravity following water deprivation for 20 dogs with normal renal function. In addition, the reliability of body weight, skin pliability, total plasma protein concentration, and packed cell volume as indices of negative water balance was assessed. Following water deprivation for periods sufficient to induce dehydration, the mean maximal urine osmolality was 2,289 mOsm/kg. The corresponding mean maximal urine specific gravity was 1.062 and ranged from 1.050 to 1.076. The ratio of mean maximal urine osmolality to mean serum osmolality at the time of peak urine concentration was 7.3. There was no detectable difference in urine concentration indices between males and females. Changes in skin pliability and packed cell volume proved unreliable as estimates of dehydration. Weight loss and increases in total plasma protein concentration proved to be more consistent indicators of hydration status. Abnormal increases in serum urea nitrogen and serum creatinine concentrations occurred rarely, even though some dogs had water withheld for periods of up to 96 hours.


Renal and ureteral duplication in a dog. Duplication of a kidney and ureter in an 18-month-old male English Bulldog was demonstrated radiographically and was confirmed surgically. Urinary tract infection had been a problem for a year. Antimicrobial therapy resolved the clinical signs of urinary infection but did not eliminate bacteriuria.


Clinical evaluation of glomerular function: 24-hour creatinine clearance in dogs. Methods of renal clearance to measure glomerular filtration rate (GFR) were compared with plasma creatinine concentration in clinically normal and partially nephrectomized dogs. Glomerular filtration rate was measured by use of a simple 24-hour creatinine clearance method in 36 normal female Beagles. Mean values were 57.6 +/- 9.3 ml/minute/m2 of body surface or 3.7 +/- 0.77 ml/minute/kg of body weight. Variability of this measurement was considerable, as determined in 4 dogs studied on 4 consecutive days. Glomerular filtration rate was measured in the same 36 dogs while they were under anesthesia, using short clearance periods to compare inulin and endogenous creatinine clearance. Mean values for inulin were 41.8 +/- 13.9 ml/minute/m2 of body surface. A close agreement with creatinine clearance was found (correlation coefficient, 0.998). Mean plasma creatinine concentration was 0.82 (range, 0.5--1.0) mg/100 ml. The value of GFR measurement compared with plasma creatinine concentration was determined in 10 dogs after 75% nephrectomy. Sixty days after partial nephrectomy, GFR was reduced to 61% of normal. Mean plasma creatinine and blood urea nitrogen were 1.2 +/- 0.14 mg/100 ml and 20.4 +/- 7.1 mg/100 ml, respectively. Thus, the detection of reduced renal function may be uncertain when plasma creatinine or blood urea nitrogen are used as a means of evaluating renal function. It was concluded that a simple method of creatinine clearance is a sensitive and useful measurement to detect early or borderline reduction in glomerular function.


Traumatic pericarditis in a dog.

Traumatic pericarditis in a dog. Signs of congestive heart faiure in a dog were caused by pericardial effusion and fibrous thickening of the epicardium and pericardium. Partial pericardiectomy and removal of the probable cause (shotgun pellets) resulted in recovery.


Primary cutaneous coccidioidomycosis in a dog and a cat. Primary cutaneous coccidioidomycosis was diagnosed in a dog and a cat examined because of lymphangitis and lymphadenitis associated with skin wounds. This benign and self-limiting form of disease was distinguished from the skin lesions associated with systemic coccidioidomycosis by means of historic, physical, and serologic criteria established in human medicine.


Feline pyothorax caused by a Borrelia-like organism and Corynebacterium pyogenes. A 2-year-old, neutered, male cat was euthanatized because of difficult repiration and inability to rise. Necropsy revealed pyothorax. Corynebacterium pyogenes was grown from the purulent thoracic exudate. A Borrelia-like microorganism was demonstrated in the exudate, using phase-contrast microscopy.


Isolation and concentration of organophosphorus pesticides from drinking water at the ng/L level, using macroreticular resin. A screening method has been developed for determining organophosphorus pesticides at ng/L levels in drinking water. Sixteen organophosphorus pesticides, diazinon, diazinon-oxon, dimethoate, ronnel, beta-phosphamidon, methyl parathion, ethyl parathion, malathion, chlorpyrifos, fenitrothion, ruelene, methidathion, ethion, EPN, phosalone, and phosmet, were extracted by Amberlite XAD-2 resin from 100 and 200 L drinking water previously spiked with these pesticides. The pesticides were eluted from the XAD-2 resin with acetone-hexane (15+85). The concentrated extract was analyzed by gas chromatography using a nitrogen-phosphorus selective detector and by gas chromatography-mass spectrometry using selected ion monitoring. Recoveries at the 10 and 100 ng/L spiking levels were greater than 90%, except recoveries for dimethoate and phosphamidon were 37 and 42%, respectively. The analysis of 300 L Ottawa tap water showed no detectable amounts (less than 1 ng/L) of any of the 16 organophosphorus pesticides.


Gas-liquid chromatographic-mass fragmentographic determination of low levels of diethylcarbonate in beverages. Sodium chloride and ethanol (omitted for samples with greater than 10% alcohol) are added to the beverage sample and the sample is allowed to equilibrate in a 30 degrees C water bath. An aliquot of the headspace is injected into a gas chromatography containing a column packed with 0.2% Carbowax 1500 on 80--100 mesh Carbopack C. During the elution of diethylcarbonate (DEC), an impurity that is present in diethylpyrocarbonate, the column effluent is vented to a mass spectrometer with a multiple ion detection system and operated in the electron impact mode. The ions at m/e 63 and 91 are monitored. Lemonade, fruit drinks, wine, and beer samples (138 total) were analyzed for DEC. Sixteen samples had greater than 30 ppb DEC. Eight analyses of a lemonade sample gave a mean of 88 ppb with a coefficient of variation of 11%.


High pressure liquid chromatographic determination of furazolidone in turkey tissue. A high pressure liquid chromatographic method for determining furazolidone in turkey tissue has been developed. Tissues are ground with methanol and centrifuged. For lower levels of furazolidone, 2--40 ppb, the supernate is evaporated to dryness and redissolved before it is injected onto the liquid chromatographic column. Using a reverse phase column and an ultraviolet absorption detector set at 365 nm, the assay is linear over the concentration range 2--400 ppb with a coefficient of variation of less than 4%. Average recovery from fortified tissues was 96% with a coefficient of variation of 6% at the 50--400 ppb level, and 105% with a coefficient of variation of 11% at the 2--40 ppb level.


Gas-liquid chromatographic determination of individual sugars in confectionery products. A gas-liquid chromatographic (GLC) procedure is presented for the separation and quantitative determination of sucrose, lactose, maltose, and glucose in commercial confectionery products. By converting reducing sugars to oximes and then forming trimethylsilyl ethers of these compounds and separating them on a 6 ft X 4 mm id glass column packed with 2% OV-17 on 100--120 mesh Supelcoport, single peaks were obtained for each of the sugars. Results for sugars present in samples at levels of 5% or more are within 2.8%, on the average, of results obtained by polarization measurements. The data also compare favorably with others in the literature on similar products analyzed by other GLC procedures that do not involve oxime formation.


Spectrofluorometric determination and thin layer chromatographic identification of tyramine in wine. A method is described for determining tyramine in wine by sand column extraction in an alkaline medium with anhydrous sodium sulfate. Tyramine is identified and quantitated by spectrofluorometry after the final extract is reacted with alpha-nitroso-beta-naphthol; identity is confirmed by thin layer chromatography. The average recovery was 98.93%. The method is applied to samples of 3 different wines obtained throughout the vinification process. Tyramine, which was not present in the must, appears in considerable quantities 15 days after the vinification process has begun.


Flotation technique for quantitative determination of mite populations in powdered and compacted foodstuffs. Samples are preheated in 25% aqueous ethanol at 65 degrees C for 5 min. A specially designed flotation flask and solvent mixture of kerosene-tetralin (1+1) is used to form an interface with 25% aqueous ethanol. Mites, insects, and insect fragments are separated from vegetative material by an overflow method. Trials incorporating a known number of mites in samples of pigmeal indicate that at an infestation level of 1000 mites/kg the method always recovered 70% of the population and at 2000 mites/kg, 75%. The method permits collection of mite specimens for microscopic examination as well as allowing differentiation between live and long-dead insects. It does not use highly volatile solvents or corrosive acids. The apparatus has been successfully tested for quantitatively determining mite populations in flour, dried skimmed milk, animal feedstuffs, soya meal, wheat, rape seed, dates, and house dust. It has also been used to estimate insect contamination in packaged dates.


Application of gel permeation chromatography and nonaqueous reverse phase chromatography to high pressure liquid chromatographic determination of retinyl palmitate and beta-carotene in oil and margarine. A high pressure liquid chromatographic method was developed using high pressure gel permeation chromatography (HP-GPC) and high pressure reverse phase chromatography (RP-HPLC) for quantitation of retinyl palmitate and beta-carotene. HP-GPC was used for fractionation of vitamin A active compounds from oil preliminary to quantitation on nonaqueous RP-HPLC. HP-GPC fractionation was completed on oil and margarine dissolved in methylene chloride by 2 elution passes through 2 muStyragel (100 angstrom) columns connected in series with methylene chloride as the mobile phase. RP-HPLC separation of retinyl palmitate and beta-carotene was achieved on muBondapak C18 (10 micrometers), using methylene chloride-acetonitrile (30+70). Based on 10 repetitive analyses, recoveries of added beta-carotene and retinyl palmitate from vegetable oils were 98.6+/-2.9 and 95.2+/-2.6%, respectively. The coefficients of variation were 2.9% for beta-carotene and 2.7% for retinyl palmitate. The determination of vitamin A activity in 7 margarine brands with label claims of 10% U.S.RDA/serving revealed that all but one of the margarines contained at least 94% of the label claim. Vitamin A activity in the margarines ranged from 90.6 to 110.8% of the label declaration.


Determination of crude protein in animal feeds, using block digestion followed by steam distillation: collaborative study. A method consisting of digesting animal feeds in a block digestor and determining ammonia by steam distillation followed by titration has been evaluated and compared with the official final action Kjeldahl method, 7.016. Fifteen laboratories analyzed 5 feed samples and lysine monohydrochloride. Statistical analysis showed that results from the 2 methods were comparable. The distillation technique has been adopted as official first action as an alternative technique for ammonia determination from the digest of the official final action block digestor method, 7.B11.


Gas-liquid chromatographic determination of morphine, heroin, and cocaine. Morphine, heroin, and cocaine are quantitatively determined with the same gas-liquid chromatographic system. The compounds are separated on a 6 ft X 2 mm id glass column packed with a 1:1 mixture of 5% SE-30 on 80--100 mesh Chromosorb W and 3% OV-17 on 80--100 mesh Varaport 30. The column is temperature-programmed. Flame ionization detector responses are measured with a computer-based data system. Heroin and cocaine are chromatographed directly; morphine is derivatized first. The procedure was evaluated with previously analyzed commercial and forensic samples. Accuracy and precision were 5 and 3%, respectively.


Screening test for amygdalin in tablets, solutions, powders, and seeds. A rapid screening test for detecting amygdalin in tablets, solutions, powders, and seeds, based on the liberation of both hydrogen cyanide and benzaldehyde as a result of enzymatic decomposition, is described. The procedure can be easily performed outside the laboratory with small amounts of sample; the samples require no special preparation or pretreatment. The test can be performed in less than or equal to 60 min and is sensitive to at least 0.2 mg amygdalin. Vicianin, lucumin, prunasin, and sambunigrin would also given positive tests, but instrumental analysis can be used to distinguish amygdalin from these compounds.


High pressure liquid chromatographic determination of the five major alkaloids in Papaver somniferum L. and thebaine in Papaver bracteatum Lindl. capsular tissue. A high pressure liquid chromatographic isocratic procedure is described for determining and quantitating the 5 major alkaloids narcotine, papaverine, thebaine, codeine, and morphine in Papaver somniferum L. and thebaine in Papaver bracteatum Lindl. Other papaveraceous alkaloids, including salutaridine, oripavine, laudanosine, isothebaine, cryptopine, alpinigenine, narceine, protopine, and gnoscopine, were also quantitated. The values for morphine, codeine, and thebaine in P. somniferum were in agreement within 5--9% with values obtained by the United Nations Narcotics Laboratory by other methods. In contrast to previously reported procedures, the advantage of this method is that no precolumn or other purification other than solvent extraction of the capsular tissue is necessary. Isocratic chromatography alone on a single column resolved the 5 major alkaloids.


Determination of vitamin B1 in pharmaceutical preparations using a lead ion selective electrode and atomic absorption spectroscopy. Simple, rapid, and accurate methods have been developed for determining vitamin B1 in pharmaceutical preparations without prior separation of the vitamin. The methods are based on the desulfurization reaction with potassium plumbite and measurement of the unreacted lead(II) ions at 217 nm using atomic absorption spectroscopy (AAS), or by titration with EDTA at pH 4.5 with the lead ion selective electrode and Gran's plot. The results compare favorably with those obtained using the USP fluorometric method. Recoveries of pure vitamin B1 averaged 99.1%, with a standard deviation of 0.8%, and 99.0%, with a standard deviation of 0.7%, for the AAS and ion selective electrode procedures, respectively.


Relative productivity of five selective plating agars for the recovery of Salmonella from selected food types. During a 3-year period, the relative productivity of brilliant green (BG), bismuth sulfite (BS), Salmonella-Shigella (SS), Hektoen enteric (HE), and xylose lysine, desoxycholate (XLD) agars for recovering Salmonella from 9 food types was determined. Following pre-enrichment, selective enrichment of food samples in tetrathionate broth followed by streaking to BS agar was the single most productive selective enrichment broth-agar combination for recovery of Salmonella in 5 of these food types. A study of the performance of these 5 agars used individually and in various combinations, showed that none of the 5 agars used individually nor any of the possible paired combinations of these agars could be used to satisfactorily detect Salmonella in the 9 food types. The use of all 5 agars was not necessary because one combination of 4 agars (BG, BS, HE, and XLD) recoverd 100% of the Salmonella isolates, as compared with the number of Salmonella isolates recovered by the 5-agar combination, in each food category. This particular 4-agar combination, along with two 3-agar combinations (BG, BS, and XLD agars, and BS, HE, and XLD agars), were each able to recover more Salmonella isolates, than the combination of BG, BS, and SS agars, the combination currently recommended by the AOAC. Finally, the relative costs of using these agars, singly and in various combinations, were determined.


The need to be held: state or trait variable. The need to be held has been observed to be related to various moods and personality dimensions. It is unclear, however, whether this need functions as a state or trait variable. Data were collected from 85 pregnant subjects at 4 months, 9 months, and 6 weeks postpartum. Measures of the need to be held, neuroticism, relationship with parents, self-concept and maternal attachment were used to determine whether the need to be held is a trait or state variable, as well as to explore the etiology of the need to be held. Data were analyzed by a trend analysis of repeated measures and analysis of variance. Results showed that the need to be held is a trait variable in that it did not change throughout pregnancy and is related to the predisposition of depression. The etiology of the need to be held remains open for investigation.


The relationship between stressful life events and hospitalized adolescent psychiatric patients. The nature and magnitude of psychosocial stresses preceding adolescent hospitalization were investigated. The findings indicate that the adolescent psychiatric patients have a greater magnitude of stressful life events when compared with general hospitalized adolescents and normal adolescents. The difference is found to be largely quantitative rather than qualitative. The results of the present study utilizing a combined version of Coddington's Junior High School and Senior High School Social Readjustment Rating Questionnaires is similar to previous findings on adult populations using the Social Readjustment Rating Scale.


Appearance of new symptoms in hospitalized patients: an instance of institutional iatrogenesis? Data from the admission and termination forms of 100 consecutively discharged psychiatric inpatients were analyzed to determine the changes that occurred in the symptom profile of each patient during hospitalization. In general, patients entered the hospitalization. In general, patients entered the hospital with severe, chronic symptoms some of which were relieved and some of which remained unchanged. A number of symptoms, however, were reported present on the termination forms that had not been noted at admission. The results of this study offer evidence that a possible effect of the therapeutic process is the occasional appearance of new symptoms in some patients.


Manic psychosis in a patient with multiple metastatic brain tumors. A 45-year-old man, with neither previous psychiatric dysfunction nor a family history of manic-depressive illness, developed a manic psychosis coincident with the growth of 4 metastatic tumors in the right cerebral hemisphere. The psychosis responded well to treatment with lithium even though the metastatic lesions could not by eradicated with surgery or radiation.


The interaction of a tyrosyl residue and carboxyl groups in the specific interaction between Streptomyces subtilisin inhibitor and subtilisin BPN'. A chemical modification study. An ultraviolet absorption difference spectrum that is typical of a change in ionization state (pKa 9.7 leads to greater than 11.5) of a tyrosyl residue has been observed on the binding between Streptomyces subtilisin inhibitor (SSI) and subtilisin BPN' [EC 3.4.21.14] at alkaline pH, ionic strength 0.1 M, at 25 degrees C (Inouye, K., Tonomura, B., and Hiromi, K., submitted). When the complex of SSI and subtilisin BPN' is formed at an ionic strength of 0.6 M and pH 9.70, the characteristic features of the protonation of a tyrosyl residue in the difference spectrum are diminished. These results suggest that the pKa-shift of a tyrosyl residue observed at alkaline pH and lower ionic strength results from an electrostatic interaction. Nitration of tyrosyl residues of SSI and of subtilisin BPN' was performed with tetranitromethane (TNM). By measurements of the difference spectra observed on the binding of the tyrosyl-residue-nitrated SSI and the native subtilisin BPN', and on the binding of the native SSI and the tyrosyl-residue-nitrated subtilisin BPN' and alkaline pH, the tyrosyl residue in question was shown to be one out of the five tyrosyl residues of pKa 9.7 of the enzyme. This tyrosyl residue was probably either Tyr 217 or Tyr 104 on the basis of the reactivities of tyrosyl residues of the enzyme with TNM and their locations on the enzyme molecule. Carboxyl groups of SSI were modified by covalently binding glycine methyl ester with the aid of water-soluble carbodiimide, in order to neutralize the negative charges on SSI. In the difference spectrum which was observed on the binding of subtilisin BPN' and the 5.3-carboxyl-group-modified SSI at alkaline pH, the characteristic features of the protonation of a tyrosyl residue were essentially lost, and the difference spectrum is rather similar to that observed on the binding of the native SSI and the enzyme at neutral pH. This phenomenon indicates that the pKa of a tyrosyl residue of the enzyme is shifted upwards by interaction with carboxyl group(s) of SSI on the formation of the enzyme-inhibitor complex.


Functioning of mitochondria-bound hexokinase in rat brain in accordance with generation of ATP inside the organelle. The function of mitochondria-bound hexokinase, the enzymatic form peculiar to the brain, in utilization of ATP generated inside the organelles, was examined by incubating rat brain mitochondrial fraction with [14C]glucose under various conditions. Addition of succinate and ADP to the incubation medium increased glucose 6-phosphate formation by the mitochondrial hexokinase and caused a smaller increase in ATP concentration in the mitochondria. The glucose phosphorylation was markedly inhibited by the addition of dinitrophenol, potassium cyanide, and oligomycin, and the ATP concentration was decreased. On the other hand, addition of atractyloside suppressed the glucose phosphorylation without affecting the mitochondrial hexokinase activity, whereas addition of antiserum against the mitochondrial hexokinase inhibited both glucose 6-phosphate formation and hexokinase activity. A part of both the glucose phosphorylation and hexokinase activities, however, remained even in the presence of the maximum dose of the anti-hexokinase serum and atractyloside. These results indicate the active utilization of intrinsically generated ATP by the mitochondria-bound hexokinase, a part of which may be located away from the surface of the mitochondrial membrane.


Bacteriophage phiNS11: a lipid-containing phage of acidophilic thermophilic bacteria. IV. Sedimentation coefficient, diffusion coefficient, partial specific volume, and particle weight of the phage. The particle weight (molecular weight) of phiNS11 was determined from the sedimentation coefficient, diffusion coefficient, and partial specific volume of the phage. The sedimentation coefficient of the phage (S(0)20, W) is 416 +/- 2.7S. The diffusion coefficient D(0)20, W), which was determined by quasielastic light scattering measurement, is (0.57 +/- 0.03) x 10(-7) cm2/s. The partial specific volume was determined by the mechanical oscillation technique to be 0.747 +/- 0.007 cm3/g. Based on these values, the particle weight of the phage was calculated to be (70.3 +/- 4.3) x 10(6) daltons, which agrees well with the particle weight (69--72 x 10(6) daltons) estimated from the molecular weight of phage DNA and the content of DNA. The Stokes radius of the phage particle was calculated to be 37.7 +/- 2 nm and hydration of the phage was estimated to be 1.18 cm3/g of dry phage. From the particle weight and the chemical composition of the phage, we estimated that one phage particle contains one double-stranded DNA molecule, 16,000 residues of fatty acid, 72 protein I molecules, 920 protein II, 42 protein III, 48 protein IV, 290 protein V molecules, and 3,700 molecules of polyamines.


Ca2+ bindings of pig cardiac myosin, subfragment-1, and g2 light chain. Ca2+ binding to pig cardiac myosin, subfragment-1 (S-1), and g2 light chain were investigated by the equilibrium dialysis method. Two different S-1s, one of which can bind Ca2+ and another which cannot, were prepared. In order to calculate the free Ca2+ concentrations adequately, the amounts of Ca2+ included in various chemicals and proteins were measured by atomic absorption spectroscopy. Ca2+ contamination was greatest in KCl among the chemicals tested. In addition, the Ca2+ strongly bound to myosin and S-1 was released in the presence of Mg2+. When Mg2+ was not added, the Ca2+-binding constant of myosin was 4 x 10(5) M-1 and the maximum binding number was 1.8 mol per mol of myosin. Cooperativity between the 2 Ca2+ bindings could not be demonstrated. Mg2+ strongly inhibited the Ca2+ binding: at a free Ca2+ concentration of 1 x 10(-5) M, 1.3 mol Ca2+ was bound to myosin in the absence of Mg2+, but 0.6 and 0.2 mol were bound in the presence of 0.3 and 4.5 mM Mg2+, respectively. The Ca2+-binding constant of S-1, which contained a 15,000 dalton component, was 8.6 x 10(5) M-1, and the maximum binding number was 0.7 mol per mol of S-1. The 15,000 dalton component could be exchanged with extraneous g2. S-1 which lacked the 15,000 component could not bind Ca2+ at free Ca2+ concentrations less than 0.1 mM. The Ca2+ binding to free g2 light chain was about 100 times weaker than the binding to myosin, as indicated previously for skeletal myosin (Okamoto, Y. & Yagi, K. (1976) J. Biochem. 80, 111--120). The Ca2+-binding constant was obtained as 4.1 x 10(3) M-1 in the absence of added Mg2+. Phosphorylation of g2 light chain did not affect the Ca2+ binding to the free g2 light chain or to myosin. Ca2+ binding to cardiac native tropomyosin was also measured.


Covalently cross-linked monovalent, divalent, and tetravalent derivatives of concanavalin A. Dimeric succinyl-concanavalin A was cross-linked with ethylenediamine using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide a condensing agent. Thus, a divalent dimer and a tetravalent tetramer composed mostly of covalently cross-linked subunits bearing altered net charges were obtained. Photoaffinity labeling of the cross-linked dimer with p-azidophenyl alpha-D-mannopyranoside resulted in a specific label for its saccharide-binding site and yielded a nonvalent dimer and a monovalent dimer (showing no subunit exchange). However, hemagglutination and glycogen precipitation data suggested that the labeled binding site is shielded but not destroyed by the label and can still bind weakly an external saccharide ligand possibly due to unsteadiness of the shielding label. Although nonvalent and monovalent derivatives were mitogenic as well as divalent and tetravalent derivatives for mouse splenic lymphocytes, binding and stimulation experiments indicated that their stimulating efficiencies after binding to the cells were far lower than those of the multivalent counterparts. Their activities were inhibited by methyl alpha-D-mannopyranoside, suggesting that the weak activities of nonvalent and monovalent derivatives were due to the labeled sites entirely and partly, respectively. We suggest that the triggering of lymphocyte mitogenesis by concanavalin A may depend on cross-linkage of cell surface receptors.


Two ribonucleases H from cultured plant cells. Two forms of enzyme with ribonuclease H (RNase H) [EC 3.1.4.34] activities, have been partially purified from cultured plant cells, strain GD-2, derived from carrot root. One is an Mn2+-dependent RNase H, and the second is an Mg2+-dependent RNase H. These enzymes degrade RNA specifically in RNA-DNA hybrid structures. They were eluted at around 0.2 M and 0.4 M potassium chloride in phosphocellulose chromatography, and were further purified using blue Sepharose. Mg2+-dependent RNase H exhibits maximal activity at pH 9.0, and requires 10 to 15 mM Mg2+ for maximal activity, whereas the Mn2+-dependent enzyme is most active at pH 8.0, is maximally active at an Mn2+ concentration of 0.4 mM, and has some activity with Mg2+. Both enzymes require a sulfhydryl reagent for maximal activity. The enzymes liberate a mixture of oligonucleotides with 5'-phosphate and 3'-hydroxyl termini. The apparent molecular weight of the Mg2+-dependent RNase H was estimated to 18--20 X 10(4) and that of the Mn 2+- dependent RNase H was estimated to be 14 x 10(4) by gel filtration.


Studies on rat liver argininosuccinate synthetase. Inhibition by various amino acids. Inhibition studies of crystallized rat liver argininosuccinate synthetase [EC 6.3.4.5] are described. 1. L-Argininosuccinate, L-histidine, and L-tryptophan inhibited the enzyme activity at saturating amounts of the substrates. 2. L-Norvaline, L-argininosuccinate, L-arginine, L-isoleucine, and L-valine competitively inhibited the enzyme activity at a low concentration of L-citrulline, with Ki values of 1.3 x 10(4) M, 2.5 X 10(-4) M, 6.7 X 10(-4) M, 6.3 X 10(-4) M, and 6.0 x 10(-4) M, respectively. 3. L-Argininosuccinate and L-arginine competitively inhibited the enzyme activity at a low concentration of L-aspartate, with Ki values of 9.5 x 10(-4) M and 1.2 x 10(-3) M, respectively. 4. The modes of inhibition by L-histidine were mixed-noncompetitive, uncompetitive, and noncompetitive types with respect to L-citrulline, L-aspartate, and ATP, respectively. 5. When the enzyme was preincubated with L-citrulline, the enzyme activity was slightly increased in the presence of a low concentration of L-histidine in the assay mixture. 6. The conformation of the enzyme was markedly changed by the addition of L-histidine as judged from the CD spectrum. This change was partially reversed by incubation with L-citrulline.


Modification of an arginine residue of a base-nonspecific ribonuclease from Aspergillus saitoi. 1. A base-nonspecific ribonuclease from Aspergillus saitoi [RNase Ms, EC 3.1.4.23; molecular weight, 12,500] was modified with phenylglyoxal (PG) and 1,2-cyclohexanedione (CHD) in order to determine whether a single arginine residue was involved in the active site of the enzyme. 2. RNase Ms was inactivated by both PG and CHD with concomitant loss of one arginine residue. A competitive inhibitor of RNase Ms, 2',(3')-AMP, protected the enzyme from inactivation by PG. These findings strongly suggest that one arginine residue is involved in the active site of RNase Ms. 3. Difference CD spectra were measured at pH 5.5 for the binding of 2'-AMP and adenosine to native RNase Ms and the CHD- and PG-modified enzyme derivatives to determine the association constants. The arginine modification brought about a marked decrease in the binding affinity of 2'-AMP for the enzyme, but only a slight decrease for adenosine, suggesting that the arginine residue had interacted with the phosphate groups of the substrate.


Orientation of the rhodopsin sugar moiety in bovine disk membrane. Rhodopsin from the bovine rod outer segment contains a covalently linked carbohydrate moiety (Heller, J. & Lawrence, M.A. (1973) Biochemistry 9, 864--868). We studied the location of this carbohydrate moiety on the disk membrane by using ferritin-conjugated concanavalin A and concanavalin A labelled with fluorescein isothiocyanate. Electron microscopic observation of sonicated disk membrane that was labelled with ferritin-concanavalin A revealed the electron-dense image of ferritin on the inner surface of the disk membrane and not on its outer surface. Intact disk membrane that was similarly treated with ferritin-concanavalin A showed a complete absence of ferritin molecules on its surface. In an independent series of experiments we confirmed that the sonicated disk membrane bound three to five times more fluorescein-labelled concanavalin A than the intact disk membrane did. From these experiments we conclude that the carbohydrate moiety of bovine rhodopsin is located on the inner surface of the disk membrane, in agreement with the report by Rohlich on the frog rod outer segment disk membrane (Rohlich, P. (1976) Nature 263, 789--791).


Rat liver glutamine 5-phosphoribosyl-1-pyrophosphate amidotransferase [EC 2.4.2.14]. Purification and properties. Glutamine 5-phosphoribosyl-1-pyrophosphate (PRPP) amidotransferase (amidophosphoribosyltransferase), [EC 2.4.2.14] was purified 1,600-fold from rat liver. The preparation gave two protein bands on acrylamide gel electrophoresis, of which only the main band showed enzyme activity. The molecular weight of the enzyme was estimated to be 215,000, 200,000, and 195,000 by Sephadex G-150 gel filtration, polyacrylamide gel electrophoresis, and sucrose density grandient ultracentrifugation, respectively. The apparent Km values for glutamine and PRPP were 1.24 mM and 0.57 mM, respectively. The concentration-activity curve for PRPP changed from a hyperbolic to a sigmoidal form on addition of AMP or GMP, and this inhibition by AMP was prevented by increasing the PRPP concentration. In the presence of high concentrations of inorganic phosphate, the catalytic activity was decreased and the sensitivity to AMP inhibition was slightly increased. The molecular size of liver amidotransferase was not changed by the addition of PRPP, AMP, or 2-mercaptoethanol. The purified rat liver enzyme has a broad pH-range of activity between 6.5 and 8.5.


Interaction of troponin-I with troponin-T and its fragment. The interaction of troponin-I (TN-I) and troponin-T (TN-T) was examined using immobilized TN-2 as an affinity adsorbent. TN-T dissolved in 0.4 M NaCl bound strongly to immobilized TN-I and required the application of 5 M urea for its elution. A chymotryptic fragment of TN-T (Ohtsuki, 1978), perhaps the C-terminal fragment, retained the ability of the original TN-T to bind to immobilized TN-I.


High performance liquid chromatography of unsaturated disaccharides produced from chondroitin sulfates by chondroitinase. High performance liquid chromatography was performed by the ion pair method on unsaturated disaccharides produced from chondroitin sulfates by the action of chondroitinase. Completely separated peaks corresponding to deltaDi-0S, deltaDi-4S, and deltaDi-6S were obtained on a column of mu-bondapak-C18 with 0.035 M tetra-butylammonium phosphate (pH 7.54) as a mobile phase. There was a linear relationship between the ratio of the peak area and the amount of each standard tested.


Cause of subunit heterogeneity in purified rat renal phosphate-dependent glutaminase. When electrophoresed on polyacrylamide gels in the presence of sodium lauryl sulfate, highly purified rat renal phosphate-dependent glutaminase exhibits subunits which range in molecular weight from 57,000 to 75,000. Peptide mapping of the separated subunits following limited proteolysis in the presence of sodium lauryl sulfate shows that all of the various subunits are related in structure. The glutaminase, immunoprecipitated from Triton X-100-solubilized mitochondria, is composed primarily of subunits which have molecular weights of 83,000. In addition, the series of smaller subunits is generated during storage of the Triton-solubilized glutaminase at 4 degrees C. These results indicate that the heterogeneity of subunit size found in the purified glutaminase results from a noninactivating partial proteolysis of the native form of the enzyme.


Purification of a hexokinase-binding protein from the outer mitochondrial membrane. Brain hexokinase (ATP:D-hexose-6-phosphotransferase, EC 2.7.1.1) binds selectively to the outer membrane of rat liver mitochondria but not to inner mitochondrial or microsomal membranes nor to the plasma membrane of human erythrocytes. A protein having subunit molecular weight of 31,000, determined by sodium dodecyl sulfate-gel electrophoresis, has been highly purified from the outer mitochondrial membrane by repetitive solubilization with octyl-beta-D-glucopyranoside followed by reconstitution into membranous vesicles when the detergent is removed by dialysis. When incorporated into lipid vesicles, the protein confers the ability to bind brain hexokinase in a Glc-6-P-sensitive manner as is seen with the intact outer mitochondrial membrane. Hexokinase binding ability and the 31,000 subunit molecular weight protein co-sediment during sucrose density gradient centrifugation. Both hexokinase binding ability and the 31,000 subunit molecular weight protein are resistant to protease treatment of the intact outer mitochondrial membrane while other membrane proteins are extensively degraded. It is concluded that this protein, designated the hexokinase-binding protein (HBP), is an integral membrane protein responsible for the selective binding of hexokinase by the outer mitochondrial membrane.


Phosphorylation of the 20,000-dalton light chain of myosin of intact arterial smooth muscle in rest and in contraction. The 20,000-dalton myosin light chain of intact pig carotid arteries was found to be rapidly labeled when the arterial muscle was incubated in physiological salt solution at 37 degrees C containing [32P]orthophosphate. Light chain phosphorylation in the intact muscle had a marked requirement for Ca2+ and was dependent upon the passive tension applied to the muscle. Norepinephrine- or KCl-induced contractures were associated with concomitant increases in light chain phosphorylation.


Carrier protein effects on DNA-cellulose chromatography of putative steroid receptors. In the absence of carrier proteins, putative androgen receptors elute from DNA-cellulose in the range of 120 to 190 mM NaCl. However, in the presence of lysozyme, most of the receptor elutes in the range of 200 to 230 mM NaCl. This is the same range in which the lysozyme itself, a basic protein, elutes after being chromatographed in the same manner. Moreover, at low ionic strength, lysozyme also increases the sedimentation velocity of both androgen and estrogen receptors. In contrast, bovine serum albumin neither adheres to DNA-cellulose nor alters the sedimentation properties of these proteins. The lysozyme effects can account for some discrepancies reported in the literature. Thus, for qualitative elution studies, the use of lysozyme as a carrier protein is not advised, although its direct interaction with receptors might facilitate quantitative fractionation.


The high potential iron-sulfur cluster of aconitase is a binuclear iron-sulfur cluster. It has been reported (Ruzicka, F.J., and Beinert, H. (1978) J. Biol. Chem. 253, 2514-2517) that aconitase in the oxidized state, as isolated, shows an electron paramagnetic resonance signal centered at g = 2.01, typical of high potential iron-sulfur proteins. Since the magnetic state corresponding to this signal has thus far only been found in tetranuclear iron-sulfur clusters in model compounds and proteins, it could be expected that aconitase also contains a [4Fe-4S] cluster. We show here that core extrusion, in the presence of hexamethylphosphoramide and o-xylyl-alpha,alpha'-dithiol and subsequent ligand exchange with p-trifluoromethylbenzenethiol yield absorption spectra typical of binuclear iron-sulfur clusters. According to the absorbance measured, the concentration of the extruded [2Fe-2S] cluster quantitatively accounts for the iron-sulfur content of the preparations examined. Preliminary studies of the 19F nuclear magnetic resonance spectrum obtained on extrusion with p-trifluoromethylbenzenethiol confirm the presence of a binuclear cluster in aconitase.


A hepatic soluble cyclic nucleotide-independent protein kinase. Stimulation by basic polypeptides. Enzymatic phosphorylation of cytoplasmic proteins by a cyclic nucleotide-independent protein kinase (casein kinase of a classical type) in rat liver is stimulated greatly, sometimes more than 10-fold, by polycations, particularly by basic polypeptides such as polylysine, histone, and protamine. These basic polypeptides themselves do not serve as phosphate acceptors but act as stimulators for the reaction by interacting with cytoplasmic proteins rather than with enzyme. The stimulatory effect varies with substrates employed; with casein and phosvitin the stimulation does not exceed 2- to 3-fold. The cytoplasmic endogenous phosphate acceptor proteins measurable in the presence of basic polypeptides are abundant for this species of protein kinase.


Structural characterization of a glycoprotein isolated from alveoli of patients with alveolar proteinosis. A soluble glycoprotein of Mr = 80,000 has been isolated from lung lavage of patients with alveolar proteinosis and found to contain 5 residues of hydroxyproline, 91 residues of glycine, 3 residues of methionine, 3.8 molecules of sialic acid, 6 molecules of mannose, 5.9 molecules of galactose, 1 molecule of fucose, and 9.1 molecules of glucosamine. Cyanogen bromide (CNBr) treatment of the glycoprotein resulted in four peptides with molecular weights of 36,000, 27,000, 12,000, and 5,000. The chemical compositions of the CNBr peptides indicated the presence of hydroxyproline and high amounts of glycine in all but one of the peptides; two of the four CNBr peptides contained carbohydrate. Limited trypsin digestion of the glycoprotein of Mr = 80,000 resulted in four peptides with molecular weights of 62,000, 36,000, 26,000 and 18,000, the latter being the NH2-terminal peptide of the native glycoprotein molecule. The peptide of Mr = 26,000 was found to be the COOH-terminal peptide.