Vitamin D status of elderly at home and institutionalised in hospital. In a study of institutionalised and non-institutionalised aged requiring medical care, the biochemical deficiency of vitamin D was noted in over 40 per cent of the subjects, while osteomalacia was diagnosed biochemically in only 5.2 per cent. The vitamin D status demonstrated no relationship to bone density.


Stereobiochemical aspects of warfarin isomers for inhibition of enzymatic alkylation of menaquinone-0 to menaquinone-4 in chick liver. The inhibition of enzymatic activity by the isomers of warfarin for alkylation of menaquinone-0 to menaquinone-4 has been studied in chick liver. Injection of (6,7--3H2)-menaquinone-0 into chicks resulted in the biosynthesis of menaquinone-4 within 3 h. Microsomes of control chicks contained 78 per cent of the total menaquinone-4. (S) (-)-Warfarin and (R) (+)-warfarin isomers inhibited the biosynthesis of menaquinone-4 of about 67 and 17 per cent respectively. Fractionation of soluble microsomal proteins lead to the the isolation of a menaquinone-4 binding protein.


Metabolism and biological activity of a thiamine new salt. I. Urinary excretion and tissue content of vitamin B1 in rats treated with thiamine chloride ester monophosphate of Bis-glucosamine. Research has been carried out on the metabolism of thiamine chloride ester monophosphate of bis-D-glucosamine (TCMPG) evaluating the urinary excretion, the blood and the hepatic content of thiamine in the rat after intraperitoneal injection of this compound. The results obtained have shown a greater retention of vitamin B1 when it is administered as glucosamine salt. In fact, the amount of thiamine excreted by the rats which have received the TCMPG is lower while the blood and liver levels of vitamin are higher, compared with the control rats treated with thiamine monophosphate only, or with thiamine and glucosamine. These results permit the conclusion that glucosamine increases the utilization of the vitamin and its penetration into the cells.


Some studies on ascorbic acid metabolism in hydrazine-treated rats. The metabolism of ascorbic acid was studied in hydrazine-treated rats. Hydrazine was administered i.p. at a dose of 1.28 mg/day (20% LD50) for each 100 g body weight for 7 days. Hydrazine administration at the present dose did not appear to have an effect on the total ascorbic acid level of liver, kidney, spleen and testis. The adrenal and plasma total ascorbic acid levels were, however, elevated. The activity of liver D-glucuronoreductase and that of liver and kidney dehydroascorbatases were diminished after hydrazine administration. The changes in the activities of liver enzymes were accompanied by a fall in the reduced ascorbic acid level and an elevation in the dehydroascorbic acid level. The uronolactonase activity of liver, on the other hand, remained independent of hydrazine treatment. It has been suggested that hydrazine treatment at the present dose reduced the biosynthesis of L-ascorbic acid from D-glucuronolactone as substrate. In spite of diminished synthesis, the normal level of total ascorbic acid in the liver of hydrazine-treated rats was maintained by reducing the degradation of L-ascorbic acid. The rise in the plasma total ascorbic acid level after hydrazine treatment was ascribed to reduced catabolism and urinary excretion of ascorbic acid, while the elevation in adrenal total ascorbic acid level might result from increased uptake of ascorbic acid by the gland from blood or from nonfunctional accumulation.


Carnitine and acetylcarnitine in skeletal and cardiac muscle. The acetylcarnitine/carnitine ratio has been found significantly increased in tibial muscles of swimming rats in comparison with normal rats. Analogous increase occurred in electrically stimulated tibial muscle in comparison with the unstimulated controlateral muscle of the same animal. Swimming also induced a statistically significant increase of the acetylcarnitine/carnitine ratio in heart, while norepinephrine administration resulted in a decrease of carnitine not accompanied by a corresponding increase in acetylcarnitine in a severe carnitine depletion of cardiac muscle. These results are discussed in terms of carnitine function in the acetyl metabolism, assuming the acetylcarnitine as a possible acetyl reservoir in the working muscle.


Liver arginase activity and plasma urea-nitrogen in steers fed diets containing different levels of protein. Seventeen Charolais-Hereford crossbred steers averaging 220.0 kg in weight initially were randomly allotted to three treatments and fed diets containing (A) 9%, (B) 11%, or (C) 13% protein ad libitum in individual pens for 238 days. Liver samples were taken by aspiration biopsy on days 47 (sampling I) and 238 (sampling II) for arginase assay. Blood samples were taken by jugular puncture at these same times for plasma urea-nitrogen analysis. Mean hepatic arginase activities (mumole urea/mg protein/hr) of steers fed the three diets at sampling I were: (A) 224.8, (B) 327.8 and (C) 333.3. The activities of B and C were significantly (P less than .05) higher than A. Plasma urea-N levels at this same time averaged: (A) 4.44, (B) 8.55 and (C) 12.22 mg/100 ml, and were significantly different (P less than .01) from each other. Arginase activities at sampling II were: (A) 240.6, (B) 305.2 and (C) 353.0. A differed significantly (P less than .05) from C. Plasma urea-N at this time averaged: (A) 8.69, (B) 10.72 and (C) 15.00 mg/100 ml, and C was significantly (P less than 5.0) higher than A and B. These data suggest that in the bovine hepatic arginase activity levels increase with increases in dietary protein and that similar patterns in activity are maintained after feeding for an extended period of time. The increased arginase activity was accompanied by increased plasma urea-nitrogen.


Classification (staging) of renal parenchymal tumours. The different classification systems (clinical, microscopic, descriptive) of renal cell tumours provide useful clues to the evaluation and prognosis of these tumours. The prognostic reliability of clinical staging as recommended by UICC is, however, questionable because the clinical parameters are not clearly defined and even the radiological features are of equivocal prognostic significance. Descriptive classification on the basis of the complete surgical specimen is regarded as fundamental to the understanding of the evolution, thus also of the prognosis, of renal cell tumours. The author describes his original staging system of renal cell tumours set forth in 1956 and its modifications in accordance with the recommendations of UICC, taking the stages of evolution (P), involvement of the regional lymph nodes (N+ or N-) and generalization ((M + or M-) into account. The significance of the intracapsular stage ("Stage A") of the tumour is stressed. Renal vein involvement (V+ or V-) is regarded as a less reliable prognostic parameter. For lymph node involvement subdivision into Stages N1-N4 is proposed. The prognostic implications of the descriptive staging system are illustrated in terms of survival figures in a series of 250 cases.


A hitherto undescribed case of "collision" tumour: liposarcoma of the seminal vesicle and prostatic carcinoma. A hitherto undescribed type of the rare "collision" tumours is reported: necropsy in a 77-year-old male patient revealed liposarcoma of the right seminal vesicle in association with anaplastic carcinoma of the prostate. The carcinoma was fused with the liposarcoma as a result of its invasive local growth and had formed deposits in the lymph nodes, bones and lungs. The left seminal vesicle showed localized amyloidosis. A review of the published cases of mesenchymal tumours of the seminal vesicles is presented.


Involuntary ureteral micturition resulting from the ectopic opening of the single ureter into the urethra. The ectopic opening of the single ureter is a rare developmental defect of the urinary system. The authors present a case of the opening displacement of the single ureter to the urethra resulting in ureteral micturition. The successful treatment is a result of the transplantation of the ureter into the bladder.


A contribution to the study of kinetics of ureteral urine passage in children. For the assessment of ureteral urine passage in children sequential scintigraphy with an analyzing system was used. Through the proper division of the time of examination into short time intervals the information on the areal distribution of urine was obtained, dependent on time. After demarcation of the spheres of interest by light-pen, the level of radioactivity was followed continually in a form of curves. Information for the assessment of etiopathogenesis of the disease was selected from the memory system of the device. Through the radioisotope examination also the dynamic changes have been recorded, not visible in excretion urograms, or only presumable from them. The method of examination described can help to assess the functional importance of morphological findings.


Cavernospongiosum shunt in management of priapism: is it a reliable method? The efficiency of cavernospongiosum shunt in the management of priapism was evaluated in 10 patients. Technical details such as the "Z" perineal incision and the opening followed by closure of the contralateral corpus cavernosum, at the time of corpora evacuation, can improve the local exposure and permits a more complete drainage of the corpus cavernosum, but did not improve our results when they were compared with other techniques for treatment of priapism. Penile flaccidity and preservation of sexual potency occurred in 6 (60%) and 5 (50%) patients, respectively, and no surgical complications were observed. Immediate penile flaccidity and regaining of physiologic erection were, however, not complete even in the successfully treated patients. The cavernospongiosum shunt does not seem to represent the definitive form of treating priapism. New knowledge about mechanisms involved in priapism has to be obtained, in order to improve the approach and the prognosis of this disabling condition.


Mesterolone treatment of patients with pathospermia. The response to Mesterolone, in doses of 25 mg/day, was examined in 42 pathospermic patients. Treatment lasted for 100 days. The pronounced response to the Mesterolone treatment was observed in hypozoo- and oligozoospermia with low initial fructose content in the ejaculate. Fructose content attained its normal range after the treatment. During the therapeutic period 11 wives became pregnant. The authors conclude that Mesterolone does not influence plasma FSH, LH and testosterone levels, it has only peripheral effects.


The pediatric tracheostomy: III). An appraisal of xerography. Comparatively new in the roentgen evaluation of infants and children, xerography would appear to be ideal for portraying the pediatric airway. Since the radiation dose of xerography is higher than conventional radiography, its image quality is the subject of careful scrutiny in a pediatric setting here described in 23 infants and children with tracheostomies. In this appraisal three pediatric radiologists independently compared each child's xeroradiograph with an accepted control, conventional radiography. The principal conclusion: the xerographic image of the pediatric airway is slightly superior to standard radiography in most studies (40 of 47 paired studies, 85.1%). In all but two comparisons, however, the tracheal morphology is correctly perceived, described, and interpreted in both the standard radiograph and the xeroradiograph. The main role of xerography, then, is its verification of tracheal detail as first depicted by the preliminary radiography. This confirmation is important in this particular pediatric problem, the tracheostomy, which has inherent difficulties in correct roentgen diagnosis.


Epinephrine enhanced knee arthrography.

Epinephrine enhanced knee arthrography. The effect of .3 cc of 1:1000 epinephrine on iodine concentration, total iodine content, fluid volume and radiographic quality was evaluated in knee arthrograms in dogs. With the addition of epinephrine the iodine concentration and total iodine content were significantly higher initially and remained significantly higher than in controls over a one hour period. The fluid volume in the knee was significantly lower with epinephrine. Initially both contrast absorption and dilution have major effects in decreasing iodine concentration. With increasing time the effect of contrast absorption becomes the major factor, having a two of three times greater effect on decreasing iodine concentration. With epinephrine radiographs were judged significantly better early and while the quality deteriorated rapidly in the controls the enhanced quality with epinephrine persisted up to one hour. Epinephrine is a very useful adjunct in enhancing the quality of knee arthrography.


A breast phantom method for evaluating mammography technique. A new breast phantom has been designed for use in evaluating mammographic system performance. This phantom incorporates simulated calcifications and fibrillar objects in fat, of graded size, to permit measurements of detail visibility. A special methodology has been developed for measuring visible object size to achieve reproducible and clinically relevant results. Materials and construction of the phantom also permit carrying out dosimetry with an appropriate ionization chamber. Dosage and detail visibility measurements are reported for the Xerox 125, Min-R and Xonics systems. In addition to providing information regarding technique and image receptors, these results demonstrate the usefulness of the basic phantom design, and suggest possible improvements.


Diameter of arterial microvessels trapping 8--10 micron, 15 micron and 25 micron microspheres as determined by vital microscopy of the hamster cheek pouch. An assumption inherent in the use of microsphere methodology for measuring regional blood flow is that microspheres are removed from the circulation by impacting in arterial vessels of approximately their own diameter. We investigated the in vivo relationship between the diameter of varying size microspheres and the calibre of trapping microvessels within the hamster cheek pouch. Intracardiac injection of 8-10, 15 +/- 5, 25 +/- 5 micron carbonized microspheres with diversion of the cardiac output into the bracheocephalic trunk provided direct, in vivo visualization of the impaction sites of these various size microspheres within the cheek pouch microvasculature. Fifteen micron microspheres usually impacted at the orifice or neck of small arterioles and protruded into the lumen of the parent vessel. Eight to ten micron microspheres lodged in vessels with a mean diameter of 11.5 +/- SD 3.4 micron indicating that they usually impacted primarily in terminal arterioles.


Microradiography of the collecting ducts in the perfusion fixed rabbit kidney: suggestions for the anatomic basis for the radiographic appearance of cortical striations and intrarenal reflux. Perfusion fixation was used to preserve the rabbit kidney, in its in vivo state (open tubules). Barium sulfate was either injected through the renal artery at a pressure sufficient to cause postulated microruptures in the glomerular tufts and opacify the tubules, or refluxed via the ureter to opacify the collecting ducts and tubules to the level of Bowman's capsule. The anatomy of the collecting ducts and their relationships to the nephrons were demonstrated, allowing suggestions for the anatomical basis for the radiographic appearances of cortical striations and intrarenal reflux.


Urographic excretion studies: preliminary results with a six-iodine, singly-ionizing sodium salt--P.286. Preliminary results of comparative studies with a new water-soluble contrast agent are described. The new agent, with the code P.286, is a molecule with 6 iodine atoms attached and only one ionizing group. The sodium salt of the new agent has been compared with sodium iothalamate and metrizamide in dogs. The results suggest that the sodium salt of P.286 is superior in terms of urinary iodine concentration to both sodium iothalamate and metrizamide.


Time-related contrast enhancement of blood, normal muscle, and V2 carcinoma in the rabbit as determined by CT scanning. After a bolus of intravenous Renografin, the delivery and washout of contrast in malignant V2 carcinoma, normal muscle, and blood using CT scanning was evaluated over a two-hour period. Diagnostically useful differences in enhancement between V2 carcinoma and muscle predictably lasted one hour, but in selected rabbits lasted longer. Delivery and washout was greater in tumor than in normal muscle, and it was these differences which accounted for the diagnostically useful differences in enhancement. After two hours, attenuation in V2 and normal muscle remained elevated, while blood returned to nearly control levels, suggesting a large amount of contrast distributed throughout the extravascular space and probably in equilibrium with excretion.


Evaluation and characteristics of currently available inulin. Inulin was not available for clinical testing in the United States between February 1973 and July 1975 because the raw material received in the United States differed from previously manufactured lots in molecular weight, glucose content, optical rotation, melting point, and water solubility. The comparability of the "new" and "old" inulin preparations was assessed in 12 adult male spinal cord injury patients, seven with normal renal function and five with compromised function. The "new" inulin proved to be as accurate and safe an agent for the determination of glomerular filtration rate as the previously available material in both groups of patients, and was made available for general clinical use in the United States in July 1975.


Elevated rate of 3H-uridine incorporation in regressing rat ventral prostate. Ventral prostates from adult Sprague-Dawley rats were removed at intervals during the first 2 weeks postcastration. Incubation of tissue with 3H-uridine and 3H-leucine was performed to determine the incorporation rate of radioactivity into the RNA and protein fractions respectively. Prostatic wet weight and 3H-leucine incorporation rate into prostate protein diminished postcastration, whereas 3H-uridine incorporation rate remained relatively high. The data suggest that RNA synthesis is relatively active in comparison with other parameters during regression.


Partial inhibition of castration induced ventral prostate regression with actinomycin D and cycloheximide. Sprague Dawley male rats were injected subcutaneously with either actinomycin D or cycloheximide in saline immediately after castration. Control animals received saline only. Treatment was repeated daily for 4 subsequent days, and 24 hr after the last injection rats were sacrificed by decapitation. Ventral prostates were dissected and weighed. Actinomycin D and cycloheximide administration significantly reduced the rate of prostatic weight loss in castrated rats; serum testosterone levels in the control and drug treated animals were comparable. Histologic studies of the prostates indicated that drug treated animals had more active epithelial cells than saline injected controls. These results indicate that the rate of prostatic regression induced by castration can be modified by drugs such as actinomycin D and cycloheximide.


Deleterious synergism of a cardiac glycoside and sodium diatrizoate. Studies were performed in mice to determine if the cardiac glycoside, Strophanthin-K, and the contrast medium, sodium diatrizoate, interact synergistically to produce death. Intravenous injections of lethal and near lethal doses of the two agents produced a significantly greater mortality than the individual agents alone. Low or near clinical doses of Strophanthin-K, when given with doses of diatrizoate in the lethal range, produced mortalities significantly greater than did the diatrizoate alone. Similarly, low or near clinical doses of diatrizoate given with doses of Strophanthin-K in the lethal range produced mortalities significantly greater than for the Strophanthin-K alone. Isotonic or hypertonic saline, when substituted for diatrizoate or Strophanthin-K did not produce synergistic increases in mortality. Thus neither the injection volume, nor agent hypertonicity or ionic strength, seem to be the primary factors in the synergism to produce death. The diatrizoate anion appears to be an important factor. Until more information is available from other animal models it appears that patients receiving cardiac glycoside should be considered to have a higher than normal risk of serious reactions to contrast media in intravenous urography.


Effect of retinoic acid on organ-cultured transitional epithelium. In view of the correlation shown in vivo between vitamin A deficiency and increased transitional cell proliferation, the ability of retinoic acid to block hyperplasia in organ-cultured bladder epithelium was investigated. Retinoic acid, at concentrations shown to block hyperplasia in other in vitro systems, was ineffective in blocking hyperplasia in bladder epithelium. The data suggest the possibility that, in vivo, vitamin A may not act directly on the transitional epithelium to control cell proliferation. The cytoplasmic volume, which normally decreases during the period of hyperplasia in vitro, was increased by retinoic acid. The increase in cytoplasmic volume stimulated by 0.3 micrometer retinoic acid was sufficient to maintain normal nuclear:cytoplasmic ratios during the hyperplastic response. The increase in cytoplasmic volume could not be correlated with increased protein, mucin, or glycogen synthesis. The data suggest that retinoic acid may have increased the permeability of the luminal membrane to water, possibly by interfering with the synthesis of the keratin component of the membrane.


Uptake of contrast materials by experimental acute myocardial infarctions: a preliminary report. The concentration of iodine within infarcted and normal myocardium after intravenous administration of contrast material was determined by fluorescence excitation analysis in seven dogs at 48 hours after coronary arterial ligation. The iodine concentration of infarcted myocardial tissue was several times greater than normal myocardium after administration of meglumine/sodium diatrizoate, iodipamide, and an experimental polymer of iothalamic acid.


Isocitrate uptake and citrate production by rat ventral prostate fragments. Prostatic aconitase was investigated utilizing fragments of rat ventral prostate. The rate of citrate formation from isocitrate was used as an index of aconitase activity. Inhibition of citrate formation from isocitrate by fluorocitrate indicated that aconitase activity was being measured. The results indicated that whereas both kidney and prostate formed citrate from isocitrate, the aconitase activity of prostate was independent of both medium isocitrate and citrate concentration. In addition the aconitase activity of prostate seemed to favor citrate formation to a much greater degree than kidney. The results showed that the aconitase catalyzed conversion of citrate to isocitrate is limited in prostate and suggested that the deficient citrate oxidation observed in prostate is the result of a unique aconitase activity.


A study of o-cresolphthalexon and its analogs as cholescintigraphic 111In carriers. 111In has been complexed to a series of metal-complexing phthaleins and sulfonphthaleins, and the hepatobiliary excretion of the compounds were compared in rats. The highest biliary excretion was obtained with 111In-o-cresolphthalexon. In a normal human subject, sequential imaging with 111In-o-cresolphthalexon visualized his gallbladder, common bile duct, and the bowel from jejunum to rectum. The potential utility and limitation of this biliary scanning agent is discussed.


Biodegradable ureteral grafts in dogs.

Biodegradable ureteral grafts in dogs. In six dogs, a segment of the right ureter was replaced by a tubular segment of poly [L(-) lactic acid], a new absorbable biomaterial, with the hope that the foreign body would be absorbed or dissolved after a new mucosally lined channel had formed. The dogs were followed with excretory urograms, blood urea nitrogens, and serum creatinines. Postoperatively, four of the six dogs had deteriorating or total loss of function of the right kidney by 8 months attributable to obstruction of the ureter by disintegrating implant.


The initiation, progress, and diagnosis of dog bladder cancer induced by 4-aminobiphenyl. Twenty-four beagles received 4-aminobiphenyl orally 5 days a week for 3 years. The dogs were examined cytoscopically at frequent intervals. Biopsy and surgery were performed according to predetermined criteria. Three basic patterns of carcinogen responses were seen. Negative or minimal disease was seen in four dogs. Of these, two remained completely free of neoplasia. Eleven dogs progressed slowly in their development of neoplasia with minor fluctuations in development. A rapidly progressive pattern was observed in the remaining nine dogs. A large variance in the time of onset of the first positive diagnosis of neoplasia was seen. Eleven dogs underwent surgery ranging from tumor sampling to cystectomy plus ureterosigmoidostomy. Histologic types and grades and patterns of metastasis as well as several other characteristics seemed to parallel closely human bladder cancer.


Comparison of arteriography, venography, and pyelography in experimental renal trauma. Experimentally produced renal injuries are shown by arteriography, venography, and retrograde pyelography using geometric magnification techniques. Arteriography is the most satisfactory modality examined. Venography is technically more difficult and generally less sensitive than arteriography, although extravasation of contrast material was shown with equal frequency. Retrograde pyelography is inferior to arteriography and correlates closely with the venographic findings.


Experimental closure of urinary bladders with surgical staples. The auto stapler, an instrument proven to be effective in other surgical procedures, was utilized to close the canine urinary bladder. The procedure was found to be simple. Anticipated postoperative complications, particularly leak, infection, or stone formation were not experienced.


The importance of renal prostaglandin synthesis in maintaining renal blood flow and glomerular filtration after renal artery stenosis in the conscious dog. We studied the effects of inhibiting prostaglandin synthesis on the renal blood flow and glomerular filtration rate of four conscious dogs with chronic unilateral renal artery stenosis. Dogs were prepared by exteriorizing the bladder trigone to allow repeated split renal function studies. Renal artery stenosis was produced with a Goldblatt clamp on one renal artery. Each of ten studies determined the function of each kidney in the conscious dog before and after indomethacin treatment. Administration of indomethacin resulted in a 90 per cent reduction of urinary prostaglandin E excretion, but no functional changes were produced in the kidney with the stenotic artery. These studies indicate that renal prostaglandins are not responsible for maintenance of creatinine and para-aminohippuric acid clearances after renal artery stenosis in the dog. However, the concentration gradient for creatinine between the stenotic and non-stenotic kidney was enhanced by indomethacin.


[Clinical aspects of pelvic stenosis syndrome (Schneider-Fischer)]. Six cases with posthrombotic obstruction of the pelvic veins are reported. These case reports demonstrate the variability of the clinical symptomatology. In addition relationships between the postthrombotic obstruction of the pelvic veins and hemorrhoids and varicocele are discussed. Therapeutical aspects are reviewed.


[Juvenile dermatofibrosarcoma protuberans]. Dermatofibrosarcoma protuberans is observed in younger or middle-aged patients after the transformation from the initial stage of a chronic fibrous plaque into a rapidly growing, sometimes painful or bleeding tumor. First manifestation in early childhood is considered in about 10 per cent of the reported patients. In the case that we observed the neoplasma was present at birth and increased to a multinodular tumor between three and four years of age. Repeated local recurrences in the scar were observed after the surgical removal. The clinical behavior of the tumor in childhood is comparable to those of adults. Because of its malignant potential extended early excision should be undertaken despite of the risk of widespread scarring.


[Triggering of immunologic reactions by Strehler's pressure wave therapy in patients with chronic venous insufficiency]. Thirteen patients with chronic venous insufficiency of the legs, mostly with ulcerations, were treated during four weeks by pressure wave therapy following the method of Strehler. Some parameters of circulation, blood, and immunology were controlled at the beginning, at the third, tenth, twentieth and fourtieth day. There was a tendency to normalization of hypo- and hypertension, of the blood calcium and kalium, and especially, there was a significant increase of IgG and C'4 during the treatment with decrease of the IgM. Other parameters did not show specific alterations. The mobilisation of edema by pressure wave therapy seems to arise an immunologic reaction, probably by activating antigens or antigen-antibody complexes.


[Contact sensitivity to lead].

[Contact sensitivity to lead]. A case of contact sensitization to lead is described which presented as an acute bullous contact dermatitis after application of an ointment containing lead oxide (unguentum diachylon Hebra). In patch testing, the patient reacted strongly to lead oxide and lead acetate. Further examination revealed a chronic lead intoxication with neurologic and labyrinthine involvement. As a painter, the patient had had professional contact with the lead paints up to 25 years ago for a period of 30 years. He had never noticed any contact allergies to his working materials.


Locus of control and the intention to lose weight. The relative importance of personal attitudes toward losing weight and the social pressures for weight loss in determining intentions to lose weight was compared for locus of control internals and externals who valued health and/or physical appearance highly. Findings consistent with the hypothesized differential importance of these two predictors of behavior intention for individuals designated internal and external by a behavior-specific locus of control scale are presented. Theoretical and practical applications of the findings are discussed.


Health locus of control and health value in the prediction of smoking reduction. Thirty-five participants in a smoking cessation program met weekly for seven weeks in small groups and then were assigned to one of four follow-up procedures for an additional eight weeks. Information on smoking levels was obtained at the beginning of the program, at the end of the seven-week treatment period, at the end of the two-moth follow-up period, and during an additional five-month extended follow-up period. As hypothesized from social learning theory, individuals who held internally-oriented health locus of control beliefs and who valued health highly were most successful in achieving and maintaining changes in their smoking behavior. Thes results are discussed in terms of their implications for the problem of maintenance of behavior changes achieved by smoking cessation programs.


A test of the construct validity of health locus of control: effects on self-reported compliance for hypertensive patients. A test of the construct validity of the Health Locus of Control (HLC) scale is presented within the larger framework of Rotter's social learning theory. Self-reported compliance behavior was predicted to relate to the subject's HLC orientation, value toward health, and level of perceived home assistance. In a sample of ambulatory hypertensive patients, a significant two-way interaction effect was calculated between the perceived level of home assistance and the patient's HLC orientation (p = .02). The more internally oriented the patient and the higher the level of perceived home assistance in complying with the prescribed medical regimen, the greater the level of self-reported compliance behavior.


Development of an instrument to measure children's health locus of control. Studies were conducted with children ages 7 to 12 years to develop an instrument to measure children's health locus of control. Findings provide evidence that the Children's Health Locus of Control scale has acceptable levels of reliability, internal consistency, and construct validity. Item analysis and factor analysis were performed to determine item effectiveness and existence of significant subscales. Implications of this research for health education of children are disscussed. Links between the theoretical basis for the locus of control construct and a more comprehensive explanation of health behavior are outlined.


Development of the Multidimensional Health Locus of Control (MHLC) Scales. The development of the Multidimensional Health Locus of Control scales is described. Scales have been developed to tap beliefs that the source of reinforcements for health-related behaviors is primarily internal, a matter of chance, or under the control of powerful others. These scales are based on earlier work with a general Health Locus of Control Scale, which, in turn, was developed from Rotter's social learning theory. Equivalent forms of the scales are presented along with initial internal consistency and validity data. Possible means of utilizing these scales are provided.


The evolution and distribution of hydroxysteroid dehydrogenase activity in human foetal skin throughout gestation. Specimens of skin from human foetuses from 6--41 weeks gestation were incubated to demonstrate the presence of hydroxysteroid dehydrogenases (HSDs) by histochemical methods. When present HSDs were noted only within the acini of the sebaceous glands and in the secretory duct. Seventeen-beta-HSD activity was first demonstrated at 16 weeks gestation, co-incident with full function in the glands. Three-beta- and 16beta-HSD activity did not appear until 22--24 weeks. There were differences between the foetal pattern of distribution of the enzymes within gland acini and that already known in skin from subjects in extrauterine life. The time of changeover to the latter pattern has been established as 38 weeks gestation. No correlation was noted between HSD activity and the sex of the foetus or body site. It is concluded that foetal skin is involved in steroid metabolism, and possible physiological roles for this activity are discussed, with speculation over skin as an excretory route or detoxication centre for steroids and the role of steriod metabolic activity in the local stimulation and functional control of foetal sebaceous glands.


Variations in sialomucins in the mucosa of the large intestine in malignancy: a quantimet and statistical analysis. The periodate-borohydride/saponification/PAS (PB/KOH/PAS) method, for the identification of sialic acid derivatives, was used to investigate possible changes in human colonic epithelium associated with carcinogenesis. The material was obtained from specimens of the large intestine resected for carcinoma and compared with normal control mucosa from biopsies of patients with no known gastro-intestinal diseases, using a Quantimet Image Analyser Densitometer to measure these staining reactions. An ordination or segregation analysis of the results revealed a marked discrimination between tumours and the normal control mucosa, with the values for the 'transitional' mucosa and mucosa remote from the tumour graded in between. Paired t-tests also showed statistically significant differences between the various types of mucosa. The fact that changes in the staining densities of the sialic acid derivatives are observed in mucosa which is normal by conventional histological criteria raises the hypothesis that malignancy in the colonic epithelium is accompanied by modifications in the sialic acid composition of the mucus secretion. Thus the PB/KOH/PAS method may be of value in the early detection of cancer in colo-rectal biopsies.


Demonstration of phosphates in calcium deposits: a modification of von Kossa's reaction. It has been suggested that in von Kóss'as technic silver cations replace calcium bound to phosphate or carbonate groups and are then reduced to black metallic silver during exposure to light. However, in test tube experiments silver phosphate retains its yellow color for days. These differences between reactions of pure calcium phosphates and calcium deposits in tissues were emphasized already by von Kóssa; he regarded only the initial yellow coloration of calcium diagnostic for calcium phosphates and deplored the subsequent blackening caused by organic compounds. Von Kóssa's experiments were easily reproducible. A review of the literature showed that reduction of silver nitrate by organic compounds was well known in the 19th century. For histochemical studies of phosphates it was deemed desirable to avoid the formation of black by-products. Sections of paraffin-embedded human tissues were exposed to solutions of silver nitrate in subdued light or darkness then treated with sodium thiosulfate. Silver phosphate was yellow to yellowish brown; other tissue structures remained colorless. No darkening was observed in sections stored for eight years. Other compounds which form yellow silver salts, e.g. iodides and periodates, are unlikely to occur in paraffin sections of human tissues.


Calcium in isolated mitochondria from the left ventricular wall. The content of calcium per mg mitochondrial protein has been measured by conventional biochemical methods in myocardial tissue of some mammalian species. In addition, a method is presented for (1) the analysis of mitochondrial volumes in the same tissues and (2) calculating the amount of calcium in units of 10(6) mitochondria. It appears that a highly significant correlation exists between the calcium content and the number of mitochondria, with a positive correlation coefficient of 0.92. The mean mitochondrial volume in fractions of the rabbit myocardium was found to be 1.3386 micron3. Electron microscopic studies demonstrate pure mitochondrial fractions and only moderate structural alterations. The method described may represent a useful supplement for the estimation of calcium fluxes in mitochondria and of alterations in their volume, number and structure under conditions of myocardial ischemia.


Quantitative evaluation of the trapping reaction of copperthiocholine histochemical procedures for localization of cholinesterases. The aim of the present study was to investigate whether the trapping reaction of the histochemical procedure for the localization of ChE of Koelle and Friedenwald (1949) and its modification by Brzin and Pucihar (1976) proceeds quantitatively. The weight of the precipitate formed in the tissue sample during the histochemical procedure was compared with enzyme activity of an equal sample. The differential magnetic microbalance was used for measurements of reduced weight and for previous determination of density of the precipitate. The evidence for the composition of the final product was drawn from the quantitative analysis of copper and iodine and from the infrared spectra. Tsuji's statement (1974) that cuprous copper thiocholine iodide is the final product of histochemical procedures investigated was confirmed. It was found that the trapping reaction of the original as well as of the modified procedure under our experimental conditions in tissue sections proceeds quantitatively which means that one of the basic conditions for reliable localization is fulfilled.


Fluorescence histochemistry of peptide hormone-producing cells: observations on the phenanthrenequinone method for the demonstration of arginine residues. Phenanthrenequinone is a sensitive and specific fluorescence histochemical reagent for monosubstituted guanidines. It is probable that it selectively demonstrates the guanidino group of arginine residues of proteins and peptides. Phenanthrenequinone induces moderate to intense fluorescence in gastric chief cells, parenchymal cells of the pancreas, and certain peptide hormone-producing cell types such as the GH cells of the adenohypophysis and the glucagon cells of the pancreatic islets. Similar fluorescence spectra were obtained from an arginine-containing peptide in a histochemical model and from the GH cells of the adenohypophysis following exposure to phenanthrenequinone. We conclude that the cells demonstrated with this reagent store peptides or proteins rich in arginine.


Histochemical studies of mucosubstances in metaplastic epithelium of the stomach, with special reference to the development of intestinal metaplasia. Processes in the development of intestinal metaplasia of the stomach were investigated from the morphological and histochemical approaches using light and electron microscopic techniques. The specimens taken from 38 gastric carcinomas and 15 gastric and/or duodenal ulcers were subjected to this study. Morphological appearances of the intestinal metaplasia observed in routine examination with hematoxylin and eosin staining was able to divided into complete and incomplete metaplasia by the light and electron microscope histochemical stainings of the mucosubstances. The columnar cells at the area of the incomplete metaplasia had both the properties of the intestinal epithelia and the gastric foveolar epithelia. The incomplete as well as the complete metaplasia arose from the generative cells at the isthmus of the gland. The generative cells, however, sometimes gradually transformed to produce the complete metaplastic cells. The two processes of the development of the intestinal metaplasia were proposed and discussed.


The localization of calcium by X-ray microanalysis in myopathic muscle fibers. An electron histochemical study was undertaken to localize calcium with ammonium oxalate precipitation technique in soleus muscle of rat in normal cases and in myopathy induced experimentally by a prolonged treatment of 2,4-dichlorophenoxyacetate (2,4-D). The calcium content of precipitates was detected by energy-dispersive X-ray microanalysis. In normal cases, the electron dense precipitates containing calcium were mainly found in the vesicles of sarcoplasmic reticulum, whereas in 2,4-D induced myopathy the deposits were shifted near the Z line into the myofibrils. Calcium, because the uptake into sarcoplasmic vesicles was inhibited by 2,4-D, could attach to other binding sites, such as to the troponin-C.A long-lasting binding of calcium might lead to a prolonged activation of the actin-myosin system.


Electron cytochemical demonstration of four hydrolytic enzymes in the rat corpora lutea at second half of gestation. Corpora lutea from rat ovaries at mid pregnancy were fixed by perfusion and studied by electron cytochemistry for localisation of four hydrolytic enzymes. Using the metal-salt methods for acid phosphatase and aryl sulphatases activity was localised in small and large lysosomes, multivesicular bodies, Golgi complex and within cisternae of endoplasmic reticulum. The azo-dye coupling method for Beta-glucuronidase was less satisfactory and gave positive results in lysosomes, lipids and in the globules within the mitochondrial matrix. The latter two localisation were probably associated with affinity of the naphthol AS-BI for lipid material. In addition to plasma membranes, the reaction product for alkaline phosphatase with the lead-salt method was seen in lysosomelike bodies, in smooth endoplasmic reticulum and in occasional Golgi elements of granulosa lutein and endothelial cells. Increased activity of lysosomal acid hydrolases occurs when regressive changes of lutein cells start at the end of gestation and this might probably reflect the initiation of lytic processes.


Oxygen and the production of formazan from neotetrazolium chloride. Electrons, generated from dehydrogenase reactions, are transferred to oxygen in preference to neotetrazolium chloride. In model systems in solution the presence of a small amount of oxygen drastically reduces the rate of formazan production. The rate of reaction in tissue sections has been followed using scanning and integrating microdensitometry. As in solution, electrons are transferred preferentially to oxygen. However, oxygen seems unable to diffuse through the incubation medium and thus the supply of oxygen at the site of the enzyme activity becomes exhausted; the time taken to use up the oxygen will depend on the rate of the enzyme activity. It is only then that electrons are passed to the tetrazolium salt and formazan is precipitated.


Electron probe X-ray microanalysis of human skeletal muscle involved in rheumatoid arthritis. Electron probe X-ray microanalysis in the scanning microscope was used to determine the elemental composition of muscle fibres from patients with rheumatoid arthritis (RA). Quantitative data concerning phosphorus, sulphur, chlorine and potassium were correlated to the fibre type by a routine method based on serial cryosectioning and histochemical staining of adjacent sections. Significantly lowered sulphur values were found in type II A and II B muscle fibres of RA patients as compared to those of healthy controls. Traces of gold were detected in muscles of two patients treated with gold salts. The basis and mechanism for the decreased sulphur content in RA muscles are so far unknown, but may depend on the decreased amount of sulphur-rich potein(s).


Studies on the optimalisation and standardisation of the light microscopical succinate dehydrogenase histochemistry. The present investigation was undertaken in order to establish an optimal tissue pretreatment and an optimal incubation medium for the histochemical demonstration of succinate dehydrogenase (E.C. 1.3.99.1). The investigations were performed on steroid producing (testicle, adrenal gland) and steroid dependent (Fallopian tube) tissues. We studied the influences fo formalin fixation, acetone, magnesium ions, cyanides, electron carries (phenazine methosulfate, menadione coenzyme Q10), osmolarity, substrate concentration and inhibitors (oxalacetate, oxalate, malonate, 4-chloromercuribenzoic acid). The following procedure yields blameless morphological integrity and enzyme localization as well as optimal SDH-activity: Freezing of tissue cubes (diameter less than 5 mm) in propane cooled with liquid nitrogen or in melting freon. Incubation of 5 micrometer cryostat sections in narrow jars in the following medium (38.5 ml):--10 ml of 0.2 M sodium phosphate buffer pH 7.6 (52 mM).--18 mg tetranitro-BT in 0.5 ml dimethylformamide and aqua bidest. ad 10 ml (0.5 mM).--2.6 mg KCN in 16 ml aqua bidest. (1 mM).--540 mg succinate (disodium salt, hexahydrate) in 2 ml aqua bidest. (52 mM).--3 mg PMS (phenazine methosulfate) in 0.5 ml aqua bidest. (0.25 mM). The incubation medium has an osmolarity of 440 mosm. The incubation is carried out for 10 min at 37 degree C in darkness. To avoid non specific formazan deposits in lipid containing tissues a preincubation of the cryostat sections in 100% acetone at--22 degree C or--40 degree C for 7--10 min and an incubation time of 20--30 min is recommended. Control incubations adduced proof at the specificity of the SDH demonstration. Parallel incubation without PMS in order to determine indirectly the content of endogenous CoQ10 is further recommended.


Cytospectrophotometric study of the kinetics of histochemical reaction of GABA-transaminase in cryostat sections of rat cerebellar cortex. A modification of the histochemical method for demonstration of GABA-transaminase is proposed. Substrate and cofactor concentrations are chosen on the basis of kinetic investigation in cryostat sections of the rat cerebellar cortex. Enzymatic reactions were measured by quantitative microspectrophotometry. Michaelis constants for alpha-oxoglutarate in the Purkinje cell layer and granular layer (Km 1.7 x 10(-3) M) and white matter (Km 3.8 x 10(-3) M) are found. It is shown that alpha-oxoglutarate in concentrations higher than 5.2 x 10(-3) M (1 mg/ml) suppresses the reaction in sections by competitive inhibition. The advisability of addition of malonate, PMS and cyanide to the incubation medium is confirmed. It is suggested that there are some isoenzymes of GABA-transaminase with predominant localization either in neurons or glia.


Morphology of nuclear ribonucleoproteins in the early period of liver regeneration. A cytochemical study on the nuclei of hepatocytes in the regenerating liver after partial hepatectomy by the method of Higgins and Anderson was carried out. The behaviour of the ribonucleoprotein components of the nucleus, using EDTA according to Bernhard, revealed altered proportions of the fibrillar and granular components of the nucleoli in the early phase of regeneration and morphologic changes in nuclear RNP, especially in perichromatin fibrils. At 6 h after hepatectomy, the reaction with perichromatin fibrils (FP) was intense, localized around fields of condensed chromatin, which suggests intense transcription of extranucleolar RNA. The behaviour of the remaining nuclear ribonucleoproteins was uncharacteristic.


A modified wire-loop method for quantitative electron microscopic radioautography. A simplified method of emulsion coating and development for quantitative electron microscopic radioautography without any special instrument in introduced. Seven grids carrying ultrathin sections are attached to a glass block with double-coated Scotch tape. They are coated with a wire-loop, 2.5 cm in diameter. Ten glass blocks are then stuck on a slide, exposed, developed, fixed and stained simultaneously. Employing this method, the emulsion coating and the following procedures are easily accomplished under an identical condition.


Cytophotometric study of nuclear differentiation during pollen development in Tradescantia paludosa. During pollen development the dry weight, total protein, histone, DNA, arginine, and lysine content were analysed by cytophotometric methods in partially isolated nuclei. The amount of analysed substances increased from the end of the meiosis to the mitosis of the microspores to the double of the initial values. After mitosis the ratio histone/DNA remained almost unchanged in both vegetative and generative nuclei. On the other hand a large difference in the ratio non-histone protein/DNA could be observed, the vegetative nucleus containing more non-histone protein than the generative nucleus. The rate of RNA synthesis being higher in the vegetative nuclei, these non-histone proteins may have some function in nuclear activation. The DNA of the generative nucleus is duplicated before anthesis, whilst in the vegetative nucleus the DNA content remains constant.


Separation and purification of immunoglobulins M,A and G from small volumes of human sera by a continuous, inline chromatographic process. A continuous, in-line chromatographic process for the separation of immunoglobulins M, A and G from small volumes of human sera is described. The process involves the sequential and uninterrupted application of molecular sieve, ion-exchange, immunoadsorbent-affinity and de-saltin-concentrating chromatographic methodologies. From 1-3 ml of starting serum it yields 45-55%, by weight, of the three classes of immunoglobulins in 36 h. Each immunoglobulin fraction is free of contamination by immunoglobulins of the other two classes and retains full biological activity as measured by a sensitive immune lysis assay.


Studies of microhigh-performance liquid chromatography. IV. Application of the micro pre-column method to the analysis of corticosteroids in serum. The micro pre-column method, which is a useful method for the pre-treatment of samples, was applied to the analysis of corticosteroids in serum. Corticosteroids were concentrated in a micro pre-column packed with porous styrene-divinylbenzene copolymer particles and separated in a micro separation column packed with porous silica particles. Concentrations of corticosterone, cortisone and cortisol in the 20-130 ppb range were determined individually by pre-treatment of only 0.2 ml of serum.


Quantitative high-pressure liquid chromatographic procedure for the determination of plasma and tissue levels of 2,4-diamino-5-(3,4-dichlorophenyl)-6-methylpyrimidine (metoprine) and its application to the measurement of brain capillary permeability coefficients. A high-pressure liquid chromatographic assay for metoprine levels in aqueous media, serum, and tissue extracts has been developed that can measure concentrations of the drug in the 25-100 ng range with accuracy, reproducibility, and ease. The half-time for metoprine disappearance from rat plasma and capillary permeability coefficients for metroprine in rate brain tissue determined with this method are in excellent agreement with values determined for related drugs using labeled compounds.


Detection of estrogen-like compounds by thin-layer chromatography. There are different national food regulations concerning the use of hormoneactive substances in the breeding of domestic animals that serve as human food. This situation requires specific and sensitive methods for the detection of any residues in food. The hormones used in practice can be determined with improved sensitivity by coupling to the diazonium compound Fast Dark Blue R Salt. After thin-layer chromatography on high-performance thin-layer chromatographic plates with two different solvent mixtures, zeranol, estradiol, estriol and estrone are analysed as azo dyestuffs. The detection limit of these hormones is below 10 ng.


Detection of preservatives in pharmaceutical and cosmetic products using agar gel electrophoresis. A method is described using high voltage agar gel electrophoresis for the separation and microbiological detection of preservatives in pharmaceutical and cosmetic products. The positions of the preservatives were detected as zones of inhibition of a test organism within the nutrient agar gel and can be characterised by their migration, shape and presence of a halo. A wide range of preservatives commonly used in pharmaceutical and cosmetic preparations could be detected in low concentrations. The method however was not suitable for certain volatile preservatives and many primarily antifungal compounds could be detected above their use concentration.


Protein-losing enteropathy secondary to intestinal lymphangiectasia in a dog. Protein-losing enteropathy secondary to intestinal lymphangiectasia was diagnosed in a 1 1/2 -year-old female Doberman Pinscher. Poor growth rate, intermittent diarrhea, ascites, edema, hypoproteinemia, grossly dilated intestinal lymphatics, and impaired fat absorption characterized the disease. There was histopathologic evidence of dilatation of lacteals and excessive chromium (51Cr)-labeled human albumin in the feces following its intravenous administration. Sixteen weeks of a special diet (high in carbohydrates and medium-chain triglycerides; low in long-chain triglycerides) led to remission of clinical signs. Serum albumin increased over its initial value of 1.09 to 1.7 g/dl, while serum globulin increased from 1.71 to 1.9 g/dl. Following 9 months of dietary therapy, serum albumin increased to 2.1 g/dl and serum globulin increased to 2.6 g/dl.


Diskospondylitis in the dog: 27 cases.

Diskospondylitis in the dog: 27 cases. A retrospective study was undertaken to provide information about diskospondylitis in the dog. This disease has not been well recognized and documented in the veterinary literature. Diskospondylitic lesions were confirmed radiographically in 27 dogs (20 males and 7 females). Pain, inappetence, and recurring fever were the prominent signs in animals that had failed to respond to short-term therapy by referring clinicians. Clinical pathology did not yield consistently diagnostic results. Radiography, which was regarded as essential, revealed typical involvement of adjacent vertebral bodies, their end plates, and the joint space represented by their intervertebral disks. Diskospondylitis occurred as multiple and solitary lesions; it was also associated with myelopathy.


Fleabite allergic dermatitis: a review and survey of 330 cases. Clinical signs of acute fleabite allergic dermatitis (FAD) in dogs included intense pruritus and erythema. Dogs with chronic FAD had diminished pruritus. The primary lesion of FAD was a papule. Secondary lesions (hyperkeratosis and hyerpigmentation) were common. Diagnosis of FAD was based on history of flea infestation and on type and location of lesions. Intradermal testing with glycerinated flea antigen was of little diagnostic value. Treatment of FAD included (1) breaking the flea life cycle in the indoor and kennel environment by vacuuming and washing bedding as well as by the use of aerosol insecticides for fumigation, (2) minimizing flea infestation on the dogs by using insecticidal dips, baths, and flea collars, and (3) hyposensitization with flea antigen.


Acute necrotizing myelopathy from nucleus pulposus embolism in dogs with intervertebral disk degeneration. Two dogs that suddenly developed neurologic signs indicating a severe cervical spinal cord lesion were found at necropsy to have necrotizing myelopathy secondary to nucleus pulposus embolism. One dog had embolism of arteries and veins; in the other dog, veins alone were embolized. Each dog had early degeneration and extrusion of the C6--7 disk, from which emboli originated. In one dog, arterial emboli apparently entered inflammatory vasculature in the anulus fibrosus and were forced by extrusion back through the arterial system to the radicular artery and then to the cord. Venous embolism occurred secondarily to arterial embolism. In the other dog, venous embolism resulted from direct herniation of nucleus pulposus into the longitudinal venous sinus. A comparison of the pathogenesis of embolization in each case and a survey of other reported cases suggested that necrotizing myelopathy is a complication of early degeneration of intervertebral disks in large dogs.


Comparison of collection techniques for quantitative urine culture in dogs. The degree of bacterial contamination of 75 urine samples collected by voluntary voiding, catheterization, and cystocentesis was studied in 25 clinically normal dogs (14 females and 11 males) to evaluate the suitability of quantitative urine culture for diagnosis or urinary tract infection. Significant bacteriuria (greater than 100,000/ml) was not observed. Insignificant bacteriuria presumably caused by urethrogenital contaminants was detected in 44% of the urine samples collected by voluntary voiding, in 20% of the samples collected by catheterization, and in 12% of the samples collected by cystocentesis. The urine was sterile in 40% of the samples collected by voluntary voiding, in 80% of the samples collected by catheterization, and in 84% of the samples collected by cystocentesis. A bacteria count suggestive of bacteriuria (10,000--100,000/ml) was obtained in 1 sample collected by cystocentesis but was attributed to inadvertent penetration of a loop of intestine. Low bacteria counts were obtained in 4 (16%) samples collected by voluntary voiding, presumably as a result of urethrogenital and integumentary bacterial contaminants. It was concluded that urine samples properly collected from dogs by voluntary voiding, catheterization, or cystocentesis are suitable for detection of significant bacteriuria.


Features of cystic calculi in cats in a hospital population. The case records of 131 cats undergoing surgery for cystic calculi at The Animal Medical Center during the period of June 1969 to January 1975 were reviewed. Data for 16 intact females, 63 spayed females, 14 intact males, and 38 castrated males were analyzed. The mean age of first occurrence was 4.9 years. Meaningful conclusions regarding breed predisposition could not be made. Phosphate calculi were the predominant (124/128 analyzed) type. Swab specimens were taken from the bladder wall at time of surgery in 86 cases; only 28 (32.8%) were positive for bacteria.


Ammonia tolerance test in clinically normal dogs and in dogs with portosystemic shunts. The oral ammonia tolerance test was investigated in 20 clinically normal dogs and in 6 dogs with naturally occurring portosystemic shunts. The dogs with portosystemic shunting had a marked rise in venous blood ammonia following the administration of ammonium chloride, as compared with the control dogs. Fasting venous blood ammonia content was not uniformly reliable in separating the dogs with portosystemic shunting from the clinically normal dogs.


Cold-agglutinin hemolytic anemia and Haemobartonella canis infection in a dog. Cold-agglutinin hemolytic anemia in a dog was manifested by weakness, progressive hemolytic anemia, hematuria, and hemoglobinuria. Corticosteroid therapy failed to alleviate the anemia. The condition became complicated by Haemobartonella canis infection berminally. Prominent postmortem findings included disseminated thrombosis and infarction, glomerulonephritis, and thickened alveolar membranes.


Contagious equine metritis: clinical description of experimentally induced infection. Contagious equine metritis was reproduced experimentally in 6 pony mares. The disease was characterized by rapidly developing cervicitis and by varying amounts of exudate. The exudate, apparent as early as 48 hours after inoculation, drained from the cervix as a tenacious, mucopurulent discharge for several days, then rapidly disappeared. In all mares, the clinical disease cleared within several weeks of inoculation, without treatment.


Geometric anatomy of the aortic--common iliac bifurcation. Increased haemodynamic forces in the arterial wall may be among the factors responsible for atherogenesis and for determining the severity of the pathological process at a particular site. The present work was designed to help explain apparent asymmetries in the distribution of atheroma at the aorto-iliac bifurcation. A technique was devised for the quantitative evaluation of the geometric anatomy of this region. Measurements were made on 12 female and 14 male cadavers. In all cases there was an asymmetrical left lateral orientation of the abdominal aorta which resulted in a longer right common iliac artery, smaller left common iliac take-off angle and larger left radius of curvature at the aortic-common illiac bifurcation. These differences may explain, through variations in the local haemodynamics, the predilection of the disease for the left iliac artery, particularly in males with their relatively narrow pelvis. Knowledge of both the exact and the average numerical values associated with the local geometry are essential for a detailed haemodynamic study of the effect of these variations on atherogenesis.


The organization of neocortical projections from the ventroposterior thalamic complex in the marsupial brush-tailed possum, Trichosurus vulpecula: a horseradish peroxidase study. Retrograde transport of horseradish peroxidase (HRP) was used to determine the extent and some of the organizational details of the cortical projection of the ventroposterior thalamic complex (VP) in the marsupial brush-tailed possum, Trichosurus vulpecula. The cortical projection field of VP is coincident with SI as determined by electrophysiological methods, and would appear not to overlap fully the primary motor cortex. Thus, in Trichosurus it appears that the motor and somatic sensory cortical regions are not fully congruent, unlike those of the American opossum, Didelphis, which are. Each division of VP projects discretely, in a non-overlapping manner, to various regions within SI. The ventrolateral subdivision or VPL projects medially and in a strict homotypic manner, though the proportion of VPL cells projecting to cortex is subject to a large amount of variation. The dorsomedial division of VP or VPM projects uniformly to cortex from all areas of that subnucleus, but the strict homotypy characteristic of VPL's projection was not as apparent. VPM also projects to two distinct regions within its cortical field. The posteromedial division of VP or VPP projects to an area of cortex that receives no other VP input but, on the basis of cortical mapping studies, appears to belong to SI. Projections from VPL (and presumably from VPM) to a small area of cortex in the extreme posterolateral part of the VP field correspond to the position expected for, and electrophysiologically confirmed to be, SII.


Ultrastructure of the filiform papillae on the tongue of the hamster. The fine structure of the filiform papillae on the hamster tongue is described level by level from the basal layer to the surface. We did not observe two distinct types of cells with different morphology or components which could be held responsible for the production of two different types of keratin as have been described in other animals, but rather a uniformity of cell structures in each layer and only the so-called "smooth" type of keratin. However, keratin granules were more abundant in the anterior part of the papilla.


Effects of blood, bile and starch in the peritoneal cavity of the rat. The reaction of rat peritoneum to the intraperitoneal injection of rat blood, human bile, starch glove powder, and a suspension of starch in blood or bile was examined by histological section and en face microscopy of the peritoneal surface (Häutchen technique). Haemoperitoneum causes little destruction of the peritoneum, but bile produces some peritonitis with a purulent exudate and destruction of subperitoneal muscle. Starch granulomas form following intraperitoneal injection of glove powder, though 0.1 g is insufficient to produce adhesions. The addition of starch to blood or bile enhances the peritonitis and delays healing. These results indicate that bile and starch should not be left in the peritoneal cavity at the end of surgical procedure.


Quantitative studies on the maturation of central and peripheral parts of individual ventral motoneuron axons. I. Myelin sheath and axon calibre. The entire lengths of central and peripheral myelin sheaths on the same ventral motoneuron axons of young rats were studied during the rapid early phase of myelination which occurs during the first 2 weeks after birth. Sheath thickness varied along central internodes at all ages, though this variation was less in the more mature sheaths. Usually there was a single level of maximum thickness, most often near the middle of the internode, but frequently close to one end. Some sheaths varied little in thickness along their lengths; a few were bimodal. The number of turns decreased on either side of the maximum as the sheath was traced towards the bounding nodes. Comparison of serially adjacent sheaths along the same axon showed no relationship between their mean thickness or patterns of longitudinal thickness variation. Patterns of thickness variation in peripheral sheaths were very similar to those found centrally. At a given age, however, peripheral sheaths were less variable along more than twice that of central internodes belonging to the same fibre. At first, the unrolled glial unit was oval in outline while the unrolled Schwann cell was trapezoidal. Subsequently, both eventually became trapezoidal. The calibre of central and peripheral stretches of the same ventral motoneuron axons were also compared with one another in young rats during the first 2 weeks after birth. At each age, the great majority of axons were thicker peripherally than centrally. The mean circumference of the peripheral segment exceeded that of the central by a factor which averaged 1.3 over the period studied. Circumference varied from one level to another along both central and perpheral stretches of the same axons. The degree of this variation differed considerably from one axon to another. In many central fibres, and in all peripheral stretches of fibres, calibre variation followed no particular pattern. However, the calibre of some central axon segments gradually increased as the fibre was traced distally towards the cord surface. A proportion of axons was dilated just deep to the cord surface. These dilatations were frequently myelinated. No association was found between patterns of longitudinal variation in axon calibre and in sheath thickness along the same fibre.


The topographical anatomy and blood supply of the carotid body region of the domestic fowl. The topographical anatomy of the carotid body region was investigated on both sides in 30 adult domestic fowls (Gallus gallus domesticus). On each side of the body the two parathyroid glands always adhered to each other, the caudal gland being the smaller. The right cranial parathyroid usually touched the thyroid gland, but on the left side the two glands were nearly always separate. The ultimobranchial gland was usually attached to the parathyroid glands on the left side, but was always well separated on the right side. The carotid body, the identity of which was checked by light and electron microscopy, was ovoid in shape, whitish in colour, and about 0.8 x 0.6 x 0.5 mm in size. It was almost always in direct contact with the medial surface of one or both parathyroid glands have commonly been portrayed astopographically separate from each other. The oesophagotracheobronchial artery, the artery to the carotid body, and the caudal thyroid artery arose close together, though in variable positions, from the lateral aspect of the common carotid artery. The artery to the carotid body was always the middle of the three. The venous drainage of the carotid body was associated with the veins of the parathyroid and ultimobranchial glands. The carotid body was innervated by a filament from the distal vagal ganglion. The topographical observations on the carotid body region have been compared with those of other authors. The cranial and caudal parathyroid glands have commonly been portrayed as topographically separate from each other. The carotid body has seldom been depicted in direct contact with the parathyroid glands. The size of the carotid body has invariably been shown far too large. These discrepancies are discussed and are attributed largely to the reliance by other authors on serial reconstruction rather than microdissection.


The fibrous growth plate of the rat tibia: tritiated thymidine autoradiographic study. Tritiated thymidine autoradiographic studies have demonstrated that the intermediate fibrogenic zone of the fibrous growth plate at the upper end of the rat tibia is a site of intense cellular proliferation, the resulting cells differentiating into osteoblasts which manufacture the bundle bone at the distal end of the growth plate.


The velocity field of growing ear cartilage. The velocity vector field of the growing rabbit ear cartilage has been investigated between 12 and 299 days. Empirical curves have been computed for path lines and for velocities between 12 and 87 days. The tissue movement has been found to behave as an irrotational flow of material. Stream lines and velocity equipotential lines have been calculated and provide akinematic description of the changes during growth. The importance of a knowledge of the velocity vector in physical descriptions of growth and morphological differentiation at the tissue and cellular levels is emphasized.


Electron microscope observations on human fetal striated muscle. The superior rectus oculi muscle from human fetuses of 5, 9.2, 12 and 24 cm crown--rump length (of ages estimated to be 10, 12, 15 and 23 weeks respectively) have been examined by electron microscopy. "Myotube satellite cells" closely associated with myotubes and myocytes were present in all specimens, but their relative numbers declined with advancing age. Some were small with scanty cytoplasm containing few organelles. Others were rich in organelles, including Golgi apparatus, granular endoplasmic reticulum, comma and dumb-bell shaped dense bodies and centriole or basal body: these cells were numerous in the three smaller specimens but almost absent from the largest. Seemingly active "myotube satellite cells" often extended cytoplasmic processes beyond their confining basal laminae into the endomysial space to contact freelying cells of similar appearance, as well as axon-associated Schwann cells, often to form an extensive network. These "myotube satellite cells" resembled Schwann cells in all respects save association with axons, and it is suggested that they are, indeed, Schwann cells so disposed as to promote axonal growth towards differentiating, but as yet uninnervated, myocytes. Neuromuscular contacts were increasingly numerous with advancing age, usually where several or many axonal terminals contacted a relatively mature (myofilament-rich) muscle cell. Immature myotubes seldom made contact with axonal terminals, even when a closely adjacent (and "coupled") mature muscle cell did so. A sequence of axonal growth and retraction has been proposed which reconciles accounts of early but temporary polyneuronal innervation with commonly accepted ideas regarding the scattered distribution of the muscle components of motor units.


Stereological analysis of the duct system of the rabbit parotid gland. The duct system of the rabbit parotid gland constitutes about 5% of gland tissue volume and is divisible into intercalated and striated ducts in a volumetric ratio of about 3:2. The intercalated duct consists of low cuboidal epithelial cells (375 micrometer 2) surrounded by a myoepithlium, and the cells contain a few small secretion granules, particularly at the proximal end of the duct. The cells of the striated duct are larger (531 micrometer 3), columnal in shape, and show a mitochondrial compartment three times that of intercalated duct cells (16.5% of cell volume). These mitochondria are concentrated in a basal and perinuclear position, but they are largely absent from the apical cytoplasm, which is permeated with microfilaments and contains numerous small smooth membraned vesicles, but no clearly recognisable secretion granules. The lateral plasmalemma of these cells is complexly folded, and basal processes interdigitate with those of adjacent cells. This results in an increase in the ratio of apical to lateral/basal plasmalemma from 1:5 in intercalated cells to 1:24 in striated cells. Some slight changes in cell morphology were detected following isoprenaline-induced secretion of the gland in vivo. These included a small increase in the volume fraction of nuclei and mitochondria in intercalated duct cells, and depletion of their secretion granules. Change in striated cells was confirmed to a small increase in the volume of smooth membraned cytoplasmic vesicles. The structure of the duct and changes wrought by isoprenaline are discussed in the context of the role of the duct in the production of saliva.


The structure of aminoglycoside antibiotic 66-40G produced by Micromonospora inyoensis. Aminoglycoside antibiotic 66-40G is a minor component produced in the fermentation of Micromonospora inyoensis. Its structure has been established as 3''-de-N-methyl-sisomicin (4) by spectroscopic means and by direct comparison with an authentic sample obtained from photochemical oxidative de-N-methylation of sisomicin (1).


Isolation of tridecaptins A, B and C (studies on antibiotics from the genus Bacillus. XXIII). Three new antibiotics, tridecaptins A, B and C, were isolated from culture broths of strains of Bacillus polymyxa AR-110, B-2 and E-23, respectively. All are acyl tridecapeptides differing from each other in the fatty acid components and amino acid residues. They are weakly active against Gram-negative and Gram-positive bacteria in vitro and in vivo.


The structure of tridecaptin A (studies on antibiotics from the genus Bacillus. XXIV). On examining the structure of the antibiotic tridecaptin A, the constituent amino acids were determined to be: 2,4-diaminobutyric acid(2D, 1L), Ser(1D, 1L), Glu(1L), Gly(1), Ala(1L), Val(1D, 1L), aIle(1D), Phe(1L) and Trp(1D). The constituent fatty acid was identified as beta-hydroxy anteisononanoic acid by gas chromatography and mass spectrometry. Cleavage reaction with N-bromosuccinimide, sequential analysis by EDMAN degradation, partial acid hydrolysis and some additional evidences clarified the structure of tridecaptin A.


Aminoglycoside antibiotics. XII. Effect of N-alkylation in kanamycin antibiotics. Four N-ethyl derivatives of kanamycin A and five N-ethyl derivatives of kanamycin B were synthesized. The antimicrobial activity of N-ethyl kanamycin derivatives was determined against aminoglycoside-sensitive and resistant organisms. The structure-activity relationship of these compounds is discussed with reference to the activity of N-acyl kanamycin derivatives.


The syntheses and biological properties of 1-N-(S-4-amino-2-hydroxybutyryl)-gentamicin B and 1-N-(S-3-amino-2-hydroxypropionyl)-gentamicin B. The syntheses of 1-N-(S-4-amino-2-hydroxybutyryl)-gentamicin B and 1-N-(S-3-amino-2-hydroxypropionyl)-gentamicin B, designated sch 20287 and Sch 21420, respectively, by procedures similar to those developed by KAWAGUCHI and co-workers for the transformation of kanamycin A to amikacin are described. The in vitro microbiological properties of Sch 20287 and Sch 21420 are compared with amikacin, gentamicin and tobramycin.


Biological activity of SCH 21420, the 1-N-S-alpha-hydroxy-beta-aminopropionyl derivative of gentamicin B. Sch 21420, the 1-N-HAPA derivative of gentamicin B, has been compared to gentamicin, tobramycin and amikacin in a variety of in vitro and in vivo tests. Based on studies with a large number of sensitive and resistant bacteria, it was shown that Sch 21420 has a spectrum and potency essentially the same as that of amikacin. Serum levels in mice, rats, and dogs are similar to those of other aminoglycosides. Sch 21420 was found to be markedly less toxic than amikacin in chronic renal function tests in rats and thus appears to have an advantageously improved therapeutic index compared to amikacin.


Studies on the ionophorous antibiotics. XV The monovalent cation selective ionophorous activities of carriomycin, lonomycin and etheromycin. The cations selectivity profiles of the carboxylic ionophores, carriomycin, lonomycin and etheromycin, have been investigated by measuring the complexation affinities for metal cations and the cation transport activity through an organic barrier. In a two-phase partition study, carriomycin and lonomycin formed complexes more readily with K+ than with NH4+, Rb+ or Na+, but not with Li+ or Cs+. On the other hand, etheromycin exhibited a great preference for K+ or NH4+ over Na+, Li+ or Rb+, but displayed no binding affinity for Cs+. The alklaine degradation product of lonomycin exhibited a preference for K+ or Na+, but its complexation affinities were much lower than those of the parent compound. Carriomycin, lonomycin and etheromycin efficiently transported K+, Rb+ and Na+ through a CCl4 barrier. But did not carry Ca2+. These antibiotics caused a massive release of K+, Rb+ or Na+, but not of Li+ and Cs+, from mitochondria previously loaded with these cations by valinomycin or monazomycin. Thus, it is concluded that carriomycin, lonomycin and etheromycin are monovalent cation selective ionophores.


In vitro activity of tiamulin (81.723 HFU), a new pleuromulin derivative, against clinically significant anaerobes. The susceptibility of more than 40 strains of Gram-negative and Gram-positive anaerobes to tiamulin (Sandoz 81.723 hfu), a new pleuromulin (pleuromutilin) derivative, was determined by broth dilution and agar dilution tests. The influences of density of the inoculum upon MICs was studied by a specially designed pour plate-technique. Bacteroides fragilis, B. vulgatus, B. splanchnicus, B. oralis, B. asaccharolyticus, B. melaninogenicus, Fusobacterium fusiforme (F. nucleatum), Sphaerophorus necrophorus, Clostridium perfringens, C. fallax, Propionibacterium acnes and several species of Peptococcaceae showed broth dilution MICs of 0.03 similar to 1 microgram/ml. Members of B. thetaiotaomicron, B. distasonis and S. freundii (F. mortiferum) were inhibited by 8 similar to 32 microgram/ml and 2 strains of S. varius had a broth dilution MIC of 256 microgram/ml. With most strains, the agar dilution MICs were 2 similar to 4 similar to 8 times the broth dilution MICs. In pour plate-tests, the MICs were not considerably influenced influenced by varying initial concentrations of viable cells. With most anaerobes, the MBCs of tiamulin were more than 100-fold higher than the MICs. The results obtained indicated that, apart from S. varius, B. thetaiotaomicron, B. distasonis and S. freundii (F. mortiferum), members of 16 other anaerobic species including B. fragilis were without exception sensitive to tiamulin.


High speed liquid chromatographic determination of cephalexin in human plasma and urine. A rapid and accurate high speed liquid chromatographic method has been developed for the determination of cephalexin in human plasma and urine. The method involves micropore filtration of urine specimens and methanol extraction of plasma samples followed by HSLC separation on a bonded reverse phase column utilizing a mobile phase of methanol-water containing acetic acid. The quantitativity of UV response at 254 nm covered a wide range of cephalexin concentrations down to 0.5 microgram/ml, and no metabolite peaks were detected. The time courses of plasma level and urinary excretion were determined until 6 hours after oral administration of cephalexin capsules to healthy volunteers. The pharmacokinetic parameters were estimated using a two compartment open model.


A simultaneous comparison of three methods for language training with an autistic child: an experimental single case analysis. In a single-case, simultaneous-treatment design, three methods for experimental language acquisition in one autistic child were compared using a Latin square design and trend-line analysis. Results showed a total communication approach to be significantly superior to sign-based and verbalization approaches. The verbalization treatment resulted in decreased performance. Results indicate that use of a cross-modality inhibitory process to explain the alleged superiority of the sign-based approach is questionable. Variation among autistic children indicates a need for further research and for caution against premature acceptance of a given treatment approach or theoretical explanation.


Sensorimotor functioning and communication in mute autistic children. There were two purposes underlying this study: to describe the sensorimotor functioning of mute autistic children and to relate their sensorimotor performance to nonverbal communication. Twelve mute children, diagnosed autistic, ranging from 4 years 9 months to 12 years of age, were administered four scales of sensorimotor development from the Uzgiris and Hunt (1975) series: object permanence, gestural imitation, means for obtaining environmental events, and causality. Subjects performed most poorly on the imitation scale with 9 of 12 performing below Piaget's fifth sensorimotor stage. In contrast, performance was highest on the object permanence scale: No child scored below Stage V. Regarding the subjects' non-scales and Stage III on the imitation scale appeared to form minimal prerequisites for intentional communication in a variety of situations. Finally, none of the subjects, even those with relatively complete sensorimotor development, spontaneously used what Bates (1976) has called "protodeclarative" gestures to point out or show objects to adults. The absence of protodeclarative gestures may represent a qualitatively distinct pattern of prelinguistic development in certain autistic children.


Behavior, language, and development in three-year-old children. An epidemiological study of 3-year-old children showed there to be a marked association between behavior problems and language delay. Behavior problems were present in 14% of a random sample of 705 children, whereas 59% of 22 children with language delay had such problems. Data obtained from a battery of developmental tests were analyzed to examine the differences between children with behavior problems, matched controls, and children with language delay. It was found that children with behavior problems scored significantly lower on these tests, particularly those concerned with language. There were no significant differences in test scores between children with language delay only and those with combined behavior problems and language delay.


Handedness in autistics, retardates, and normals of a wide age range. Tests of handedness were carried out with 34 autistic children aged from 4 years 10 months to 18 years 11 months, and with sex-, age-, and IQ-matched retardates and sex- and age-matched normals. There were no significant differences between the groups on frequency of handedness, degree of righthand usage, or degree of dominant-hand usage. There was however a significant increase in the variance of dominant-hand usage from normals and retardates to autistics. These results were taken to indicate that earlier reports of handedness differences in young autistic children may reflect a developmental lag rather than a specific etiology of autism. Some support for this hypothesis was found from a comparison of age-trends between the groups. The relationship of such a developmental delay to the etiology of autism was discussed.


Disruptive behavior: a dietary approach. The effect of particular foods on levels of hyperactivity, uncontrolled laughter, and disruptive behaviors was studied in an 8-year-old autistic boy. The floor of the child's room was taped off into six equal-sized rectangles to measure general activity level. Frequency data were recorded on screaming, biting, scratching, and object throwing. A time-sample technique was used to record data on laughing. Data were gathered during four phases. During an initial 4-day period the child was fed a normal American diet. A 6-day fasting period followed, during which time only spring water was allowed. The third phase lasted 18 days and involved the presentation of individual foods. During the final phase of the study the child was given only foods that had not provoked a reaction in the third phase. Results showed that foods such as wheat, corn, tomatoes, sugar, mushrooms, and dairy products were instrumental in producing behavioral disorders with this child.


Cerebral asymmetry and the development of early infantile autism. Two experiments involving listening preferences of autistic and normal subjects were conducted to test the hypothesis that the right cerebral hemisphere is more active than the left hemisphere in autistic children. Results showed that when given a choice between verbal and musical material, the autistic children preferred music, while normal children showed no preference. Secondly, autistic children listened to both types of material predominantly with the left ear. Although normal subjects showed greater variation among themselves, they tended to listen to music more often with the left ear and to listen to verbal material more often with the right. These results support the notion that some autistic children are predominantly right-hemisphere processors.


Using self-stimulation as reinforcement for autistic children. This experiment examined the effects of using self-stimulatory behavior as reinforcement for spontaneous appropriate sentences in two autistic children. The children were put on a token system and always received one token for every spontaneous appropriate sentence they made. An ABABA design was employed. In condition A, the opportunity to self-stimulate was contingent on the payment of tokens (two tokens for 2 minutes of self-stimulation). In condition B, no tokens were required for self-stimulation. The results showed that both subjects exhibited a much higher rate of spontaneous appropriate sentences during the contingent self-stimulation (A) condition, demonstrating that self-stimulation functioned as an effective reinforcement. The possibility of using self-stimulation as reinforcement in the treatment of autistic children is discussed.


Caulobacter flagellar organelle: synthesis, compartmentation, and assembly. In Caulobacter crescentus biogenesis of the flagellar organelle occurs during one stage of its complex life cycle. Thus in synchronous cultures it is possible to assay the sequential synthesis and assembly of the flagellum and hook in vivo with a combination of biochemical and radioimmunological techniques. The periodicity of synthesis and the subcellular compartmentation of the basal hook and filament subunits were determined by radioimmune assay procedures. Unassembled 27,000-dalton (27K) flagellin was preferentially located in isolated membrane fractions, whereas the 25K flagellin was distributed between the membrane and cytoplasm. The synthesis of hook began before that of flagellin, although appreciable overlap of the two processes occurred. Initiation of filament assembly coincided with the association of newly synthesized hook and flagellin subunits. Caulobacter flagella are unusual in that they contain two different flagellin subunits. Data are presented which suggest that the ratio of the two flagellin subunits changes along the length of the filament. Only the newly synthesized 25K flagellin subunit is detected in filaments assembled during the swarmer cell stage. By monitoring the appearance of flagellar hooks in the culture medium, the time at which flagella are released was determined.


Membrane phospholipid composition of Caulobacter crescentus. The phospholipid composition of Caulobacter crescentus CB13 and CB15 was determined. The acidic phospholipids, phosphatidylglycerol and cardiolipin, comprise approximately 87% of the total phospholipids. Neither phosphatidylethanolamine nor its precursor phosphatidylserine was detected. The outer and inner membranes of C. crescentus CB13 were separated, and phospholipid analysis revealed heterogeneity with respect to the relative amounts of phosphatidylglycerol and cardiolipin in the two membranes. As has been shown to be the case for other bacterial membranes, the concentration of cardiolipin increases and phosphatidylglycerol decreases as cell cultures enter stationary phase.


Selection for nonbuoyant morphological mutants of Caulobacter crescentus. Morphological mutants of Caulobacter crescentus were isolated by selecting for cells that did not possess normal, buoyancy-conferring stalks. The most prevalent phenotype enriched by the selection was structurally deficient stalks (designated "Cds"), observable as crumpled or flattened stalks. The second phenotype ("Abs") was observed as spontaneous shedding of normal-appearing stalks. Third, one mutant ("Ecs") was isolated that sheds not only stalks, but also miniature stalked and nonstalked cells.


Effect of growth temperature on membrane fatty acid composition and susceptibility to cold shock of Bacillus amyloliquefaciens. We investigated the fatty acid composition of the membrane of Bacillus amyloliquefaciens grown at different temperatures. A decrease in growth temperature was accompanied by an increase in the ratio of branched- to straight-chain fatty acids and a marked increase in the level of unsaturation of branched-chain fatty acids. When cells of this organism grown at 30 degrees C were cold shocked, viability and ability to secrete extracellular protease were lost. Growth of this organism at lower temperatures or addition of Tween 80 to cells caused the critical temperature zone for cold shocking to be lowered significantly. These results suggest a direct correlation between membrane fluidity and the susceptibility to cold shock.


Stimulation of tetrapyrrole formation in Rhizobium japonicum by restricted aeration. Cultures of Rhizobium japonicum were grown with vigorous aeration to stationary phase and were then incubated under restricted aeration for several days. Under these "microaerobic" conditions, cellular heme content increased 10-fold, and visible amounts of porphyrins were released into the culture medium. The two predominant porphyrins produced were identified, on the basis of their spectrophotometric and chromatographic properties, as protoporphyrin and coproporphyrin. The cytochrome complement of microaerobic cells partially resembled that of the symbiotic bacteria in that cytochromes alpha-alpha3 were absent and a CO-binding cytochrome 552 was present. During the period of restricted aeration, at the time that the heme content was increasing, there was a similar 10-fold increase in the activities of the first two enzymes of heme biosynthesis, delta-aminolevulinic acid synthase and delta-aminolevulinic acid dehydrase. However, during the same period, the activity of succinyl thiokinase (an enzyme that is required in large amounts whether or not heme is being produced) increased only twofold. These results suggest that reduced oxygen tension may play a role in inducing heme synthesis necessary for leghemoglobin formation and bacterial differentiation in soybean root nodules.


Ligninolytic enzyme system of Phanaerochaete chrysosporium: synthesized in the absence of lignin in response to nitrogen starvation. The relationship between growth, nutrient nitrogen assimilation, and the appearance of ligninolytic activity was examined in stationary batch cultures of the wood-destroying hymenomycete Phanerochaete chrysosporium Burds. grown under conditions optimized for lignin metabolism. A reproducible sequence of events followed inoculation: 0 to 24 h, germination, linear growth, and depletion of nutrient nitrogen; 24 to 48 h, cessation of linear growth and derepression of ammonium permease activity (demonstrating nitrogen starvation); 72 to 96 h, appearance of ligninolytic activity (synthetic 14C-lignin leads to 14CO2). Experiments with cycloheximide demonstrated that appearance of ligninolytic activity occurs irrespective of the presence of lignin; lignin did not induce additional activity. Addition of NH4+ to cultures immediately prior to the time of appearance of the ligninolytic system delayed its appearance, suggesting that the NH4+ led to interference with synthesis of the enzyme system. Addition of NH4+ to ligninolytic cultures resulted in an eventual, temporary decrease in ligninolytic activity. The results suggest that all or essential protein components of the ligninolytic enzyme system are synthesized as part of a series of physiological ("secondary metabolic") events that are initiated by nutrient nitrogen starvation.


Isolation and characterization of the pesticide-degrading plasmid pJP1 from Alcaligenes paradoxus. A strain of Alcaligenes paradoxus, unable to degrade phenoxyacetic acid, was shown to degrade two synthetic derivatives of this molecule, the herbicides 2,4-dichlorophenoxyacetic acid and 2-methyl-4-chlorophenoxyacetic acid. The ability to degrade these pesticides is encoded by a 58-megadalton conjugal plasmid, pJP1.


Newly discovered polyamine, 2-hydroxyspermidine, in Pseudomonas acidovorans. A previously unknown hydroxylated polyamine has been recovered from Pseudomonas acidovorans 29. It has been identified as 2-hydroxyspermidine, N4-(3-aminopropyl)-1,4-diaminobutane-2-ol, by its chromatographic behavior, electrophoretic mobility, and reaction with metaperiodate. It can be synthesized enzymatically from 2-hydroxyputrescine by cell-free preparations from Escherichia coli or P. acidovorans 29 which contain propylamine transferase. It is interesting to note that the naturally occurring compound is the 2-hydroxyspermidine and not the 3-hydroxyspermidine, N1-(3-aminopropyl)-1,4-diaminobutane-2-ol, indicating that the propylamine transferase reacts preferentially with the amine distal to the hydroxyl group. A mixture of 2- and 3-hydroxyspermidines and hydroxyspermine was synthesized by reacting acrylonitrile with 2-hydroxyspermidine and catalytic reduction of the products with hydrogen. N-(gamma-aminopropyl)-beta-alanine, used to help identify the hydroxyspermidines, was synthesized from N-(3-aminopropyl)-3-aminopropanenitrile by hydrolysis with 10% NaOH.


Mycoplasma growth factors in bovine serum fraction. Mycoplasma growth factors in bovine serum fraction were separated by Sephadex G150 column chromatography and density ultracentrifugation. The major growth factor of bovine serum fraction eluted from the Sephadex column in the void volume. Its growth-supporting activity was greatly enhanced by the presence of bovine serum albumin in the mycoplasma culture media. Other investigators had previously identified the major growth factor in serum as an alpha-lipoprotein. Although density ultracentrifugation revealed the presence of traces of a high-density lipoprotein in bovine serum fraction, another, less dense component, isolated by ultracentrifugation (component 3) and containing cholesterol, cholesteryl esters, free fatty acids, triglycerides, and protein, but no lipoprotein, exhibited considerably more growth-supporting activity than did the high-density lipoprotein, thus indicating that at least two mycoplasma species do not require intact serum lipoprotein for growth. Both the high-density lipoprotein and component 3 exhibited maximum activity only in the presence of bovine serum albumin. A chloroform extract containing component 3 lipids combined with bovine serum albumin to form an effective, partially defined, less complex substitute for serum in mycoplasma culture media.


Evidence for a functional glyoxylate cycle in the leishmaniae. Isocitrate lyase (EC 4.1.3.1) and malate synthase (EC 4.1.3.2), the two enzymes characteristic of the glyoxylate cycle, were demonstrated in promastigotes of five species of Leishmania (L. brasiliensis, L. donovani, L. mexicana, L. tarentolae, and L. tropica). Both enzymes were present in cells grown in a medium containing 10 mM glucose. Substitution of glucose with 20 mM acetate did not enhance enzyme levels. Acetate was readily taken up and metabolized by the cells. The distribution of label from acetate into various intermediary metabolites indicates a functional glyoxylate cycle and its role in gluconeogenesis/glyconeogenesis. The glyoxylate cycle in conjunction with alanine-glyoxylate aminotransferase and glyoxylate-aspartate aminotransferase could also be important in providing glyoxylate, the precursor for glycine biosynthesis.