Water soluble proteoglycans from bovine duodenal mucosa. Glycosaminoglycan-protein complexes were extracted from bovine duodenal mucosa with distilled water, resulting in solubilization of a fraction of the total proteoglycan of the tissue. The extracted material was purified by anion exchange chromatography on DEAE-Sephadex A-25, and then characterized by chemical analysis and by fractionation on Dowex 1. By using these procedures, two major fractions were identified, which were eluted from Dowex with 1.0-1.25 M NaC1 and with 1.5-1.75 M NaC1 respectively. Analyses showed that both fractions were mainly composed of glucosamine-containing, hyaluronidase-resistant polysaccharides, which were identified by their N-sulphate: D-glucosamine and total sulphate: D-glucosamine ratios as heparan-sulphate in the less acidic fraction, and as heparin in the more acidic fraction. Dermatan sulphate molecules were also present in both preparations, with an approximate ratio 1:3 to the glucosamine-containing polysaccharides. Solubility behaviour of the complexes formed by the isolated polyanionic molecules with cetylpyridinium chloride was strongly modified by papain digestion of the duodenal material. This reduction of molecular size of papain treatment suggests that the molecules extracted with water from duodenal mucosa are complex proteoglycans, perhaps in the native state.


Pattern of prophylactic use of antibiotics in six surgical departments of a teaching hospital in Jerusalem. The prophylactic use of antibiotics for various surgical procedures was studied in six surgical departments of the Hadassah University Hospital over an 18-month period between January 1972 and June 1973. A total of 7,339 patients was discharged from the six departments during this period. Antibiotics had been used prophylactically in 17.7% of these patients and in 20.4% of the 6,313 patients in whom there was no evidence of infection on admission. The rate of prophylactic use varied from 12.8 to 47.8% in different departments. Different prophylactic regimes were used in the various departments, but there was a tendency for the same regime to be used within one department, irrespective of the nature of the surgical procedure. The antibiotics were usually started postoperatively. The duration of the prophylactic courses varied but there was a general tendency to continue for more than five days, often up to 10 or more days. The frequency of use seemed to be related to the presence of factors regarded as carrying a high risk of infection, such as advanced age, operation through a contaminated field and the presence of malignancy. The association between prophylactic use and subsequent wound infection was examined. The importance of continuous surveillance of the prophylactic use of antibiotics and the role of the infection-control unit in this surveillance is stressed.


Purification and properties of rat liver L-threonine deaminase. Rat liver threonine deaminase has been partially purified. The enzyme deaminates L-threonine and L-serine, is not affected by isoleucine, nor by AMP and ADP. L-cysteine and analogues are inhibitors of threonine deaminase and it is very likely that the inhibition is due to the formation of a thiazolidine ring with PLP bound to the enzyme. However, the simple formation of this ring does seem to explain completely the different degree and type of inhibition shown by L-cysteine and analogues. The hypothesis that the different behaviour of L- and D-cysteine is due also to interactions independent of the formation of thiazolidinic ring is discussed.


Cyclic combination chemotherapy for metastatic breast cancer. Forty-four patients with metastatic breast cancer were treated with monthly courses of cyclophosphamide, methotrexate and 5-fluorouracil. Complete remission was achieved in five of the 44 patients (11.4%), partial remission in 17 (38.7%) and stabilization in 19 (43.2%). Progression of the disease was seen in three patients (6.7%). Forty patients are still alive and continue to receive chemotherapy. The main side effects were leukopenia, thrombocytopenia, weakness, nausea and vomiting; all were mild and transient, and were treated symptomatically.


Characterization of the polymer formed from methylglyoxal in the presence of L(+)-lysine. L(+)--lysine reacts with methylglyoxal in aqueous solution to give a yellow polymer. This polymer has been subjected to some chemical and physico-chemical characterizations. The gross structure of the polymer is probably formed by 3-hydroxypyrrolic nuclei, bridged together by vinylenic groups or -CH(OH)-CH2-CO-CH(OH)-groups.


Tinea inguinalis treated with miconazole cream: a double-blind study. A double-blind study on the effect of 2% miconazole cream and its vehicle was conducted on 84 men with tinea inguinalis. Relief of itching and improved clinical and mycological cure rates were noted after 14 and 28 days of treatment with miconazole in patients in whom the causal agent was Trichophyton rubrum. No conclusion about the efficacy of the active substance could be drawn in patients in whom the causal agent was some other dermatophyte or Candida.


Infectious hepatitis and glucose-6-phosphate dehydrogenase deficiency. The occurrence of infectious hepatitis in patients with coexistent glucose-6-phosphate dehydrogenase (G6PD) deficiency may present a clinical picture similar to that of fulminant hepatitis. To determine the factors which enable a rapid diagnosis of this disease combination, the clinical and biochemical findings in 14 patients were compared with those in 50 patients with uncomplicated hepatitis and those in 14 patients with fulminant hepatitis. Similarities with the latter group included persistence of fever, tachycardia, leukocytosis and hyperbilirubinemia of greater than 340 mumol/liter (20 mg/dl). A rise in bilirubin values of more than 50 mumol/liter (3 mg/dl) per day in patients with viral hepatitis strongly suggested the presence of G6PD deficiency. Despite the severity of the illness, prothrombin activity was well maintained in G6PD-deficienct patients.


Renal failure and interstitial nephritis due to trichloroethylene anesthesia and high-dose penicillin. An unusual case of acute renal failure is described. The patient, who had no previous history or signs of renal impairment, underwent lumbar laminectomy under general anesthesia with trichloroethylene (Trilene) and nitrous oxide. On the fifth postoperative day i.v. administration of 12 million units of penicillin was started. Within 16 h the patient developed oliguria and a clinical picture of acute renal failure. The course of the disease was prolonged and necessitated four hemadialysis treatments. The patient recovered only about 50% of his renal function. A biopsy performed two months after the onset of the acute renal failure was interpreted as indicating tubulo-interstitial nephritis. The role of trichloroethylene in the etiology of acute renal failure is discussed.


Study on protein content of rabbit reticulocyte ribosomal subunits: correlation with functional activity. The correlation between chemical composition (protein content) of native and derived ribosomal subunits and their activity in a cell-free system, programmed with haemoglobin messenger RNA, has been investigated. On the basis of isodensity sedimentation of formaldehyde fixed particles in CsC1 and of two-dimensional gel electrophoresis of ribosomal proteins, it appears that the native ribosomal subparticles contain a large protein complement than the derived subparticles and, correspondingly, they are less dependent on an external supply of initiation factors for haemoglobin synthesis in vitro.


Renal tubular acidosis due to the milk-alkali syndrome. A 60-year-old man with a history of excessive ingestion of calcium carbonate presented with azotemia, hypercalcemia and hyperphosphatemia. His acid-base status was initially normal. Following the cessation of calcium carbonate treatment, the hypercalcemia and azotemia disappeared, and the patient was found to be in metabolic acidosis with blunted acid excretion and a urine pH of 6.1. Kidney biopsy showed focal tubular calcification; the tubular damage was apparently caused by hypercalcemia and had resulted in renal tubular acidosis. During the three months of observation since that time there has been a tendecy for spontaneous remission of the renal tubular acidosis. Impaired renal hydrogen ion excretion prevented the development of metabolic alkalosis despite ingestion of alkali initially, and was later responsible for the metabolic acidosis. Renal tubular acidosis occurring as a sequel to the milk-alkali syndrome may aggravate the danger of nephrocalcinosis in this syndrome.


[Etiopathogenesis and epidemiology of candidiasis and related yeast mycoses]. The clinically and histologically well differentiated clinical picture of candidosis (CA) is heterogenous with regard to etiology: Only about 95 per cents of the cases are caused by candida species, the rest by other cryptococcaceae species. Formal pathogenetically the CA may be primary as well as secondary. Which kind of pathogenicity is predominating depends first of all on the momentary ecological, civilisational and medical conditions. At present in unselected dermatological cases at least 90 per cents are secondary; the severe systemic cases without exception. Primarily the pathogenesis of CA is determined by certain disposing factors, being partly exogenous, partly endogenous and/or constitutional, conditional or accidental, the knowledge and consideration of them representing the basis for an effective therapy. Particular CA-forms are largely characteristic for the kind of "underlying disease". Beside the disposing factors in the pathogenesis of CA also the quantity of the germs is of great importance whereas their virulence has only a minor meaning.


[Mycoplasma infections of urogenital tract]. Our investigationes during the last seven years stress the importance of mycoplasma hominis and ureaplasma on the so called abacterial inflammation of the urogenital tract. Because of diagnostic problems and differential diagnoses only specialized centers should explore these inflammatory conditions. Therapy is possible, however, by dermatologists, urologists, gynecologists, and general practitioners.


[Mycoplasma induced damage to sperm head in infertile men]. The heavy contamination of T-mycoplasmas in ejaculates of infertile men can be found in coiled forms as well as in swollen necks of the spermatozoa. The analysis of the sperma of one infertile man shows significant changes of the head of numerous spermatozoa possible caused by mycoplasmas. Our findings support the assumption of a correlation between T-mycoplasmas in ejaculates and the infertility of men.


[Verrucous type of Thomson's syndrome. A contribution to the congenital poikilodermias]. A case of a verrucous type of Thomson's syndrome in a 12 year old boy is presented. The verrucous type is discussed as a prognostically serious form of Thomson's syndrome, because of tendency to malignant degeneration. A review of the literature is given.


[Antibiotics resistance of staphylococci in skin diseases]. Cutaneous infection due to bacteria are predominantly caused by staphylococci. The sensitivity of 1036 staphylococcal strains towards various topical and systemical antibiotics was investigated over a period of three years (1973-1975). Comparing the present results with results from previous investigations revealed that staphylococcal sensitivity towards common antibiotic drugs remained nearly unchanged. A comparatively high proportion of strains has acquired resistance against multiple antibiotics. The importance of routinely determining bacterial sensitivity in antibiotic therapy is emphasized.


[Granulomatous cheilitis and Crohn's disease]. A case of Cheilitis granulomatosa is described. The various etiologic hypotheses concerning the disease are summarized and a relation with Crohn's disease is emphasized in this case. The Cheilitis granulomatosa Miescher is to be considered as a cutaneous symptom which can be observed in various diseases (e.g. sarcoidosis, herpetic infections. Crohn's disease, panniculitis).


[Lymphangiosis melanoblastomatosa].

[Lymphangiosis melanoblastomatosa]. After endolymphatic therapy and following surgical excision of lymphnodes in a patient with malignant melanoma the involved leg became edematous. In the edematous tissue there was a wide and diffuse spreading of the melanoma cells. Besides numerous cutaneous and subcutaneous metastatic nodules there were areas of blue-black coloring which showed a fine and reticular pattern. Histology confirmed that melanoma cells had filled up predominantly the superficial lymphatic vessels. Fluorescence histochemical investigations showed that these cells were viable and pigment-producing melanoma cells. There were also single enlarged melanocytes in the basal part of the epidermis with a high amount of fluorophores.


Facility location: a review of context-free and EMS models. EMS location models are those formulated to address specific problems of emergency medical services systems; context-free location models are those developed without reference to particular applications. The literature on these two types of public facility location models is reviewed, and the development of the maximal covering model from several earlier context-free models is described, with emphasis on problem statements and articulation of service objectives. An application of the maximal covering model to fire truck location points up the ability of this model to handle multiple objectives; its ability to compare alternative solutions gives it great utility for planning and evaluating EMS systems of a wide range of complexity. Potential applications of the maximal covering model are discussed regarding EMS problems involving multiple time standards and service objectives, location of special equipment, and siting of fixed facilities.


Are "medically underserved areas" medically underserved? A comparison of medically underserved areas (MUAs) and adequately served areas (ASAs) is presented. Nonmetropolitan areas represented in the Health Interview Survey (HIS) are classified as MUAs or ASAs by the official criterion of their scores on the Index of Medical Underservice (IMU), and HIS data from the two types of areas are examined for differences. Standard metropolitan statistical areas are also compared with the nonmetropolitan MUAs and ASAs. Results show no difference between MUA and ASA residents in number of physician visits per year or proportion with at least one visit in the past year, although MUA residents reported poorer health status, used some preventive services less, and used nonsurgical hospitalization more than did ASA residents. In gereral, most MUA-ASA differences tend to be similar in size to differences between ASAs and SMSAs. An alternative to the IMU, using HIS data to identify underserved areas, is discussed.


Size and performance of planning agencies. A preliminary analysis of the 1973-74 national assessment of comprehensive health planning [P.L. 89-749, sections 314 (a) and (b)] agencies is reported. The analysis suggests that agency staff size and budget are more strongly related to high performance scores than is size of population served. Complete analysis of the 1973-74 assessment data was not possible because of time constraints; it is argued that plans should be made for detailed analysis of data from future assessments of planning agencies under P.L. 93-641 so that factors relevant to performance may be identified more precisely and incorporated into agency structure.


Substance P and 5-HT in granules isolated from an intestinal argentaffin carcinoid. Intestinal argentaffin carcinoids, thought to originate from enterochromaffin cells, occasionally contain large amounts of substance P-like immunoreactivity in addition to 5-HT. The cytoplasmic granules of one such tumour were isolated. The granules, which in the electron microscope were shown to be argentaffin, contained both substance P-like immunoreactivity and 5-HT. The results support the view that substance P is localized in a population of enterochromaffin cells where it is stored in the cytoplasmic granules together with 5-HT.


Magnesium ions in catecholamine fluorescence histochemistry. Application to the cryostat and vibratome techniques. The effects of high concentrations of magnesium ions in the cryostat and Vibratome procedures for visualization of catecholamine fluorescence in the central nervous system have been investigated. In cryostat sections, obtained from specimens perfused with a formaldehyde and glyoxylic acid containing buffer, the addition of high concentrations of MgSO4 to the perfusion solution enhances the fluorescence intensity and reduces the unspecific background fluorescence and the diffusion of the catecholamine fluorophore. This improves the visualization of all portions of the central catecholamine-containing neurons. Similar effects are obtained in the formaldehyde-Vibratome technique by the introduction of an immersion bath containing MgSO4 after the sectioning procedure. The use of the magnesium perfusion or immersion steps furthermore increases the reproducibility of the Vibratome and cryostat techniques. The paper describes the improved Vibratome and cryostat techniques used in our laboratory.


[A new image-processing system designed for densitometry and pattern analysis of microscopic specimen. Application to the automated recognition and counting of cells in the various phases of the mitotic cycle (author's transl)]. A new image analysing system, designed for microphotometric measurement and pattern recognition has been applied in the discrimination of cells from the various phases of the mitotic cycle. The data acquisition procedure is controlled by a programmable electronic unit and involves the combination of the shifting of the microscope moving stages and the scanning of the successive fields by a mechanical device. The data processing is achieved by a computer. The preliminary results we obtained have shown that such a system allows the automatic recognition and counting of the M, G1, S and G2 cells as also the G0 resting cells. The most useful parameters of the cell proliferation kinetics are thus obtained from a single specimen of a cell population.


[Suitability of naphthyl-alpha-L-fucosides for the investigation of alpha-L-fucosidases (author's transl)]. In comparison with 1- and 2-naphthyl beta-D-glucoside, beta-D-galactoside, beta-D-glucuronide, beta-D-N-acetylglucosaminide, alpha-D-glucoside, alpha-D-galactoside and alpha-D-mannoside 1- and 2-naphthyl alpha-L-fucoside are hydrolyzed more quickly or to the same extent by homogenates prepared from freeze-dried cryostate sections of various rat organs. Nevertheless, when the fucosides are employed for the histochemical demonstration of alpha-L-fucosidase mostly negative data were obtained independent on the method used, whereas all other naphthyl glycosies deliver positive results. The reasons for these discrepancies are the marked inhibition of alpha-L-fucosidase by aldehyde fixation and diazonium salts. Then, alpha-L-fucosidase activity is suppressed to 90% and between 85 and 98% respectively; the inhibition of alpha- and beta-D-glucosidase, alpha- and beta-D-galactosidase, alpha-D-mannosidase, beta-D-glucuronidase and beta-D-N-acetylglucosaminidase by the fixative or coupling reagent does not exceed 70%. Therefore 1- and 2-naphthyl alpha-L-fucoside cannot be recommended in general for histochemical purposes. Small amounts of dimethylformamide do not influence the activity of most of the glycosidases investigated. For biochemical measurements, however, especially 1-naphthyl alpha-L-fucoside represents a suitable alternative in a fluorometric procedure instead of p-nitrophenyl alpha-L-fucoside used for the photometric evaluation of alpha-L-fucosidase. With the fluorometric method the enzyme was measured in rat organs, which posses remarkably different activities of alpha-L-fucosidase.


Localization of eosinophils in the thymus by the peroxidase reaction. Thymuses of human fetuses and infants and of young mice were investigated histochemically for peroxidase. Eosinophils were shown to be the only peroxidase-positive cells in the thymus. In human thymuses the eosinophilic cells were predominantly localized in medullar areas, with concentration of cell clusters at the cortico-medullar junction, around or inside Hassall's bodies and occasionally in high numbers in the intraseptal vessels of the cortex. In the normal mouse the eosinophils were evenly distributed throughout the medulla. Treatment with corticosteroids or X-rays produced a severe involution of the thymus with concommitant change in cellular pattern. The central areas of the thymus residue contained lymphocytes while the peripheral regions consisted of reticuloepithelia, macrophages and numerous eosinophils. Azathioprine did not change the morphology of the thymus. The numbers of eosinophils were slightly reduced, the distribution pattern remaining unchanged.


Cytofluorometric analysis of cell proliferation and differentiation of the human erythroblasts. The method of cytofluorometric measurement of the contents of Hb and nuclear DNA on a single erythroid cell (Fukuda et al., l975; 1977a) was used for the quantitative analysis of the erythropoiesis in normal bone marrow. The intracellular Hb in an erythroid cell was converted to fluorescent porphyrin after removing the Giemsa staning by irradiation with violet light in the presence of SH-donor (mercaptoethylamine hydrochloride, MEA) and its nuclear DNA was subsequently stained with pararosaniline Feulgen staining. With the two quantitative parameters, Hb content and DNA amount, the erythroid cells in normal human bone marrow were classified into 6 classes of different maturation stages (EI-EIV). The morphological characteristics of the most primitive erythroblast (EI cells) were described. The "proerythroblasts" which were identified on the bases of morphological criteria had in general aneuploid amounts of nuclear DNA with disproportional contents of Hb, thereby indicating that they are rather aberrations from the normal steps of cell maturation. The DNA amounts of "orthochromatic erythroblasts" (EV cells) showed continuous decrease from diploid range to almost zero suggesting that the removal of nuclear DNA from the erythroblast is not exclusively due to mechanical explusion of a whole intact nucleus.


High-speed autoradiography of 3H-thymidine-labelled nuclei. High-speed autoradiography with stripping film of 3H-thymidine-labelled cells was tested. The tests involved: (a) various times of immersion of emulsion-covered cell preparations in the mixture of dioxane-PPO-POPOP, at 20 degrees C, (b) exposure of cell preparations and blanks for various times at either -70 degrees C or +20 degrees C, with different humidity levels. Autoradiographs of good quality could be produced by 2-min immersion in the scintillator, exposure time greater than or equal to 1 h at either temperature and relative humidity 20--30%. A linear relationship between autoradiographic efficiency and exposure time of 1--7 h was found at either temperature, although the efficiency of autoradiographs exposed at -70 degrees C was by approximately 30% higher than that of autoradiographs exposed at +20 degrees C. Background values of autoradiographs dried with a fan and exposed for 1/4--7 h at either -70 degrees C or +20 degrees C were 0.6--0.8 grain/100 micron2. Theoretical calculations and experimental data showed that high-speed autoradiographs are 30--50 times more efficient as compared with conventional stripping film autoradiographs, thus allowing a shortening of the respective exposure time. Theoretical aspects of efficiency and resolution of high-speed autoradiography are considered.


[Microsurgery of the temporal bone (author's transl)]. Microsurgery of the temporal bone has opened a new frontier in otology, neurosurgery and head and neck surgery. The operative approaches used to reach the most hidden supra-, retro-, infra- and premeatal regions of the temporal bone are described and their surgical uses are illustrated. Particular attention has been paid to the subtotal petrosectomy with permanent anterior transposition of the facial nerve and is indicated for the extirpation of large infra- and premeatal tumors as well as for the repair of sub- and infratemporal aneurysms of the internal carotid artery.


[The treatment of carcinoma of the tongue (author's transl)]. In the past 18 years 34 patients (21 males and 13 females) with carcinoma of the tongue were treated in the author's clinic. Patients with T1N0 lesions were treated satisfactorily only with irradiation. Patients with advanced disease, such as T2-T4 with metastases, were classified into 2 groups. One group was treated surgically (hemiglosectomy and radical neck dissection) and postoperative irradiation (about 6000 rads). The other group was treated with preoperative irradiation (about 4000 rads) and the same operation as the first groups. The results are significantly different. In the first group, 15 patients out of 17 died from local or cervical recurrences, whilst in the latter group 12 patients of 14 are in good health. As two patients died from unknown disease the 5 year survival of this group is 100%.


[A case report of an esophageal fibrolipoma (author's transl)]. A case of a rare, large pedunculated esophageal fibrolipoma is reported. Early symptoms involved regurgitations into the oral cavity. Correction was accomplished by endoscopic removal of the growth.


[Manifestations of paranasal sinus malignant lymphoma (author's transl)]. Malignant lymphomas rarely arise initially in the paranasal sinuses, although these may become involved at a later stage of the disease. Such cases have rarely been reported because the paranasal sinuses are seldom subjected to routine postmortem examination, and the clinical signs often resemble those of suppurative sinusitis. The present paper reports 3 patients in whom the paranasal sinuses were affected by a malignant lymphoma (chronic lymphatic leukaemia). The histological findings are illustrated and the pathogenesis discussed.


[Observations on the accent-method of Smith in the management of voice and speech disturbances. Part II (author's transl)]. In an earlier paper [HNO 25, 102 (1977)], the accent-method for the treatment of voice disorders has been described in detail. The rhythmical pronunciation of stressed ("accentuated") syllables with a physiologically correct phonatory respiration and a loose articulation form the basis for this method, and allow treatment of voice disorders as well as problems related to abnormal speech behavior. Further discussion is based on present theories of the physics and the physiology of voice function as well as on concepts from modern psychology and pedagogics.


[The results of comparative examinations in patients with grass pollen allergies (author's transl)]. Detailed histories of 20 patients with suspected hay fever were prepared from questionnaires. These patients were skin-tested with allergens prepared form four different grass mixtures, and a nasal provocative test with a fifth grass preparation was utilized. This latter test was evaluated by the clinical reaction of the nasal mucous membrane, by measuring the temperature variations in the nasal lumen, and by determining the nasal resistance with the use of "passive anterior rhinomanometry" (P.A.R.). In addition, the total serum IgE level and three grass-specific IgE concentrations were determined. The following results were then found: eight patients were consistently positive and four patients were consistently negative in all tests for allergens. Among the remaining eight patients, two false-positive and six false-negative patients by skin tests were uncovered by the additional use of history, IgE determinations and the nasal provocative test with rhinomanometric evaluation.


Branchial cyst in a heifer.

Branchial cyst in a heifer. A branchial cyst in a heifer was removed surgically. Diagnosis was based on clinical signs, analysis of cyst contents, and histologic examination of the cyst wall. Biochemically, the cyst fluid resembled a transudate. The cyst lining consisted of nonciliated, pseudostratified columnar epithelium and pigmented, keratinized, stratified squamous epithelium. The embryologic origin was thought to be endoderm of the 2nd pharyngeal pouch and adjacent ectoderm.


Depressed thyroid function in two tetraplegic dogs. Rapidly progressing tetraplegia developed in two 6-year-old dogs, both of which had eaten decayed meat sometime during the 24 hours prior to the onset of paralysis. Both dogs were found to have depressed thyroid function, as indicated by extremely low triiodothyronine (T3) and thyroxine (T4) values. Electromyographic studies revealed some abnormal insertional activity but no abnormal potentials when the fore- and hindlimb muscles were at total rest. Nerve conduction velocity was slightly depressed in 1 of the dogs and severely depressed in the other. Recovery was complete in less than 3 weeks in 1 dog, but the other dog was not completely recovered when placed under the owner's care 2 1/2 months after admission.


Surgical approach to the equine brachial plexus. Eleven ponies were used to perfect a surgical approach to the brachial plexus that would offer maximal exposure to the plexus, with minimal trauma. One pony was euthanatized to determine whether surgical exposure to the plexus was feasible. By approaching the plexus from the prescapular region, the only muscle that was found necessary to incise was the cutaneus omobrachialis. The rest of the procedure required only blunt dissection. In the other 10 ponies, the wounds healed by first intention, and the gait was not affected by the surgery.


Fibrous foreign body impaction colic in young horses. OF 207 horses with colic seen over a 36-month period, 10 were determined to have impaction colic caused by ingestion of synthetic fencing material. In 6 cases, there was history of exposure to rubberized fencing products. All horses affected were less than or equal to 3 years of age, had signs of mild to moderate abdominal pain, and were unresponsive to usual symptomatic therapy. At surgery, each horse was found to have an impaction involving the distal right dorsal colon, transverse colon, or small colon, and in some cases, all 3 bowel segments. In 9 cases, the involved segment of bowel could not be brought out of the abdominal cavity, and in each of these cases, the foreign bodies were removed through one or more enterotomies. Five of the 10 horses survived.


Clinical application of cerebrospinal fluid creatine phosphokinase determination. Creatine phosphokinase activity was determined in cerebrospinal fluid samples submitted for analysis from 126 animals suspected of having central nervous system disease. Values less than 1 sigma unit/ml were obtained on 32 samples and values less than or equal to 1 sigma unit/ml were obtained on 94 samples. The creatine phosphokinase values were increased in all cases of feline toxoplasmosis and feline infectious peritonitis. From other cases, especially seizure disorders and suspected poisonings, and apparent aid in prognosis for future cases was identified.


Electron microscopic and enzyme cytochemical studies on granules of mature chicken granular leucocytes. Detailed morphologic and enzyme cytochemical analysis was carried out by electron microscopy on granules of mature granulocytes obtained from the circulating chicken blood. Heterophils possessed three types of granules: large, rod-shaped, dense (Type I); medium sized, oval, light (Type II); and small-core (Type III). Acid phosphatase activity was present in Type I granules, but peroxidase and alkaline phosphatase were not demonstrable. The cytochemical nature of Types II and III granules remains unknown. Eosinophils contained only one type of granule, which was circular and had electron-opague contents. Both peroxidase and acid phosphatase, but not alkaline phosphatase, were present, indicating that the granules are lysosomes like the granules of mammalian eosinophils. Basophils possessed two types of granules, the characteristic large basophilic granules (Type I) and small dense granules (Type II). Acid phosphatase activity was found in only a small proportion of Type I granules: peroxidase and alkaline phosphatase were not demonstrable.


The early innervation of the developing deciduous teeth. Ingrowth of dental nerve fibres into the mesenchyme of the tooth-bearing areas of the mouth takes place at an relatively early stage, when the dental laminae are forming but the tooth buds are undeveloped. By the time a developing tooth has reached the cap and bell stages small nerve bundles have begun to enter the mesenchyme from which the dental papilla and sac arise.


Numbers and diameters of motoneurons and myelinated axons in the facial nucleus and nerve of the albino rat. The number and diameter of the motoneurons in serial sections of the facial nucleus in the albino rat were estimated using a photograph-camera lucida technique for counting and measuring cells in conjunction with a systematic section method for sampling the population. The mean diameter of facial motoneuron nucleoli was also estimated. Total cell counts were estimated using two formulas, one a basic count and the other the basic count corrected for split nucleoli errors. The mean motoneuron diameter is 33-93 micronm (+/- 6-18 micronm S.D.). The number and diameter of myelinated axons in both the facial nerve and its cutaneous auricular branch at the level of the stylomastoid foramen were estimated. There are 5353 myelinated axons in the facial nerve, with a mean diameter of 2-74 micronm, and 627 myelinated axons in the cutaneous auricular branch with a mean diameter of 1-47 micronm. The accuracy and reproductibility of results using the photograph-camera lucida technique for counting and measuring cells and the systematic section method of sampling are compared with those of the methods used by previous investigators. The accuracy of results obtained for estimates of the total cell count using the formulas for the basic count and the basic count corrected for split nucleoli are also compared.


Testis differentiation in the fetal and postnatal ferret. Testis development has been examined in a series of 59 fetal and 9 postnatal ferrets from day 22 of the 40-42 day pregnancy, to 12 days after birth. Developing seminiferous tubules and interstitial cells were first seen on day 26, and were well established one day later. Differentiation was associated with cell enlargement and cell division, and consequently the testes grew distinctly more rapidly at this time than the corresponding ovaries. Up to the end of pregnancy the epithelioid interstitial cells, derived from stromal tissue, formed a large proportion of the testis. Lipid was distinguishable in them, histologically, from about day 30. As in various other mammals, there appeared to be some regression of the interstitial Leydig cells around the time of birth, when the seminiferous tubules resumed their growth. The intra-gonadal rete was present from day 22 onwards; it established connexions with the seminiferous tubules through the small tubuli recti by day 32.


The developmental changes in the placenta of the guinea-pig. The placental disc and the yolk sac endoderm of intact guinea-pigs from day 14 of gestation to term, and of ovariectomized and fetectomized animals at 35 days of pregnancy, were observed by histological means. The chorionic giant cells were the first to attain maximal development (between days 17-35) and they had begun to degenerate by day 40. The spongy zone syncytiotrophoblast was well established and vascularized by day 18, and the maximal size of the zone was maintained between days 30 and 64. The spongy zone then dwindled in size just before parturition. Labyrinthine tissue appeared on day 18, but rapid growth and complete vascularization was not attained until as late as day 40. However, its continued increase in size almost up to term closely paralleled the growth of the fetus. If the placenta secretes progesterone from day 18 to parturition, the spongy zone is the only tissue sufficiently well established to carry out this activity throughout this period. The compensatory hypertrophy of the spongy zone syncytiotrophoblast in 35 day pregnant animals after ovariectomy and fetectomy supports the hypothesis that the spongy tissue secretes progesterone.


The growth patterns of three hindlimb muscles in the chicken. This study was designed to investigate the growth patterns of three hindlimb muscles of the chicken relative to the functional-biomechanical demands of increasing body size. The biceps femoris, a bipennate non-postural muscle, grew relatively faster in terms of wet and dry weight than did the parallel-fibred adductor superficialis or the unipennate adductor profundus, both postural muscles. All three muscles exhibited positive allometry (relative to body weight) in muscle length but only biceps femoris and adductor profundus showed positive allometry in cross sectional area adductor superficialis having isometric growth in this parameter. In biceps femoris and adductor superficialis the lengths of the longest and shortest fasciculi grew at equal rates, whereas in adductor profundus the shortest fasciculi grew faster than the longest. We conclude that muscle weight alone is an insufficient indicator of changing function in growing muscle. Hence, growth studies should include other functionally relevant parameters such as cross sectional area, which is proportional to the force-producing capabilities of the muscle, or fibre (fasciculus) length, which is indicative of the absolute amount of stretching or shortening that is possible and of the contraction velocity.


Morphological effects of chronic tracheal ligation and drainage in the fetal lamb lung. The relationship between lung liquid flow and fetal lung development has been studied at the cellular level using ultrastructural techniques. Continuous in utero tracheal ligation and drainage (over a period of 21-28 days) both result in malformations of the developing fetal lamb lung. Ligated lungs are larger, and drained lungs are smaller, than normal lungs at a similar gestational age. These changes are not merely due to altered lung liquid volume, but actual tissue growth thas been affected. Future alveolar wall thinning is enhanced in ligated lungs and inhibited in drained lungs, whilst the presence of differentiated alveolar type II cells (probably related to surfactant production) is decreased in ligated lungs and markedly enhanced in drained lungs. These results indicate the importance of fetal lung liquid in the regulation of pulmonary development in the fetus.


Experimental study of the development of the truncus and the conus in the chick embryo. The development of the truncus and the conus was studied in the chick embryo by in vivo labelling techniques. The earliest labels were placed at the stage of fusion of the myocardial troughs (stage 9-) and they were traced until the mature heart stage (stage 35). Microdissections and light microscopic studies were also carried out. The results are discussed in relation to the human heart. Our experiments permit the following conclusions: (1) At stage 9- fusion of the myocardial troughs takes place at the level of the primordium of the trabeculated portion of the right ventricle, when neither the conus nor the truncus are present. (2) At stage 12 (loop stage) there appears the caudal portion of the conus, which constitutes the cephalic end of the cardiac tube. (3) The truncus appears between stages 13 and 22. (4) At stage 22 angular junction between the conus and the truncus is the area where the semilunar valve cusps of the great arteries will develop and that, at this same stage, the junction between the conus and the trabeculated portion of the right ventricle seen from the right surface corresponds to the inferior edge of the crista supraventricularis. (5) It was confirmed that the pulmonary semilunar valve cusps originate from the walls of the truncus. (6) The development of the conus and truncus are similar in chick and man. (7) Histologically, in the chick, the wall of the truncus and the conus contain cardiac muscle as late as stage 28, but from then on the walls of the truncus are transformed into connective tissue and plain muscle.


The fine structure of the rat metrial gland in relation to the origin of the granulated cells. The fine structure of the metrial gland of the rat was examined at days 12, 14, 16, 18 and 20 of pregnancy and at all these stages two well differentiated cell types were seen. A relatively pale-staining cell which contained numerous dense granules was readily distinguished from a darkly stained fibroblast-like cell. Numerous cells could be identified, however, which did not fit either of these categories. On the basis of their morphological appearances these cells were placed in a series, with the members of the series showing a transition from a small cell with a high nuclear/cytoplasmic size ratio to a readily identifiable metrial gland cell containing a few typical granules. The small cell, apparently the precursor of the granulated metrial gland cell, was characterized by a markedly heterochromatic nucleus and few cytoplasmic organelles, although there were occasional mitochondria and Golgi bodies. On morphological grounds this cell appeared to be from the lymphocyte series and was probably a small lymphocyte.


The avian bronchial arteries: species variations. The bronchial arteries arose from the oesophagotracheobronchial branches of the common carotid arteries. In the goose, duck, Muscovy duck, and turkey several small bronchial arteries supplied the whole length of each primary bronchus, including the orifices of the secondary bronchi. In the guinea-fowl and quail similar bronchial arteries supplied only the extrapulmonary part of the primary bronchus. In the pigeon a single true bronchial artery supplied the extrapulmonary part of each primary bronchus; this pair of bronchial arteries arose asymmetrically from a common bronchial trunk derived from the left oesophagotracheobronchial artery only. In this species, and in the guinea-fowl and quail, the intrapulmonary part of the primary bronchus was supplied by the pulmonary artery. There were no branches to the exchange tissue in any species. In all species the bronchial veins of the extra-pulmonary part of the primary bronchus drained via oesophageal veins, whereas those of the intrapulmonary part emptied into branches of the pulmonary vein.


Fine structure of squamous epitheilum and submucosal glands of human oesophagus. Normal squamous epithelium and submucosal glands of the human oesophagus have been studied in biopsy and resection specimens. The squamous epithelium has a structure similar to that seen in other histologically comparable sites, and contains Langerhans cells. Features of note within the squamous cells include microfibrillar intranuclear bodies and occasional intracytoplasmic desmosomes. The submucosal glands are comparable in many respects to the mucous-secreting minor salivary glands. Myoepithelial cells are observed within the acini and smaller ducts; occasional oncocytic cells are identified at the junction between duct and acinus. The innvervation of these glands appears to be indirect.


The structure of the cells lining the stomach of the tammar wallaby (Macropus eugenii). The stomach of the tammar may be divided into cardiac, "oesophageal", fundic and pyloric regions. In the cells of the cardiac region (which occupy 65% of the stomach) at least three types of mucous droplet are present. The stratified squamous "oesophageal" region occupies 20% of the stomach and the fundic and pyloric regions make up the remaining 15%. The cellular structure of the three last named regions seems to be similar to that described for other monogastric mammals. Endocrine cells were found in all glandular regions.


Pre-elastic (oxytalan) fibres in the developing elastic cartilage of the external ear of the rat. During chondrogenesis in the external ear of the rat oxytalan fibres precede the appearance of mature elastic fibres by 6-7 days. The spatial distribution of oxytalan fibres in fetal and neonatal pre-cartilage corresponds to that of the elastic fibres in mature cartilage. These findings support the hypothesis that oxytalan fibres in the pre-cartilage of the external ear of the rat are pre-elastic in nature.


A histochemical study of cholinesterases in arm and forearm muscles of the pigeon and fowl. Various arm and forearm muscles of the pigeon and fowl were studied histochemically. Fibres of the muscles from both birds hydrolysed acetyl, butyryl and propionyl thiocholine iodide. With the exception of M. brachialis, all the fibres of the muscles had "en plaque" motor end plates. M. brachialis had "en grappe" endings on all its fibres. The possibility of M. brachialis being a physiologically "slow" muscle is discussed.


Neurons in the mouse anterior commissure. A light microscopic, electron microscopic and autoradiographic study. Both limbs of the anterior commissure of the mouse brain were examined to find the number, distribution, times of origin and structure of the neurons present, and also the number of synapses within the commissure. Neurons form between 0-1 and 0-4% of the total cell population and are produced between the twelfth and fourteenth days of gestation. It seems likely that the neurons within the anterior commissure are derived from adjacent septal nuclei, with the bed nuclei of the anterior commissure mainly contributing to the posterior limb and the nucleus triangularis septi mainly contributing to the anterior limb. The neurons are almost certainly functional, and distribute to the nuclei from which they are derived. There are probably also other connexions between these nuclei and both limbs of the anterior commissure through dendrites from the septal nuclei which ramify throughout the commissure. The large number of synapses scattered throughout the anterior commissure suggests that the neurons within the commissure, and dendrites entering it, may contribute substantially to the pathways between the anterior and posterior limbs and the septal nuclei without diminishing the number of axons in the commissure.


The structure and composition of peripheral nerves and nerve roots in the Sprawling mouse. Peripheral nerves and lumbar nerve roots of Sprawling, a neurological mutant mouse, were examined with light and electron microscopy. The peripheral nerves and the dorsal roots were thin and grey and were composed predominantly of small myelinated and unmyelinated axons. No evidence of axonal or myelin degeneration was found. Quantitative studies showed a marked reduction in the total number of myelinated axons with preponderance of those of 2-5 micron in diameter or less, most marked in the dorsal roots in which there was also an increase in the proportion of axons which were unmyelinated. In the ventral roots there was a deficiency in the contribution formed by myelinated axons of small calibre, probably indicating a deficiency of gamma fibres. Examination of the myelinated axons in nerves and roots showed a normal relationship between fibre size and internodal lengths and number of myelin lamellae. The findings suggest that the genetic defect in Sprawling is responsible for a failure of myelination of sensory axons. The deficiency of large sensory axons and of small motor axons can be correlated with the deficiency of muscle spindles.


A radiographic study of the human fetal spine. 1. The development of the secondary cervical curvature. The present study has shown that the secondary curvature in the cervical spine of the human fetus develops at a much earlier age than has been thought. It may, in fact, develop soon after the embryo first acquires a neck and begins to uncurl. The early appearance of this curvature may be related to the early development of function in the muscles responsible for head extension, this movement being a basic component of the primitive "gasp" reflex. It is further suggested that, although the embryo develops in flexion, the traditional assumption that such flexion is a consequence of fetal musculoskeletal development must be questioned, and the expression "normal fetal flexed position" is probably misleading, since it does not take into account the wide range of movements of which the fetus is capable.


Regional and segmental characteristics of the human adult spinal cord. Ten human adult spinal cords were assessed for weight, length, thickness, cholesterol, water and DNA-P content on both a regional and a mean segmental regional basis. The mean segmental DNA-P content, unlike the other parameters, was exceptional in being constant in the cervical, thoracic and lumbar regions. It is concluded that the cellularity of the cord between C1 and L5 is determined segmentally, but is reduced in the segments of the sacrococcygeal cord because of embryonic atrophy.


The effects of age on mucosal morphology and epithelial cell production in rat small intestine. Six groups of male Wistar rats were used, with mean weights of 29, 63, 97, 161, 249 and 399 g. Pieces of small intestine from three sites were examined after staining in bulk with the Feulgen reaction. Crypt/villus ratio (the number of crypts per villus) rose with age at all three sites, Villus height and crypt depth were measured on microdissected specimens. Villi in the proximal intestine were always taller than those distally. Proximal villi increased in height in successively older rats, except in the oldest group. Villi at the two distal sites tended to be tall in the youngest group of rats, but suffered a temporary reduction in height in the next two age groups. Crypt depth increased markedly within the first three age groups, and more slowly thereafter. Colchicine-metaphase accumulation rate was estimated from counts on microdissected intact crypts. The rate was low in the youngest group (8 cells/crypt/hour) but about 30 cells/crypt/hour in all other groups. After the changes during the early phase of rapid growth, no marked changes were seen during later life. The significance of these findings is discussed.


Diameter distribution spectral of myelinated axons in the median and ulnar nerves of the Soay sheep. An ulnar and a median nerve were taken from each of three Soay sheep, two aged 13 months and one aged 6 years. The axon diameter distribution spectra in these six nerves were determined by measurement of transverse sections stained by the Weigert-Pal technique. The histograms of myelinated axon diameters, between 1-17 micron, were unimodal in all the ulnar nerves. The mode lay between 4-5 micron in the nerves from the young sheep and between 6-7 micron in the old animal. The histograms of the median nerve axon diameters, between 1-18 micron, were all tri-modal. The nerves from the young sheep had modes between 2-3, 4-5 and 9-11 micron, and in the nerve from the old animal the modes were between 3-4, 6-7 and 12-13 micron. These findings are correlated with the conduction velocities recorded from these nerves in an earlier series of experiments.


Semisynthetic beta-Lactam antibiotics. II. Penicillins from alpha-hydrazinoarylacetic acids. A number of penicillins (2) have been synthesized from the alpha-hydrazinoarylacetic acids (4) via the activated chloride hydrochlorides (5) or via the mixed anhydride of the corresponding N2-benzyloxycarbonyl derivatives (6). The penicillins, 2b, e, j, show good activity against gram-positive and gram-negative bacteria and enhanced penicillinase resistance in comparison with ampicillin.


Amino acids and derivatives of thiazole-4-carboxylic acid as constituents of thiopeptin B. L-Alanine, L-threonine, L-valine, D-cystine and three derivatives of thiazole-4-carboxylic acid (thiostreptin, 1-aminomethylthiazole-4-carboxylic acid and thiostreptoic acid) were isolated from the acid-hydrolysate of thiopeptin B. In addition, the presence of dehydrobutyrine and dehydroalanine residues in the antibiotic was determined. Other components remain unidentified.


The determination of cephradine and cephalexin by reverse phase high-performance liquid chromatography. The application of reverse phase high-performance liquid chromatography to the separation and analysis of cephradine and cephalexin is demonstrated. The procedure has been applied to chemicals, pharmaceutical formulations and reaction solutions. The preparation of samples is simple and rapid. Chromatographic conditions are described for both pellicular and small particle columns. The feasibility of determing cephradine and cephalexin in physiological fluids has also been demonstrated.


Effects of ethidium bromide and acriflavine on streptomycin production by Streptomyces bikiniensis. The treatment of Streptomyces bikiniensis with ethidium bromide or acriflavine resulted in the loss of the ability to produce streptomycin in 2 approximately 16% of the colonies isolated from the treated spores. These isolates had also lost their resistance to streptomycin. Treatment with the dyes caused partial to total loss of the ability to produce aerial mycelium by S. bikiniensis, but the isolates regained this ability upon repeated transfer to fresh medium. The dyes did not appear to effect pigment production by S. bikiniensis.


A new antibiotic, platenocidin.

A new antibiotic, platenocidin. A new antibiotic named platenocidin was isolated from the culture broth of Streptomyces H 273 N-SY2 and the producting microbe was classified as Streptomyces platensis. Platenocidin gives 5-hydroxymethyluracil by acid or alkaline hydrolysis and can be considered to belong to one of the nucleoside antibiotics. The antibiotic inhibits the growth of certain species of yeasts, but not that of bacteria.


Macrolide antibiotics M-4365 produced by Micromonospora. I. Taxonomy, production, isolation, characterization and properties. A series of basic 16-membered macrolide antibiotics, M-4365 A1, A2, A3, G1, G2 and G3, were isolated from the culture broth of strain MCRL 0940 which is assigned to be a new species of Micromonospora and for which the name Micromonospora capillata sp. nov. is proposed. Among these antibiotics, M-4365 A2 and G2 showed strong inhibitory activity against Gram-positive and Gram-negative bacteria.


Macrolide antibiotics M-4365 produced by Micromonospora. II. Chemical structures. By physiochemical analyses and chemical procedures, the structures of a series of basic 16-membered macrolide antibiotics, M-4365, A1, A2, A3, G1, G2 and G3 were elucidated, and it was found that M-4365 A1, G1 and G2 were novel, while M-4365 A2, A3 and G3 were identical with rosamicin, juvenimicins A4 and B1, respectively.


G7063-2, a new nitrogen-containing antibiotic of the epoxydon group, isolated from the fermentation broth of a species of Streptomyces. Antibiotic G7063-2, isolated from a Streptomyces species, is a new nitrogen-containing analogue of phyllostine and terreic acid. In vitro G7063-2 is moderately active against both Gram-positive and Gram-negative bacteria and weakly active against fungi. It is toxic to mice.


Melinacidins II, III and IV. Structural studies. The structures of melinacidines II, III and IV were determined by physicochemical methods. Melinacidine IV is considered to be identical to 11alpha,11alpha'-dihydroxychaetocin while melinacidins II and III are isomeric to chaetocin and verticillins A and B.


Microbial transformation of antibiotics. Clindamycin ribonucleotides. Addition of clindamycin to whole-cell cultures of Streptomyces coelicolor Müller resulted in the loss of in vitro activity against organisms sensitive to clindamycin. Incubation of such culture filtrates with crude alkaline phosphatase generated a biologically active material identified as clindamycin. Fermentation broths containing inactivated clindamycin yielded clindamycin 3-ribonucleotides and clindamycin 3-phosphate the structure of which was established by physicochemical and enzymatic means. Attempts to transform clindamycin to clindamycin 3-ribonucleotides by lysates or partially purified enzyme preparations from S. coelicolor have failed.


Biosynthetic studies on the macrolide antibiotic turimycin using 14C-labeled precursors. Using a strain of Streptomyces hygroscopicus JA 6599 several 14C-compounds were investigates as potential precursors of the macrolide antibiotic turimycin followed by partial degradation to localize the radioactivity. L-Methionine-14C-methyl and n-butyrate-1-14C were incorporated exclusively and in a specific manner. The incorporation ratios were dependent on the addition time of the precursors. Studies of the incorporation of acylmycaroses and demycarosyl turimycin into the antibiotic are also reported.


Effect of rifampicin derivatives on the ion compartmentation of biological membranes. Based on the experimental results that 3-formyl rifamycin SV acts as an uncoupler in vitro on rat liver mitochondria, the effect of some rifampicin derivatives on the K+ and H+ compartmentation of biological membranes was examined to obtain a chemiosmotic hypothesis for oxidative phosphorylation. The K+ release from mitochondria was remarkably stimulated by 3-formyl rifamycin SV in accordance with uncoupling of the oxidative phosphorylation. 3-Formyl rifamycin SV also stimulated the K+ release from red blood cells, though its action was not as effective as in mitochondria. It can be suggested that 3-formyl rifamycin SV interacts with biological membranes, causing a change in permeability to ions, especially of K+ and H+ through the mitochondrial membrane, resulting in uncoupling of the oxidative phosphorylation.


Development and evaluation of treatment paradigms for the suppression of smoking behavior. A multiple-baseline component-analysis design was employed to assess the effectiveness of three treatment programs for suppressing the cigarette smoking behavior of 24 subjects. Sartiation, cognitive control, and continger shock procedures were evaluated. The results demonstrated a consistent relationship between contingent shock and suppression of smoking. It was further indicated that subjects should be exposed to the number of sessions necessary to achieve total suppression in order to gain maximally from treatment and to avoid relapse. Neither the satiation component nor the cognitive control component was correlated with clear, permanent decrements in smoking frequencies. To date no other treatment program has demonstrated the dramatic effects ofethe contingent shock procedures used in the present study.


Social-skills training for unassertive children: a multiple-baseline analysis. The effects of social-skills training consisting of instructions, feedback, behavior rehearsal, and modelling were examined in a multiple-baseline analysis in four unassertive children. The treatment was effective in that the behaviors selected for modification changed markedly. The effects of treatment generalized from trained to untrained items (interpersonal situations requiring assertive responding) and gains were maintained at the two-and four-week followup probe sessions. In addition, overall assertiveness in all subjects increased from baseline assessment to the conclusion of treatment and into followup.


Assessing and training teachers in the generalized use of behavior modification with autistic children. This study investigated the feasibility of developing reliable, valid criteria for measuring and training the skills necessary to teach autistic children. The behaviors of 11 teachers and 12 autistic children were recorded in a series of different teaching situation. Teacher-training was initiated at different times for different teachers. The results showed: (1) it was possible to asses empirically whether a teacher was correctly using defined behavior-modification techniques; (2) generally, for any given session, systematic improvement in the child's behavior did not occur unless the teacher working in that session had been trained to use the techniques to a hight criterion; (3) all 11 teachers were rapidly trained to use these techniques; and (4) the teachers learned generalized skills effective with a variety of children and target behaviors.


The effect of modelling on drinking rate. Three male college seniors were asked to drink beer at their normal rate in a simulated tavern setting. Each was paired with a confederate, also a male college senior, in an ABACA single subject design. In the baseline conditions, the confederate matched the drinking rate of the subject. Baseline and all subsequent conditions were continued in 1-hr sessions until a stable drinking rate was achieved. In Condition B, the confederate drank either one third more or one third less than the subject's baseline rate. In Condition C, the direction was reversed. All three subjects closely matched the confederate's drinking rate, whether high or low. All subjects reported they were unaware of the true purpose of the study.


Stimulus selection and tracking during urination: autoshaping directed behavior with toilet targets. A simple procedure is described for investigating stimuli selected as targets during urination in the commode. Ten normal males preferred a floating target that could be tracked to a series of stationary targets. This technique was used to bring misdirected urinations in a severely retarded male under rapid stimulus control of a floating target in the commode. The float stimulus was also evaluated with nine institionalized, moderately retarded males and results indicated rapid autoshaping of directed urination without the use of verbal instructions or conventional toilet training. The technique can be applied in training children to control misdirected urinations in institution for the retarded, in psychiatric wards with regressed populations, and in certain male school dormitories.


The reduction of stealing in second graders using a group contingency. Three clases of second graders served as subjects in this study of the effects of two types of intervention programs upon stealing: (1) an antistealing lecture with no specific contingency implied, and (2) a direct group contingency applied, whereby children were rewarded with (a) extra free time for no thefts, (b) allowed normal free time if stolen items were returned, and (c) punished with lost of free if stolen items were not returned. A multiple-baseline design across the three classes showed the group contingency to be effective in reducing stealing behavior; the anti-stealing lecture was ineffective.


The modification of leisure behavior in a half-way house for retarded women. A multi-element baseline design combined with a multiple-baseline design, was used to investigate the effect of availability of materials, prizes for participation, and instruction on the leisure behavior of 14 mentally retarded women in a half-way house. A leisure program was conducted on weekday evenings, during which residents could choose to participate in any of six activities offered: puzzles, card games, clay, painting, weaving and rug making. It was found that instruction in weaving and rug making significantly increased the percentage of residents participating in these activities, and that following instruction, prizes were not necessary to maintain high levels of participation. In contrast, prizes were more effective than mere availability of materials in maintaining participation in the other activities.


Stimulus factors effecting peer conversation among institutionalized retarded women. The effect of a series of stimulus changes, including attendant present or absent and agent of token reinforcement in a sorting task, on peer interaction among retarded women was examined. Conversation was not differentially directed toward the agent of reinforcement when that agent was a peer, nor did a peer agent increase the overall rate of peer interaction. Peer interaction did increase when no attendant was present, and during the final phase of the experiment, the high rate of peer interaction was maintained while the attendant was faded back into the situation in a series of discrete steps. The procedure was found to be efficient in terms of staff time and did not adversely affect task performance.


The influence of behavior preceding a reinforced response on behavior change in the classroom. The influence of behavior that immediately precedes a reinforced target response on the effectiveness of a reinforcement contingency was examined in two experiments with mentally retarded children in a special-education classroom. Two reinforcement schedules were examined in each experiment. For each schedule, a prespecified period of attentive behavior served as the target response. The schedules differed in whether inattentive or attentive behavior was required immediately to precede the target response. These schedules were examined with one child in a simultaneous treatment design using praise as the reinforcer (Experiment I), and with two children in separate reversal designs using tokens as the reinforcer (Experiment II). While attentive behavior increased under each schedule, the increase was greater when attentive rather than inattentive behavior preceded the reinforced response. The results indicated that the effect of a contingency may be determined not only by the specific response reinforced but also by the behavior that immediately precedes that response.


Oxalate, formate, formamide, and methanol metabolism in Thiobacillus novellus. Thiobacillus novellus was able to grow with oxalate, formate, formamide, and methanol as sole sources of carbon and energy. Extensive growth on methanol required yeast extract or vitamins. Glyoxylate carboligase was detected in extracts of oxalate-grown cells. Ribulose bisphosphate carboxylase was found in extracts of cells grown on formate, formamide, and thiosulfate. These data indicate that oxalate is utilized heterotrophically in the glycerate pathway, and formate and formamide are utilized autotrophically in the ribulose bisphosphate pathway. Nicotinamide adenine dinucleotide-linked formate dehydrogenase was present in extracts of oxalate-, formate-, formamide-, and methanol-grown cells but was absent in thiosulfate- and acetate-grown cells.


Effect of light on Bdellovibrio bacteriovorus. Bdellovibrio underwent photooxidation by visible light in the presence of exogenous photosensitizer and by near-ultraviolet light (325 to 400 nm) in its absence. The colorless, host-dependent wild type was more sensitive to the lethal effect of light than was its pigmented, facultative parasitic mutant. The latter's ability to form colonies was much more sensitive to light than was its plaque-forming capability. The biosynthesis of the mutant pigment was inhibited by diphenylamine, though this inhibition did not result in additional sensitivity to photokilling.


Characterization of lysocardiolipin from Acinetobacter sp. HO1-N. Triacyl-lysocardiolipin (triacyl-LCL) and diacyl-LCL were isolated from Acinetobacter sp. HO1-N, and their structures were determined by chemical, physical, and enzymatic procedures. Deacylation of triacyl-LCL and diacyl-LCL yielded bis-glycerylphosphorylglycerol. Periodate oxidation of both lysolipids was negative. Diglyceride and 2-monoglyceride resulted from the acetic acid hydrolysis of triacyl-LCL, whereas 2-monoglyceride was the sole product obtained from diacyl-LCL. Cardiolipin (CL)-specific phospholipase D treatment of triacyl-LCL yielded lysophosphatidylglycerol and phosphatidic acid. Pancreatic lipase treatment of CL yielded triacyl-LCL and diacyl-LCL. 31P nuclear magnetic resonance spectrometry showed two resonance peaks separated by 40 HZ for CL, two overlapping peaks separated by 14 HZ for triacyl-LCL, and one peak for diacyl-LCL. The proportion of lysocardiolipin increased as a function of cell age, representing 2 to 3% of the total phospholipids in early- and mid-exponential growth, 5 to 7% in late-exponential growth, and 12% in the stationary growth phase.


Source of energy for gliding motility in Flexibacter polymorphus: effects of metabolic and respiratory inhibitors on gliding movement. The effects of selected metabolic and respiratory inhibitors on the gliding motility of Flexibacter polymorphus were examined. Motility and oxygen consumption were quantitatively inhibited in a reversible manner by specific respiratory poisons, suggesting that gliding velocity was linked to electron transport activity. Arsenate had little influence on the number or rate of gliding filaments, despite a 95% decrease in the concentration of intracellular adenosine 5'-triphosphate (ATP). At concentrations of cyanide or azide that abolished gliding movement, cells possessed a level of ATP that should have been sufficient to allow motility. Proton-conducting uncouplers of oxidative phosphorylation, such as carbonylcyanide m-chlorophenylhydrazone (CCCP) and tetrachlorosalicylanilide, strongly inhibited locomotion yet did not suppress respiratory activity or intracellular ATP sufficiently to account for their effect on movement. Inhibition of motility by CCCP (but not by tetrachlorosalicylanilide) was partially reversed by sulfhydryl compounds. However, unlike CCCP, inhibition of motility by p-chloromercuribenzoate, a known sulfhydryl-blocking reagent, was associated with a corresponding reduction in respiratory activity and ATP content of cells. Protein synthesis was not blocked by concentrations of CCCP inhibitory for motility, indicating that utilization of existing ATP in this energy-requiring process was not impaired. These data suggest (but do not unequivocally prove) that ATP may not function as the sole energy donor for the gliding mechanism, but that some additional product of electron transport is required (e.g., the intermediate of oxidative phosphorylation).


Occurrence of a class II restriction endonuclease in Staphylococcus aureus. The occurrence of class II restriction endonucleases (enzymes that both recognize and cleave a specific nucleotide sequence in deoxyribonucleic acid [DNA] in Staphylococcus aureus has been investigated by analysis of crude extracts obtained from different propagating strains of the International Phage Typing System. Of the four main groups of strains in the International System, only extracts of group II strains were found to contain class II restriction endonucleases. The identical cleavage patterns obtained by incubation of different DNAs with cell extracts of group II strains suggest that these enzymes all recognize and cleave the same nucleotide sequence. This recognition site has been determined to be 5'-G-A-T-C-3'-3'-C-T-A-G-5' for the prototype of these enzymes, Sau3AI (J. S. Sussenbach et al., Nucleic Acids Res. 3:3192-3202, 1976). Evidence is presented that the classification of group II strains is based on restriction modification and is correlated with the presence of a class II restriction enoduclease that recognizes and cleaves the above sequence.


Photodynamic induction of a bacterial cell surface polypeptide. The photodynamic action of several dyes on cells of a bacterium, tentatively identified as a species of Arthrobacter, resulted in remarkable stimulation of synthesis of a polypeptide 21,000 daltons in mass. This polypeptide resides on the cell surface and can be solubilized by sodium dodecyl sulfate without lysis of the cells. Chlorophyllin and rose bengal are effective in inducing synthesis of the polypeptide in proportion to their ability to sensitize the photooxidation of histidine. Etiolated cells of the alga Chlamydomonas reinhardtii y-1 excrete a substance into the medium that also sensitized the photoinduction of the polypeptide.


Polyamines in trypanosomatids.

Polyamines in trypanosomatids. Polyamines were determined by n-butanol extraction and thin-layer chromatography in four trypanosomatids: Trypanosoma brucei (rat infection) and cultures of Crithidia fasciculata, Leptomonas sp., and Trypanosoma mega. All had putrescine and spermidine but no detectable spermine. Putrescine and spermidine levels were quantitated for extracts of leptomonas during the normal growth cycle. Spermidine values peaked 18 h before peak cell populations. Spermidine-putrescine ratios for all organisms were related to the presumed phylogeny of the group.


Ultrastructure of lipid bodies in Tilletia caries teliospores. The ultrastructure of lipid bodies within developing, dormant, and germinating Tilletia caries (DC). Tul. (race T-16) teliospores was studied by freeze-etching and thin-sectioning techniques. When teliospores were prefixed in sodium cacodylate-buffered glutaraldehyde-acrolein for 24 h before further processing, most of the lipid bodies appeared to have a uniformly osmiophilic matrix. Some of these lipid bodies were surrounded by thin electron-dense lines that appeared to be half-unit membranes. Occasionally this membrane seemed to be absent, allowing for a direct interface between lipid and cytosol. Irregular electron-dense patterns were occasionally observed in lipid bodies of developing, dormant, and germinating teliospores. A lamellar substructure with 6- to 10-nm center-to-center spacing was visible in the electron-dense patterns at high magnifications. Irregular fracture patterns were visible in freeze-etch replicas.


Reversible activation of hepatic adenylate cyclase by guanyl-5'-yl-(alpha,beta-methylene)diphosphonate and guanyl-5'-yl imidodiphosphate. Guanyl-5'-yl-(alpha,beta-methylene)di[gamma-32P]phosphonate (Gp-(CH2)pp) is not hydrolyzed by rat liver membranes under conditions in which GTP and guanyl-5'-yl imidodiphosphate (Gpp(NH)p) are hydrolyzed. Gp(CH2)pp activates adenylate cyclase in hepatic membranes with characteristics similar to those of Gpp(NH)p activation but with lower potency and effectiveness. The analogs, although with lower potency than GTP, also share the ability to change the glucagon receptor from a high to a low affinity state. Both Gp(CH2)pp and Gpp(NH)p stimulate adenylate cyclase activity following a lag period of about 1 min addition of GTP after steady state rates are achieved results in reduction in the rate following a lag period of 6 min from the time of addition of GTP. Pretreatment of the enzyme with Gpp(NH)p or Gp(CH2)pp, followed by washing the membranes, leads to a high activity state of the enzyme which slowly decays in rate unless the analogs are continuously present in the medium. These data suggest that the guanosyl nucleotide analogs act on the enzyme system by a slowly reversible process that possibly reflects slow binding and dissociation from different transition states of the enzyme system and suggest that activation of adenylate cyclase by GTP, Gpp(NH)p, and Gp(CH2)pp does not involve covalent modification of the enzyme.


alpha-Hydroxylation of fatty acids in brain. Substrate specificity and deuterium isotope effect. The alpha-hydroxylation of fatty acids with various chain length by a rat brain postnuclear preparation was investigated. The maximum velocity of the alpha-hydroxylation were higher for lignoceric (tetracosanoic), tricosanoic, docosanoic, and heneicosanoic acids and lower for hexacosanoic, pentacosanoic, eicosanoic, and nonadecanoic acids. Stearic palmitic, and phytanic (3,7,11,15-tetramethylpalmitic) acids were not hydroxylated by this preparation. Apparent Km values were lower for fatty acids with the higher Vmax and higher for those with the lower Vmax values. The relative order of alpha-hydroxylation strongly resembles the relative distribution of these fatty acids in brain cerebrosides, and we suggest that alpha-hydroxylation plays an important role in determining this distribution. An improved synthesis of [U-14C]phytanic acid is described. The rate of alpha-hydroxylation with [2-2H2]lignoceric acid was one-fifth of nondeuterated lignoceric acid and one-third of [3-2H2]lignoceric acid, while apparent Km values for these acids were similar. The reduced rate with [2-2H2]lignoceric acid indicates that the cleavage of the C-H bond is the rate-limiting step of the alpha-hydroxylation.


L-Histidinol inhibits specifically and reversibly protein and ribosomal RNA synthesis in mouse L cells. The addition of the histidine analogue L-histidinol to mouse L cells in suspension culture results in an abrupt inhibition of protein and ribosomal RNA synthesis with relatively little effect on messenger RNA production. Messenger RNA synthesized in the presence of L-histidinol associates with ribosomal subunits to form initiation complexes whose ability to commence elongation appears impaired. As a result of this mode of action, L-histidinol treatment results in a dramatic reduction of polyribosomes and a concomitant accumulation of 80 S ribosomes. The inhibition of both protein and ribosomal RNA synthesis mediated by L-histidinol is rapidly reversed by the addition of L-histidine to treated cultures.


Isolation and recombination of ovine lutropin subunits with complete recovery of biological activity. Based on the results obtained in the previous paper (Liu, W.-K., Ascoli, M., and Ward, D. N. (1977) J. Biol. Chem. 252, 5274-5279) we have devised a method that permits the isolation of the subunits of ovine lutropin (oLH) in their "native" form. The hormone was dissociated by guanidine HCl, and the subunits separated by salt precipitation (Sairam, M. R., and Li, C. H. (1974) Arch. Biochem. Biophys. 165, 709-714). The present method differs significantly from that of Sairam and Li in that all operations are carried out at pH 5.0 or higher, a condition we have shown to be crucial to maintain the integrity of the isolated subunits. The yield of subunits was 75 to 85%. The isolated subunits exceed 95% purity (i.e. contamination with the other subunit or intact lutropin is less than 5%), migrate as single bands in sodium dodecyl sulfate-polyacrylamide gels, and show only the NH2-terminal heterogeneity expected from that of the native hormone. Recombination of the subunits can be accomplished with 75 to 85% yield (with respect to mass) and with full recovery of biological activity.


Purification and properties of a soluble protein activator of rat liver squalene epoxidase. A soluble rat liver protein, termed "supernatant protein factor" (SPF), that stimulates microsomal squalene epoxidase has been purified approximately 11,000-fold. The most highly purified preparation obtained by isoelectric focusing shows a single coincident peak for activity and protein (the isoelectric point, pI, was 6.74). SPF is about 95% pure, judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it migrates to a position corresponding to an apparent molecular weight of 47,000. An amino acid analysis of SPF is presented, and the properties of SPF and of the various soluble protein activators of microsomal sterol biosynthesis described by other laboratories are compared.


Isolation and amino acid sequence of COOH-terminal fragments from the beta subunit of human choriogonadotropin. The amino acid sequence of the unique COOH-terminal region of the beta subunit of human choriogonadotropin has been reinvestigated. The desialylated subunit was digested with thermolysin and a 27-residue peptide from positions 115 through 141 isolated in a high yield. Quantitative Edman sequence degradation of this peptide, of another peptide produced by thermolysin digestion containing residues 142 to 145, and of two tryptic peptides (residues 123 to 145, 134 to 145) has established that the amino acid sequence of this region is: (formula: see text). In addition, the positions of attachment of the carbohydrate moieties to serine residues was established by a direct procedure using alkaline elimination and 35S-labeled sulfite addition, which yields [35S]-cysteic acid residues at the site of a substituted serine. Carbohydrate side chains in the COOH-terminal region have been shown to exist at residues 121, 127, 132, and 138. These studies have also resulted in the development of improved methods for the purification of COOH-terminal peptides of the human choriogonadotropin beta subunit.


Oligosaccharides of human milk. Structural studies of two new octasaccharides, difucosyl derivatives of para-lacto-N-hexaose and para-lacto-N-neohexaose. Two new octasaccharides were isolated from human milk of a nonsecretor Lewis a+b- individual. Structural studies by sequential enzymic degradation and by quantitative methylation analysis revealed that their structures were as follows: Galbeta1 leads to 3 (Fucalpha1 leads to 4) GlcNAcbeta1 leads to 3Galbeta1 leads to 4 (Fucalpha1 leads to 3) GlcNAcbeta1 leads to 3Galbeta1 leads to 4Glc, Galbeta1 leads to 4 (Fucalpha1 leads to 3) GlcNAcbeta1 leads to 3Galbeta1 leads to 4 (Fucalpha1 leads to 3) GlcNAcbeta1 leads to 3Galbeta1 leads to 4Glc. The core portions of these sugars, which are newly found linear hexasaccharides, are named para-lacto-N-hexaose and para-lacto-N-neohexaose, respectively.


Isolation and in vitro translation of the messenger RNA coding for pancreatic amylase. RNA prepared from dog pancreas polysomes or microsomes directs the synthesis of pancreas-specific proteins in heterologous cell-free translation systems. A translation product, approximately 1500 daltons larger than authentic amylase, corresponding to pancreatic amylase was identified by immunoprecipitation with anti-amylase gamma-globulin and tryptic peptide analysis. We suggest that this larger form of amylase is an amylase precursor. Using amylase immunoprecipitation of reticulocyte translation reactions as an assay, we have shown that greater than 99% of the mRNA for amylase is associated with polysomes bound to the endoplasmic reticulum. Electrophoresis of pancreatic mRNA preparations in formamide-containing polyacrylamide gels and subsequent translation of the fractions have shown that amylase mRNA is of a discrete size with a mobility equivalent to that of 18 S ribosomal RNA, and therefore significantly larger than required to code solely for the amino acid sequence of the amylase precursor.


Ca2+-dependent protease in human platelets. Specific cleavage of platelet polypeptides in the presence of added Ca2+. A pronounced Ca2+-dependent protease in lysed platelets was detected by the hydrolysis of an added substrate, azocasein. This protease also catalyzed the cleavage of endogenous platelet polypeptides which was demonstrated by the disappearance of four high molecular weight polypeptides (greater than 200,000) either by adding Ca2+ to lysed platelets or by increasing the cytoplasmic Ca2+ within intact platelets with the ionophore A 23187. In both cases, lower molecular weight cleavage products were clearly identified. The caseinolytic activity as well as the cleavage of the high molecular weight polypeptides was inhibited by sulfhydryl reagents, but not by reagents reacting with trypsin-like proteases. Because of the similarity between this Ca2+-dependent protease and myofibrillar Ca2+-dependent protease (CAF), we have termed the former "platelet CAF".


In vitro synthesis of a DNA probe for antisense globin sequences. A DNA probe specific for sequences complementary to globin RNA has been prepared in vitro from globin complementary DNA (cDNA). Globin cDNA was used as the template for the synthesis of a complementary strand (ccDNA) by avian myeloblastosis virus DNA polymerase. After treatment with single strand-specific S1 nuclease, double-stranded globin DNA was denatured in the presence of a vast excess of globin RNA. After hybridization of the cDNA with globin RNA, the ccDNA was isolated by hydroxyapatite chromatography. The ccDNA probe hybridized efficiently to globin cDNA, but not at all to globin RNA. This probe should prove useful in assessing the asymmetry of gene expression in cell-free transcriptional systems.


Cytochrome b5 and cytochrome b5 reductase-phospholipid vesicles. Intervesicle protein transfer and oreintation factors in protein-protein interactions. NADH-cytochrome b5 reductase readily binds to preformed phospholipid vesicles either below or above the phase transition temperature of the lipid and in the absence of detergents. The isolated vesicles are free of unbound reductase, and the lipid is present as small, closed bilayers (250 to 400 A in diameter) as indicated by gel filtration, density gradient centrifugation, and internal volume measurements with [3H]glucose. The order of substrate specificity of the bound reductase is: ferricyanide = cytochrome b5 bound to reductase vesicles (100%) greater than cytochrome b5 heme peptide (13%) greater than unbound cytochrome b5 (4.5%) greater than cytochrome b5 vesicles (0.1%). This indicates that a specific orientation of cytochrome b5 and reductase in the bilayer is required for optimal interaction. Protein transfer occurs between reductase vesicles and cytochrome b5 vesicles. The transfer is time-dependent (40 to 70% complete in 2 h), does not involve vesicle fusion, is most rapid at the phase transition temperature of the phospholipid, and appears to require a fluid bilayer.


Regulation of citrate transport and pyruvate dehydrogenase in rat kidney cortex mitochondria by bicarbonate. 1. Bicarbonate increased citrate and 2-oxoglutarate accumulation when rat kidney cortex mitochondria were incubated with pyruvate or L(-)-palmitoyl carnitine in the presence of L-malate. 2. Bicarbonate stimulated the exit of citrate from mitochondria. The Km for bicarbonate was 13.5 mM and the Vmax was 0.59 nmol/min/mg of protein at 10 degrees. 3. The bicarbonate-stimulated exit of citrate from the mitochondria was prevented by inhibitors of the tricarboxylate, dicarboxylate, and phosphate transport systems. 4. The activity of pyruvate dehydrogenase was significantly increased by preincubation of rat kidney mitochondria with bicarbonate. This bicarbonate-induced activation was not observed in presence of inhibitors of citrate transport. Bicarbonate did not activate pyruvate dehydrogenase in rat heart mitochondria. Bicarbonate had no effect on pyruvate dehydrogenase activity in either broken mitochondria or whole tissue preparations. 5. The mechanism of this activation is discussed in the light of the known regulatory properties of pyruvate dehydrogenase, pyruvate carboxylase, and citrate synthase.


Studies on the specificity of phosphorylase kinase using peptide substrates. The action of phosphorylase kinase on synthetic peptides is reported. These peptides are variants of the amino acid sequence. Ser-Asp-Gln-Glu-Lys-Arg-Lys-Gln-Ile-Ser-Val-Arg-Gly-Leu, found in the natural substrate, phosphorylase b. The effects of size, the cluster of basic groups at the NH2-terminal side, the phosphorylatable seryl residue, the hydrophobic groups surrounding serine, and the arginyl function at the COOH-terminal side were tested and analyzed by evaluation of the kinetic parameters, Km and Vmax. The first 6 residues were found to be nonessential, but substitution of residues in the sequence. Lys-Gln-Ile-Ser-Val-Arg, had a large effect on phosphorylation. A comparison was made between the action of nonactivated and activated phosphorylase kinase on selected peptides and phosphorylase b. Various forms of phosphorylase b were tested as substrates for cyclic AMP-dependent protein kinase in the presence of effectors and salts. Although phosphorylase would not serve as a substrate for protein kinase, the aforementioned synthetic peptide of the phosphorylase b sequence would do so, indicating that the primary sequence surrounding the phosphorylatable serine did not block phosphorylation, which suggests that higher order structural features prohibit the phosphorylation.


Incubations of testis myo-inositol-1-phosphate synthase with D-(5-18O)glucose 6-phosphate and with H218O show no evidence of Schiff base formation. myo-Inositol-1-P synthase (EC 5.5.1.4) purified from rat testis and from bovine testis was allowed to react with D-[5-18O]glucose-6-P. myo-Inositol, obtained in these reactions, retained all of the 18O originally in the glucose-6-P. When these enzyme preparations were incubated with unlabeled glucose-6-P in a medium enriched in H2 18O no uptake of the oxygen isotope occurred that could be ascribed to the myo-inositol-1-P synthase reaction. By these criteria this enzyme, which is considered to use an aldolase mechanism in the cyclization step, cannot form a Schiff base during the reaction. In addition, these enzymes are not inhibited by 10 mM EDTA. One interpretation of this evidence is that the myo-inositol-1-P synthases we have studied are neither Class I nor Class II aldolases, and simply use base catalysis in the cyclization step.


Inhibition of Azotobacter vinelandii RNA polymerase by cibacron blue F3GA. Cibacron blue F3GA is a potent inhibitor of the Azotobacter vinelandii DNA-directed RNA polymerase. Addition of 8 micrometer Cibacron blue F3GA prior to initiation results in a greater than 90% inhibition of the poly[d(A-T]-directed synthesis of poly[r(A-U)] while addition of the dye during the course of the reaction is without effect on chain elongation. Binding of RNA polymerase to [3H]poly[d(A-T)] is inhibited by only 15% in the presence of 8 micrometer Cibacron blue F3GA. Inhibition by Cibacron blue F3GA is noncompetitive with regard to ATP, UTP, or template. The poly[d(A-T)]-directed pyrophosphate exchange reaction is relatively resistant to inhibition by Cibacron blue F3GA. Rifampicin added to a similar reaction (in the presence of absence of Cibacron blue F3GA) results in 95% inhibition of the exchange reaction. The interaction of the RNA polymerase core enzyme with Cibacron blue F3GA is shown by the formation of a difference spectrum with a positive maximum at 675 nm which is not affected by the presence of a high concentration (4 micrometer) of rafampicin. The data indicate that Cibacron blue F3GA acts by binding to RNA polymerase and inhibits a step between the synthesis of the initial phosphodiester bond and formation of a stable ternary elongation complex.


Mycobacterium smegmatis fatty acid synthetase. Long chain transacylase chain length specificity. Long chain transacylase activity, acyl-CoA + enzyme in equilibrium acyl-enzyme + CoA, catalyzed by the multienzyme complex fatty acid synthetase from Mycobacterium smegmatis was measured by exchange of radioactive coenzyme A into even numbered fatty acyl-CoA substrates 14 to 24 carbon atoms long. This transacylase activity decreases sharply with increasing chain length. It is suggested that C24 transacylation may be rate-limiting in de novo fatty acid synthesis catalyzed by the myocobacterial system. Mycobacterial polysaccharides stimulate the rate of transacylation, and this enhancement becomes more marked as the chain length of the substrate increases. The magnitude of the effect is similar to polysaccharide stimulation of overall synthetase activity. It is therefore proposed that terminal transacylation is the specific and perhaps only partial reaction catalyzed by the M. smegmatis fatty acid synthetase which is facilitated by polysaccharide. The product distribution of the synthetase is distinctly bimodal, with peaks for acyl chains 16 and 24 carbon atoms long. A scheme based on nonoverlapping unimodal chain length specificities for the rates of two activities, elongation and terminal transacylation, is offered to explain this bimodal distribution.


Mycobacterium smegmatis fatty acid synthetase. Polysaccharide stimulation of the rate-limiting step. An initial activity burst lasting 5 to 10 s is observed for both de novo synthesis with acetyl-CoA as primer and for elongation of palmitoyl-CoA catalyzed by the multienzyme complex fatty acid synthetase from Mycobacterium smegmatis. After the initial burst, synthetase activity slows at least 6-fold to the steady state rate. The size of the initial burst is proportional to the amount of synthetase protein and corresponds to the synthesis of a small number C three to five) of C24 or C26 acyl chains per mol of enzyme. During the initial burst, C24, C26 acyl enzyme is formed and can be isolated by ammonium sulfate precipitation. On incubation with CoA, enzyme-bound acyl chains undergo transacylation to form the corresponding CoA derivatives. Diffusion of C24-CoA and C26-CoA from the enzyme is slow and rate-limiting for overall fatty acid synthesis. Mycobacterial polysaccharides markedly accelerate this rate-determining step but bovine serum albumin does not. This facilitation of product diffusion accounts for the large stimulation of de novo synthesis and of elongation of mycobacterial polysaccharide. It is also shown that the high apparent Km for acetyl-CoA (approximately 400 micrometer) in the steady state reflects the substrate concentration required to shift the product pattern in favor of shorter chain fatty acids (C16,C18). These conditions circumvent the slow, rate-limiting diffusion of C24-CoA and C26-CoA.


Mycobacterium smegmatis fatty acid synthetase. A mechanism based on steady state rates and product distributions. The initial steady state rate and product distribution of fatty acid synthesis catalyzed by Mycobacterium smegmatis fatty acid synthetase has been investigated as a function of various concentrations of acetyl-CoA, malonyl-CoA, mycobacterial polysaccharide, and bovine serum albumin. Polysaccharide has a large effect on both rate and chain length. The steady state rate stimulation by polysaccharide is not duplicated by other acyl-CoA-binding molecules such as bovine serum albumin. It is concluded that relief of product inhibition does not adequately explain the specific effects of the mycobacterial polysaccharide. A general mechanism is presented which accounts for variations in reaction rate and produce pattern over a wide range of experimental conditions. We propose that the diffusion of long chain acyl-CoA (C14 to C24) from the enzyme is the rate-limiting step in fatty acid synthesis catalyzed by the M. smegmatis synthetase. Polysaccharide facilitates this rate-limiting step by forming a ternary complex with enzyme-bound acyl-CoA causing rapid release of product.


Biosynthesis of phytosphingosine by the rat. Labeled C18-phytosphingosine was detected in a glyco-lipid-rich fraction from rat intestine 4 and 24 h after injection of D-erythro[4,5-3H]C18-dihydrosphingosine. The criteria for identification were: (a) co-migration on gas-liquid chromatography of the trimethylsilyl ethers of the free and N-acetylated base with authentic standard, (b) co-migration on thin layer chromatography of the dinitrophenyl derivative with authentic standard, (c) labeled pentadecanol detection by gas-liquid radiochromatography after subjecting intact sphingolipids or a mixture of free long chain bases to periodate oxidation followed by sodium borohydride reduction, and (d) obtainment of the same fragmentation pattern as with authentic standard when the trimethylsilyl ethers of the N-acetylated base were subjected to gas-liquid chromatography-mass spectrometry. Smaller amounts of labeled C18-phytosphingosine were also detected in ceramide- and sphingomyelin-rich fractions of intestine and in liver and kidney. Experiments with germ-free rats strongly suggest that the intestinal flora is not the (sole) site for the biosynthesis of C18-phytosphingosine.


Solubilization and resolution of thromboxane synthesizing system from microsomes of bovine blood platelets. The thromboxane synthetase system of the microsomes of bovine blood platelets was solubilized by the treatment with Triton X-100. The solubilized preparation was separated into two enzyme fractions by DEAE-cellulose chromatography. One catalyzed the formation of prostaglandin H2 from arachidonic acid in the presence of heme and tryptophan. The other fraction converted prostaglandin H2 to thromboxane B2 and 12L-hydroxy-5,8,10-heptadecatrienoic acid. However, incubation of the latter fraction with prostaglandin H2 at lower temperature produced an unstable compound with platelet-aggregating activity, which was presumably thromboxane A2 and which decomposed readily to thromboxane B2 and 12L-hydroxy-5,8,10-heptadecatrienoic acid.


Degradation of L-[3H]fucose-labeled plasma membrane components from regenerating rat liver. The rates of degradation of liver cell plasma membranes from control and partially hepatectomized rats were investigated by a single isotope administration procedure utilizing L-[3H]fucose as a precursor. The label was incorporated maximally into normal plasma membranes at 2 to 4 h after intravenous injection. At 4 h, L-[3H]fucose activity was 8-fold more abundant in membrane protein than in total liver protein. Paper chromatography of neutral sugars liberated by acid hydrolysis of the plasma membranes indicated that greater than 90% of the recovered radioactivity was in the form of fucose. After partial hepatectomy, the wet weight of the right lateral liver lobe increased linearly during a 4-day postoperation interval. In addition, the specific activity of plasma membrane alkaline phosphatase was elevated to a level that remained 3 to 4 times higher than that of the control membranes. However, there was not much difference between the mean rates of degradation of the labeled plasma membrane components from the control and regenerating livers. The apparent half-lives were 33 and 40.8 h, respectively. This suggests that the dissociation of fucose-containing components from the plasma membrane is not markedly altered during the process of rapid compensatory liver growth.


Possible femoral "steal" syndrome following femoral-femoral bypass. 1. The results of a clinical study of two patients who exhibited transitory donor limb ischemia without angiographic evidence of donor limb occlusive arterial disease following femoral-femoral bypass has been reported. 2. Although the circulation seemed to have been diminished in the donor limb following creation of a femoral-femoral bypass, the acute development of ischemia, there may have been other causes for the donor limb ischemia.


Brachial artery entrapment syndrome. Intermittent arterial compression as a result of muscular hypertrophy. A description is given of an entrapment syndrome of the brachial artery in a thirty-nine-year-old muscular man. The diagnosis was made with the help of angiography. The intermittent compression was not caused by a congenital anomaly, but exclusively by muscular hypertrophy. By cutting the lacertus fibrosus the complaints and symptoms completely disappeared. No previous record of this clinical picture can be found in the world literature, but it can be assumed that this is not an isolated observation.


Aneurysm of the gastroduodenal artery.

Aneurysm of the gastroduodenal artery. A case of aneurysm of the gastroduodenal artery associated with obstruction of celiac axis, is presented. The patient underwent successful surgical correction. The clinical and surgical aspects of this patient are discussed.