Cushing's syndrome associated with a bronchial adenoma. Possible periodic hormonogenesis. Diagnostic and therapeutic problems in a patient with ectopic ACTH syndrome caused by a malignant bronchial adenoma are discussed. Persistent Cushing's syndrome was present following apparent total adrenalectomy, but radioactive scanning with 131I-19-iodocholesterol showed the presence of residual adrenal tissue in the right suprarenal bed. Amelioration of the hypercortisolism occurred after removal of the bronchial adenoma. A paradoxical elevation of adrenocortical activity followed administration of dexamethasone and data are presented which suggest that periodic secretion of ACTH accounted for this phenomenon.


Nonrheumatic mitral incompetence.

Nonrheumatic mitral incompetence. This review clearly indicates that a clinical picture of pure, isolated mitral insufficiency constitutes an interesting diagnostic challenge. In adult patients especially, this common valvular lesion is often nonrheumatic and may be found in a variety of cardiac conditions. The following provides a general orientation for their differential diagnosis. The first clue to the presence of papillary muscle dysfunction, a "spontaneous" chordal rupture, or a congenital or traumatic lesion which may account for the mitral insufficiency, may be derived from the patient's case history. A history suggesting systemic manifestations raises the possibility of atrial myxoma. When a familial incidence is reported, various syndromes or a cardiomyopathy should be considered as the etiology of the mitral incompetence. The auscultatory findings are typical in the mid-late systolic click and murmur syndrome, but recognition of this condition may require careful examination of the patient in different postures. The possibility of obstructive cardiomyopathy may be confirmed by the characteristic carotid pressure tracing. ECG findings of acute or chronic coronary heart disease favor the possibility of papillary muscle dysfunction. In addition, the ECG may support the clinical impression of a cardiomyopathy. Fluoroscopy may show calcification of the coronary arteries and/or dyskinetic left ventricular contractions in papillary muscle dysfunction, intracardiac calcifications in atrial space-occupying lesions, or calcification of a mitral annulus. Chest X-rays may contribute to the diagnosis of acute mitral insufficiency by showing a relatively small left atrium and ventricle in the presence of severe congestive failure. While echocardiography is invaluable as a noninvasive procedure and readily demonstrates the presence of a flail mitral leaflet from chordal rupture, or aids in the recognition of obstructive cardiomyopathy, an atrial space-occupying lesion, or of a billowing mitral leaflet, left ventriculography and coronary angiography constitute the procedure of choice for the fine anatomic diagnosis and functional evaluation of most cases. The accurate diagnosis of the anatomic disruption of the mitral valvular apparatus, as well as of the state of the myocardium and of the coronary arteries, is of particular importance in symptomatic patients, in order to determine the prognosis and to plan the surgical approach.


[Antipsoriatic effect of dithranol (anthralin). 1]. This review deals with the biochemical-pharmacological actions of anthralin (dithranol) and with the various possibilities of regulating the disturbed metabolism in psoriasis. A close relationship exists between enzyme inhibition, blocking of nucleic acid metabolism, and cytostatic-cytotoxic actions exerted by anthralin. Special reference is made of the inhibitory capacity on important enzymes of the pentose phosphate shunt the importance of which is briefly outlined. In vitro experiments with dithranol confirmed clinical observations such as the advantageous application of anthralin together with ultraviolet light or the decreased efficacy of anthralin incorporated in plain zinc paste.


[Entity of the Gianotti-Crosti's syndrome and its relation to hepatitis B infection]. 6 cases of acrodermatitis papulosa eruptiva infantum (acrodermatitis papulosa infantilis or Gianotti-Crostisyndrome: G.C.S.) were observed and the clinical features of this disease are discussed. 1. An erythemato-papular dermatitis mainly of the face, legs, arms, buttocks, not itching, without re-occurrence lasting on the average 20--25 days. 2. An distinct enlargement of the lymph nodes, especially of the inguinal and axillary areas (reactive reticulo-histocytic lymphadenitis). 3. Acute hepatitis, mostly without icterus. 4. Presence of HB Ag-Australia Antigen in the serum of patients a few days after the onset of the disease. Doubts concerning the entity of G.C.S. are getting irrelevant considering the distinct characteristics. The viral genesis already suggested by Gianotti and Crosti of the picture of the disease seems to be true: in all cases Australian antigens are positive. The infectiousness, but the small danger of infection, even if the disease dates back a few months, should not be underestimated. When diagnosing G.C.S. among others the akrolocalized papular-vesicular syndrome described by Crosti and Gianotti should be considered. In 2 patients we could find by electron microscopic studies microtubular aggregates having a diameter of 200 A. These aggregates were situated in the cytoplasm of endothelial cells of smaller vessels in the upper part of the corium.


[Electron microscopy of the oral mucosa in pemphigus vulgaris]. Oral lesions in a typical case of pemphigus vulgaris were investigated electron microscopically. The region of the basement membrane shows only minor changes. The basal cells have normal hemidesmosomes whereas the desmosomes disappear. Acantholytic cells sometimes reveal remnants of desmosomes. The tonofilaments show perinuclear centralization. The findings seem to indicate that the immunopathological process affects the cellular walls and thus separates existing desmosomal contacts and at the same time inhibits new desmosome formation.


[Disseminated granuloma annulare].

[Disseminated granuloma annulare]. A case of generalized granuloma annulare in a 55 year old man is reported. The disease appeared five years before the first admission to the hospital. A large number of bluish-red or skin-colour papules were scattered mainly around the earlobes, buttocks and on the extremities. Some of the lesions were lined up in rings or plaques. Small depigmented and brownish scars were present. Two biopsies revealed characteristic foci of complete collagen degeneration accompanied by a palisading infiltrate in the upper dermis. Treatment with tuberculostatics and antimalarics was without improvement. The lesions cleared after a course of prednison, but reappeared when the drug was discontinued.


[Case contribution on a group affliction with photodermatitis bullosa striata pratensis (Oppenheim)]. After duty in the open field a group of 58 soldiers were simultaneously affected by photodermatitis bullosa striata pratensis Oppenheim. Among others Heracleum sphondylium was found in the botanical inventory of the lower vegetation of the terrain in question. This plant is in our latitudes the most important trigger of phototoxic reactions of the skin.


Studies on the valine sensitivity in non-ketotic hyperglycinemia. An oral loading test with L-valine (100 mg/kg body weight) in 3 patients with non-ketotic hyperglycinemia was accompanied by drowsiness and hyperreflexia of the patients. Metabolic studies revealed a slightly lower rate of disappearance of valine from blood in 2 of the patients. In a third patient, the curve was not different from controls. Gas chromatographic analysis for short-chain fatty acids in serum carried out during the valine loading test did not show increased concentrations. Urine collected during the valine loading test did not show excretion of N-acylglycine derivatives. As neither of the branched chain amino acid transaminases proved to be inhibited by glycine, inhibition by glycine of the uptake of valine by the tissues became likely. This uptake has been measured in rat liver slices. Glycine was found to be a competitive inhibitor of valine uptake, with a Ki of 4.9 mM. It is concluded that the tendency to a decreased rate of valine disappearance from blood in non-ketotic hyperglycinemia could be due to an inhibition of valine uptake by the high plasma glycine concentration. The relationship of the inhibited valine uptake in liver slices of rats with the clinical symptoms in non-ketotic hyperglycinemia patients after a valine load remains to be established.


Ehlers-Danlos syndrome in two siblings with deficient lysyl hydroxylase activity in cultured skin fibroblasts but only mild hydroxylysine deficit in skin. Two siblings suffered from Ehlers-Danlos syndrome characterized by skin fragility, joint laxity and dermal hyperelasticity. The association with microcornea and muscle hypotonia allowed the preliminary classification into type VI according to McKusick. Ultrastructure analysis of skin biopsies revealed poor integration of collagen fibrils into fibres; accordingly, the texture of the connective tissue appeared irregular. Lysyl hydroxylase activity of cultured skin fibroblasts was markedly reduced in the cells of the two patients. Preliminary studies revealed intermediate activity in the cells cultured from the skin of the parents. This finding suggested an autosomal recessive mode of inheritance. Unexpectedly and in contrast to the 3 cases reported in the literature, the hydroxylysine deficit in the patients' skin was, for reasons not yet understood, only mild. Therefore, amino acid analysis of skin is not adequate for the diagnosis of lysyl hydroxylase-deficient Ehlers-Danlos syndrome type VI.


Testicular endocrine function and cyclophosphamide in prepubertal and pubertal patients. Testicular function was examined by means of HCG-stimulation tests in 6 prepubertal or pubertal patients with leukemia or malignant tumours during cytostatic therapy and in 5 patients with nephrotic syndrome after discontinuation of treatment. The results of urinary testosterone assays suggest no damaging effect of cyclophosphamide on Leydig cell function.


Trisomy of the distal third of the long arm of chromosome 10. Report of a new case due to a familial translocation t(10;18) (q24;p11). A new case of trisomy of the distal third of the long arm of chromosome 10 due to familial translocation t(10;18) (q24;p11) is described. The main clinical and radiological signs may be summarized as follows: growth at lower limits of normal; poor facial expression; round, flat face with high, broad forehead, fine, highly arched eyebrows, pseudohyperthelorism, microphthalmia, flat, broad bridged nose, hypoplasia of the bony structures of the central area of the face, "fish mouth", macroglossia, micrognathia; short neck; marked dextroconvex lumbar scoliosis; psychomotor delay of mild degree; selective, more pronounced speech delay. Our observation confirms the suggestion by Yunis and Sanchez that a clinical syndrome corresponds to this chromosomal alteration. However, some interesting differences from the previously reported cases, i.e., the absence of microcephaly and of severe impairment of growth and psychomotor development induce us to establish a more favorable prognosis in our case.


[Glomerulonephritis in the Schoenlein-Henoch syndrome in childhood]. Ten children suffering from anaphylactoid purpura Schoenlein-Henoch and presenting urinary abnormalities of various degrees and reported. The histological and immunohistochemical tests allowed to distinguish two types of glomerulonephritis in these patients: focal glomerulonephritis and extracapillary glomerulonephritis. The prognosis was always favorable: no relationship was found between the clinical signs and symptoms and the findings in the kidney. The authors discuss, on a practical and theoretical level, the indications for kidney biopsies in children with the Schoenlein-Henoch syndrome.


Different types of osteochondrodysplasia in a consecutive series of newborns. Among 14816 consecutive live births there were 7 cases of osteochondrodysplasia (incidence 1:2117). In addition there was 1 case of stippled epiphyses, possibly induced by anticonvulsive drugs taken by the mother during the pregnancy, and one case of cerebro-hepato-renal syndrome of Zellweger with roentgenological features similar to those of hypochondrodysplasia. None of 102 stillborns seen at the same time had osteochondrodysplasia. For genetic counselling an exact diagnosis is mandatory, radiological examination of the skeleton is often of decisive importance.


Simultaneous fluorescence histochemical demonstration of catecholamines and tryptophyl-peptides in endocrine cells. Simple and efficient fluorescence histochemical methods for the concomitant demonstration of tryptophyl-peptide-containing cells and dopamine-containing cells have been developed in this study. Combined formaldehyde and chloral vapour or solution of 5% glyoxylic acid monohydrate in n-butanol induced concomitantly strong yellow fluorescence in the tryptophyl-peptide-containing cells and moderate green fluorescence in the dopamine-containing cells in the sections of the freeze-dried adenohypophysis.


Cytochemical localization of cholinesterase activity at the giant synapse of the squid. The giant synapse of squid stellate ganglion is a chemical synapse where the transmitter substance is not known. The components of the ACh-system are present in squid nervous tissue in large quantities. However externally applied cholinergic drugs have no effect on junctional transmission. Using the Copper thiocholine method for electron microscopic cytochemistry the reaction product was found at the axolemmal surface, in the cisternae of the endoplasmic reticulum of neurons and occasionally between the infoldings of the sheat cells surounding the axons. Abundant deposits of end product are observed in the extracellular space in the proximity to junctional region. However, the localization of the cytochemical end product at the junctional region proper was observed frequently, but not consistently. Radiometric measurements of enzyme activity have revealed that neither specific inhibitors nor specific substrates generaly used for differentiation of cholinesterases in mammalian nervous tissue can be employed for differentiation of squid enzymes. Considering the permeability barriers imposed for external acetylcholine by cytoplasmic processes and the high enzyme activity of structures surrounding the giant synapse, the possibility that acetylcholine may still be a candidate for the missing transmitter is discussed.


Histochemical study of monoamine oxidase latency in the liver of the rat. In a histochemical test system with adrenaline as substrate and nitroblue tetrazolium (NBT) as electron acceptor, an increase of NBT reduction in rat liver sections was found microspectrophotometrically following short hypotonic treatment. Investigations with iproniazide, a monoamine oxidase inhibitor, and non-enzymatic NBT reduction showed that the increased formazan formation was related to the presence of monoamine oxidase. It is suggested that the reason for the observed increase of formazan formation is due to increased permeability of the inner mitochondrial membrane to NBT. Consequently, the increase of monoamine oxidase observed in the histochemical test system does not represent mobilization of a latent activity, but rather complete assessment of activity that is normally present.


Studies on 5'-nucleotidase histochemistry. III. 5'-Nucleotidase activity in smooth muscle cells of the rat's gastrointestinal tube. The distribution pattern of histochemically detectable 5'-nucleotidase (5'-Nase) activity is described in smooth muscle cells of the rat's gastrointestinal tube (esophagus, stomach, small intestine, large intestine). Both, light and electron microscopic methods are used. Faint positive 5'-Nase activity is observed on smooth muscle cells of the lamina muscularis mucosae in the thoracal esophagus whereas it is completely absent from smooth muscle cells of the abdominal esophagus and the stomach. In the small and large intestine strong positive 5'-Nase reaction is found on smooth muscle cells of the lamina muscularis mucosae and the innermost part of the lamina muscularis externa. In the circular and longitudinal layer of the lamina muscularis externa a slight increase in 5'-Nase activity is observed from the proximal to the distal segments. The reaction product is restricted to the outer cell surface of smooth muscle cells. In the small intestine the strong enzymatic activity in the innermost part of the muscularis externa is found to be localized at small and dense muscle cells (sd-cells). Common morphological and histochemical characteristics of sd-cells and smooth muscle cells of the lamina muscularis mucosae are emphasized. Hypothetical functions e.g. uptake of precursors of nucleosidephosphates, possible functional connection to a high glycogen content, correlation between 5'-Nase activity and proliferation capacity and local vasodilatory effect are discussed.


A comparison of various procedures for fine grain development in electron microscopic radioautography. Fine grain development for electron microscopic radioautography was investigated with two types of radioactive specimens: sections of tritiated methacrylate, which provide a homogeneously labeled source for quantitative evaluation of the radioautographic reaction, and sections of 125I-labeled thyroid. Radioautographs were prepared with Ilford L4, Sakura NR-H2, Agfa-Gevaert NUC 307 or Kodak NTE emulsions. The radioautographs were developed with one of several "solution physical" development procedures (Agfa-Gevaert, phenidone-ascorbic acid, p-phenylenediamine developers) or with arrested "direct" developments (D-19b, Elon-ascorbic acid developers). By arresting each development at an early stage of the reaction and at progressively longer time intervals, it was possible to examine the sequence of shapes in the growth of developed silver deposits for each emulsion-development combination. Thus, conditions which resulted in the development of small, round, compact silver deposits were defined for each emulsion. These developments were used in conjuction with gold latensification, a treatment which increases the sensitivity of the emulsions and thus compensates for the lowered sensitivity of fine grain development procedures. The location of the silver deposits in relation to the silver bromide crystals from which they derive was investigated. The emulsion gelatin surrounding the crystals was stained whereas the spaces, which remained after the crystals were dissolved in the photographic fixer, appeared transparent. This analysis permitted the selection of development procedures in which the single or multiple round silver deposits originating from a single crystal will remain within or on the boundary of this crystal. By this method, quantitation of radioautographic reactions composed of small, round silver deposits was studied by using the uniformly labeled 3H-methacrylate sections as a standard source of radiation. The conditions under which grain counting is feasible are discussed.


[The development of the guinea pig gallbladder epithelial cells. II. Electron microscopical and enzymhistochemical investigations (author's transl)]. The development of the guinea pig gallbladder epithelium follows a distinct time schedule. During the first phase (up to 30th day of intrauterine life) the epithelial cells increase in number. They remain small and undifferentiated. In the second phase, from the 30th to the 44th day, cytodifferentiation is a dominating feature. The epithelial cells increase in height, the nuclei become more basal, the cells acquire their final zonal structure and the cell organelles exhibit their characteeristic appearance. Weak enzyme activities can be observed. In the third phase, from the 45th day until birth, there is functional differentiation and the adult pattern of enzyme distribution is established. Glycogen appears first on day 29, increases in amount and then disappears by day 57. There is a marked development of the Golgi apparatus associated with increased synthesis of secretory material. Between the 59th embryonic day and the 6th day of life mucoid cells with different functional states appear in the gallbladder epithelium. Later on these cells can be visualized only in the depths of the invaginations. After birth the epithelial cells become more columnar and by 10 days after birth the adult appearance of the epithelium is fully established.


Fine structural localization of arylsulfatase B activity in the rabbit blood platelets. Fine structural localization of arylsulfatase in the rabbit blood platelets has been investigated in this study. Among many cell organellae, reaction products were exclusively observed in the alpha granules of the platelets. Within the alpha granules, arylsulfatase activity appeared to localize in variable patterns, i.e. reaction products confined mainly at the peripheral region in many granules, while they deposited heavily throughout the granule matrices in some others. In a blood platelets, each alpha granule showed the different staining pattern which indicated more variable functional heterogeneity in the granules.


Motor vehicle accidents in urban dogs: a study of 600 cases. The records of 600 dogs injured in motor vehicle accidents and admitted as emergencies to the Trauma Emergency Service of the University of Pennsylvania Veterinary Hospital were studied. Young males were the most frequently injured dogs. The motor vehicle accidents resulted in a wide variety of injuries. Superficial wounds were diagnosed as the only manifestation of trauma in 190 (31%) dogs. Among the remaining 410 dogs, skeletal injury was diagnosed in 87% and soft organ injury in 27%. Multiple region injury occurred in 36% of all dogs. Among specific injuries, it was found that the pelvis was the most frequently injured skeletal structure and that the liver was the most frequently injured abdominal organ. Almost all deaths resulted from intrathoracic and intraabdominal injuries, and over 50% of the dogs euthanatized had central nervous system injury. The overall fatality rate, including euthanasias, was 12.5%.


Pharmacokinetics and dosage of aspirin in cattle. Sodium salicylate was given intravenously to clinically normal cows, and aspirin (acetylsalicylic acid) was given orally to arthritic and clinically normal cattle. Despite slow absorption (half-time of absorption, 2.91 hours) of orally administered aspirin and rapid elimination (biologic half-life, 32 minutes) of salicylates, oral dosage of 100 mg/kg every 12 hours maintained serum salicylate concentration greater than 30 mug/ml, which was considered to be therapeutically effective.


Perirenal edema and toxic nephrosis in cattle, associated with ingestion of pigweed. Twenty-two young cows died or were euthanatized after intoxication associated with ingestion of redroot pigweed (Amaranthus retroflexus) growing in marginal grass pasture. After several days of weakness and posterior incoordination, the cattle became recumbent but remained alert. Pertinent clinical laboratory findings included increased blood urea nitrogen content and marked proteinuria. At necropsy, perirenal edema and toxic tubular neprosis were seen.


Empyema of the equine paranasal sinuses. A survey of 28 cases of chronic purulent infection of the sinuses in the horse showed a relationship between incidence and age, and a probable relationship with breed. Dental disease was apparently the cause of half the cases. The conventional treatment of trephination followed by irrigation with antiseptic solutions resulted in freedom from recurrence of the disease for over 1 year in 17 of the 28 horses. Antibiotic therapy appears to be useless in chronic empyema. The prognosis is adversely affected by the development of facial swelling, a sign seen usually only in long-standing cases.


An expansion and compression technique for reducing and stabilizing proximal epiphyseal fractures of the tibia in foals. Proximal tibial epiphyseal fractures in 2 foals were reduced and stabilized by expansion and compression between 2 transversely placed Steinman pins, one on each side of the fracture. A Charnley apparatus or turnbuckles placed between the pins on each side of the fracture provided the mechanical advantage for repositioning the fracture fragments and achieving rigid fixation during healing. A light cast was applied over the fixation  apparatus, keeping the limb in an extended position. The cast and fixation apparatus were removed after 4 weeks. The technique allowed rapid healing and adequate ambulation. The foals had functional, straight hindlimbs 6 months after the fractures occurred.


Muscular cysticercosis from Taenia ovis in feedlot lambs. From a Colorado enterprise annually fattening about 50,000 lambs, 3,664 animals were examined for muscular cysticercosis. Of the total number, 406 (11.1%) were parasitized, wheras an estimated 0.1% of Colorado lamb carcasses are infected. Of the infected lambs, 60% had cardiac lesions. Two types of cysticerci-viable and degenerated-were found. The lambs probably acquired infection from grazing summer ranges and pastures where coyotes or dogs infected with Taenia ovis contaminated the forage.


Hydrocephalus associated with the noneffusive form of feline infectious peritonitis. The noneffusive form of feline infectious peritonitis was diagnosed in a cat with progressive vestibular and cerebellar deficits. Histopathologic findings included generalized pyogranulomatous lesions of the meningeal, ependymal, and subependymal areas of the brain. Hydrocephalus was associated with the lesions.


Leiomyosarcoma of the urinary bladder in a cat. An 81/2-year-old female domestic short-haired cat was admitted with signs of chronic cystitis. Radiography revealed marked and irregular thickening of the bladder wall. Biopsy revealed the thickening to be a malignant tumor of muscle origin and treatment with proteolytic enzymes was instituted. After one month, the cat was euthanatized because of uremia associated with progressive tumor growth. At necropsy the bladder was grossly thickened. The histopathologic diagnosis was leiomyosarcoma.


Human psittacosis associated with commercial processing of turkeys. Between May 1 and Aug 1, 1974, 114 of approximately 1,000 employees in 7 turkey processing plants in Texas (5), Missouri (1), and Nebraska (1) had an illness compatible with psittacosis. The illness was characterized by fever, severe headache, and pneumonia. Treatment with tetracycline was effective in most cases. Psittacosis was confirmed in 41 (36%) of the cases by a fourfold change in complement-fixation titer to the Chlamydia group antigen. Investigations implicated several turkey flocks from Texas as the source of the disease. Control measured, including screening turkey flocks before slaughter and assuring treatment of infected birds, were instituted by a special task force of state and federal veterinarians.


An epizootic of bovine tuberculosis in Georgia. A tuberculous beef animal was detected Aug 29, 1973, by a veterinary medical officer in a Georgia slaughter establishment. Animal identification and traceback eventually led to a purebred beef herd in Decatur County, Georgia. Investigation and tuberculin testing of 23,000 cattle in 350 herds revealed 11 Mycobacterium bovis-infected beef herds and resulted in the complete depopulation of 5 herds. The epizootic was confined to Georgia and was eradicated by April, 1975.


Epizootic of anthrax in Falls County, Texas. An epizootic of anthrax in Eastern Falls County, Texas, killed at least 238 animals during a 6-week period ending July 31, 1974. Infection appeared to be caused by the ingestion of contaminated soil and grass in the drought-stricken central Texas area. The participation of the Texas Department of Agriculture, the Texas National Guard, and other state agencies was of great assistance to the Texas Animal Health Commission in handling the epizootic and panic stricken public. Use of the unencapsulated Sterne strain spore vaccine was credited with preventing many livestock losses in the area. Contaminated city water was detected during the epizootic but human disease did not result.


The histology of epiphyseal union in mammals. Epiphyses of man and dog in various stages of union are described. As union approaches, new cartilage is added to the epiphyseal surface of the cartilaginous epiphyseal plate by chondrification of the epiphyseal marrow. Before and during early union the cellular arrangement of the cartilaginous epiphyseal plate is well preserved, with good cartilage columns. Mineralization of the cartilage, demarcated by tide-lines, spreads from both surfaces, engulfing the cartilage columns. The first union of an epiphysis is by such mineralized tissue, a form of metaplastic bone. In the smaller and earlier-uniting epiphyses, the mineralized cartilage and the sheets of lamellar bone that cover its surfaces are first removed in one restricted area and replaced by new bone and marrow. The original perforation is later enlarged until all the cartilage is destroyed and union in complete, leaving no scar. In the larger and later-uniting epiphyses there are multiple perforations in the epiphyseal plate and remnants of epiphyseal bone often persist as an epiphyseal scar. In both types of union remnants of the peripheral parts of the plates may be found for a while with the cartilage columns set at right angles to the axis of the bone as a whole, an unexplained peculiarity. At full union all such "residual" cartilage is destroyed.


Gland ultrastructure in human gall bladder. The ultrastructure of two gland cell types in the chronically inflamed human gall bladder is described. The lumina of the glands in the neck region are wide and the gland cells are cuboidal in shape. The nuclei are round and relatively large and there are only a few secretory granules in the scanty cytoplasm. Glands elsewhere resemble those in the pyloric antrum of the stomach in their ultrastructure, and therefore the former may be called "antral type" glands. They contain abundant mucin granules in their cytoplasm and the Golgi apparatus is prominent. There are rather tall irregular microvilli on the apical cell surface and the nuclei are located close to the basal plasma membrane. Enterochromaffin-like cells are to be found in the antral type glands, but not in the glands of the neck region.


Factors involved in the nutrition of the human lumbar intervertebral disc: cellularity and diffusion of glucose in vitro. Post-mortem specimens of the human lumbar (L4-L5) intervertebral disc have been studied histologically and physico-chemically. Blood vessels were found only at the margin of the anulus fibrosus and in the vertebral marrow spaces. Contact between disc tissue and marrow spaces occupied about 10% of the bone-cartilage interface. The disc was most cellular at the periphery of the anulus fibrosus and in the hyaline cartilage next to the vertebral bone. Cellularity declined towards the nucleus pulposus where it achieved a low constant value. The cell density of the disc as a whole was about 60000 cells/mm3. For glucose, the diffusion coefficient of the anulus fibrosus and hyaline cartilage end plate was 2.5 cm2/sec and 2.4 cm2/sec respectively at 37 degrees C, comparable to that of cartilage elsewhere. The permeability of the bone-cartilage interface was low, particularly in the peripheral part. Calculations, based on the present findings and derived values for glucose utilization in disc tissue, indicate that nutritional conditions in the intervertebral disc are more critical than, for example, in articular cartilage.


Structure and innervation of extraocular muscles of Carassius. The extraocular muscles of the carp Carassius contain two types of muscle fibre. Large white fibres have ribbon-shaped peripheral myofibrils and triads located at the Z line. Small red fibres, rich in mitochondria, have polygonal-shaped myofibrils and triads at the A-I junction. Silver- and cholinesterase-stained preparations show that the large fibres are innervated by axons which spiral around them and exhibit intense cholinesterase activity over long distances. Axons supplying small muscle fibres run across bundles of fibres, making one contact with each fibre. By electron microscopy the nerve endings on each fibre type appear identical, both having a smooth post-junctional muscle membrane. The differences in structure and innervation pattern of the two fibre types are discussed in relation to their possible functional roles.


The dentinal tubule and odontoblast process in the cat. The odontoblast process and dentinal tubule of the cat have been examined after vascular perfusion of aldehydes for periods of 20-60 minutes. The odontoblast process is apparently confined to the inner half of a dentinal tubule. Its structure consists of microtubules, microfilaments and a variable population of vesicles, all surrounded by a plasma membrane. The apparent limited peripheral extent of the process is perhaps an artefact due to incomplete penetration of the fixatives.


The ultrastructure of the adrenal cortex of the Mongolian gerbil (M. unguiculatus). Ultrastructurally, the adrenal cortex of the Mongolian gerbil can be divided into two main regions and one narrow interposed border zone. The outer region corresponds to the zona glomerulosa and zona fasciculata of the rat adrenal cortex, whereas the inner region corresponds to the rat zona reticularis. The mitochondria are variable in shape, size and internal structure but generally lamelliform or tubulo-vesicular with a dense matrix in the outer region and plate-like and tubular in the inner region. Some of the mitochondria in the border region are of the polylaminar membranous type. The endoplasmic reticulum is abundant and smooth in the outer region but less prominent in the inner regions, where it is both smooth and rough. The concentric whorled membranes of rough endoplasmic reticulum are a characteristic feature of the border zone. Lipid vacuoles are abundant in the outer region. Lysosomes are numerous in the inner region and tend to form groups of 4 or 5. Aminoglutethimide causes a less conspicuous alteration in the adrenal cortex of the Mongolian gerbil than in the rat. The main alterations consist of a profound increase of lipid and lysosomes, a decrease in the number of SER profiles, and complete disappearance of the whorled membranes of RER.


The postnatal development of the liver in a marsupial, Didelphis virginiana. 2. Electron microscopy. The postnatal development of the liver has been examined with the electron microscope in 15 groups of young opossums. At birth, hepatic cells show little organization and large islands of haemopoietic elements occur between clusters of hepatic cells. Hepatocytes appear small and contain an abundance of granular endoplasmic reticulum. The cytoplasm is also characterized by scattered lipid droplets of varying sizes. Canaliculi occasionally are found between hepatocytes. Only a few scattered lining cells are found in relation to the sinusoids and extensive areas appear to lack a lining endothelium. By the 9th postnatal day large accumulations of glycogen are present in the majority of hepatic cells and large areas of the sinusoids remain devoid of a lining endothelium. Glycogen and lipid droplets remain abundant in hepatocytes of the 15 day old opossum and later stages. By the 69th postnatal day, hepatocytes are relatively mature in appearance and the principal cytological features appear similar in this and subsequent stages. The cytoplasm is characterized by the presence of glycogen, granular endoplasmic reticulum and numerous small, spherical mitochondria. A heterogeneous population of hepatic cells was noted in the majority of stages examined. This ultrastructural study confirmed the tendency for haemopoietic cells to remain in discrete foci of like kind, both with regard to cell lineage and stage of maturation.


Electron microscopic observations on the repair of perforated tympanic membranes in the guinea-pig. Experimental perforations of the tympanic membrane in the guinea-pig have been examined electron microscopically. The perforation site became filled with maturing granulation tissue, covered on its external (meatal) and internal (middle ear) surfaces by cells derived from the epithelia of the respective surfaces of the normal membrane. The fibrils and interfibrillar matrix of the normal tympanic membrane in this species have a unique ultrastructure. The fibrils are quadrangular, apparently composed of four subunits, and the surrounding matrix is more electron-dense than in most connective tissues. During repair this type of fibril and matrix was not reconstituted within the 6 month period examined. In its place were formed fibrils of varying diameter, probably collagenous, and differing from those of normal membrane both in size and in the lack of characteristic interfibrillar matrix. The fibroblasts of the wound site, even if of local origin, thus do not appear to be capable of synthesizing the local type of connective tissue.


Growth of human intervertebral discs and vertebral bodies. In normal children, who usually learn to walk unaided before the age of two, the resultant change from a primary to secondary lumbar curvature is accompanied by a change in the relative position of the nucleus pulposus within the intervertebral disc, from a predominantly posterior situation to a central situation. During the third and fourth years of life, an increase in the rate of vertical growth of the L 4-5 "total disc", as measured at its centre, is associated with the gradual change from convexity to concavity in the shape of the bony vertebral end surfaces bounding the disc. In non-ambulant spastic children, the increased vertical growth of L 4-5 discs and the change in shape of the lumbar vertebral bodies, described in normal children, are not seen. In these non-ambulant children, though normal vertebral body heights are attained, the heights of the L 4-5 discs are severely reduced compared with normal heights for corresponding ages, and the anteroposterior diameters of lower lumbar vertebral bodies and discs are also more than two standard deviations below the corresponding norms. It is suggested that while vertical growth of the central region of lumbar vertebral bodies may be genetically determined and independent of mechanical factors, vertical growth of lumbar intervertebral discs and anteroposterior growth of lumbar vertebral bodies and discs are dependent on the activity associated with weight-bearing in the erect posture.


Observations on the structure and autonomic innervation of the guinea-pig seminal vesicle and ductus deferens. A combination of light microscopic, neurohistochemical and electron microscopic techniques were used to study the distribution of autonomic nerves in the seminal vesicle, and in the submucosa of the ductus deferens, in the guinea-pig. In routine histological preparations, the muscle coat of the seminal vesicle consisted largely of circularly orientated smooth muscle cells with an additional longitudinal layer confined to its urethral end. Using fluorescence microscopy, a rich plexus of noradrenergic nerves was demonstrated throughout the muscle coat of the seminal vesicle and this was confirmed electron microscopically. Numerous noradrenergic terminal regions were frequently observed 10-100 nm from adjacent smooth muscle cells, and catecholamine-containing nerve cell bodies were observed in the adventitia of the urethral end of the seminal vesicle. With the light microscope, a rich plexus of AChE+ve nerves was found in the circular muscle layer but the outer longitudinal layer contained few similar nerves. AChE+ve ganglion cells were confined to the adventitia of the urethral end of the seminal vesicle. Electron microscopy revealed a second type of terminal region containing small agranular vesicles, the distribution of which corresponded to that of the AChE+ve nerves observed by light microscopy. In both layers, these terminal regions were associated with other axons, although the circular muscle layer also contained single axons of this type in close association with adjacent smooth muscle cells. Very few noradrenergic nerves were found in the submucosa of the seminal vesicle and ductus deferens and those which were observed usually accompanied the vascular supply. In contrast, a rich AChE+ve nerve network was observed in the submucosa of both the seminal vesicle and the ductus deferens. Electron microscopy revealed numerous groups of submucosal axons, many of which appeared varicose and contained small, agranular vesicles. Similar intra-epithelial axons were occasionally observed in both organs. These findings have been discussed in relation to the possible functional significance of this extensive autonomic innervation.


N-acetyl-L-phenylalanyl-L-phenylalaninol a metabolite of Emericellopsis salmosynnemata. A new metabolite N-acetyl-L-phenylalanyl-L-phenylalaninol was isolated from culture filtrates of Emericellopsis salmosynnemata which produces zervamicins I and II. The structure was assigned from spectral properties and degradative studies.


The structure of a glyoxalase I inhibitor and its chemical reactivity with SH-compounds. The structure of a glyoxalase I inhibitor (I), isolated from a cultured broth of Streptomyces griseosporeus, was found to be 2-crotonyloxymethyl-4,5,6-trihydroxy-cyclohex-2-enone by chemical studies. Stereochemistry and absolute configuration were determined to be 4R, 5R and 6R by X-ray crystallographic analysis of a bromine-containing crystalline derivative. The crotonyloxy group of I shows a surprising proclivity to be displaced by SH-compounds. This property is shown to be the basis for its biological activity.


Studies on the biosynthesis of basic 16-membered macrolide antibiotic, platenomycins. I. Selection of and cosynthesis by non-platenomycin-producing mutants. In a search for blocked mutants which may produce a biosynthetic intermediate, mutation by N-methyl-N'-nitro-N-nitrosoguanidine treatment and/or ultra-violet irradiation were performed on platenomycin-producing Streptomyces platensis subsp. malvinus MCRL 0388 (NRRL 3761). Twenty four non-platenomycin-producing stable mutants were thus obtained and tested for cosynthesis ability. Antibiotic cosynthesis with a pair of these mutants made it possible to detect the producer of an intermediate. Among these mutants which were classified into eight groups (A to G and doubtful groups), mutants of groups A and B appeared from their complementation pattern to be the useful producers of biosynthetic intermediates of platenomycin.


Studies on the biosynthesis of basic 16-membered macrolide antibiotics, platenomycins. II. Production isolation and structures of 3-O-propionyl-5-O-mycaminosyl platenolides I and II, 9-dehydro demycarosyl platenomycin and demycarosyl platenomycin. Four basic glycosides have been isolated from the fermentation broth of the blocked mutants of Streptomyces platensis subsp. malvinus MCRL 0388. These compounds isolated and purified by solvent extraction and column chromatography were identified as 3-O-propionyl-5-O-mycaminosyl platenolides I (PPL-I-MC) and II (PP-II-MC), 9-dehydro demycarosyl platenomycin (DDM-PLM) and demycarosyl platenomycin (DM-PLM).


Studies on the biosynthesis of basic 16-membered macrolide antibiotics, platenomycins. III. Production, isolation and structures of platenolides I and II. Two neutral macrocyclic lactones designated platenolides I and II have been isolated as the major products from the fermentation broth of the blocked mutants of Streptomyces platensis subsp. malvinus. These two compounds were isolated by solvent extraction and purified by column chromatography. Both platenolides [PL-I: C20H32O6, PL-II: C20H34O6] are closely related to the platenomycin aglycone.


Studies on the biosynthesis of basic 16-membered macrolide antibiotics, platenomycins. IV. Biosynthesis of platenomycins. To elucidate the biosynthetic pathway of platenomycin (PLM), biosynthetic relationships of platenolides I (PL-I) and II (PL-II), 3-O-propionyl-5-O-mycaminosyl platenolides I (PPL-I-MC) and II (PPL-UU-MC), 9-dehydro demycarosyl platenomycin (DDM-PLM) were examined with growing cultures or the washed mycelium of blocked mutants of Streptomyces platensis subsp. malvinus MCRL0388, a  platenomycin-producing organism. As a result, it was revealed that PLM was biosynthesized from PL-I via DM-PLM and DA-PLM along the pathways shown in Chart 1. 4''-Isovaleroyl unit of PLM-A and 4''-propionyl unit of PLM-B were respectively derived from L-leucine and L-isoleucine.


New isoflavones, inhibiting catechol-O-methyltransferase, produced by Streptomyces. In the screening of catechol-O-methyltransferase inhibitors in streptomyces culture filtrates, three new isoflavones were isolated. Their structures were shown to be 3',5,7-trihydroxy-4',6-dimethoxyisoflavone (I), 3',5,7-trihydroxy-4',8-dimethoxyisoflavone (II), 3',8-dihydroxy-4',6,7-trimethoxyisoflavone (III). I and II inhibited both catechol-O-methyltransferase and dopa decarboxylase, and showed hypotensive action. III was a specific inhibitor of catechol-O-methyltransferase, and showed no hypotensive action.


A new antibiotic, fumaramidmycin I. Production, biological properties and characterization of producer strain. A new antibiotic, fumaramidmycin, has been isolated from a streptomycete NR-7GG1 which was characterized and named Streptomyces kurssanovii. The strain produced the antibiotic only when grown on agar plates but not in the submerged culture broth, where the contact with the vegetative mycelia appears to cause the inactivation of the antibiotic. The antibiotic shows an antimicrobial activity against both Gram-positive and Gram-negative bacteria.


A new antibiotic, fumaramidmycin. II. Isolation, structure and syntheses. A new antibiotic fumaramidmycin produced by Streptomyces kurssanovii NR-7GG1 was isolated as colorless crystals. The structure was shown to be N-(phenylacetyl) fumaramide. Starting from fumaramic acid, fumaramidmycin has been synthesized in good yield, in which the key stage involves N-acylated imino ether formation followed by mild acid hydrolysis. Five analogues of fumaramidmycin have also been prepared.


A behavioral-educational alternative to drug control of hyperactive children. A behavioral procedure for controlling hyperactivity without inhibiting academic performance is described. Using a time-sample observational method, the hyperactivity displayed by three school children was recorded during math and reading classes. Concurrently, math and reading performances were measured. The study consisted of two baselines, one while the children were on medication and the second while they were off medication. A multiple-baseline design across the two academic subject matters was used to assess the behavioral intervention, which consisted of token reinforcement for correct academic responses in math and subsequently math and reading. Discontinuation of medication resulted in a gross increase in hyperactivity from 20% to about 80%, and a slight increase in math and reading performance. Introduction of a behavioral program for academic performance, during no medication, controlled the children's hyperactivity at a level comparable to that when they were on drugs (about 20%). At the same time, math and reading performance for the group jumped from about 12% during baseline to a level of over 85% correct. Each child performed behaviorally and academically in an optimal manner without medication. Contingency management techniques provided a feasible alternative to medication for controlling hyperactivity in the classroom while enabling the children to grow academically.


Modification of human gastric acid secretion with operant-conditioning procedures. In an attempt to control gastric acid secretion with operant-conditioning techniques, four normal women were given visual feedback on gastric pH plus money reinforcers. When money was made dependent on increased secretion in a differential-reinforcement-of-high-rates schedule, the rate of secretion of three of the four subjects increased to three times baseline. When money was then made dependent on decreased secretion in a differential-reinforcement-of-other-behaviors schedule, the rate of secretion of these three subjects returned to baseline levels. Heart rate, respiratory frequency, abdominal electromyographic activity, and stomach motility (measured by the electrogastrogram method) were not consistently correlated with acid secretion across subjects, although individual subjects showed substantial correlations between acid secretion and one or more other physiological response.


Developing generalized behavior-modification skills in high-school students working with retarded children. Seven high-school trainees each conducted training sessions with two profoundly retarded children. Each trainee was asked to teach one child to follow the instruction "Bring ball" and the other child to follow the instructions "Sit down" and "Come here". During baseline sessions, before the trainees had been instructed in behavior-modification techniques, no trainee successfully taught either child to follow the instructions. After differing numbers of baseline sessions, trainees were exposed to training procedures designed to teach them to teach one child to follow the instruction "Bring ball". The training procedures consisted of videotaped modelling, rehearsal, and corrective feedback and praise. Following the training procedures, four of the seven trainees successfully taught their child the instruction "Bring ball". Further, all trainees were able to teach their other children to follow the instructions "Sit down" and "Come here", even though they had received no modelling, rehearsal, or feedback on how to teach the children to follow these instructions. The ability of the trainees to teach new behaviors to different children indicates the development of generalized skills in behavior modification.


Generalizing the use of descriptive adjectives through modelling. Two retarded children were exposed to daily imitation training in which teacher or nurse modelled and instructed each child to imitate 12 sentences containing one of six animal names. The subjects were praised for correct verbal imitation. Across three phases of a multiple-baseline design, sentences varied as to the presence or absence of size and/or color adjectives describing the animals. In probe sessions at another time of day and in a different setting, the experimenter twice asked each subject to dfescribe 12 different animal pictures. The subjects' use of descriptive adjectives (color and/or size) greatly increased during probe sessions as a function of the sentence content (presence or absence of color and/or size adjectives) in modelling and imitation training sessions. Generalization to descriptions of animals also not used in imitation training sentences was also obtained..


The impact of applied behavior analysis on diverse areas of research. The impact of applied behavior analysis on various disciplines and areas of research was assessed through two major analyses. First, the relationship of applied behavior analysis to the general area of "behavior modification" was evaluated by examining the citation characteristics of journal articles in JABA and three other behavior-modification journals. Second, the penetration of applied behavior analysis into diverse areas and disciplines, including behavior modification, psychiatry, clinical psychology, education, special education, retardation, speech and hearing, counselling, and law enforcement and correction was assessed. Twenty-five journals representing diverse research areas were evaluated from 1968 to 1974 to assess the extent to which operant techniques were applied for therapeutic, rehabilitative, and educative purposes and the degree to which methodological desiderata of applied behavior analysis were met. The analyses revealed diverse publication outlets for applied behavior analysis in various disciplines.


Setting generality and stimulus control in autistic children. This study was designed to assess the transfer of treatment gains of autistic children across settings. In the first phase, each of 10 autistic children learned a new behavior in a treatment room and transfer to a novel extra-therapy setting was assessed. Four of the 10 children showed no transfer to the novel setting. Therefore, in the second phase, each child who failed to transfer participated in an analysis of stimulus control in order to determine the variables influencing the deficit in transfer. Eachof the four children who did not transfer were selectively responding to an incidental stimulus during the original training in the treatment room. Utilizing a reversal design, each of the four children responded correctly in the extra-therapy setting when the stimulus that was functional during training was identified and introduced into the extra-therapy setting. The extreme selective responding and the resulting bizarre stimulus control found are discussed in relation to the issue of setting generality of treatment gains.


Identifying specific erotic cues in sexual deviations by audiotaped descriptions. Using audiotaped descriptions of sexual experiences and a direct measure of penile erection, it is possible to specify more precisely erotic cues in sexual deviates. Results indicated that such cues are highly idiosyncratic. Some tentative conclusions and suggested application for the method are discussed.


Treating overweight children through parental training and contingency contracting. Fifteen overweight girls aged 5 to 11 yr were randomly assigned to one of two weight-reduction treatments: response-cost plus reinforcement, response-cost only, or a no-treatment control group. In the response-cost plus reinforcement group, parents contracted to facilitate their child's weight loss by carrying out reinforcement and stimulus control techniques, completing weekly charts and graphs, and encouraging their child to exercise. The response-cost only group parents did not contract to reinforce their child's performance. The response-cost program applied to both experimental groups was conducted in weekly meetings in which parents lost previously deposited sums of money. Twenty-five per cent was deducted for missing the weekly meeting, 25% for failing to fill out charts and graphs, and 50% if their child failed to meet her specified weekly weight-loss goal. At the end of the 12-week treatment period, both experimental groups had lost significantly more weight than the control group. After an eight-week, no-contact follow-up, some of the lost weight was regained. The response-cost plus reinforcement group was still significantly below the controls. The response-cost group just missed significance. A 31-week, no-contact follow-up failed to show a treatment effect, but did show a trend towards slower weight gain by the response-cost plus reinforcement group.


The use of prompts to enhance vicarious effects of nonverbal approval. The effect of nonverbal teacher approval (physical contact in the form of patting approvingly) delivered to target subjects on the attentive behavior of adjacent peers was examined in a special-education classroom. In a reversal design, two pairs of moderately retarded children were exposed to nonverbal approval, with only one subject in each pair receiving approval. In different phases, nonverbal approval was delivered alone or in conjunction with a verbal prompt directed to the adjacent peer or to the class as a whole. The prompt was designed to make salient the target subject's attentive behavior and the nonverbal reinforcing consequences that followed. Providing contingent nonverbal approval alone consistently altered attentive behavior of the target subjects but did not alter the attentive behavior of adjacent peers. However, accompanying nonverbal approval with a verbal prompt did increase attentive behavior of nonreinforced peers.


Replication of the Achievement Place model in California. Attempting to replicate procedures from Achievement Place, token reinforcement procedures were used to modify savings, conversational interruptions, and table-setting of delinquent boys residing in a home-style, community based, treatment setting. The tokens (points) were redeemable for various privileges and could be earned for specified appropriate behaviors and lost for specified inappropriate behaviors. Contingent point fines reduced the frequency of interruptions. Point rewards improved table-setting, but even large point rewards did not substantially increase savings. Baseline data indicated that lateness to dinner was not a problem, as it was in Achievement Place. Withdrawal of contingent points and back-up rewards did not disrupt the clean-up behavior of two boys.


Training mentally retarded adolescents to brush their teeth. The need for self-care by retarded individuals in behaviors such as brushing teeth led to the development and evaluation of a comprehensive toothbrushing program that included a task analysis and training procedure specific to each component of the task analysis. Eight mentally retarded adolescents, in two groups, individually received acquisition training that included scheduled opportunities for independent performances, verbal instruction, modelling, demonstration, and physical assistance. The first group of four subjects received token plus social reinforcement; the second received only social reinforcement. All eight subjects showed improved toothbrushing behaviors when compared to baseline. Six of the eight subjects correctly performed all toothbrushing steps in two of three consecutive sessions. The study emphasizes the need for systematic program development and evaluation.


Teaching generative use of sentence answers to three forms of questions. Three retarded and four economically disadvantaged children were taught, through modelling and reinforcement procedures, to produce complete sentences in response to three types of questions involving changes in verb inflections. To evaluate generalization of training, new but similar questions were periodically asked, answers to which were never modelled or reinforced. Modelling and reinforcement effectively taught correct sentence answers to training questions and produced new sentence answers to questions for which no specific training had been given.


Do elephants ever forget?

Do elephants ever forget? Three adult female elephants (Elephas maximus) were tested on a light-dark discrimination problem with an 8-yr intertrial interval. The first subject took only 6 min to reach criterion and made only two errors, suggesting remarkable retention. The other two subjects were found to have visual anomalies that would have gone undetected without this research.


Effects of expiratory threshold loading during steady-state exercise. Increases in functional residual capacity (FRC) decrease inspiratory muscle efficiency; the present experiments were designed to determine the effect of FRC change on the ventilatory response to exercise. Six well-trained adults were exposed to expiratory threshold loads (ETL) ranging from 5 to 40 cmH2O during steady-state exercise on a bicycle ergometer at 40-95% VO2max. Inspiratory capacity (IC) was measured and changes of IC interpreted as changes of FRC. ETL did not consistently limit exercise performance. At heavy work (greater than 92% VO2max) minute ventilation decreased with increasing ETL; at moderate work (less than 58% VO2max) it did not. Decreases in ventilation were due to decreases in respiratory frequency with prolongation of the duration of expiration being the most consistent change in breathing pattern. At moderate work levels, FRC increased with ETL; at maximum work it did not. Changes in FRC were dictated by constancy of tidal volume and a fixed maximum end-inspiratory volume of 80-90% of the inspiratory capacity. When tidal volume was such that end-inspiratory volume was less than this value, FRC increased with ETL. Mouth pressure measured during the first 0-1 s of inspiratory effort against an occluded airway (P0-1) was increased by ETL equals 30 cmH2O, in spite of the fact that ventilation was decreased. We concluded that changes in FRC due to ETL had no effect on the ventilatory response to exercise and that changes in P0-1 induced by ETL did not reflect changes of inspiratory drive so much as changes of the pattern of inspiration.


Localization of the effective thermosensitive site in the preoptic region of the ox. These studies were carried out using chronically implanted thermode probes in conscious oxen at 15 degrees C air temperature. The probes were perfused with water either 4 degrees C above or 5 degrees C below resiting hypothalamic temperature and the thermoregulatory response measured by the change in auditory meatal temperature during the perfusion periods of 25 or 40 min duration, respectively. The anatomical position of the probes was known relative to the third cerebral ventricle in each animal. The thermosensitive area of the ox anterior hypothalamic/preoptic region was shown to be highly localized in a position corresponding to the posterior part of the area preoptica. The same area was responsible for both warm and cold sensitivity, the warm sensitivity being approximately three times greater than the cold.


Effect of chest strapping on regional lung function. We studied lung mechanics and regional lung function in five young men during restrictive chest strapping. The effects on lung mechanics were similar to those noted by others in that lung elastic recoil increased as did maximum expiratory flow at low lung volumes. Chest strapping reduced the maximum expiratory flow observed at a given elastic recoil pressure. Breathing helium increased maximum expiratory flow less when subjects were strapped than when they were not. These findings indicated that strapping decreased the caliber of airways upstream from the equal pressure point. Regional lung volumes from apex to base were measured with xenon 133 while subjects were seated. The distribution of regional volumes was measured at RV, and at volumes equal to strapped FRC and strapped TLC; no change due to chest strapping was observed. Similarly, the regional distribution of 133Xe boluses inhaled at RV and strapped TLC was unaffected by chest strapping. Closing capacity decreased with chest strapping. We concluded that airway closure decreased during chest strapping and that airway closure was not the cause of the observed increase in elastic recoil of the lung. The combination of decreased slope of the static pressure-volume curve and unchanged regional volumes suggested that strapping increased the apex-to-base pleural pressure gradient.


Tetanic force development of adductor pollicis muscle in anesthetized man. The tetanic force development of the human adductor pollicis muscle was studied under light anesthesia with nitrous oxide, oxygen, and Demerol, by the use of tetanic stimulation of the ulnar nerve at frequencies ranging from 10 to 100 Hz. The time necessary for the tetanic contraction to reach a plateau was longest at frequencies between 15 and 20 Hz. Fusion of tetanus occurred between 40 and 45 Hz. The mean maximal force of 6.92 kg was developed at a mean frequency of approximately 75 Hz. The maximal force was well maintained up to a stimulation frequency of 100 Hz. The results indicate that in lightly anesthetized man, the maximal force is developed at higher stimulation frequencies than those observed in conscious man and that it is well sustained at higher frequencies.


Substrate utilization by the inactive leg during one-leg or arm exercise. Substrate utilization by the nonexercising leg was studied in healthy subjects during one-leg exercise at an average work load of 105 W for 40 min (n equals 8) or during arm exercise at 65 W for 20 min (n equals 5). During one-leg exercise both the blood flow and the A-FV difference of oxygen for the non exercising leg rose, resulting in an approximately five fold increment in oxygen uptake. EMG activity of the leg was increased above basal. Despite unchanged or falling arterial levels of insulin, the A-FV difference for glucose across the nonexercising leg rose during exercise and the estimated glucose uptake increased approximately fourfold. Release of lactate in the basal state reverted to a significant net uptake of lactate by the nonexercising leg. During arm exercise there was a 20-70% rise in leg blood flow and the leg oxygen uptake rose 25-45% in spite of minimal EMG activity from the thigh muscles. There was a large uptake of lactate  by the legs during arm exercise. We conclude that several important metabolic alterations take place in the nonexercising leg tissues during physical exertion: 1) blood flow and oxygen uptake rise, partly as a consequence of motor activation; 2) substrate utilization shifts from a predominant FFA uptake in the basal state to a greater utilization of carbohydrate; 3) nonexercising muscle, and possibly adipose tissue, play an important role in the removal of lactate during exercise.


Regional vascular influences on tail-cuff measurements of drug-induced changes in systolic pressure. When drug effects are quantified using the tail-cuff method, changes in systemic arterial pressure are extrapolated from those occurring in the caudal artery. The validity of this extrapolation was tested in anesthetized rats by recording drug-induced changes in phasic arterial pressure simultaneously from catheters inserted into the lower abdominal aorta, carotid, and caudal arteries. Pressor responses to norepinephrine or angiotensin were of equal magnitude at all three sites, but phentolamine reduced systolic pressure in the aorta or caudal artery more than that in the carotid artery. Unlike previous discrepancies between carotid and tail-cuff systolic pressures, aortic hypotension caused by injections of phentolamine or pentolinium in awake normotensive or spontaneously hypertensive rats was accurately predicted by the tail-cuff method. Because drug-induced changes in diastolic pressure always varied much less than those in systolic pressure, should indirect measurement of diastolic pressure become technically feasible, it might be preferable for assessing drug effects on blood pressure.


Physiological evidence consistent with the presence of a specific O2 carrier in the placenta. In 30 experiments we perfused the fetal side of the sheep placenta with fluids which had different solubilities for argon, N2, and O2 (dextran, blood, and fluorocarbon emulsions). In some of the experiments we partially exchange-transfused the ewe with the fluorocarbon emulsion. By these procedures we were able to change the physical solubility of argon and N2 severalfold in the fetal perfusion fluid and maternal blood. We found that the diffusing capacity for argon and N2 did not increase with increases in physical solubility in the fetal perfusion medium or in maternal blood. This indicated that the rate-limiting step in the placental transfer of these gases is the small diffusing capacity of the placenta. In contrast, O2 diffusing capacity increased markedly with increased solubility in the fetal perfusion medium. Also the Po2 was frequently the same in the venous blood leaving both sides of the placenta. This indicates that O2 may reach equilibrium between maternal and fetal capillaries in one pass through the placenta. The results are compatible with the presence of specific O2 carrier in the placenta.


Properties of lung parenchyma in distortion. This study offers a basis for evaluating and developing models of stress-strain behavior of the lung in distortion. Tensile forces were applied along three axes to cubes of dog lung parenchyma. With axially symmetrical force-loading, expansion was reasonably symmetrical and pressure-volume relationships were reasonably conventional in range, hysteresis, and time-dependent behavior. When the force load was changed on one axis only, that axis appeared more compliant than it did during symmetrical loading and the other axes changed length in the opposite sign. Similar distortion was apparent at the alveolar level. Data for five specimens over a range of applied loads are filed with the National Auxiliary Publications Service; graphical examples are presented herein. Relationship among the compliances for symmetrical and asymmetrical loadings were consistent with elastic theory. We derived the elastic coefficients, bulk and Young's moduli, and Poisson's ratio from the data. Poison's ratio was about 0.30 in air-filled specimens, but was lower (0.16-0.24) and increases with stress in saline-filled specimens.


Effects of hypoxic and CO hypoxia on isolated hearts. Isolated perfused dog hearts were made hypoxic by respiring the support dog with low oxygen (hypoxic hypoxia) or with carbon monoxide (CO hypoxia). Each heart was exposed to both types of hypoxia, separately. Effects on coronary flow (Qt), coronary vascular resistance, cardiac oxygen consumption (Vo2), and contractility (%deltadP/dt) were studied. Two series of experiments were done. Series I: At constant perfusion pressure. As oxygen content (Cao2) was lowered from 20 to 5 vol%, Qt doubled with hypoxic hypoxia and almost tripled with CO hypoxia (P less than 0.01). Vo2 and contractility increased with both types of hypoxia. Beta-adrenergic blockade eliminated the increase in VO2 and contractility but not the difference in Qt increase between hypoxic and CO hypoxia. Series II: At constant Qt (with beta-blockade), vascular resistance decreased more with CO than hypoxic hypoxia. Finally, alpha-blockade eliminated the difference in vascular resistance and thus with complete (alpha and beta) blockade, the two types of hypoxia have the same effect and are indistinguishable.


Cellular distribution and clearance of aerosolized dipalmitoyl lecithin. Wistar-Lewis rats were anesthetized anc connected to a 3-MHz nebulizer which aerosolized 250 muCi l-alpha-1-palmitoyl-2palmitoyl-[9-10-3H]phosphatidylcholine ([3H]DPL) for 3 min. Appleton frozen-section autoradiographs showed greater than 4 times background radioactivity in approximately 30% of alveoli at 1 min and 2 h after aerosol. As tritium content in the lung decreased, it increased in liver, spleen, kidney, blood, and urine. Percentage of radioactivity from [3H]phosphatidylcholine in the liver declined with time, while [3H]phosphatidylethanolamine doubled between 2 and 12 h. One minute postaerosol 2,500 +/- 500 (SE) type I cells/mm3 lung and 2,500 +/- 750 type II cells/mm3 lung had greater than 20 times background radioactivity; 2 h later only 950 +/- 250 type I cells/-m3 lung still had levels of radioactivity greater than 20 times background while 3,150 +/- 600 type II cells/mm3 lumg now had this level of 3HIDPL. Corresponding numbers of alveolar macrophages were 450 +/- 250 1 min postaerosol and 1,100 +/- 200 after 2 h. Aerosolized DPL as a synthetic surfactant is hampered by significantly faster clearance from the alveolar surface as compared with normal in vivo DPL.


Slow respiratory stimulant effect of hyperoxia in chemodenervated decerebrate cats. A direct stimulating action of oxygen on the CO2 respiratory control system was determined from steady-state and dynamic observations in unanesthetized decerebrate cats. In peripheral nerve-intact animals, inhalation of oxygen (1 atm) produced a small but significant shift to the left as well as a decrease in slope in the steady-state VT vs. log PACO2 relationship. Carotid sinus neurotomy more than doubled the shift, to the extent that the mean PACO2 apneic point was lowered by 6.5 mmHg. Neither vagotomy nor chronic ablation of the area postrema had any detectable influence on the stimulating effect of oxygen on CO2 responsiveness. The arterial-alveolar PCO2 difference, prior to and following carotid chemo-denervation, remained unchanged or was increased by a negligible amount during oxygen inhalation. The oxygen threshold for respiratory stimulation, obtained isocapnically, occurred between 115 and 200 mmHg; VT then increased exponentially tending to level off as PAO2 approached 1 atm. The dynamic response to sudden presentation of oxygen after carotid chemodenervation consisted of a monotonic rise in VT, starting after 20-30 s with a t 1/2 of about 75 s.


Thermal responses of unclothed men exposed to both cold temperatures and high altitudes. Six resting men were exposed to three temperatures (15.5, 21, 26.5 degrees C) for 120 min at three altitudes (sea level, 2,500 m, 5,000 m). A 60-min sea-level control at the scheduled temperature preceded the nine altitude episodes. Comparison of the base-line results at any one temperature showed no differences between rectal temperatures (Tre) or mean weighted skin temperatures (Tsk). After 120 min, Tre and Tsk not only depended on ambient temperature but also altitude. The initial rate of fall in Tre increased with altitude and equilibrium occurred earlier. At 15.5 degrees C, Tre was 0.3 degrees C lower at 5,000 m and 0.2 degrees C lower at 2,500 m than at sea level. Tsk was almost 2 degrees C higher at 15.5 degrees C at 5,000 m and 1 degrees C higher at 2,500 m than at sea level. Similar, smaller differences were observed at 21 degrees C. Mean weighted body temperature showed no change with altitude, but, since the gradient between core and shell was reduced, a shift of blood toward the periphery is implied.


Respiratory sinus arrhythmia: noninvasive measure of parasympathetic cardiac control. The degree of parasympathetic heart rate control, PC, was defined as the decrease in average heart period (RR interval) caused by the elimination of parasympathetically mediated influences on the heart while keeping sympathetic activity unchanged. By reviewing published results on the interaction of sympathetic and parasympathetic heart rate control, the prediction was made that PC should be directly proportional to VHP, the peak-to-peak variations in heart period caused by spontaneous respiration. In sevel chloralose/urethan-anesthetized dogs the vagi were reversibly blocked by cooling, and PC (the difference between average heart period before and after cooling) and VHP (without cooling) were determined under a variety of conditions that included a) increasing vagal activity by elevating the blood pressure b) sympathetic blockade, and c) parasympathetic blockade. The relationship between VHP and PC was linear with an average correlation coefficient of 0.969 +/- 0.024 (SD) and a PC-axis intercept of 15.2 +/- 25.9 ms. In each dog the correlation coefficient between VHP and PC was higher than between VHP and the average heart period (avg correlation coef: 0.914 +/- 0.044). These results suggest that the degree of respiratory sinus arrhythmia may be used as a noninvasive indicator of the degree of parasympathetic cardiac control.


Menstrual variations in thermal properties of the human breast. Menstrual variations in breast skin temperature (Tbr), heat flow (HF), volume (Vol), surface area (A), and core temperature (Tre) have been measured in 13 women. Measurements were made twice weekly on resting subjects throughout one cycle. Twelve of thirteen subjects showed increased Vol and A during the second half of the menstrual cycle; six also showed a minor peak prior to the midcycle rise in Tre. Eight of thirteen subjects had significant peaks in Tbr which preceded the estimated day of ovulation. Breast HF was generally low during the first half of the cycle, rising following estimated ovulation. Analysis of core-to-breast conductance indicates that a decrease in resistance to heat transfer occurs both during the luteal phase and at the Tbr peak. It is hypothesized that the control mechanism underlying the Tbr peak is independent of that controlling the postovulatory increases in Vol, HF, and Tre.


Localization of the medullary respiratory neurons in rats by microelectrode recording. Neuronal activity has been recorded extracellularly from the medulla of anesthetized rats. Units whose discharge frequency varied in phase with respiratory airflow were located bilaterally between 1.5 and 2 mm lateral to midline, extending from 1 mm caudal to 1.5 mm rostral to the obex, in the ventral two-thirds of the medulla. Expiratory units predominated and were intermingled with inspiratory units. Ten different patterns of discharge were distinguished, varying from a short burst at the beginning of expiration to a resting discharge which increased in frequency during either inspiration or expiration. Evidence was also obtained that fiber tracts from other areas of the brain cross midline just caudally to the obex and pass to the respiratory centers on which they apparently exert and excitatory action.


Longitudinal measurements of blood volume and essential body mass in human subjects. Repeated determinations of blood volume and body density were made on 34 Chinese subjects (28 men and 6 women) in Taiwan over a period of 12 yr, as the mean age increased from 31 to 43 yr. Essential body mass calculated from body density and body weight showed no significant change over the 12-yr period. Changes in body weight (mean gain equals 6.0 kg) were attributable to alterations in adipose tissue weight, the density of which was found to be 0.948 g/cm3. In two-thirds of the subjects the second blood volume increased by more than 5% over the first determination, and the mean blood volume for all subjects increased by 7.5% (P less than 0.01). Correlation of the blood volume data with the findings on essential body mass and adipose tissue mass suggests that blood content per unit tissue mass increased in the second determination. This interpretation is supported by the increase in nutrient availability in Taiwan over the 12-yr period, and it may explain the lack of blood volume increase in an earlier longitudinal study on American subjects with stable nutrient availability.


Longitudinal studies on adipose tissue and its distribution in human subjects. On 27 men and 6 women, total body density and 10 skinfolds were measured 12 yr apart, with the mean age increasing from 31 to 43 yr. The increase in skinfold thickness was found to be related to the increase in total body adiposity, calculated from hydrostatic weighing. The external adipose tissue was calculated from the mean skinfold thickness and body surface area. Variations in total adiposity among the population studied as well as changes in total adiposity with age showed a characteristic distribution with approximately two-thirds on the surface and one-third in the interior. The essential body mass or total adipose mass determined by hydrostatic weighing was compared with the values obtained by water-immersion volumetry, total body potassium counting, and skinfold measurements. Teh volumetric and skinfold determinations gave better estimates of these parameters than total body potassium counting.


Myocardial infarction in closed-chest dogs: a simplified method for production. Due to our need, we sought a simple method to reliably create myocardial infarction in the closed-chest dog. Previous techniques were dangerous, time consuming, unreliable, and costly. Here we described a new coaxial catheter method by which occluded catheter plugs are embolized selectively to branches of the left coronary artery in closed chest dogs anesthetized with sodium pentobarbital (10 mg/lb). Infarcts varying in size from 3 to 27 g, 2-27% of the left ventricle, were reliably created in dogs weighing 26-70 lb. Complications were rare with only a single fatality in the last 15 procedures. The method proved safe, simple, quick, versatile, reproducible, and inexpensive.


Glottal aperture during panting with voluntary limitation of tidal volume. A disadvantage of the forced oscillatory technique for measuring total respiratory resistance (namely, that it is usually done during quiet breathing or breathing holding--breathing patterns where the glottic aperture may be highly variable) was overcome by making the measurement during panting. The imposed forced oscillations (Hz) were distinguished from the spontaneous quiet breathing and panting frequencies by ensemble averaging. However, when panting was voluntary restricted so as to standardize the quiet breathing and panting flow amplitudes, resistance values frequently increased. The suggestion that partial glottal closure occurred during voluntarily restricted panting was confirmed by simultaneous inspection of the glottis with a fiberoptic bronchoscope. Thus, maximal opening of the glottis is assured only during unrestricted panting.


Alveostat, an alveolar PACO2 and PAO2 control system. In the study of the physiological regulation of respiration through a control system model it is necessary to test the ventilatory response to various forcing functions of either the parrtial pressure of alveolar carbon dioxide (PACO2) or oxygen (PAO2). Since PACO2 and PAO2 are both functions of alveolar ventilation and metabolic rate, such a result cannot be obtained by merely changing the composition of the inspired gases without a feedback control. Thus a servomechanism is necessary. The input to the servomechanism is an instantaneous determination of PACO2 and PAO2. This is accomplished by using the criterion of equality of the exchange ratio in mean alveolar gas and mean expired gas. The servomechanism described has three essential characteristics: rapidity, accuracy, and stability. In experiments of step, ramp, and sinusoidal forcing functions, variations of PACO2 have been obtained without change in PAO2, and step variations of PAO2 have been obtained without change in PACO2.


Suturable gastrointestinal cannulas for animals of various sizes. A new procedure in which gastrointestinal cannulas are prepared from suturable material improves in vovo methods and requires little machining. Substitution of a suturable material for rigid collars, previously used, prevents leakage, infection, and loss of the cannulas into the gut lumen. The technique has been successfully applied to rats, rabbits, and dogs; consequently it should be useful in animals of almost any size. Two designs of cannulas with descriptions of their implantation are presented. The first version fits into each end of a gut loop and is useful for perfusion, digestion, and secretion studies. The second version forms a sidearm through which feeding or sampling can be carried out at any point in the intestine or the stomach.


An automatic, closed-circuit oxygen consumption apparatus for small animals. An apparatus suitable for the continuous measurement of oxygen consumption of rats and mice is described. The system uses a motorized syringe dispenser to deliver fixed volumes of oxygen to a closed animal chamber. The dispenser is controlled by a micro-differential pressure switch to maintain chamber pressure slightly above ambient. The rate of oxygen consumption is determined by timing the interval between successive operations of the dispenser. The system has proved suitable for a range of experimental conditions and treatments.


Continuous estimation of adipose tissue blood flow in rats by 133Xe elimination. A method is described which permits continuous estimation of adipose tissue blood flow (ATBF) in anesthetized female rats. The method is basesd on continuous monitoring of the elimination of 133Xe after labeling of the animal by intraperitoneal injection. From 2 to 6 h after the beginning of the elimination period close to 100% of the measured activity is shown to be located in adipose tissue, mainly in the parametrial fat. About 18% of the elimination is by way of intertissue diffusion, 82% of the perfusing blood. Changes in ATBF can readily be detected. The coefficient of variation for ATBF determinations is 9-11%. Changes in ATBF can be determined with great accuracy. Average ATBF per g tissue for fed and 48-h fasted rats were 0.105 and 0.122 ml-g-1-min-1, respectively. Total ATBF was lower in fasted than in fed rats (1.05 vs. 1.43 ml-min-1 for parametrial plus retroperitoneal fat). Intravenous administration of glucose (250 mg/h) decreased ATBF significantly in fed but not in fasted rats.


Luft's syndrome: O2 cost of exercise and chemical control of breathing. The oxygen cost of exercise and chemical control of breathing were studied in a subject with Luft's syndrome, a disorder in which skeletal muscle mitochondria have a high "resting" O2 consumption which is imcreased only slightly by stimulation with excess phosphate acceptor, but a normal P/O ratio. The O2 consumption was more than three times normal (1.05 1/min) at rest but could be doubled when stimulated by maximal exercise. The O2 cost of exercise was similar to that of normal subjects. At rest, arterial blood PCO2 and ventilatory response to CO2 were normal, while ventilatory response to hypoxia was four times the predicted value. The data 1) confirm, in vivo, the normal respiratory efficiency of skeletal muscles in this disorder; 2) suggest that in vitro estimates of the extent to which mitochondrial respiration can be stimulated may not correlate with in vivo determinations; and 3) suggests that hypermetabolism per se can cause the ventilatory adjustments which are associated with exercise in normal subjects.


A technique for measurement of retrograde coronary blood flow in intact anesthetized dogs. A technique for measurement of retrograde coronary blood flow in intact anesthetized dogs is described. Occlusion of the coronary artery is produced by the inflation of a small rubber balloon at the tip of a no. 9 cardiac catheter placed under fluorescopy in a branch of the left coronary artery. Blood which bleeds back from the occluded coronary artery through the no. 9 catheter is diverted into a small reservoir of 1-ml capacity. The time to fill this reservoir is recorded electrically. Retrograde coronary blood flow is calculated from the time required to fill this reservoir. Results indicate good repeatability of meadurements. The technique seems to be a simple, adequate, and convenient means for assessing agents for possible vasodilator action on the collateral circulation in intact animals.


A device for testing mechanical properties of biological materials--the "Biodyne". An electromechanical servo-controlled device has been developed. This device can be used to test the mechanical behavior of a wide variety of biological soft tissues. Control and execution of material testing procedures such as stress-strain, vibration, relaxation, creep etc. can be performed by manual operation of the device or by interfacing it with a laboratory type minicomputer. Experiments on excitable tissues such as muscle can also be executed. The design details and system performance are discussed.


A design improvement in continuous blood sampling and analysis for glucose in rest and exercise. In the past 15 yr attempts to sample blood continuously for glucose analysis with the AutoAnalyzer (Technicon Corp.) have been reasonably successful in resting subjects. However, they have sometimes required heparinization of the subjects so as to avoid clotting in the tubes of the system. To avoid the hazards of heparinization during exercise, a method is described for sampling and analyzing blood glucose continuously with the AutoAnalyzer, using a specially designed male Luer adaptor which fits into disposable Teflon catheters.


Growth of Pseudomonas C on C1 compounds: a correction. On reexamination Pseudomonas C was found to be incapable of growth on formaldehyde or formate as a sole carbon source and to contain a hexose phosphate synthase activity when grown on methanol.


Induction of morphogenesis in the genus Arthrobacter. Thirty-eight different compounds were used with 17 species of Arthrobacter to determine their ability to support growth and to induce the morphogenic cycle. In most cases, when a compound supported growth, it also induced the rod phase of growth. However, in a few cases, a compound would support growth with cells remaining in the coccoidal phase throughout the growth cycle. Arthrobacter crystallopoietes was unique in the most compounds that supported growth did not induce the rod phase of the morphogenic cycle.


Recombinant plasmid obtained from two different, compatible staphylococcal plasmids. From two different, compatible staphylococcal plasmids that determine streptomycin and chloramphenicol resistance, respectively, a recombinant plasmid was obtained. This plasmid can be transduced with a rather high frequency (10(-4)/plaque-forming unit) to plasmid-negative strains, the linkage of the two markers being 100%. The maintenance of the recombinant plasmid in the host cell seems to be controlled by the chloramphenicol resistance plasmid. The recombinant plasmid proved to be incompatible with both parental plasmids, which are unrelated. The relationship between the chloramphenicol resistance plasmid and the recombinant plasmid was the same as the between genetically marked derivatives of the recombinant plasmid, whereas the relationship of the streptomycin resistance plasmid to the recombinant plasmid was of a different, asymmetrical type.


Formation of cell wall polymers by reverting protoplasts of Bacillus licheniformis. The biosynthesis of peptidoglycan and teichoic acid by reverting protoplasts of Bacillus licheniformis 6346 His-, in cubated at 35 C on medium containing 2.5% agar, is detectable after 40 min. The amount of N-acetyl-[1-14C]glucosamine incorporated into peptidoglycan and teichoic acid on continued incubation doubles at the same rate as the incorporation of [3H]tryptophan into protein. At the early stages of reversion the average glycan chain length, measured by the ratio of free reducing groups of muramic acid and glucosamine to total muramic acid present, is very short. As reversion proceeds, the average chain length increases to a value similar to the found in the wall of the parent bacillus. The extent of cross-linkage found in the peptide side chains of the peptidoglycan also increases as reversion proceeds. At the completion of reversion the wall material synthesized has similar characteristics to those of the walls of the parent bacilli, containing peptidoglycan and teichoic and teichuronic acids in about the same proportions. Soluble peptidoglycan can be isolated from the reversion medium, amounting to 30% of the total formed after 3 h of incubation and 8% after 12 h. This amount was reduced by the presence in the medium of the walls of an autolysin-deficient mutant; they were not formed at all by reverting protoplasts of the autolysin-deficient mutant itself. Analysis of the soluble material provided additional evidence for their being autolytic products rather than small unchanged molecules. When protoplasts were incubated on medium containing only 0.8% agar, 53 to 67% of the peptidoglycan formed after 3 h of incubation was soluble, and 21% after 12 h. Fibers that appeared to be sheared from the protoplasts at intermediate stages of reversion on medium containing 2.5% agar were similar in composition to the bacillary walls.


Metabolism of naphthalene, 2-methylnaphthalene, salicylate, and benzoate by Pseudomonas PG: regulation of tangential pathways. Naphthalene is metabolized by Pseudomonas PG through 1,2-dihydroxynaphthalene and salicylate to catechol, which is then degraded by the meta pathway. 2-Methylnaphthalene, but not 1-methylnaphthalene, also serves as a growth substrate and is metabolized by the same route, through 4-methylcatechol. The same nonspecific meta pathway enzymes appear to be induced by growth on either naphthalene or 2-methylnaphthalene. The level to which 2-hydroxymuconic semialdehyde hydrolase is induced is low and probably of no metabolic significance. Growth on salicylate or catechol, both intermediates of naphthalene degradation, or benzoate results in induction of the ortho pathway, the alternative route for catechol dissimilation. No induction of 1,2-dihydroxynaphthalene oxygenase was found in salicylate-grown cells. Anaerobic growth on a succinate-nitrate medium in the presence of various inducers indicates that cis, cis-muconate, or one of its metabolites is the inducer of the ortho pathway enzymes. The inducer or inducers of the early enzymes of naphthalene degradation and of the meta pathway enzymes must be an early intermediate of the naphthalene pathway above salicylate.


Cell wall composition of novobiocin-resistant pleiotropic mutant staphylococci. Physically purified cell walls were prepared from selected pleiotropic novobiocin-resistant staphylococcal strains. The quantitative amino acid, amino sugar, and phosphorus contents of these walls are reported. This pleiotype was culturally diagnosed by its inability to support the growth of typing phages, inability to release latent bacteriophage, failure to elaborate coagulase, altered sugar catabolic pattern, and resistance to novobiocin. The strains were divided into two groups on the basis of wall composition. The walls of both groups of strains appeared to possess at least two phosphorus-containing polymers. On group of strains contained elevated levels of phosphorus in the cell walls. The second group contained the novel amino sugar galactosamine in the cell walls. This amino sugar is probably associated with one of the phosphorus-containing wall polymers of this group. On the basis of the data presented, it is suggested that the pleiotropy of these strains is the result of genetic change in the control of the biosynthesis of teichoic acids.


Ultrastructural study of the reversion of protoplasts of Bacillus licheniformis to bacilli. The reversion of protoplasts of Bacillus licheniformis 6346 His- on a medium containing 2.5% agar has been studied in sectioned material after reaction with a ferritin-conjugated antibody specific to the peptidoglycan isolated from the walls of the bacilli. Freeze etching has also been used. Fibrils of material reacting with the antibody have been detected emerging from isolated areas of the protoplasts after 3 h of incubation. This material gradually covers the cell and can eventually (at 6 h) be seen in freeze-etched preparations as a fringe of up to 400 nm around the cells and covering the surfaces with particles that can be removed by lysozyme. At later stages the wall begins to take on a compact, well-defined appearance that can be seen in sections; however, the cells are still grossly deformed. A transitory emergence, beyond the wall of long fibers of 6 nm in diameter, takes place after about 12 h of incubation. These fibers react with the conjugated antibody and after freeze etching show a regular banded structure. They are probably indentical with the fibers isolated elsewhere (Elliott et al., 1975) and shown to contain all the wall constituents (i.e., peptidoglycan, teichoic acid, and teichuronic acid). These fibers are not detectable in the final stages of reversion.


Ultrastructural studies of the mycelium-to yeast transformation of Sporothrix schenckii. Fine details of the internal and external morphology of the in vitro mycelial phase (MP) to yeastlike phase (YP) transition of the dimorphic fungal pathogen Sporothrix schenckii are shown in electron micrographs of ultrathin sections. Morphological transformation at the ultrastructural level was observed to occur by direct formation of budlike structures at the tips and along the hyphae and by oidial cell formation. Direct budding of yeast from conidiospores was not observed. Early transitional forms arising by direct blastic action from the MP possessed conspicuous electron-dense microfibrillar material at the outer limits of the cell wall. The electron density of this microfibrillar material was enhanced by staining with acidified dialyzed iron. It is believed that this extracellular material may be composed in part of an acid mucosubstance. No acid phosphatase activity was associated with this microfibrillar material. This substance was found to be a characteristic of the outer limits of the cell wall of the YP of S. schenckii. Oidial YP cell formation occurred later during the transition. The cell wall of the developing oidial YP transitional form arose from an inner layer of the converting hyphae. No consupicuous alterations of the cytoplasmic content of the parent MP cell was observed during MP-to-YP transition. It is suggested that the MP-to-YP transition of S. schenckii may be regulated by at least two mechanisms involving alterations of the biochemical and/or biophysical nature of the cell wall of the MP cell in response to the conversional stimuli.


The possible role of palmitoyl-CoA in the regulation of the adenine nucleotides transport in mitochondria under different metabolic states. I. Comparison of liver mitochondria from starved and fed rats. It has been shown that KM values for ADP when rat liver mitochondria oxidized succinate were strictly dependent on the values of the respiratory control ratios. The Ki values for palmitoyl-CoA inhibition of the ADP-stimulated succinate oxidation and the inhibition of the uncoupler-stimulated ATPase activity were equal to 0.5 muM. Mitochondria from livers of starved rats showed 30% inhibition of the state 3 respiratory rate (compared to the uncoupled respiratory rate) which was abolished by addition of carnitine. It was supposed that this inhibition was due to the influence of acyl-CoAs bound to the inner mitochondrial membrane on the adeninenucleotide translocase. Mitochondria from livers of fed rats showed a strong inhibition of succinate oxidation both in state 4 and state 3, although the rate of uncoupled respiration was normal. It was assumed that in this case the changes in mitochondrial behaviour was caused by the decrease in the concentration of ADP and ATP in the matrix space of mitochondria.