Epidemiology of hyperuricemia in an ambulatory elderly population.

The relationship of serum uric acid concentration to age and sex was assessed in 1,701 participants in a geriatric multiphasic screening clinic. Included were 1,067 women and 634 men. The mean uric acid level for men was 6.35 mg/100 ml compared to 5.44 mg/100 ml for women. No statistically significant increases in serum uric acid between age groups were apparent, except for women over 84 years of age. The mean serum urate concentrations found in this geriatric study are higher than those reported for other population groups, suggesting a higher normal value for the elderly.


Importance of a regular source of medical care among the elderly.

In recent years health services researchers have become increasingly aware of the importance of a regular doctor to whom the patient usually goes when sick or in need of advice about health, as a means of providing a point of entry to needed medical care. For example, persons who have a regular source of care seem more likely to receive an annual physical examination, to experience continuity of care when needed, and to be satisfied with the care received. Yet, despite the importance of an established tie to a regular treatment source, evidence is scanty with respect to the aged. To address the issue, this study employs data from a national probability sample of 11,619 persons, of whom 1,506 were aged 65 or older. The findings indicate that several factors (e.g., sex, race, education, and community size) which traditionally have served as important indicators of the use of health services fail to emerge as strong determinants regarding the regular source of care among the aged. The consequences of these findings and implications for future research are discussed.


Avoiding diagnostic errors in admitting patients to a nursing home.

Sometimes the members of the medical profession have been castigated for unpreparedness and shortcomings in the management of nursing home populations because their pre-admission diagnoses often have not agreed with the post-admission evaluations. In contrast, the authors' survey of a local situation revealed a high degree of diagnostic accuracy in admissions to a nursing home. Discussed are appropriate steps that nursing-home admission evaluators can take to ensure the avoidance of errors in the entrance diagnosis.


Some observations on not telling the truth.

Lying, in the analytic situation of essentially neurotic patients, is proposed to represent a re-enactment within the analysis and transference neurosis of a particular aspect of the oedipal conflict. These lies permit the partial recovery of old memories and perceptions; the emergence of certain unconscious wishes and fantasies, while simultaneously continuing to protect those wishes by a variety of mechanisms which are best conceptualized as "screen functions"; and the expression of resentment over being lied to by one or both of the oedipal objects. Connections are also made between the oedipally determined lie and an infantile primal-scene trauma.


Child abuse and deprivation: soul murder.

Soul murder involves the deliberate traumatization or deprivation by an authority (parent) of his charge (child). The victim is robbed of his identity and of the ability to maintain authentic feelings. Soul murder remains effective if the capacity to think and to know has been sufficiently interfered with--by way of brainwashing. Questions are raised about pathogenesis. Some suggestions are made about treatment of those whose pathogenic fantasy life has been so influenced by traumatic reality. The need to identify with and to maintain the illusion of a good parent enforces the difficult resistance of denial (brainwashing becomes self-enforced). Paradoxically, in order to survive and adjust, some of these people so traumatized as children develop unusual strengths and gifts.


Transference in the rat man case: the transference in transition.

This study has concentrated on the use of the transference concept in the Rat Man case as a method for revealing repressed memories and to gain conviction about these repressed memories. In this treatment-transference model, transference interpretations focusing on the analyst are not germane, since the transferences are merely the vehicle by which memories are uncovered. Thus, Freud's transference work in 1907 revealed a transitional phase of his clinical activity soon to be supplanted by later insights into the curative aspects of transference analysis.


Unconscious fantasy and theories of cure.

It has long been known that patients' wishes for cure by analysis give expression to unconscious wishes for instinctual gratification which originate in childhood mental life. Patients sometimes develop theories about how they believe analysis attains its ends; these are likely to affect the way they behave in the analytic situation, and analysis of them may therefore constitute an important contribution to progress in the analysis. Two illustrative cases are presented; in both, the unconscious determinants of the patients' theories turn out to be infantile sexual fantasies connected with their uncon scious wishes for instinctual satisfaction. It is further suggested that analogous unconscious fantasies also influence the theories of analysts and other therapists about how analysis works. In some instances these factors, while not affecting theory formation explicitly, may, without being recognized as doing so, contribute to decisions regarding the modification of analytic technique. Awareness of that possibility may aid analysts in assessing the indications for such proposed modifications.


Teratologic assessment of maleic hydrazide and daminozide, and formulations of ethoxyquin, thiabendazole and naled in rats.

Teratogenicity studies were conducted in rats treated orally from days 6-15 of gestation with single daily doses of 400-1600 mg/kg of maleic hydrazide, 300-1000 mg/kg daminozide, 125-500 mg/kg ethoxyquin or thiabendazole, or 25-100 mg/kg naled. Dams were killed on the 22nd day of gestation, and fetuses were evaluated by routine teratologic methods. No adverse effect was related to any treatment other than an increased incidence of anomalous fetuses at the highest dose (500 mg/kg) of thiabendazole.


Divergent toxicity of parathion in two freshwater invertebrates, orconectes rusticus and viviparus malleatus.

The toxicity of parathion and its metabolite paraoxon was examined in the crayfish Orconectes rusticus and the snail Viviparus malleatus and attempts were made to relate this to parathion uptake and metabolism. The crayfish was found to be very susceptible with all animals tested dying at 0.1 ppb of parathion in 48 hours. The snail was resistant to exposure to 1000 ppm. The absence of lethality even to injected (50 mg/kg) parathion and paraoxon suggests an innate lack of susceptibility. Metabolism of the compounds could not be detected in vitro or in vivo in either species.


A study on the lethal toxicity of aminocarb to freshwater crayfish and its in vivo metabolism.

Adult crayfish (Orconetes limosus) were exposed to 0.1, 1.0, 5.0, 10.0, 25.0, 30.0, 40.0, 50.0 and 60.0 ppm of aminocarb in water at 15 degrees C under laboratory conditions for 144 h. No apparent behavioral changes were observed in crayfish exposed to 0.1, 1.0 and 5.0 ppm of aminocarb during the experiment. Symptoms of acute toxicity were apparent at concentrations greater than or equal to 10 ppm, and mortality occurred at and above 25 ppm. The LC50, 96 h, to adult crayfish was about 33 ppm. The parent compound and its metabolites, MA (4-methylamino-m-tolyl N-methylcarbamate) and AM (4-amino-M-tolyl N-methylcarbamate), were detected in crayfish 96 h after exposure to various concentrations of aminocarb. The primary metabolite detected was MA which accounted for 75% to 95% of the body residue. The highest total residue (Aminocarb + MA + AM) was 40 ppm detected in crayfish exposed to 60 ppm of aminocarb for 96 h.


Effect of chronic exposure to cadmium on hepatic drug metabolism.

In an attempt to examine the chronic effect of low levels of cadmium on hepatic drug-metabolizing enzyme system, an experiment was carried out in which growing male rats were given 0, 5, 10, and 20 ppm of cadmium in drinking water for a period of 8 weeks. An ip administration of a hypnotic dose of pentobarbital to the cadmium-treated and the control rats 24 hr following the termination of the experiment exhibited that there was no significant difference in the drug metabolism in control and any of the treated groups. Next, liver microsomes were isolated from animals in all groups to study their ability to metabolize drugs in vitro. The results indicated that the activity of benzphetamine N-demethylase and aniline hydroxylase, and the concentration of microsomal cytochrome P-450 were almost identical in the control and treated groups. On the other hand, a single ip dose of cadmium (2 mg/kg) caused significant decrease in the vivo and in vitro microsomal drug metabolism. These results suggest that although a single ip dose of cadmium (2 mg/kg) causes significant inhibition of drug metabolism, chronic exposure to cadmium up to 20 ppm in drinking water over a period of 8 weeks is unlikely to affect hepatic drug metabolism.


Influence of pesticides on acetylene reduction and growth of microorganisms in an organic soil.

The effects of 32 pesticides at two concentrations on acetylene reduction (non-symbiotic nitrogen fixation), nitrogen fixers, bacteria and fungi in an organic soil were assessed. None of the pesticide treatments suppressed C2H2 reduction as compared to controls. No significant inhibition of the population of non-symbiotic nitrogen fixers occurred. However, stimulatory effects were observed with treatments of fensulfothion, fonofos, oxamyl, DDR, TeloneR and Telone CR. Bacterial and fungal populations showed temporary declines but all recovered within 7 days to levels similar to or higher than those in the controls.


The aqueous solubility of twenty-seven insecticides and related compounds.

The aqueous solubilities of 27 insecticides and related compounds were determined. Diazinon, fensulfothion and paraoxon had solubilities greater than reported in the literature. The presence of impurities and/or additional components in the mixture altered the measured solubility values. Addition of acetone in amounts up to 1% (volume) produced increases in parathion solubility up to 11%. The pH values of the equilibrated solubility systems were, in most instances, acidic and, in several instances, were in the pH 3-4 range.


Carbofuran residues in organic soils in Southwestern Ontario, 1977.

Organic soils from 22 farms with a history of carbofuran use for soil insect control were sampled in November, 1977. Analysis for carbofuran was by electron capture gas chromatography of the heptaflurobutyric derivative. Nineteen of the 22 soils contained detectable (sensitivity 0.02 ppm) carbofuran residues. However only 8 of the soils contained greater than 0.5 ppm total carbofuran. The highest total carbofuran residue was 1.5 ppm, of which 0.31 ppm was 3-ketocarbofuran. In other soil samples 3-ketocarbofuran comprised 7-50% of the total carbofuran residue. No 3-hydroxycarbofuran was detected. The order of persistence of granular application of 3 insecticides as seed-furrow treatments was ethion greater than fonofos greater than carbofuran.


Loss of synapses in the cerebellar cortex of the senescent rat.

Numbers of synapses were compared in the cerebellar cortex of adult (12 months of age) and senescent (25 months of age) male rats of the Fisher-344 strain. The total number of axodendritic synapses was found to be 24% lower in the senescent rats as compared with adults. A differential analysis of synapses involving dendritic shafts and spines showed no significant change in numbers of synapses involving shafts, but a highly sigificant 33% decrease in numbers involving spines in senescent rats. These data suggest that the selective age-related loss of synapses involving dendritic spines (but not shafts) in the cerebellar cortex results from the impairment with advanced age of specific afferent neurons and/or a selective age-related vulnerability of dendritic spines.


Quantitative morphological analysis of age-related changes in flight muscle of Musca domestica L.

Electron micrographs of dorsal indirect flight muscle of 37 (adult) and 68 (old) day old female houseflies were analyzed by morphometric techniques. In the muscle of the old female housefly there is no significant atrophy and the fine structure is well preserved. There is a small loss in both the myofibril and mitochondrial volumes, an increase in average myofibril size, and a 28% decrease in myofibril number. The thin/thick filament ratio and sarcomere structure are not altered. There is little decrease of particulate glycogen. The fractional volume occupied by SR at the dyads is increased. The most marked change is in the density of mitochondrial cristae which declines by 55% in old muscle and could account for the functional loss. Cristal anomalies and SR changes indicate that the membranes may be the specific target of deleterious aging effects.


Age-related differences in salt taste acuity.

Salt taste detection thresholds were measured in 76 adults aged 23 to 92 years using a forced-choice staircase procedure, with distilled water rinses between all stimuli. Although an age-related decrease in salt taste acuity was observed, the decrement was smaller than previously reported in other studies. Past reports have probably exaggerated taste deficits in the elderly, because adequate rinse procedures and controls for subject response biases have not been incorporated in threshold measures.


Stress and adaptation in later life.

The effects of five major life events, and of three types of resources, on the physical and social-psychological adaptation of 375 participants in a longitudinal study were examined. As expected, medical events had the most impact on physical adaptation, but they had surprisingly little impact on social-psychological adaptation. Retirement had the most negative social-psychological effects, but had little effect on physical adaptation. The other three events had even less effects, although multiple events tended to cumulate in impact. Better physical resources helped only physical adaptation, and better psychological and social resources mainly helped satisfaction. It appears that most of these potential stressors have less serious long-term outcomes than the crisis orientation would suggest.


Doctor patient relationships and the older patient.

The authority of physicians in patient encounters is posited by the sicrole and the professional-client models in sociology. Yet, public challenges to physician authority are currently occurring, as shown by self care movements, and by some empirical data. Whether the elderly share in such challenges is, however, an unresolved question. Will longer experience with health services produce skepticism about medicine's claims, or conversely will the aged's probable more authoritarina upbringing and concern about failing health produce acceptance of physician authority? Data from an interview survey of 640 randomly sampled persons in a midwestern state, reveal that the 153 respondents aged 60 and over are more likely to accept physician authority, both in terms of attitudes and behaviors, than are the younger groups. Moreover, the findings show that the level of older persons' challenging attitudes is affected by their health knowledge and general views on authority, while their challenging behaviors are influenced by belief in right to medical information and an experience factor. Implications of the findings for appropriate modes of doctor-patient relationships for the elderly are discussed.


The never-married in later life.

Sources or consequences of singlehood are analyzed for the 162 never-married persons aged 50 and over from the six years of the NORC General Social Surveys. Highly-educated older women are most likely to remain single, but family background was not a predictor. Although the never-married find life more exciting than other marital statuses among younger (25 to 49) respondents, this reverses in later life. The never-married are also less happy than the married, and only slightly happier than the widowed and divorced. The characteristic that best explains their relative unhappiness is greater dissatisfaction with family life. The lower well-being of the never-married is attributable either to changes accompanying aging which lessen the viability of single life styles, or to less support of single living among current older cohorts.


Life changes and perceptions of life and death among older men and women.

Although social scientists have suggested that feelings about life and death may be related, for the most part, theories of social gerontology have developed independently of conceptions of death and finitude. This study examines whether life changes are associated with concurrent life and death attitude types among older men and women. Data were analyzed from structured interviews conducted with 214 men and 354 women during the second phase of a longitudinal study of the aged in small towns. Individuals were identified as positivists, negativists, activists, and passivists. Controlling for sex and income, changes in four life areas were examined in relation to these perceptions of life and death. Some of the patterns varied by sex, suggesting that the same changes have different implications for the well-being of men and women in late life. In general, those who had experienced discontinuity were more frequently negativists or passivists, while continuity tended to characterize positivists and activists.


Pectoralis major transplantation to restore elbow flexion to the paralytic limb.

In four patients, two with brachial plexus palsy and two with arthrogryposis, the entire pectoralis major muscle was transplanted to act as an elbow flexor, rotating it on its neurovascular pedicles and attaching the tendon to the acromion and the muscle origin to the tuberosity of the radius. Excellent motion and power was restored in three patients. One procedure failed due to fibrofatty degeneration of the transplanted muscle.


A cause of painful clicking wrist: a case report.

In a patient with symptomatic unilateral clicking of the wrist, a partial tear of the scapholunate ligament with subsequent scarring of the proximal third of the dorsal portion of the ligament had occurred. At operation the dorsomedial edge of the proximal pole of the scaphoid had snapped over the dorsal edge of the lunate as the palmar-flexed wrist was being returned to a neutral position. Release of the scarred portion of the scapholunate ligament was associated with widening of the scapholunate joint space to normal dimensions and cessation of the clicking phenomenon. Fourteen months after operation there was complete relief of symptoms, a full range of wrist motion and no radiographic evidence of widening of the scapholunate space, ligamentous instability, or rotatory subluxation of the scaphoid.


Rheumatoid synovitis of the volar compartment of the wrist joint: its radiological signs and its contribution to wrist and hand deformity.

The radiological signs that suggest involvement of the volar compartments of the wrist include grooving of the scaphoid, pseudocysts of the distal radius, and scaphoid-lunate dissociation. These indicate involvement of the radiocarpal ligamentous support. This leads to loss of stability of the scaphoid, which then goes into a volar-flexed position, contributing to shortening the radial carpal height and favoring rotation of the carpus into supination, radial deviation of the metacarpals, and ulnar drift of the fingers.


Hamate erosion: an unusual result of ulnar artery constriction.

An erosive lesion of the hamate hook in a patient with symptoms of arterial insufficiency was the result of a pseudoaneurysmal formation of the ulnar artery proximal to an obstruction by cicatrical bands.


Carpal tunnel release under local anesthesia: evaluation of the outpatient procedure.

In 100 consecutive cases of carpal tunnel release done under local anesthesia in an outpatient ambulatory care operating room, 93 had satisfactory results at 6 months without any complications. Two patients developed a neuroma of the palmar cutaneous branch of the median nerve, and five showed early signs of reflex sympathetic dystrophy. These complications are discussed, as well as the prevention of other complications of this procedure.


Experimental studies in chickens on the initial nutrition of tendon grafts.

A study of nutrition of various tendon graft preparations in adult chickens (up to 2 weeks after grafting), using tritiated proline and a trichloracetic acid extraction technique which separated the free and metabolized amino acid fractions, suggests that diffusion of nutrients is an important process in the initial nutrition of tendon grafts, that tendon grafts are metabolically active and viable structures, that adhesions which are frequently associated with tendon grafts do not appear to be essential to the nutrition of grafts, and that tendon grafts within fibrous pseudosheaths are nourished as effectively as grafts within synovial sheaths.


Fibular autografts for distal defects of the radius.

A protocol for osteoarticular grafting was established to avoid fracture, nonunion, and loss of motion when replacing the distal radius. Proximal fibular autografts were used and stabilized proximally by compression plating and, at the wrist, by ligamentous reconstruction. Postoperative splinting and therapy were coordinated with graft healing, which was monitored by bone scans and roentgenograms. Graft incorporation in three patients appeared to be well-established within 1 year, but functional use of the extremity and return to duty were achieved much earlier.


Scintillation fluid shortens exposure times in autoradiography.

Premixed, commercially available scintillation fluids were used to reduce exposure time of tritium (3H) and iodine-125 (125I)-labeled whole cells, and of 3H-labeled Giemsa-banded chromosome preparations. Emulsion-coated slides were dipped into scintillator for no longer than 2 min and exposed in the dark at 4 degrees C. Maximal values for percentage of human diploid cell (WI-38) nuclei labeled with 3H-thymidine of moderate specific activity were obtained in 12 hr. Without scintillator the exposure time was 4 days. Exposure time for cells labeled with 125I-serum was reduced from over 90 days to 14 days. The shortened exposure time for banded chromosomes permitted successful prestaining with Giemsa, a sequence that is not possible without scintillator.


Adenylate cyclase activity in the parotid gland of the mouse after isoproterenol stimulation.

Previous studies have described a decrease in the activity of adenylate cyclase in the parotid gland of isoproterenol-treated rats. In the present studies, a similar decrease was observed in mice treated with isoproterenol. Studies on the subcellular distribution of adenylate cyclase after isoproterenol stimulation of the parotid gland showed that enzyme activity was increased in the lysosomal fraction and decreased in the cellular membrane fractions. Cytochemical studies on the localization of adenylate cyclase in stimulated gland showed an increase in vesicles which contained enzyme activity and a decrease in activity at the luminal and plasma membranes. It is suggested, based on the present findings and results reported by other investigators, that after isoproterenol stimulation of the parotid gland, adenylate cyclase (along with excess membrane) is degraded by lysosomes. If this suggestion is true, then the observed decrease in adenylate cyclase would have a molecular explanation.


Localization of adenylate cyclase in unfixed sections of cardiac muscle.

Previous investigators have shown that prefixation and lead staining completely inhibit the activity of adenylate cyclase. Lead has also been shown to stimulate the nonenzymatic hydrolysis of AMP-PMP (the substrate for adenylate cyclase) after 30 min incubation. The present studies were performed to determine if the omission of prefixation would provide a better method for localizing adenylate cyclase in cardiac muscle. These studies were also performed to determine the effect of short incubation on the lead-induced nonenzymatic hydrolysis of AMP-PNP. In prefixed sections the reaction product was diffusely localized over the section. However, in unfixed sections the reaction product appeared only on the sarcolemma and sarcotubule system. Results are presented showing that short incubation (i.e., 5 min) prevents the nonenzymatic hydrolysis of AMP-PMP by lead. In biochemical studies lead (10(-3) M) was shown to completely inhibit the activity of this enzyme. However, in the presence of 4 micrograms phosphatidylinositol, lead inhibition of this enzyme was reduced to 50% of the control value. Based on this observation, it is suggested that approximately 50% of adenylate cyclase is present in sections of cardiac muscle exposed to 2 x 10(-3) M lead, which is presumably enough activity for demonstration of adenylate cyclase activity.


Lead aspartate, an en bloc contrast stain particularly useful for ultrastructural enzymology.

Lead aspartate is a new en bloc stain for electron microscopy. Its predictable staining depends on chelation that results from the interaction of the two stain components, lead nitrate and aspartic acid, which must be present in a specific ratio. Lead aspartate stain is 0.02 M in lead nitrate and 0.03 M in aspartic acid, adjusted to pH 5.5. Cells or tissues are stained at 60 degrees C for 30 to 60 min. Cells stained en bloc with lead aspartate closely resemble cells stained on grids by lead citrate, except that the former seldom have contamination. En bloc staining with lead aspartate bypasses the grid-staining step so that samples can be viewed and photographed immediately after they are thin-sectioned. The lower pH of the lead aspartate solution allows counterstaining of enzyme reaction products that dissolve in the highly alkaline lead citrate stain. Lead aspartate en bloc staining to enhance contrast should especially benefit studies of ultrastructure requiring a clean and predictably lead stain.


Horseradish peroxidase as a tracer for capillary permeability studies.

The permeability of capillaries was investigated utilizing an in vivo injection of horseradish peroxidase (HRP) and an in situ perfusion of a balanced salt solution containing HRP and lanthanum chloride. In the continuous capillaries of heart and muscle, HRP diffuses mainly through intercellular junctions, while in testicular capillaries, the transport is via micropinocytotic vesicles. The diffusion and micropinocytotic transport of HRP was demonstrated in both directions, i.e. from the capillary lumen to the interstitium and vice versa. Lanthanum can be used as a bidirectional inhibitor of micropinocytosis. The transport of HRP is then almost completely hindered in testicular capillaries. In heart muscle, the effect on HRP transport is not significant, due to second transport pathway, i.e. intercellular cleft passage.


Reproductive tract peroxidases as endproducts of estrogen-specific gene expression.

Estrogen is an essential requirement for the postpubertal trophic development and maintenance of the differentiated state of the oviduct, uterus, cervix, vagina and mammary glands of mammals. Estrogen, apparently functioning through its specific cytoplasmic receptor protein via a multistep interaction pathway induces gene expression of specific biochemical events leading to growth and differentiation of target tissues (Jensen et al., Proc Natl Acad Sci, 59:632, 1968; Gorski et al., Recent Prog Horm Res 24:45, 1968). One biochemical expression of the estrogen gene is the synthesis of specific mRNA transcripts for certain specific marker proteins, including ovalbumin, lysozyme and ovomucoid in the chick oviduct (O'Malley and McGuire, Proc Natl Acad Sci 60:1527, 1968; Palmiter and Schimke, J Biol Chem 248:1502, 1973), tubulin in the mammalian oviduct (Brenner and Anderson, Handbook of Physiology 7(2):123, 1973; Brenner et al., Endocrinology 95:1094, 1974) and peroxidase (EC 1,11.1.7) in the rodent uterus (Brockelmann and Fawcett, Biol Reprod 1:59, 1969; Churg and Anderson, J Cell Biol 62:449, 1974; Anderson et al., J Cell Biol 64:668, 1975).


Pituitary adenomas. Clinico-morphologic correlations.

Immunohistochemistry and electron microscopy allow new insights into the biology and natural history of the proliferative processes of the hypophysis. Hyperplasia can be diffuse or focal. Its function and/or growth are controlled by hypothalamic factors. Hyperplasia may precede the formation of an adenoma. Function and/or growth of adenomas is partially or completely independent of hypothalamic regulation. Adenomas are divided according to their functional activity into an endocrine active and inactive group. A further subdivision is made according to the secreted hormone. Inactive adenomas may have lost the ability to produce hormones, may secrete hormones at a very low rate, or may secrete abnormal substances.


The diffuse neuroendocrine system. Studies of this newly discovered controlling system in health and disease.

Numerous peptides (neuropeptides) have been recently found to be present in both the nervous and endocrine systems composing what is now known as the diffuse neuroendocrine system. Two immunological methods, radioimmunoassay and immunocytochemistry, have been used here in combination to study their distribution and cellular localization. A number of these neuropeptides have recently been found to be abnormal in disease state, thus providing further information as to their role in normal and pathological conditions.


Membrane differentiation of developing hemic cells of the bone marrow demonstrated by changes in concanavalin A surface labeling.

The concanavalin A-gold labeled horseradish peroxidase (Con A-HRP-G) method has been employed in the ultrastructural localization of Con A surface receptor sites on glutaraldehyde-fixed normal human and guinea pig bone marrow cells. The number of gold particles per micron of cell surface was counted and data subjected to statistical analysis. All cells of the bone marrow exhibited Con A binding; however, the extent of surface labeling was dependent both on cell type and stage of differentiation. Distinctive modifications in mean surface density correlated with specific periods during the maturation of the erythrocytic, neutrophilic, eosinophilic and monocytic cell series. In several instances, the differentiative changes in surface Con A labeling proved to be species dependent. These observations are discussed in relationship to methodology and to potential changes in number and/or spatial arrangement of Con A receptor sites, primarily attributable to mannosyl and/or glucosyl residues associated with membrane glycoproteins and/or glycolipids of developing neutrophilic and erythrocytic cells.


A fluorescent assay of proteinases in cultured mammalian cells.

We have demonstrated proteinase activity in unfixed cells grown on tissue culture plates with a technique using 5-nitrosalicylaldehyde and peptide derivatives of 4-methoxy-2-naphthylamine. The 4-methoxy-2-naphthylamine liberated by proteinase activity reacts with 5-nitrosalicylaldehyde to form a fluorescent product. The substrates CBZ-alanyl-arginyl-arginyl-4-methoxy-2-naphthylamine and lysyl-alanyl-4-methoxy-2-naphthylamine, were used for the direct visual detection of two arylamidase activities in BALB/c 3T3 and C3H 10T 1/2 cells. With low magnification these enzyme activities can be detected in single clones; with higher magnification the fluorescent product can be seen within the cytoplasm of single cells.


Cytochemical localization and biochemical characterization of dipeptidyl aminopeptidase II in macrophages and mast cells.

Dipeptidyl aminopeptidase II (DAP II) was demonstrated cytochemically at light and electron microscope levels in rat macrophages and mast cells using Lys-Ala-4-methoxy-2-naphthylamide as a specific substrate. The enzyme which was found to be lysosomal in both cell types, was analyzed biochemically in extracts by measuring fluorometrically the liberated naphthylamine, and was visualized in sections microscopically using azo-coupling methods. DAP II was further characterized by isoelectric focusing techniques. Macrophage DAP II was found to be typical of that found in other rat tissues in terms of its structural latency, substrate specificity, inhibitor sensitivities, and pH activator requirements. Addition DAP II isozymes, not previously recognized, were observed.


A study of acid phosphatase and dipeptidyl aminopeptidase II in monodispersed anterior pituitary cells using flow cytometry and electron microscopy.

A brief historical review of cytoenzymology is presented from the time of introduction into electron microscopy to the present, where the direction for quantification of an enzyme in single cells appears most promising by fluorescent staining. First attempts are reported to quantitate acid phosphatase (AcPase) and dipeptidyl aminopeptidase II (DAP-II) in monodispersed anterior pituitary cells from lactating and postlactating rats by flow cytometry, fluorescent, and electron microscopy. 3-Hydroxy-flavone is introduced as a new fluorescent cytochemical stain for AcPase, useful in flow cytometry but of only limited use in fluorescent microscopy. Histograms for AcPase indicate a single peak of cells staining more intensely in cell preparations from postlactating over lactating animals. Histograms for DAP-II staining indicate two distinct populations of cells present in the lactating and only one in the postlactating rat anterior pituitary gland. The application of dual laser staining indicates that not all cells stain for both enzymes. Electron microscopy shows the subcellular localization of DAP-II to be limited to lytic bodies and in mammotrophic cells to some secretion granules.


Freeze-fracture autoradiography. Progress towards a routine technique.

Freeze-fracture autoradiography was introduced in 1976 as a new technique for the autoradiography of diffusible compounds at the electron microscope level. With the original approach coating of the frozen replicated specimens was performed in a cryostat at atmospheric pressure. Ice contamination of the specimen surface acting as an outstanding source of artifacts was thereby not excluded. With the use of a specially designed coating device and volatile spreading substances it was made possible to coat the frozen replicated specimens in the maintained vacuum of the freeze-fracture plant. In this complicated technique we have recently extended the freeze-fracture autoradiography to labeled frozen-dried "half" membranes of red blood cells.


Freezing of tissue-limits for the autoradiographic localization of diffusible substances.

Frozen thin sections and sections from freeze-dried and embedded tissue are used for the autoradiographic localization of diffusible substances at the electron microscope level. The presence of ice crystals in such sections may limit the autoradiographic resolution. Ice crystals are formed during freezing and may grow during subsequent processing of tissue. The contribution of ice crystal growth to the final image was estimated by measuring the distribution of the ice crystal sizes in freeze-etch replicas and in sections from freeze-dried and embedded tissues. A surface layer (10-15 mu) without visible ice crystals was present in both preparations. Beneath this surface layer the diameter of ice crystals increased towards the interior with the same relationship between crystal size and distance from the surface in the freeze-etch preparation as in the freeze-dry preparation. Ice crystal growth occurring during a much longer time during freeze-drying compared to freeze-etching does not significantly contribute to the final image in the electron microscope. The formation of ice crystals during freezing determines to a large extent the image (and therefore the autoradiographic resolution) of freeze-dry preparations and this probably holds also for thin cryosections of which examples are given.


High resolution scintiautoradiography.

An electron microscopic scintiautoradiographic method for studying histological sections is reported. An increase in the silver grain yield is attained by using a scintillator in embedding medium. The amplification effect for 3H-labeling reaches 3-fold and for 14C reaches a 15-fold. No lessening of autoradiographic resolution was observed. On the basis of these preliminary results only qualitative evaluation of data is discussed.


Photometric methods in quantitation of light microscope radioautography.

Photometric evaluation of autoradiographic grain densities can be performed according to various optical principles. In all instances the amount of light recorded should be a measure of the radioactive substance amount in the specimens. It is shown that grain densities suitable for light microscopic autoradiography are indeed linearly related to radiation exposure. Dependent on the photometric system used, there is a larger or smaller section of linear correlation of the photometric response with grain density. The advantages of incident light bright-field illumination for silver grain counting are discussed. Quantitation of substance amounts in autoradiographs depends on the use of radioactive standard sources. Two different approaches of quantitation are discussed. In 14C-autoradiography which can be applied for cell-kinetic studies, standard plates are used consisting of 14C-polymethylmethacrylate. Allowance has to be made for the different condition of radiation geometry of the cells and the standards. In 125I-autoradiography, 125I-labeled red cells are used as standards. This technique allows for quantitating the number of antibodies bound to individual cell surfaces.


Quantitation and resolution in electron microscope radioautography.

Electron microscopy can resolve structures with accuracy of the order of 0.2-0.3 nm, but the radioautographic technique is able to locate radioactive label with much less certainty, at best 50 nm. This difference in resolution is what creates the problem of quantitation in electron microscope radioautographs.


Morphometry of the chinchilla organ of Corti and stria vascularis.

This research describes a procedure for a morphometric analysis of the organ of Corti and stria vascularis in the chinchilla. In nine normal cochleae the length of the basilar membrane and the stria vascularis measured 18.47 and 25.22 mm, respectively. An average of 1910 inner and 7501 outer hair cells were present while an average of 15 inner and 90 outer hair cells were absent. In all cochleae examined there were always some missing hair cells in varying numbers even though the animals had no known ototoxic exposure. Stria area, width and thickness increased from the cochlear apex toward the base. Consistency of changes in stria dimensions among animals was enhanced by expressing position in terms of percentage stria length rather than distance as such. Total stria volume was estimated at 0.15 microliter.


Applications of stereology to the mechanics of soft tissues.

Quantification of microstructure in conjunction with mechanical testing and modeling may provide new insight into problems of soft tissue mechanics with potential clinical significance. Some general observations concerning this as well as brief discussions of specif examples are presented.


Morphometry of the effect of increased experience and training on synaptic density in area CA3 of the rat hippocampus.

A morphometric analysis of the effect of differential experiences on synapses was performed in area CA3 of the rat hippocampus. Two methods of quantitation that used a superimposed lattice grid were employed. In the first method squares overlying synapses were measured as a proportion of the total number of squares scanned; in the second method intersections of lattice lines with synapses were counted as a proportion of the total length of lines. Both measures showed that rats which were housed in an enriched environment and received 10 weeks of training had a significant increase in total area of synaptic contacts compared to both nonenriched controls and a croup with increased motor activity.


Numerical density of convex, nonbranching organelles in anisotropically oriented cells. Cilia in tangential chondrocytes.

A stereological expression relating the mean number of organelle profiles per cell profile, Np, to the mean number of organelles per cell, Nc, is first derived and then tested for cilia in tangenital articular chondrocytes where Nc is known to be 1.0. Nc derived stereologically for this organelle-cell model is 1.19. This close approximation of the estimate to a known value provides empirical evidence of the usefulness of the derived stereological expression.


Influenza vaccination with live-attenuated and inactivated virus-vaccines during an outbreak of disease.

Immunization procedures with live attenuated and inactivated vaccines were carried out on a group of young recruits at the beginning of an outbreak of infection due to an A/Victoria/3/75-related virus strain, which occurred in February 1977 in a military camp. A retrospective investigation on protection from clinical influenza was then performed in order to investigate whether immunization with live virus vaccines, administered at the beginning of an epidemic, could provide early protection from the disease. In the course of the two weeks following vaccination, laboratory-confirmed clinical influenza cases occurred in 4 subjects among the 110 volunteers of the control group which received placebo, and in 8, 7 and 4 subjects respectively of the 3 groups of about 125 individuals, each of which received one of the following vaccine preparations: (a), live attenuated A/Victoria/3/75 influenza virus oral vaccine, grown on chick embryo kidney culture; (b), live attenuated nasal vaccine, a recombinant of A/Puerto Rico/8/34 with A/Victoria/3/75 virus; and (c), inactivated A/Victoria/3/75 virus intramuscular vaccine. These data do not support the hypothesis that, during an epidemic of infection, early protection from clinical influenza can be achieved through immunization with live attenuated or inactivated influenza virus vaccines, in spite of the high immunizing capability of the vaccine preparations.


Seroepidemiology of rubella infection in Chinese and Caucasians in Hong Kong.

The seroepidemiology of rubella infection in Hong Kong Chinese was examined by haemagglutination inhibition of normal sera and a comparison made where feasible with Caucasians living in Hong Kong. Taking reciprocal titre of 20 as a baseline, the incidence of maternally acquired antibody was 84% for Chinese and 90% for Caucasians. In babies up to 2 months this incidence was maintained in Caucasians but declined to 54% in Chinese. High titre antibody was more frequently detected in Caucasians generally, including women of child-bearing age. The detection of rubella-specific IgM in the Caucasian babies was suggestive of recent maternal infection. However, fewer Caucasian (20%) than Chinese (36%) women of child-bearing age (15-40 years) appeared unprotected against rubella. When all age groups were considered, 88% of Caucasians and 53% of Chinese were seropositive. The occurrence of a rubella outbreak during the study did not give rise to a significant increase in the incidence of seropositivity in Chinese 19-25 years, the only age group able to be evaluated in this manner. The ethnic differences in seroepidemiology are considered in the light of known HLA-1 and HLA-8 antigen distributions in Caucasian and Mongoloid people and the apparently low incidence of congenitally acquired rubella in Chinese.


HLA antigens and responses to rubella vaccination.

Attempts were made to correlate virus excretion, joint symptoms and antibody response with human leukocyte antigens (HLA) in seronegative adult women given attenuated rubella vaccine. No association was shown between HLA antigens of the A and B loci and excretion of either high or low titres of RA27/3 vaccine among 26 volunteers. However, virus excretion was influenced by such factors as the time of day at which specimens were collected and the method of virus isolation. Our study therefore failed to confirm the hypothesis that certain persons are good 'spreaders' of rubella virus and that this capacity is associated with HLA-A1 and B8. The study of joint symptoms following vaccination with Cendehill, HPV77.DE-5, RA27/3 or To-336 vaccines showed no association between such symptoms and HLA antigens. However, joint symptoms occurred within 7 days of the onset of menstruation in 33 of 47 (70%) vaccinees (P less than 0.01) and it is therefore suggested that hormonal factors must play a role. No association between HLA antigens and haemagglutination inhibition (HAI) antibody titres, 8 weeks after vaccination with RA27/3, was found amongst 34 volunteers.


Antibody studies in natural bovine cowpox.

Serological studies on cows recovered from natural cowpox indicated that Haemagglutinin-Inhibiting (HAI) antibody persisted for at least 27 weeks, and Virus Neutralizing (VN) antibody persisted for at least 98 weeks.


Comparison of selenite F, Muller-Kauffmann tetrathionate and Rappaport's medium for the isolation of salmonellas from sewage-polluted natural water using a pre-enrichment technique.

Three enrichment broths, selenite F, Muller-Kauffmann tetrathionate and Rappaport, were examined for their efficiency in salmonella isolation. The three media, prepared from single ingredients in the laboratory, were compared with their commercial equivalents. Laboratory-prepared media were more efficient for isolating salmonellas from sewage-polluted natural water samples. A pre-enrichment stage using buffered peptone water was employed throughout the investigation. The size of inoculum from the pre-enrichment medium was relevant to successful salmonella isolation. Inocula studied were 1 ml and one loopful (3 mm diameter loop). The smaller inoculum gave better results with Rappaport, the larger with selenite and tetrathionate. Using the optimal inocula, Rappaport was the most efficient enrichment broth of the three fluid media in this study.


Potential risk of salivary-mediated viral hepatitis type B transmission from oral exposure to fomites.

Twelve grade school and junior high school students had oral exposures to hepatitis B surface antigen (HBsAg) positive saliva via contact with contaminated musical instruments. The 12 exposed students and 18 students who served as age and sex matched controls were tested for the presence of HBsAg and antibody to the hepatitis surface antigen (anti-HBs) at 2 weeks, 8 weeks and 6 months after exposure. All students were negative for HBsAg and anti-HBs on all dates tested and reported no illness during that time suggestive of viral hepatitis. There was no evidence of viral hepatitis, type B transmission from the exposure. The students probably experienced the maximum possible oral exposure from direct or fomites contact, since there was no cleaning of the musical instruments between use by the students and teacher. Based on these results, the risk of transmission of viral hepatitis, type B from oral contact with fomites is unlikely.


Laboratory breeding of the European rabbit flea, Spilopsyllus cuniculi (Dale).

A method is described for the laboratory breeding of the rabbit flea in which the immature stages are reared at constant temperature and humidity. Eggs are obtained by confining fleas taken from a rabbit and her nest shortly after parturition with two of her nestlings in an incubator for 24 h. The eggs are transferred to an artificial diet medium on which the immature stages are reared. On average a female flea produces 50 eggs during the first six days post-partum. At 25 degrees C, 95% of eggs hatched at 79% RH and 98% at 84% RH. Most eggs hatched on the third day after laying and hatching was completed by the fourth day. Significantly more fleas of both sexes were obtained when larvae were reared at 25 degrees C on a medium containing powdered 41B rodent diet than on one containing terrier meal. Both diets also contained yeast and dried rabbit blood. There was no significant difference between the numbers of fleas obtained at 79% RH and 84% RH. Significantly more fleas were also obtained when larvae were reared at 27 degrees C, 84% RH, than at 25 degrees C. Female fleas emerged sooner than males at both 27 degrees C and 25 degrees C. Fleas from the laboratory culture were heavier than those from wild nests. Female fleas were heavier than male fleas in both cases.


Leukocyte chemotaxis under agarose: manipulations of serum and plasma before incorporation into agarose can influence cell movement.

In the migration under agarose assay for measurement of polymorphonuclear leukocyte chemotaxis and random migration, the agarose gel ordinarily contains 10% human serum or plasma. Influences of serum and plasma on cell movement using this assay were defined. Utilizing plasma as protein source allowed more random migration and chemotaxis than when serum was used. Heat inactivation of both serum and plasma decreased random migration and chemotaxis. If heating at 56 degrees C was prolonged beyond 15--30 min, leukocyte movement was augmented. Excessive concentrations of platelets or platelet products, or heparin in agarose, also augmented migration, but storage of plasma or sera at various temperatures for 24 h did not appreciably affect migration. Large amounts of hemoglobin in agarose decreased chemotaxis, but the effect of trace amounts of hemolysis was negligible. Handling plasma or sera in standard ways prior to use in the under agarose assay may make this assay more reproducible and sensitive.


Transfer plate radioassay using cell monolayers to detect anti-cell surface antibodies synthesized by lymphocyte hybridomas.

A solid-phase [125I]Protein A radioassay for anti-cell surface antibodies is described which employs target cell monolayers cultured on fenestrated polyvinyl chloride 96-well plates ('transfer plates'). The calibrated aperture in the bottom of each well is small enough to retain fluid contents by surface tension during monolayer growth, but also permits fluid to enter the wells when transfer plates are lowered into receptacles containing washing buffer or test sera. To assay for antibodies directed against target cell surface antigens, transfer plates bearing monolayers are inserted into microculture plates with corresponding 96-well geometry, thereby simultaneously sampling 96 wells. This assay allows rapid screening of hundreds of hybird cell colonies for production of antibodies with desired tissue specificity.


A general affinity chromatographic method for preparing monospecific antibody to mammalian serum haptoglobin.

A general affinity chromatographic method for preparation of monospecific antibody to serum haptoglobin of any species is described. Hemoglobin prepared from the species to be immunized is coupled to an organomercurical substituted agarose gel support (Affi-Gel 501). The immobilized hemoglobin binds haptoglobin with great affinity and allows removal of other serum proteins by extensive washing. The haptoglobin-hemoglobin complexes are then specifically eluted by buffers containing dithiothreitol or other thiols and are further purified by chromatography on concanavalin A-agarose and Sephacryl S-200 columns. The pure complexes are very effective immunogens. Potent monospecific antisera to rabbit and to human haptoglobin have been prepared. The potential usefulness of affinity chromatography support media with specifically cleavable ligand sites in studies of haptoglobin and in other biological studies is discussed.


Radioimmunoassay of carcinoembryonic antigen (CEA) without extraction and dialysis using solid-phase antibody.

A modified radioimmunoassay has been developed for plasma CEA levels. This procedure used immobilized, solid-state antibodies, does not require perchloric acid extraction of the plasma samples and purification of the extract by dialysis. The accuracy, sensitivity and precision are equal to those of the most widely used procedure and the laboratory effort is greatly reduced.


A sensitive assay for the detection cytotoxic antibodies to mammalian cell surface antigens.

A microcytotoxicity assay for the detection of complement-dependent serum toxicity against adherent cells has been modified to include post-labeling of the target cells with [3H]leucine or [3H]thymidine. The adaptation represents a significant improvement over the visual counting of residual cells. The technique is more objective and less tedious than the original procedure, while retaining the features of economy of cells and sera, reproducibility, and sensitivity. Examples are given which demonstrate the suitability of the modified assay for the analysis of human and mouse tumor antigens using autologous, allogeneic, and syngeneic sera.


Metabolic studies of C3 in man.

The results of metabolic studies with radioactively labelled C3 in 14 healthy individuals are presented. Plasma volume determined by labelled C3 was generally higher than when determined by labelled albumin, and different batches of C3 behaved differently during the early part of the experiments. These phenomena are probably caused by C3 inhomogenities which are difficult to detect by usual biochemical and immunological methods. We have, therefore, excluded the first 24 h of the experiments from the mathematical analysis. In this way, rather narrow limits for the metabolic parameters have been obtained.


Crossed radioimmunoelectrophoresis for the analysis of lymphocyte surface antigens.

A sensitive crossed radioimmunoelectrophoreses technique is described as a model tool for studying lymphocyte surface antigens, using Triton X-100 solubilized sufface radiolabeled human peripheral blood lymphocytes as the antigen source and experimental antiserum produced  by hyperimmunization of rabbits as the antibody source. The technique seems to have potential for qualitative as well as quantitative analyses of lymphocyte surface antigens.


C3b binding immune complexes and immunoconglutinins in human sera. Detection by enzyme-linked immunosorbent assay (ELISA).

An enzyme-linked immunosorbent assay (ELISA) designed to measure autoantibodies against C3b (immunoconglutinins: IK) also detects immune complexes (IC). Solid phase C3b, in addition to binding IK of IgG, IgM and IgA classes, bound aggregated human IgG, IgA and aggregated immunologically purified anti-tetanus toxid antibodies as well as model complexes of tetanus toxoid-human anti-toxoid. Significant C3b binding IgG, IgM and IgA activities were seen in the sera of 20 SLE patients but not in sera from healthy blood donors. Ultracentrifugation analysis of two SLE sera revealed C3b binding IgG and IgA activities in both light (7S) and heavy (greater than or equal to 11S) fractions. This indicates simultaneous presence of IK and IC in these sera. On the basis of the known relationship between IK and IC formation we suggest that the solid phase C3b ELISA may be of value in evaluating immune reactions in patients.


Anomalous reactions in the haemagglutination assay for anti-collagen antibodies: studies on patients with rheumatoid arthritis or chronic low back pain.

Sera from 23 patients with chronic low back pain, 20 rheumatoid patients and 16 normal controls were tested for antibodies to collagen types I, II and III, both native and denatured, by haemagglutination. Weak reactions against denatured collagen types I and II were found in 30-40% of the sera. Sera from individuals with rheumatoid arthritis or chronic low back pain behaved similarly, while only one normal serum showed any positive reaction. Reactions to denatured collagen type III and to native collagen of all 3 types were largely negative. Non-antibody serum components were thought to be responsible for these haemagglutination reactions since weakly positive reactions were abolished by cryoprecipitation and could not be confirmed by a solid-phase fluorimetric assay. Using the latter technique sera from 62 rheumatoid patients were screened for antibodies to type II collagen (native and denatured) and only one positive serum found. We conclude that haemagglutination is subject to false positive reactions and that the incidence of anticollagen antibodies in sera from patients with rheumatoid arthritis or chronic low back pain is low.


Quantitative estimation of IgA in rats: a comparison of two methods.

Two methods for estimating IgA in body fluids of rats were devised and tested for their accuracy and reliability. Specifically purified monomeric and polymeric IgA was prepared so that results obtained could be properly assessed. The two systems tried were prepared so that results obtained could be properly assessed. The two systems tried were rocket immunoelectrophoresis, carried out after reduction of samples with dithiothreitol and using monomeric IgA as standard, and a radioimmunoassay utilising a double antibody precipitation method and polymeric IgA as standard. Rocket immunoelectrophoresis was found generally unsuitable for IgA estimation: the reduction methods employed were found to be unreliable or unsuitable for some samples, and the estimation of IgA in bile by this method was further complicated by the presence of IgA fragments. The radioimmunoassay system, however, could be used to estimate IgA to a lower limit of 1 microgram/ml, and accurate results could be obtained almost irrespective of the molecular weights of the IgA in the sample.


Intraperitoneal systemic anaphylaxis in the mouse. I. Age-dependence of fatal anaphylactic shock.

Following the i.p. challenge of a shocking dose of BSA from 9 days up to 133 days after the s.c. injection of BSA in CFA, fatal anaphylaxis was induced regularly in female ICR mice that had been given the immunizing antigen when 8 weeks old. These immunized mice provided an antiserum to BSA that had the capacity to transfer fatal shock to normal recipient mice at a minimum Ab-N dose of 8 microgram when the i.p. route for challenge was employed. The optimal dose-range of antigen and antibody in order to elicit fatal shock following the i.p. challenge was much broader than that obtained by i.v. injection. Age is critical in producing fatal shock in mice; a 100% fatal anaphylaxis never occurred in groups of 6- and 7-week-old recipient mice although those at 8 weeks and older were sufficiently sensitized by the amounts of antibody given.


The evaluation of a radioimmunoassay for phenothiazines and thioxanthenes using an iodinated tracer.

Antisera have been raised in rabbits to the immogen 2-trifluoromethylphenothiazine-10-beta-propionate bovine serum albumin. An [125I]-labelled tyrosine methyl ester derivative of the immunogen precursor has been synthesised and used with the antisera to develop a simple, precise and sensitive radioimmunoassay for phenothiazines and thioxanthenes bearing 2-trifluoromethyl substituents. The assay can detect 0.4 ng/ml of fluphenazine, trifluorperazine or (Z)-flupenthixol in 100 microliter of human serum without interference from their sulphoxide or 7-hydroxylated metabolites. An acceptable correlation between this assay and an established fluphenazine radioimmunoassay using commercial [3H]fluphenazine has been obtained for plasma samples.


Dependence of the guinea pig IgE and IgG1 immune responses on the inclusion of potassium in the preparation of alum adjuvant.

Primary immunization with alum prepared using AlK(SO4)2 and adjuvant enhanced IgE production in the guinea pig. Alum prepared from Al2 (SO4)3 showed greatly reduced IgE and IgG1 anti-EA titers. This variance in immunoglobulin titer was observed only in the guinea pig. Both rats and mice respond to alum preparations prepared from either AlK(SO4)2 or Al2(SO4)3 equally as well.


Radiotoxicity of 111indium.

Bone marrow cells were labelled with various concentrations of 111Indium-oxine and their capacity to form colonies (CFU-s) in an adoptive transfer was investigated. Labelling with more than 0.1 muCi/ml/107 cells impaired colony formation. It is concluded that the 111Indium-oxine complex is detrimental to cell proliferation.


Inactivation of the mitogenic property of pokeweed mitogen by erythrocytes.

In whole blood culture, pokeweed mitogen (PWM) is unable to stimulate human lymphocytes. This is because the mitogenic property of PWM is inactivated by erythrocytes, presumably due to absorption. The inactivation was observed with as few as 5 x 10(6)/ml of human erythrocytes. Therefore, when PWM is used to study the functions of human lymphocytes, especially of suppressor T lymphocytes, erythrocytes should be removed from lymphocyte preparations for accurate analysis.


The effects of psoralen and UVA (PUVA) on epidermal melanocytes of the tail in C57BL mice.

The morphological and numerical changes in epidermal melanocytes in tail skin of C57BL mice after exposure to psoralen followed by UVA irradiation were studied. The dorsal aspect of the tail was painted daily for 5 days a week with 0.1 ml methoxsalen (8-methoxypsoralen) in acetone, followed by UVA irradiation (peak 365 nm) after 2 hr with increasing energies up to a maximum of 2.0 J/cm2 daily. Control animals received UVA or acetone treatment only. The total duration of the experiment was 8 weeks. Biopsies taken after 1 and 3 days, 1, 2, 4, 6 and 8 weeks were studied by the combined skin-splitting DOPA techniques. After 2 weeks of PUVA treatment the original brick like arrangement of melanocytes was obliterated and replaced by a confluent layer of melanocytes, and after 4 weeks the number of melanocytes had tipled and these cells also appeared on the epithelium covering the pilosebaceous units. UVA treatment alone did not show these effects. Inflammation and denuding of the skin after 4 weeks was followed by regeneration of the epidermis and the melanocyte system, in spite of continuing PUVA treatment.


Abnormal proliferation of uninvolved psoriatic epidermis: differential induction by saline, propranolol, and tape stripping in vivo.

Cell proliferation of the uninvolved psoriatic skin was compared with normal skin of volunteers following stimulation by the intradermal injections of saline or propranolol, or stripping of the stratum corneum with pressure-sensitive tape. Initially, no significant in vivo difference in tritiated thymidine labeling indices were observed between normal and uninvolved epidermal cells. However, 48 hr after each stimulus more DNA synthesizing cells were found in the uninvolved psoriatic than in normal epidermis. Of these 3 stimuli, propranolol was the most effective in distinguishing between uninvolved psoriatic skin and skin from normal control subjects. Thus, uninvolved psoriatic skin appears to possess an abnormal regulation of epidermopoiesis that may be amplified by propranolol injection.


Multiple forms and glycoprotein nature of acid phosphatase, alpha-fucosidase and alpha-mannosidase of psoriatic scales.

Isoelectric focusing and concanavalin A-Sepharose chromatography were used to study the multiple forms and glycoprotein natures of so-called lysosomal hydrolases from psoriatic scales. Acid phosphatase appeared as 5 different forms with pI values of 6.5, 6.1, 5.8, 5.6 and 5.45. Seven isoenzymes of alpha-fucosidase were identified with pI values of 6.4, 6.2, 5.9, 5.75, 5.65, 5.4 and 5.2. Acid alpha-mannosidase activity appeared as one peak with pI value of 6.75 and a weak activity of neutral alpha-mannosidase was present with pI value of 6.7. Incubation of the extract with neuraminidase increased their pI values of acid phosphatase, alpha-fucosidase and alpha-mannosidase to the more basic forms. This finding suggests that epidermal acid phosphatase, alpha-fucosidase and alpha-mannosidase have some N-acetylneuraminic acid residues. In addition concanavalin A-Sepharose column chromatography was also performed to confirm the glycoprotein nature of acid phosphatase. This enzyme was bound to the column and not released from the column even with the treatment of 0.5 M NaCl, but the enzyme was eluted from the column with the treatment of alpha-methyl-D-glucoside.


Surface area measurements of psoriatic corneocytes: effects of intralesional steriod therapy.

Quantitative measurements of surface areas of corneocytes from the desquamating portion of psoriatic lesions in 11 patients showed that psoriatic corneocytes were much smaller (27%) than corneocytes from noninvolved normal skin in the same patients. Corneocyte surface areas gradually increased in size following one intralesional injection of triamcinolone acetonide suspension and were almost normal approximately 6 weeks after treatment. These findings support other observations suggesting that rapidly proliferating epidermal cells have smaller surface areas than normal cells.


Kinetics of epidermal Langerhans cells.

Langerhans cells are considered to play an important role in the initiation of the immune response. This study was performed in order to analyze the kinetics of the Langerhans cell population under different experimental conditions. Using tritiated thymidine, the number of labeled Langerhans cells (demonstrated by the Leucinaminopeptidase reaction), and of labeled basal keratinocytes was investigated by autoradiography in guinea pig skin in vivo, before and 2, 5 and 8 days after stripping and before and 2, 5 and 8 days after initiation of repeated topical exposures to a 0.25% solution of dinitrochlorobenzene (DNCB). In addition the total number of Langerhans cells per mm2 was determined before and after DNCB treatment of epidermal guinea pig sheet preparations using the ATPase reaction. A total of more than 100,000 cell as of basal keratinocytes was stimulated significantly (by statistical analysis), both by stripping and by application of DNCB. After stripping, however, the increase of the Langerhans cell turnover was found to be secondary to the turnover of basal keratinocytes, whereas after DNCB application the increase in the proliferative activity of Langerhans cells appeared as the primary event in epidermal cellular kinetics.


Wheat protein antibodies in dermatitis herpetiformis.

An enzyme-linked immunosorbent assay was used to detect class-specific antibodies to wheat protein antigens. Antibodies which we detected by this technique reacted indistinguishably with antigens prepared from crude gluten, crude gliadin, alpha-gliadin, Frazer fraction III, and subfraction B and B3 of Frazer fraction III. No sera reacted with a human serum albumin control antigen. The prevalence of IgG antibodies to wheat protein antigens was significantly greater in patients with gluten sensitive enteropathy, 12 of 17, (p = .00011) and in patients with dermatitis herpetiformis, 5 of 14, (p = .046) than in normal control subjects. Strongly positive reactions for IgG antibodies were present only in patients with gluten sensitive enteropathy or dermatitis herpetiformis. IgA antibodies to wheat protein antigens were found only in gluten-sensitive enteropathy patients. We have found this to be a sensitive, precise technique for measurement of antibodies to wheat protein antigens and feel that it will prove useful in evaluation of the role of immune complexes involving wheat protein antigens and their antibodies in the pathogenesis of dermatitis herpetiformis.


Appearance of "pemphigus acantholysis factor" in human skin cultured with pemphigus antibody.

These studies deal with the mechanism of pemphigus IgG-induced epidermal acantholysis. When normal human skin was culted with defined medium containing IgG from pemphigus serum, extensive epidermal acantholysis developed and heat-labile proteolytic enzyme(s) were recovered in the culture medium. The enzyme(s) displayed maximal activity at pH 6.5 when a 3H-amino acid-labeled, insoluble epidermal cell material was used as substrate. The enzyme activity increased during the first 3 days of culture and the appearance of maximal activity coincided with the time of onset of acantholysis. Acantholysis did not occur in control cultures incubated with normal IgG and the enzyme did not appear in the medium or in aqueous extracts of cultured tissues. The enzyme(s) is probably not of lysosomal origin because low pH-active proteases characteristic of these organelles remained within the cells. The effects of puromycin on appearance of enzyme activity, acantholysis and cell viability was studied. At cytotoxic concentrations, the appearance of the enzyme(s) and acantholysis were prevented, whereas at less toxic concentrations enzyme activity and acantholysis were not prevented. Because inhibition of protein synthetic rates by puromycin could not be dissociated from the cytotoxic effects, it is uncertain whether enzyme appearance and acantholysis were dependent upon living tissue or on specific protein synthesis. After pemphigus IgG was removed from the conditioned medium by DEAE cellulose and affinity column chromatography, the remaining material contained enzyme activity and caused acantholysis in fresh skin explants. Similar activities were not present in normal IgG-containing conditioned medium or unfractionated epidermal extracts from normal skin. These data indicate that when the pemphigus IgG autoantibody interacts with epidermal cell surface antigens, the cell responds by synthesis or activation of a non-IgG "pemphigus acantholysis factor" (PAF) which may be a nonlysosomal proteolytic enzyme. It is suggested that PAF causes loss of adhesion between keratinocytes and ultimately produces the characteristic acantholytic cells of pemphigus.


Diagnosis of bacterial meningitis by gas-liquid chromatography. II. Analysis of spinal fluid.

The feasibility of gas-liquid chromatographic analysis of spinal fluid from dogs with experimental meningitis and from patients with meningitis was evaluated. Normal canine and human spinal fluid contained consistent levels of palmitic and stearic acids, glucose, and trace amounts of glycerol. Pneumococcal meningitis in dogs and humans was characterized by a complex fatty-acid pattern in spinal fluid similar to that seen in pneumococcal cells. Spinal fluid from dogs with Haemophilus influenzae meningitis yielded several saturated fatty acids as well as rhamnose, xylose, and glucosamine; four of five spinal fluid specimens from patients with H. influenzae meningitis gave similar results. Fatty-acid profiles of spinal fluid from dogs with staphylococcal meningitis were complex, with branched-chain and straight-chain fatty acids. Spinal fluid specimens from Neisseria meningitidis-infected dogs yielded 16- and 18-carbon fatty acids and sialic acid. Gas-liquid chromatographic analysis of spinal fluid proved valuable as a tool for identifying bacterial infections and differentiating between common spinal fluid pathogens.


Transmission of hepatitis B virus in clinical laboratory areas.

The transmission of hepatitis B virus (HBV) in clinical laboratory areas was delineated by the use of hepatitis B surface antigen (HBsAg) as presumptive evidence for the presence of the infective agent. Twenty-six (34%) of 76 environmental surfaces sampled were positive for HBsAg. The outer surfaces of blood- and serum-specimen containers had HBsAg contamination rates of 55% (six of 11) and 44% (four of nine), respectively. Subsequent handling of pipetting aids, marking devices, and other items led to their contamination and further dissemination of HBsAg. An assay instrument for complete determinations of blood cell counts was observed to splatter and drip blood during its operation. The contamination rate for environmental surfaces associated with this instrument was 15%. The data indicate that transmission of HBV in the clinical laboratory is subtle and mainly via hand contact with contaminated items during the various steps of blood processing. These data support the concept that the portal of entry of HBV is through inapparent breaks in skin and mucous membranes.


Altered neutrophil migration during bacterial infection associated with a serum modulator of cellular motility.

The motility of polymorphonuclear leukocytes (PMNLs) from 25 patients with bacterial infection was assessed by using the migration-under-agarose assay. A spectrum of responses was observed. Random and directional motility stimulated by C5a or pressed in parallel. These responses contrasted with those of normal PMNLs preincubated with either FMLP or C5a; preincubation with one chemotaxin inhibited a subsequent response to the same stimulus but did not inhibit the response to the other. The parallel responses of patient PMNLs to C5a and FMLP suggest that the altered motility reflects a perturbation independent of a specific chemotactic receptor. Heat-inactivated serum from patients with bacterial infection, when preincubated with normal PMNLs, resulted in significant enhancement of both random and directional migration. The data indicate that bacterial infection is associated with appearance of a modulator of neutrophil motility.


Dopamine and saralasin antagonism of renal vasoconstriction and oliguria caused by amphotericin B in dogs.

Amphotericin B (2.5 mg/kg, administered intravenously) increased vascular resistance (renal more than pulmonary more than systemic) and decreased glomerular filtration and urine flow 94% in 16 anesthetized female mongrel dogs. Dopamine decreased renal vascular resistance 31% in 14 dogs; when amphotericin B was given with dopamine, there was partial antagonism of amphotericin B-induced renal vasoconstriction. Saralasin partially antagonized amphotericin B-induced renal vasoconstriction in seven dogs. When amphotericin B was given during combined infusion of dopamine and saralasin in eight dogs, renal blood flow remained at initial control levels, urine flow increased above initial levels, and glomerular filtration decreased only 21% from initial values. Amphotericin B increased renal vascular resistance 296% when given alone but only 41% in dogs during injection of both dopamine and saralasin (P = 0.002). The antagonism of amphotericin B-induced renal effects by the combination of dopamine and saralasin was significant and specific for the renal vascular bed.


The influence of dosage regimen on experimental gentamicin nephrotoxicity: dissociation of peak serum levels from renal failure.

Peak serum levels of gentamicin were varied in rats by administering a standard nephrotoxic dosage of 40 mg/kg per day in one (QD), two, or three (TID) daily doses. The QD animals had the highest peak serum levels but showed no appreciable increase of serum creatinine concentrations over a 10-day treatment period. The TID rats had the lowest peak serum levels, but, after 10 days of drug administration, the serum creatinine concentration (2.8 +/- 0.2 mg/100 ml, mean +/- SE) was significantly higher than in control rats (0.6 +/- 0.01 mg/100 ml) (P less than 0.001). After two days of gentamicin treatment, the renal concentration of gentamicin was 269 +/- 77 micrograms/g in the QD rats and 820 +/- 29 micrograms/g in the TID rats (P less than 0.001). In this rat model, the frequency of doses was a more important factor in the development of nephrotoxicity than the peak serum concentration of gentamicin. The results suggest that dose frequency should be considered when data from different laboratories are compared.


Structure-function relationship: biological activities of the lipopolysaccharides and lipid A from Vibrio cholerae.

The biological activities of lipopolysaccharides and the lipid A moiety from five different strains of Vibrio cholerae have been investigated. Lipid A was biologically the most active component of lipopolysaccharides from these bacteria. Immunization of rabbits with lipid A from Inaba (66/64) strain led to the production of specific antibodies to lipid A. On fractionation of the immunoglobulins with Sephadex G-200 column chromatography, antibodies against lipid A were detected in the IgM and IgG fractions.


Viral shedding patterns of children with influenza B infection.

During an epidemic of influenza B, 43 ambulatory children were prospectively followed to determine the quantitative shedding patterns of influenza B viral infection, because these have not been previously described. The spectrum of illness included 74% with a typical influenzalike illness, 7% with an afebrile infection of the upper respiratory tract, and 19% with croup. Mild myositis occurred in 21%. For the first three days of illness, greater than or equal to 93% of the children shed virus, and 74% shed on day 4. The average peak quantity of virus shed in the nasal wash was 4.0 log10 50% tissue culture infective doses/ml(range, 1.5-6.0), which gradually declined over four days to 2.4 log10 50% tissue culture infective doses/ml. The quantities of virus shed correlated significantly with severity of illness and fever score, but not with sex, type of illness, or occurrence of myositis. These results suggest that the degree of clinical illness may be directly related to the cytotoxic effects of the virus and to the transmissibility of the disease.


Persistent urethral leukocytosis and asymptomatic chlamydial urethritis.

Chlamydia trachomatis was isolated from 47% of asymptomatic, sexually active men whose urethral smears contained four or more polymorphonuclear leukocytes (PMN) per high-power field (hpf) one week after their sexual activity was restricted. C. trachomatis was not detected in any of 23 asymptomatic, sexually active men having less than 4 PMN/hpf in two urethral smears obtained one week apart. Semiquantitation of urethral PMN in the urethral Gram-stained smear is useful in the evaluation of asymptomatic men without gonorrhea who are concerned about the possibility of harboring a urethral pathogen.


Experimental plague in the California ground squirrel.

In experiments to determine the current susceptibility to plague, it was demonstrated that 256 Yersinia pestis (a local strain of high virulence) were required to produce a 50% infectious dose (ID50) in California ground squirrels (Spermophilus beecheyi) that had been collected in southeastern Monterey County, California; 6,070 Y. pestis were required to produce a 50% lethal dose (LD50). (The LD50 was about 24 times the ID50.) The frequency of serologic response to the specific fraction 1 antigen of Y. pestis and mortality were dose-related. Approximately half of the squirrels inoculated with six to 6,070 Y. pestis survived without seroconversion, whereas antibody to specific fraction 1 antigen was always observed in squirrels that survived challenges of greater than or equal to 60,700 Y. pestis. However, titers never exceeded 1:64. The implications of these data for enzootic and epizootic transmission of plague in resistant squirrel populations are examined.


[Condylo-cephalic nail fixation for trochanteric fractures of the femur (author's transl)].

Condylo-cephalic nailing which has been developed by Ender is described. Three elastic pins are inserted at the proximal part of the medial femoral condyle and passed through the femur across the fracture into the head of the femur. The operative procedure is very simple and not hazardous to elderly patients. A special advantage is early weight-bearing. One hundred and three patients with intertrochanteric and subtrochanteric fracture were treated by this method. The average age was seventy-six years. Our own experiments have shown that this method can allow immediate weight-bearing.


[Studies of femoral neck fractures--biomechanical experiments with various devices for internal fixation with follow-up study on clinical cases-- (author's transl)].

The Tri-cancellous compression screw method was developed in order to overcome some defects which we feel exist in other methods of fixation presently in use. The author has conducted studies on the stability of the Tri-cancellous compression screw method and on the results of its clinical use. The conclusions are as follows: 1) In compression and torsion tests, the Tri-cancellous compression screw method showed greater strength than other methods. 2) The holding strength in cancellous bone of the screw is determined solely by its outside diameter and is never influenced by its thread profile and pitch. 3) The stability of the Tri-cancellous compression screw method decreases every hour postoperatively. 4) Clinically, when the Tri-cancellous compression screw method was correctly used, we found no evidence of the development of non-union. However, late segmental collapse has remained a problem of the same level as when other methods are used.


[Experimental and clinical studies concerned with the Jikei femoral head prosthesis (author's transl)].

For experimental purposes, femoral head prostheses for dogs, using both high density polyethylene (HDP) or titanium as the material for the head, were developed. Periodic observations of the acetabular cartilage influenced by the prosthetic head were done. The results showed that HDP was not suitable as a material for the artificial head because of rapid degeneration of the acetabular cartilage and marked wear and deformity of the HDP femoral head as soon as the surgery was done. On the other hand, the titanium femoral head was seen as causing much less damage to the acetabular cartilage in those cases where the diameter of the prosthetic head was quite adequate in its size. These findings indicated that the metal prosthetic head was useful for clinical cases. It was also clarified that the fixing of the stem to the bone could be achieved with our self-locking design instead of the use of bone cement. In follow-up studies of clinical cases used by the Jikei titanium femoral head prosthesis based upon our experimental results, it was found that better results could be expected when care is taken to use a femoral head which is not larger against the acetabulum. In addition, it was proved the procedure with the femoral head prosthesis is not indicated in cases where degeneration in acetabular cartilage has already occurred.


[An experimental study on the pathogenesis of osteoarthritis--histological and biochemical changes of proteoglycan in the osteoarthritic cartilage of rabbit in the early stage-- (author's transl)].

Osteoarthritis is generally a slowly progressive destructive disease of joints. As the disease is asymptomatic in its initial phases, it was detectable only at the later stage when radiological changes and clinical symptom had developed. The biochemical and cellular changes that had been recorded were those of the later clinically recognizable stage of the disease. Because the time of onset in not known, the initial events which might be the most important to investigate the pathogenesis of osteoarthritis, are virtually impossible to investigate except in experimentally induced models of osteoarthritis. For the purpose of investigating the pathogenesis of degenerative osteoarthritis, experimental osteoarthritic model was prepared. Section of the medial collateral and both cruciate ligaments combined with resection of the medial meniscus in adult rabbit knees showed progressive histological changes similar to those in human osteoarthritis. Therefore, qualitative and progressive alteration of the cartilage matrix proteoglycan in experimental osteoarthritis of the early stages were investigated with time. Biochemical analysis of the cartilage from experimental osteoarthritic model demonstrated a decrease in the content of glycosaminoglycan and increase in the rate of the synthesis of glycosaminoglycan as compared to the normals. The analysis of disaccharide chain revealed a decrease in the concentration of kelatan sulfate and chondroitin-6 sulfate, along with an increase in the concentration of chondroitin-4 sulfate. These findings, which were quite consistent with those in human osteoarthritis and were contrary to those in aging, indicate the attempts of cells to compensate for the proteoglycan deficiency in osteoarthritic cartilage by synthesizing large quantities of glycosaminoglycans of immature pattern. On the other hand, synthesized aggregated proteoglycans in arthritic cartilage had shown a polydispersive pattern in CsCl density gradient compared to that of normal one, which had shown a single curved pattern. Further, newly synthesized proteoglycan in osteoarthritic cartilage seemed to be polydispersive compared with control proteoglycan in molecular sizes on Sepharose 2B gel chromatography under non-aggregated dissociative condition. Large molecular size proteoglycan composed of small molecular size glycosaminoglycan on its polysaccharide chain and small molecular size proteoglycan composed of large molecular size glycosaminoglycan. This might represent the expression of repair mechanism in the damaged cartilage, or phenotypic transformation of extracellular macromolecule in osteoarthritis.


Functional anatomy of the elbow joint and three-dimensional quantitative motion analysis of the elbow joint.

This work consists of two parts. Part One is a three-dimensional study of the passive motion of the elbow joint and revealed that the elbow joint was not a true hinge joint. Part Two is a three-dimensional quantitative motion analysis which was undertaken to compare shoulder and trunk motion in normal subjects with those with contracted elbows. This analysis revealed that shoulder rotation, shoulder abduction, trunk flexion and trunk rotation compensates for a contracted elbow.


Radiation-induced fibrosarcoma of the mandible following treatment for bilateral retinoblastoma.

A case of fibrosarcoma of the mandible following radiotherapy for bilateral retinoblastoma and occurring in an 11-year-old female child is described. After a clinical description of the case, reported with histological documentation, problems connected with the pathogenesis of the malignancies are dealt with. Besides irradiation, genetic mutation as a carcinogenetic co-factor is taken into consideration.


True carcinosarcoma of the larynx: a case report.

A case of true carcinosarcoma of the larynx occurring in a 55-year-old male is reported. The two components of this very rare tumour are pleomorphic rhabdomyosarcoma and squamous cell carcinoma. A review of the literature has revealed only two authentic cases of true carcinosarcomas.


Chondrosarcoma of the larynx.

A chondrosarcoma arising in the posterior cricoid plate is presented. The tumour gave rise to increasing inspiratory stridor: laryngectomy was performed. The tumour consisted of loose cartilaginous tissue with great predominance of highly differentiated cartilage cells and only small areas with nuclear polymorphism. This complicated the differential diagnosis between chondroma and highly differentiated chondrosarcoma. From the fact that the patient died from massive pulmonary metastases 3 1/2 years later, it is evident that the degree of malignancy of cartilaginous tumours should be determined even on small polymorphic foci. The DNA histogram for the foci of the laryngeal tumour with atypia differed distinctly from those for benign chondroma and normal cartilage, but resembled those of the pulmonary and splenic metastases and of a nasal chondrosarcoma. Photometric examination may provide an aid in the difficult differential diagnosis between chondroma and highly differentiated chondrosarcoma.


The effects of vascular occlusion on the human inner ear.

Occlusion of the anterior vestibular artery has resulted in severe degeneration and new bone formation limited to the utricle, saccule, and superior and lateral semicircular canals. Depriving the inner ear of its main blood supply, i.e. the internal auditory artery, has resulted in severe degeneration and ossification of the entire membranous labyrinth, except the endolymphatic duct and sac. A more severe cochlear sclerosis was seen when both arterial and venous blood supplies to the cochlea were occluded. The implications of these findings on the etiology and management of inner ear disorders are emphasized.


Use of middle ear ventilation tubes in recurrent acute otitis media.

A method of treatment of recurrent acute otitis media with polythene middle ear ventilation tubes in young children is presented. 56 ears in which infection had persisted for one to six months despite antibiotic therapy and multiple myringotomies were included in the study. The tubes were inserted under topical anaesthesia and cure within one week was obtained in 73 per cent of ears. In another 9 per cent the otorrhoea stopped within 3 weeks, but in 18 per cent reinsertion of the tube became necessary and in this latter group 11 per cent resolved in two weeks following adenoidectomy as well. In 7 per cent the problem remained unresolved. Therefore with this regimen a total of 93 per cent of recurrent infections settled. It is emphasized that the ventilation tube should remain 'in situ' for over 3 months.


Domestic pets and perennial nasal allergy in children.

Domestic pets form an important etiological factor in the occurrence of perennial allergic rhinitis. In their management, patients responded well to animal exclusion and hyposensitization, with good amelioration of all their symptoms; these measures form an effective treatment for this type of disease.


Hearing loss in perforations of the tympanic membrane.

70 patients with dry central perforations have been studied to assess the hearing loss. The hearing loss is greater at the lower frequencies and increases with the size of the perforation. Malleolar perforations cause more hearing loss than the non-malleolar unless the perforation involves less than 10 per cent of the tympanic membrane surface area. Perforations of the postero-inferior quadrant cause more hearing loss than those in the antero-inferior quadrant.


Sequential audiometric evaluations of cats with regrowing tympanic membranes.

Audiometric evaluations were made for cats which were undergoing TM regeneration. For a one month period of regrowth and testing, there was an improvement in thresholds over the range of frequencies which were tested and the final evaluations (complete TM regrown) indicated greater auditory deficits in the high frequencies. The greater high frequency losses started at 1 to 2 kHz. and increased with frequency. The pattern of greater high frequency loss is thought to result from the fact that the regrown TMs did not adhere to the manubrium, but rather grew straight from the fibrous annulus and formed a flat rather than conical membrane.


Ameloblastoma of the maxilla. Report of three cases.

Three cases of ameloblastoma of the maxilla are presented. Symptoms, diagnostic procedures and therapy are discussed in relation to these cases. It is stressed that surgery of a maxillary ameloblastoma must be radical from the beginning. Otherwise the tumour may spread to the pterygopalatine fossa, temporal fossa or base of skull. Such tumour extensions are not resectable. The tumour is slowly-growing and a careful follow-up is mandatory.


Nasal metastasis from primary cancer of the kidney.

A case of nasal metastasis, indicating primary cancer of the kidney, and occurring in a 67-year-old man, is described. Differential diagnosis and pathogenesis are discussed. In particular, stress is laid on the problems concerning diagnosis when the nasal metastasis is the first clinical symptom. A clinicosemeiologic study of the patient is mandatory, as histology alone is not always sufficient to settle all doubts, even with the support of ample histochemical investigations.


The radioallergosorbent test (RAST) in nasal polyposis.

The incidence of atopy among 64 adults undergoing polypectomy was 25 per cent. Among these atopic patients it was usual for the polyp IgE level to exceed that in the serum. Similarly, D. pteronyssinus-specific IgE was more commonly identified in polyp fluid by the RAST technique than in serum. However, this was not so for grass pollen-specific IgE, where serum and polyp fluid were equally positive. We conclude that there are atopic and non-atopic polyps.


Lennert's lymphoma (lympho-epithelioid cellular lymphoma).

The authors describe a case of Lennert's lymphoma occurring in a 64-year-old male patient. After a comprehensive examination of the world literature on the disease, clinical and pathologic problems, as well as differential diagnoses from other malignant lymphoid proliferations, are discussed. This is the first observation to be reported in the otorhinolaryngologie literature.


Sjögren's syndrome involving the larynx: report of a case.

Sjögren's syndrome is a presumably autoimmune disease with pathologic findings of lymphocytic infiltration of the involved structures. Although the salivary and lacrimal glands are the most commonly involved sites, extraglandular lymphoproliferation also occurs. Upper and lower respiratory tract lymphoproliferation is known, but only one case of Sjögren's disease of the larynx has been previously reported. We present a case of laryngeal involvement in which the gross pathologic appearance is different from that in the earlier case report. The otolaryngologist should consider Sjögren's disease in the differential diagnosis of diffuse laryngeal lesions.


A contemporary view of the ventricular gradient of Wilson.

We have derived quantitative expressions for QRS, T, and QRST areas of the scalar electrocardiogram. The QRST area, or ventricular gradient, is seen to be essentially independent of the activation sequence and to reflect recovery properties of the tissue as weighted by the vector lead field of a given lead. The results are derived for uniform isotropic conditions and under the assumption that the temporal waveforms everywhere are identical except for possible variations in the duration of the plateau. However, it is noted that the results are, probably, valid under anisotropic conditions as well. The examination of ventricular gradients from epicardial and intramural leads should reflect local recovery properties and be a useful tool in study of the physiology of recovery, as well as the study of arrhythmias.


An approach to direct detection of sinus nodal activity in man.

We are reporting a series of 13 out of a total of 16 patients in whom sinus nodal activity was recorded with the use of intra-atrial electrodes, during diagnostic His-bundle electrocardiography investigations. The protocol consisted of the simultaneous monitoring of the following intra-atrial points: high right atrium, low lateral right atrium, left atrium and the His-bundle electrogram (HBE), which accompanied the detecting sonde placed as a loop in the superior vena cava, with the electrodes facing the sinus nodal area. This permitted an adequate assessment of the sequence of the depolarizing activity. All patients showed sinus nodal potentials preceding the initiation of the P-wave in the surface-EKG or the atrial potential in the high right atrial lead, respectively, in the range of 60 to 270 msec depending upon the functional status of the sinus node in each patient. Such waves were clearly separated from the initiation of the atrial potential by an isoelectric line and showed functional traits compatible with the S-A nodal functions, as many authors have previously reported in experimental animal models. The text contains a review of the relevant literature and discussion upon the diagnostic significance of this method in the assessment of sinus nodal disease.


Effects of dobutamine on electrophysiological properties of the specialized conduction system in man.

The electrophysiological effects of Dobutamine, a new beta adrenergic drug, were investigated using intracardiac electrograms and the extrastimulus method, in 19 patients with 1:1 AV conduction and in 10 other patients, five with second and five with third degree AV block. The electrophysiological effects were studied at three concentrations of the drug: 5, 10 and 15 micrograms/Kg/m'. Dobutamine induces: 1) an enhancement of SA node automaticity, showed by a decrease of the sinus cycle length (P less than 0.001 at the first concentration) and by a decrease in the corrected sinus node recovery time (P less than 0.001 at the second concentration); 2) a decrease in the effective atrial refractory period (P less than 0.001 at the first concentration); 3) an improvement of AV conduction, showed by a decrease in AH interval (P less than 0.001 at the first concentration) and by a shortening of functional and effective refractory periods (P less than 0.001 at the first concentration); 4) no change in the HV interval; 5) an improvement of conduction in patients with second degree AV block proximal to the His bundle; and 6) a minimal increase in the heart rate in patients with complete AV block distal to His bundle.


Carotid sinus massage induced elimination of rate related bundle branch block during paroxysmal atrial tachycardia: a simple method of proving bypass tract participation in the tachycardia.

Four cases of paroxysmal atrial tachycardia are described in whom rate related left bundle branch block (LBBB) was often present which persisted indefintely and showed no signs of spontaneous disappearance. Transient slowing of the tachycardia by carotid sinus massage in each case eliminated LBBB and this led to tachycardia acceleration. The tachycardia acceleration was traceable to a shortening in ventriculoatrial conduction. These observations proved the participation of a left sided bypass tract in the tachycardia circuit in each of these cases.


Spectral analysis of the normal electrocardiogram in children and adults.

We performed spectral analysis of the X-lead Frank VCG from six normal adults (Group A) and eight normal children (Group B) using a Fast Fourier Transform. The Group A magnitudes were consistently and significantly lower than Group B in the 20--90 Hz region. Similarly, the -40 dB point was 75 Hz for Group A and 107 Hz for Group B. The P waves of the two groups had similar spectra, as did their T waves. When the entire cardiac cycle was analyzed, there was again a consistent and significant difference in the frequencies from 25--80 Hz. Despite the significant differences in the Group A and B spectra, the vast majority of the information contained in the ECG signal was confined to frequencies below 100 Hz. Therefore, a bandwidth of 100 Hz is probably adequate for accurate recording of the ECG signal from children, as well as from adults.