An autoradiographic study of the efferent connections of the ventromedial nucleus of the hypothalamus.

Efferent projections from the ventromedial nucleus of the hypothalamus (VMN) were traced using tritiated amino acid autoradiography in albino rats. Ascending fibers passed through the anterior hypothalamus. Labelled fibers and terminal fields were seen in the preoptic area, bed nucleus of the stria terminalis, substantia innominata, the anterior amygdaloid area, diagonal bands of Broca and lateral septum. Fibers also projected laterally from VMN and entered the supraoptic commissures and zona incerta. These lateral projections were responsible for the fibers observed in the cerebral peduncle, the amygdala, the thalamus and the reticular formation. Fibers descending in a medial position projected through the posterior hypothalamus and then swept dorsally to terminate in the mesencephalic and pontine central grey. A projection from VMN into the median eminence was noted. The overall patterns of projection from different parts of VMN were similar; differences that existed were primarily in the relative strengths of the different projections. The efferent projections from VMN are extensive, well organized, and would appear capable of supporting significant physiological actions on extra-hypothalamic structures.


Substrate pathways which guide growing axons in Xenopus embryos.

Optic axons from eyes transplanted to hindbrain and spinal cord (i.e., non-optic areas) in Xenopus embryos reproducibly follow a particular pair of continuous tracts, the external sensory tract and the internal sensory tract. The course of the external sensory tract may coincide with the course of the general somatic afferent tracts (i.e., the ascending and descending tracts of nerve V, the tract of Lissauer, and the Rohon-Beard cell tracts). The course of the internal sensory tract parallels that of the external sensory tract but runs more medially. Transplanted optic axons follow the same tracts regardless of where along the neural axis the axons have entered the CNS. What kind of developmental cues guide these optic axons? It is unlikely that the guidance cues are organized as spatial coordinates, rather it appears that the optic axons of these two tracts are guided along pre-existing substrate routes. We have called these routes substrate pathways. Substrate pathways may be a device normally used for organizing fiber tracts in the developing nervous system.


"Extra" optic fibers exclude normal fibers from tectal regions in goldfish.

A small group of selected optic fibers were surgically deflected from one tectum into the other, thus creating a novel additional projection originating from a small area of ipsilateral retina. The normal fibers to this "recipient" tectum were also severed so that both the deflected and the normal fibers regrew into this tectum at about the same time. The reinnervation pattern was analyzed by autoradiography and electrophysiologic mapping. Both techniques showed that the deflected fibers and the "normal" fibers failed to intermix. The deflected fibers typically formed several well-defined patches of innervation in roughly the appropriate region of denervated recipient tectum. The normal fibers filled in the remaining uninnervated tectal areas and were completely or nearly completely exculded from the patches occupied by the deflected fibers. This segregation was often quite sharp having an apparent average overlap less than 25-50 micron. The electrophysiology indicated that the projections of both deflected and normal fibers were retinotopically organized but that the mapping by the normal fibers was compressed. This compression, an apparent consequence of being squeezed onto a smaller than normal region of tectum, was similar to that previously observed following ablations of part of the tectum. The negligible surgical damage in the present experiment, however, excludes the kind of cytochemical reorganization previously suggested to produce compression. The findings also provide evidence for a competitive type of interaction between optic fibers.


Proprioceptive reflex connections of head musculature and the mesencephalic trigeminal nucleus in the carp.

The location of the sensory cells concerned with the proprioception of respiratory and extraocular musculature in the carp was studied by retrograde transport of horseradish peroxidase. Sensory cell labeling after intramuscular HRP injection was exclusively found in the trigeminal-facial-anterior lateral line ganglion complex. The muscles innervated by the trigeminal system are represented in the more rostral ganglion areas, the muscles innervated by the facial system in the more caudal ganglion parts. Nearly all labeled cells were situated on the ipsilateral side. Sensory cells labeled after extraocular muscle injection were also found all over the V-VII ganglion, however, to a considerable degree also on the contralateral side. All muscle injections failed to give mesencephalic trigeminal cell labeling. The resluts of intranerve HRP injections in peripheral trigeminal nerve branches strongly suggests a perioral mechanoreceptive function for the mes.V neurons. A bisynaptic proprioceptive reflex model is described for respiratory musculature consisting of a sensory cranial ganglion component connected to the descending trigeminal nucleus, which on its turn links the proprioceptive ganglion cells to the trigeminal and facial motorneurons. Monosynaptic proprioceptive reflex circuits are discussed on neurophysiological grounds.


A neurophysiological determination of the vertical horopter in the cat and owl.

We have undertaken a determination of the vertical horopter in two species by simultaneously mapping the receptive field positions of binocular cortical neurons at various elevations along the zero azimuthal meridians. In the paralyzed cat our recordings show that the zero meridians of the two eyes are parallel and vertical under paralysis. Slit-pupil photographs demonstrate that paralysis induces an average net intorsin of 9 degrees between the two eyes. Correction back to the unparalyzed state results in the zero meridians themselves being out-torted with respect to each other. Since the two eyes' zero meridians define physiologically the positions of corresponding retinal points, this out-torsion results in a vertical horopter in the mid-sagittal plane which is tilted away from the alert, unparalyzed cat. The limited eye movements of the owl permit the use of an unparalyzed preparation; it is therefore possible to avoid the problem of the cyclotorsion under paralysis which occurs in the cat. The results of our physiological analysis in the burrowing owl (Speotyto cunicularia) also reveal a tilted horopter in this terrestrial avian species.


Synaptic contacts of the horizontal cells in the retina of the marine teleost, Callionymus lyra L.

The horizontal cell system in the retina of the fish Callionymus lyra L. was investigated light microscopically and electron microscopically. One type of rod horizontal cells, which are only found in the mixed ventral part, exclusively contacts spherules with lateral endknobs in the triads. Three types of cone horizontal cells occur. The first type, c-H1, contacts pedicles of pale and dark double cones and single cones. The processes always have a lateral position in the triads. This type has a constant contact pattern with the photoreceptor cells. The second type, c-H2, selectively contacts pedicles of pale double cones and the endbuds occupy a central position in the triads. In the single cone pedicles in the dendritic field of the c-H2 horizontal cell, the endbuds never seem to reach the triads. The third type, c-H3, only sends processes to pedicles of single cones where the endbuds occupy a central position in the triads. The synaptic connections of the horizontal cells of Callionymus differ from those observed in Nannacara anomala and Carassius auratus. The difference from the results obtained from Carassius is such that the information transfer model proposed for Carassius cannot apply to Callionymus.


Branching of sensory axons in the dorsal root and evidence for the absence of dorsal root efferent fibers.

The present study demonstrates that there are more dorsal root axons than dorsal root ganglion cells in the L6-S1 dorsal roots of the rat. The excess fibers do not come from aberrant dorsal root ganglion cells and our control procedures indicate that there are no extraneous fibers in these dorsal roots. Accordingly many dorsal root axons must branch in the dorsal root.


The organization of afferents to the cerebellar cortex in the cat: projections from the deep cerebellar nuclei.

The topography of the cerebellar nucleo-cortical projection was investigated in the cat by experiments employing the horseradish peroxidase (HRP) technique or by combined HRP-autoradiographic methods. The results of the HRP studies extend previous findings showing that neurons in the deep nuclei project to the cerebellar cortex in an orderly way. Thus, it appears that the cortex of the vermis-proper receives projections from neurons located predominately in the fastigial nucleus. Intermediate and lateral zones of mid-vermal cerebellar cortex are projected on by neurons located in the interposed and dentate nuclei. Crus II receives input from neurons located predominately in the dentate nucleus, while the paramedian lobule is projected on by neurons located in a large postero-dorsal sector of the interposed nucleus and in a smaller medial strip of the dentate nucleus. Neurons in the ventral part of the dentate nucleus and the lateral part of the interposed nucleus send fibers to the paraflocculus. The nucleo-cortical pathway to the flocculus and nodulus arises largely from a population of neurons located in a ventral region stretching from the medial border of the dentate nucleus to the lateral border of the fastigial nucleus. The results of experiments using the combined HRP-autoradiographic method show that clusters of neurons in the deep cerebellar nuclei project back to the cerebellar cortical areas from which they receive input, establishing a fairly precise feedback loop between the cerebellar cortex and deep nuclei.


Conduction velocity distribution of the retinal input to the hamster's superior colliculus and a correlation with receptive field characteristics.

Cells driven reliably by shocks delivered to the optic nerve or optic chiasm were encountered throughout the depth of the colliculus. The incidence of such cells, however, decreased markedly in the laminae ventral to the stratum opticum. The distribution of conduction velocities for the retinal afferents to the tectum was quite broad (range: 1.7-25.5 m/sec) and clearly biomodal with peaks at about 6 and 12 m/sec. A small number of cells were innervated by rapidly (greater than 15 m/sec) conducting axons. No evidence of an indirect-fast pathway from the retina to the colliculus via the lateral geniculate nucleus and visual cortex was obtained. Afferent conduction velocity was not correlated with retinal eccentricity, collicular depth or speed selectivity. It was, however, clearly related to directional selectivity. Ninety percent of the tectal neurons receiving inputs from axons having conduction velocities of less than 5 m/sec were directionally selective while only 41% of those neurons innervated by more rapidly conducting fibers (greater than 5 m/sec) exhibited selectivity. One hundred and sixteen cells in the anterior portion of the colliculus were tested with shocks delivered to the ipsilateral optic nerve and photic stimulation of the ipsilateral eye. Of these, 11% exhibited some degree of binocularity and only 6% were responsive to optic nerve shocks. These electrophysiological findings were correalted with the limited nature of the retinal input to the ipsilateral superior colliculus.


The cerebellar projection of the vestibular nerve in the cat.

The projection of the vestibular nerve to the cerebellum of the cat was examined with silver degeneration methods after complete lesions of the vestibular ganglion. The majority of the primary vestibular afferents were traced to the cortex of the ipsilateral nodulus and uvula, relatively fewer entering the ipsilateral flocculus. Fibers were not traced to the paraflocculus, lingula or lateral cerebellar nucleus. A sparse projection to the ipsilateral fastigial nucleus may exist, but it remains equivocal until confirmed with additional methods. Light microscopic examination of plastic sections confirmed these observations and showed further details of the organization of the primary vestibular projection to the nodulus and uvula. These results show that the region of the cerebellum densely innervated by primary vestibular afferents is smaller than previously believed.


The brainstem projection of the vestibular nerve in the cat.

The projection of the vestibular nerve to the brainstem of the cat was re-examined with silver degeneration methods after complete lesions of the vestibular ganglion. Whereas previous investigations emphasize that only parts of the major vestibular nuclei are innervated by the vestibular nerve, the present investigation shows that only the dorsal division of the lateral vestibular nucleus is uninnervated. Thus, the terminal field of the vestibular nerve extends to the cytoarchitectonic boundaries of the superior, medial, descending, and the ventral division of the lateral vestibular nuclei. Vestibular nerve fibers were also traced to discrete terminal fields in the reticular formation lateral to the abducens nucleus and in the rostral, lateral parts of the accessory cuneate nucleus. These observations indicate a more widespread distribution of vestibular nerve fibers in the brainstem of the cat than previously believed.


Sources of subcortical projections to the superior colliculus in the cat.

A comprehensive search for subcortical projections to the cat superior colliculus was conducted using the retrograde horseradish peroxidase (HRP) method. Over 40 different subcortical structures project to the superior colliculus. The more notable among these are grouped under the following categories. Visual structures: ventral lateral geniculate nucleus, parabigeminal nucleus, pretectal area (nucleus of the optic tract, posterior pretectal nucleus, nuclei of the posterior commissure). Auditory structures: inferior colliculus (external and pericentral nuclei), dorsomedial periolivary nucleus, nuclei of the trapezoid body, ventral nucleus of the lateral lemniscus. Somatosensory structures: sensory trigeminal complex (all divisions, but mainly the gamma division of nucleus oralis), dorsal column nuclei (mostly cuneate nucleus), and the lateral cervical nucleus. Catecholamine nuclei: locus coeruleus, raphe dorsalis, and the parabrachial nuclei. Cerebellum: medial, interposed, and lateral nuclei, and the perihypoglossal nuclei. Reticular areas: zona incerta, substantia nigra, midbrain tegmentum, nucleus paragigantocellularis lateralis, and the hypothalamus. Evidence is presented that only the parabigeminal nucleus, the nucleus of the optic tract, and the posterior pretectal nucleus project to the superficial collicular layers (striatum griseum superficiale and stratum opticum), while all other afferents terminate in the deeper layers of the colliculus. Also presented is information concerning the rostrocaudal distribution of some of these afferent connections. These findings stress the multiplicity and diversity of inputs to the deeper collicular layers, and more specifically, identify multiple sources of the physiologically well-known representations of the somatic and auditory modalities in the colliculus.


Remodelling of optic nerve myelin sheaths and axons during metamorphosis in Xenopus laevis.

Whole mounts and transverse sections of Xenopus optic nerves were examined with the light and electron microscopes before, during, and after metamorphosis. In stage 52--58 tadpoles, almost all myelin sheaths were circular in transverse sections. Early in metamorphosis (stages 60--61) large redundant myelin loops surrounded many large axons in central regions of the nerve. The loops subsequently were broken down into ovoids and lamellar segments that remained mostly within oligodendrocytes. These myelin changes were not observed in the chiasm or next to the eye. They were not associated with significant axonal degeneration and were no longer apparent in optic nerves of young frogs. Xenopus optic nerves also became shorter during metamorphosis. We therefore suggest that myelin sheaths with redundant loops which degenerate and disappear are being remodelled as the nerve decreases in length.


Modification of neuron numbers in the visual system of the rat.

After unilateral lesions of the posterior cerebral cortex at birth, the ipsilateral dorsal lateral geniculate nucleus disappears within 96 hours. Several of the remaining visual centers may be distorted, atrophic or hypertrophic. Examples of the last are the ipsilateral nucleus of the optic tract (NOT) and superficial layers of the superior colliculus (SC), which usually show a volume increase. In order to determine changes in neuron numbers in these two regions, we exposed rats to 3H-thymidine on specific embryonic days. Some of these rats received poterior cortical lesions on the day of birth and others served as controls. Counts from adult rats exposed to 3H-thymidine on embryonic days 13 and 14 reveal a significant increase in the number of labeled neurons in both the NOT and SC on the side of the lesion. The retinal projections in rats with posterior lesions at birth were determined by the application of silver degeneration methods after eye removal. Atypical optic projections are found to the lateral posterior nucleus and the NOT on the side of the lesion. We conclude that the redistribution of optic collaterals after early destruction of the dorsal lateral geniculate nucleus serves to increase neuron numbers in the NOT and SC. Therefore, the present findings are comparable to the results in nonmammals after transplantation of additional peripheral organs.


Scalpel excision of primary cutaneous malignant melanomas without metastasis.

Views and opinions about surgical treatment of primary malignant melanomas that are without local spread or metastasis are presented. An attempt is made to determine the limitations of office-based dermatologists in the treatment of malignant melanomas of this stage. Using accepted criteria for clinical recognition of malignant melanomas early, it is suggested that surgical excisions of small malignant melanomas that are judged to be complete obviate preliminary biopsies and furnish surgical specimens that can be read histopathologically for confirmation of clinical diagnosis and adequacy of excision.


Electro-excision of melanomas.

Excision of primary malignant melanomas by a cautery (actual heat) or by endothermy (electric cutting current) is a simple method of management. It consists of excision of a disc of tissue that contains the evident malignancy plus some surrounding normal-appearing skin.


Surgical management of advanced cutaneous malignant melanoma.

In the past, surgical treatment of cutaneous melanoma has been determined largely by assessment of the regional nodes by clinical palpation. More recently, an increasing number of reports indicate that measurement of the histologic thickness of primary melanomas and evaluation of the level of penetration of tumor cells into the dermis can predict the likelihood of development of nodal metastases. In addition performance of fine needle aspiration of questionably involved regional nodes has provided cytologic data regarding nodal metastases. The role of certain procedures, such as prophylactic (elective) node dissection remains clouded because adequate numbers of properly controlled studies have not been reported. However, increasing knowledge of important prognostic factors, cytologic findings from needle aspiration of regional nodes, as well as the clinical assessment of regional nodes, can all aid formulation of more rational management recommendations.


Chemotherapy of unresectable or recurrent metastatic malignant melanomas: an update.

Satisfactory chemotherapeutic management of malignant melanomas in advanced stages must await development of more active agents and combinations of them than are presently available. The development of entirely new means of treatment by hormones, heat, and adjuvants like amphotericin B may improve the efficacy of currently available agents.


Chemosurgery for familial malignant melanomas.

Three patients with numerous melanomatous lesions of the B-K mole syndrome are reported who were the first to be treated chemosurgically. The fixed-tissue chemosurgical technique provided a microscopically controlled, safe, effective, and conservative means of removing the melanomas. Atypical moles suspected of malignant change were removed by biopsy-excision.


Does immunotherapy have any value in the treatment of malignant melanomas?

To date, immunotherapy has not proven to be curative of malignant melanomas. Some immunologic procedures have produced regressions of some recurrences and perhaps have prolonged life by eliminating some dispersed malignant cells that would have gone to metastasis, but have not affected established, disseminated metastases to a meaningful degree.


Surgery by laser for malignant melanoma.

Lasers have been used for photocoagulation and photoexcision of malignant melanomas when conventional instruments could not be used. Lasers are useful for destruction of cutaneous metastases from malignant melanomas especially when numerous and dispersed. The color of melanomas makes for selective absorption of energy from lasers in the visible light range. Surgery by lasers is still an investigative form of surgery with current research studies on immediate placement of grafts, lymphatic migration of malignant cells after laser surgery, and relevant immunologic studies.


Prognosis of primary malignant melanomas.

In all probability, a patient with a primary malignant melanoma, but without clinical, radiological or other laboratory evidence of metastases, has a 75 to 80% chance of living five years and 55 to 65% of surviving for ten years regardless of other prognostic factors and the skill with which surgical therapy is applied. It has not been established whether or not malignant melanomas are absolutely curable.


Mangement of malignant melanomas: an overview.

This paper presents an overview of the management of malignant melanoma. It considers the value of wide reexcision relative to the depth of invasion of the melanoma. It considers the indications for elective lymphadenectomy and presents a critical review of chemotherapy, immunotherapy and other procedures, such as X ray. The conclusion is that surgery, wherever feasible, is still the best approach.


Nutritional realities--where does technology fit?

Food technology involves much more than fortification. It encompasses processing, i.e., a means of maintaining food in an acceptable, safe, and nutritious form for an extended period. Preservation processes free us from dependence on solely fresh foods. Thus, technology provides us with a stable food supply the year round and removes the threat of seasonal starvation. By increasing food acceptability through technology, the nutritional status of a population can be raised since the food will actually be eaten. To make technology the servant, rather than the master, however, more research is needed--for example, to establish the bioavailability of fortification nutrients.


The relationship of family structure and interaction to nutrition.

The relationship of nutritional adequacy to family structure and interaction is reviewed in light of previous research. Family structure is described by demographic data and measures of organization and status discrepancy of family members. Measures of family interaction include parent-child interactions, shared activities, and child-rearing habits. Nutritional research and educational programs need to give more attention to family units rather than individual respondents or clients. The review presents research findings and raises questions in order to generate ideas for future research.


Caffeine content of common beverages.

Tea, coffee, carbonated and chocolate beverages were analyzed for caffeine, and results compared in terms of usual serving sizes. Significant differences in caffeine levels were found to result from the preparation method of coffee or brewing time of tea. It is possible for a cup of tea, instant coffee, or can of cola beverage to have similar caffeine content (55 to 65 mg.); however, the mean values per cup of black tea (28 to 46 mg.) are considerably lower than for brewed coffee (107 to 151 mg.). Caffeine is readily absorbed and can have pharmacologic effects on adults or on children who consume quantities of cola beverages or chocolate. Both preparation method and quantity of beverage consumed should be considered in taking dietary histories or estimating caffeine intake.


Nutritional status of white college students in Virginia.

The nutritional status of the fifty men, fifty-three women, and forty-seven women taking oral contraceptives--all white--between the ages of seventeen and twenty-two years--as reflected by height-weight measurements, hemoglobin-hematocrit values, and dietary records, was similar or slightly better than reported by other investigators. Low hemoglobin values were observed in 2 to 4 per cent of these students, and low hematocrits, in 8 to 17 per cent. Similar hemoglobin and hematocrit values were observed in all women, regardless of whether they were taking oral contraceptives. Exercise patterns; use of vitamin-mineral supplements, "pot," alcoholic beverages, special diets; whether subjects ate alone; where subjects ate; snacking; number of meals consumed daily; whether subjects had ever been diagnosed as being anemic; and what subjects thought of their food habits did not affect hemoglobin or hematocrit. Iron intakes of many of the women students were below the allowance.


Fructose: comparison with sucrose as sweetener in four products.

The relative sweetness and acceptability of sucrose and fructose were determined at various levels in sugar cookies, white cake, vanilla pudding, and lemonade. Because of the reported increased sweetness of fructose and greater tolerance in individuals with diabetes mellitus for fructose, the study was designed to investigate its potential as an alternative sweetener. Results indicate that sucrose was both preferred and considered sweeter than fructose in sugar cookies, white cake, and vanilla pudding; however, the reverse was true in lemonade. On the basis of these results, the authors do not recommend the substitution of fructose for sucrose.


A low-cholesterol menu in a steak restaurant.

A twelve-month pilot project was conducted to test public reaction to a special restaurant menu identifying food choices low in cholesterol and saturated fat. The Houston steak house in which the "Help Your Heart" menu was tested prepared foods according to guidelines from dietitians of the Diet Modification Clinic, Baylor College of Medicine. Two newspaper articles, a brief magazine feature, and regularly scheduled radio advertising spots publicized the special menu. At the end of each month of the test period, the number of orders from the special menu was tallied for fourteen randomly selected days. Monthly sales from the special menu ranged from 2.5 to 5.1 per cent, with an overall average of 3.4 per cent. There was no statistically significant change from the mean of the percentage of total sales from the special menu over the months. Despite the relatively low percentage of total sales, restaurant executives were pleased with the results of the study and planned to offer the low-cholesterol menu indefinitely, eventually in an expanded form.


Nutrition component in a comprehensive child development program. I. The home visitor's role in the prenatal intervention phase.

Nutritional services were integrated in a comprehensive child development program for sixty-eight disadvantaged urban families in an attempt to promote maximum cognitive and psychosocial functioning in their children. Child development trainers made weekly home visits beginning in pregnancy, which combined data gathering, direct nutritional counseling, and early sensory excercises for infants. This paper describes the prenatal home visit program, the training of the home visitors, how the 24-hr. food recall method was adapted for their use, the nature of the home visits, and the complex role of the home visitor. The group whose families participated in the prenatal home visits and were also followed for six months after the infants' birth scored higher on six-month Cattell Scales than did a control group who entered the program at six months of age. There were no stillbirths or neonatal deaths. The incidence of low-birth-weight infants was lower than the national averages. Participation in the program was high.


Nutrition component in a comprehensive child development program. II. Nutrient intakes of low-income, pregnant women and the outcome of pregnancy.

Twenty-four-hr. food recall records taken by home visitors in an intervention program for disadvantaged families yielded data similar to three-day records and 24-hr. recalls taken by nutritionists. The young, urban, low-income expectant mothers studied had intakes below the recommended allowances for all nutrients studied except ascorbic acid. Energy, calcium, and iron were particularly low. The use of energy- and/or sodium-restricted diets during pregnancy was associated with lower intakes of other nutrients. Recent studies showed that low energy intake appears common among low-income, young, pregnant women.


Effect of supplements on the nutrient intake of children.

Daily intakes of nutrients were calculated from records of food eaten at home, as school lunches, and morning milk, of boys and girls in second and sixth grades. Vitamin/mineral supplementations were also calculated. Comparisons were made (with and without supplementation) with the Recommended Dietary Allowances (RDAs). Analyses of variance between sexes and between grade levels showed second graders consumed better diets than sixth and boys better diets than girls. Although many received supplements, intakes of several nutrients were below the 67 per cent level. The milk program played a significant role in calcium consumption. Excessive amounts of vitamin A and ascorbic acid were consumed by some children.


Development of an instrument to evaluate critical incident performance.

A performance evaluation instrument was developed for the clinical components of a course in a coordinated undergraduate program (CUP) in dietetics. The critical incident technique was chosen as being most appropriate. Twenty-six students in the "Foodservice Systems" course of the CUP at Kansas State University comprised the research population. The course instructor, clinical instructors, and dietitians in the affiliations served as observers. The performance evaluation instrument, as developed consisted of ten categories of behavioral activities. Entries into the form were either behaviors to be encouraged or suggestions for improvement. Students' reactions were extremely favorable because of the feedback element. The instrument should be an effective tool and worthy of adaptation for other courses.


A nutrition attitude instrument for nursing students.

This communication has focused on selected aspects of attitude measurement used in developing an attitude instrument, composed of one scale, appropriate for nursing students. The study generated a scale with a Coefficient Alpha of r = 0.87 and a split-half reliability estimate of r = 0.84. Due to the dependence of these reliability estimates on statistical correlations, the size and/or the manner of the selection of the sample is an important consideration. The study was discussed in relation to the present status of attitude measurement in nutrition education research.


Vietnam, 1978: crisis in food, nutrition, and health.

Evidence of shortages of food, medical equipment, and supplies and indications of malnutrition were observed by an American Study Mission to Vietnam in the summer of 1978. Recommendations of humanitarian aid were made by the study team and presented as promoting peace and stability in Southeast Asia.


Dietetic audit--a giant step for nutritional care. The approach at Brooke Army Medical Center.

Issues which must be addressed by clinical dietitians as participants in their hospitals' quality assessment programs (audits) are defined. Solutions and approaches used at various phases of the audit processes by the clinical dietitians at Brooke Army Medical Center are described. Outcome and process criteria for nutritional care of the patients with maturity onset diabetes mellitus are presented, along with samples of the clinical dietitian's documentation in the patient's record. The impact of "audits" on the quality of nutritional care is exemplified by results of a series of audit studies on diabetes mellitus conducted over the past three years. Other benefits which have accrued from participation in the hospital's audit programs are enumerated.


Changes in nutrient intake during a behavioral weight control program.

Behavioral modification holds promise of being an effective mode of therapy for obesity. It focuses on changing clients' eating habits and does not always include specific dietary guidance. This study was conducted to learn whether such behavior modification training results in the adoption of nutritionally sound dietary patterns. Originally, the diets of fifteen individuals met two-thirds of all Recommended Dietary Allowances. After a twenty-week program, the only nutrients below this level were iron, thiamin, and calcium. Other changes involved an increase in nutrient density for protein, fiber, phosphorus, iron, vitamin A, thiamin, riboflavin, niacin, ascorbic acid, and cholesterol. Nutrient density for carbohydrates, fat, and calcium decreased.


Attitudes toward obesity and the obese among professionals.

Negative attitudes toward obese patients by health professionals have been attributed to termination of weight reducing attempts. This study measured attitudes concerning the obese held by professional participants in a continuing education conference on causes and treatment of obesity. Substantial variation was found on items comprising obesity-related belief dimensions: (a) Disparaging image of the obese; (b) causes of obesity; and (c) ways to lose weight. Evidence is presented for the association among measures of these dimensions. While background and educational characteristics and conditions of practice were not related to such attitudes, the professional's personal experience with successful weight reduction was the best predictor of favorable attitudes.


Dietetic services in a cross-modality system.

The nutrition component of a cross-modality training program is described. This training specifically enabled non-professionals to provide selected dietary and clinical evaluations and basic nutrition education. The role development of a dietitian, either functioning in a C-M system or facilitating the introduction of such a system, is discussed. The dietitian's contributions and limitations are presented from the viewpoint of the dietitian as a member of a C-M training team. Implications are drawn from utilization of the C-M system in institutions.


Approaches to wage and salary determination. Implications for hospital dietitians.

Although many types of rewards are to be found in work, salaries and benefits remain primary concerns. Thus the compensation policy of the employer is of great importance to the employee. Described here are current salary administration approaches and practices, i.e., market pricing and internal equity systems. Also explained are components of wage and salary plans, including job evaluation, labor market surveys, and job pricing. A detailed discussion of methods of job evaluation--qualitative and quantitative--is presented. The paper concludes by reporting figures from a recent salary survey of four hospitals in the Chicago area.


A social policy directed toward the health and welfare of the aged in the Soviet Union.

Dealt with in detail are the factors which have determined the characteristics and peculiarities of medical and social care for the aged in the USSR. The forms of geriatric care, personnel training and delivery of social and domestic services to the aged are also presented. Facts are given about the state policy directed at improvement of well-being of the aged as well as at drawing these people into public life and preventing their "disadaptation". The importance of the UN- and WHO-sponsored international programs in this field is emphasized.


Systemic lupus erythematosus in the older age group: computer analysis.

A computer analysis was made of the data from a prospective study of the clinical course of systemic lupus erythematosus (SLE) in 234 patients followed for an average of 46 months. All fulfilled four ARA criteria for the diagnosis of SLE. Sixteen of the 234 patients were aged 51 or older. They were compared with the 218 younger SLE patients to determine the influence of age on the signs and symptoms of the disease, the organ systems involved, the laboratory data, amount and duration of corticosteroid or azathioprine therapy, and the prognosis. The older group showed more discoid lupus, photosensitivity and pulmonary fibrosis than did the younger group, but a similar incidence of malar rash, alopecia, arthritis, arthralgia, myalgia and serositis, and a lower incidence of oral ulcers, Raynaud's phenomenon, cutaneous vasculitis, neuropsychiatric manifestations, leukopenia, hypocomplementemia and profus proteinuria. The older patients needed a lower dosage of corticosteroids, and a shorter course of azathioprine therapy. These findings suggest a milder form of SLE with better response to therapy in the older group.


Sexuality in nursing homes.

Since so little has been written on the subject of sexuality among the elderly in nursing homes, a study was conducted in some nursing home populations in Wisconsin. Resistance was encountered in certain areas but finally 63 residents (27 men and 36 women) agreed to be interviewed. A recently designed questionnaire was used, followed by a relaxed conversational period. Our findings indicate that the aged interviewees believed that sexual activity was appropriate for other elderly people in the homes; they personally were not involved, chiefly because of lack of opportunity. Most of them admitted having sexual thoughts and feelings. Medical and behavioral personnel showed great reluctance to discuss the subject. It would seem that, if the quality of life in old age is to be improved, there should be some provision in nursing homes for those who desire appropriate sexual activity.


Dihydroergotoxine: 6-mg versus 3-mg dosage in the treatment of senile dementia. Preliminary report.

In 14 patients with senile dementia, a study was made of two dosage levels (3 mg or 6 mg daily) of dihydroergotoxine mesylate (DEM, Hydergine). Only a nonstatistically significant trend was found for superiority of the higher dosage. However, one patient showed remarkable clinical improvement during the 6-mg period; the mechanism remains unexplained. Further studies are needed with this higher dosage in less impaired patients and in those with well-defined cerebral pathologic changes.


Drug usage and health characteristics in noninstitutional retired persons.

A study was made of total drug usage (prescription, over-the-counter, social), health characteristics, and life satisfaction in 55 noninstitutional retired men and women whose average age was 70 years. The sample was purposively selected on the basis of involvement in peer group interaction networks. Among those using drugs, the average number taken per person was 2.0 prescription, 1.8 over-the-counter, and 1.8 social (alcohol, caffeine, nicotine), for a total of 5.6 drugs. For men the average was 7.5, and for women 4.7 drugs. Although drug interaction hazards from prescription drugs alone appeared to be minimal, examination of total drug usage indicated potential interactive hazards. Despite the number of chronic illnesses and physical impairments (5 and 1 per person respectively) and the number of drugs ingested, the respondents usually expressed overall satisfaction with their life condition. A commonly expressed dissatisfaction (males 75 percent, females 35 percent) was with government officials. Respondents had confidence in both their physicians' diagnoses and the effectiveness of the drugs prescribed. The findings that women reported more chronic diseases and physical impairments, and were relatively more dissatisfied with their life condition than were men, may be associated with their greater acquiescence with self-disclosure.


Prosopo-affective agnosia associated with chronic organic brain syndrome.

Impairment of the ability to recognize facially expressed emotions was studied in 14 chronically disoriented patients with chronic organic brain syndrome (CBS). This impairment was named prosopo-affective agnosia (PAA). A diagnostic requirement was relatively intact neurologic functioning in underlying perceptual-verbal-motor processing. A test was designed for facial-affect recognition in the accurate differentiation of normal persons from chronically disoriented CBS patients. No normal subject made any errors in this test. Despite decades of illness and hospital living, patients with a history of schizophrenia or major affective disorders scored almost at a normal level (95 vs. 100 percent) in this test, and significantly higher (95 vs. 66 percent) than did the disoriented CBS patients. The social and therapeutic implications of the findings are stressed. CBS patients may be impaired with respect to receiving and appreciating elementary aspects of social communications such as recognizing a smile, anger, sadness or disapproval on the faces of people who surround them. This disability requires understanding and a special attitude on the part of the therapeutic team toward such patients.


Visual training, the alpha activation cycle and reading.

Two patients with visual problems and reading difficulties originally unable to suppress alpha have been shown to demonstrate alpha rhythm attenuation during the course of and after a program of visual training with simultaneous improvement in reading performance. No direct training of alpha attenuation was engaged in. No other therapy directed toward solution of the patients reading or behavior problems was attempted during the course of visual training. It is concluded that the change in alpha activation and the improvement in reading were both brought about by the combined visual therapy and reading glasses. Further research concerning the number and proportion in the population of learning disabled who respond similarly to the cited cases is presently being undertaken by the author.


Measuring contrast sensitivity with gratings: A new simple technique for the early diagnosis of retinal and neurological disease.

The measurement of contrast sensitivity using gratings is a new clinical procedure which yields information about the functioning of the visual system. Previously, this type of information has not been readily obtainable in the clinical environment. This article includes the basic concepts of measuring contrast sensitivity with gratings and preliminary clinical results.


The NIH-ERG ganzfeld stimulator.

The design specifications of a NIH-ERG ganzfield stimulator is discussed. Based on a proposed standard for clinical electroretinography, the stimulator uses stroboscopic stimuli as a whole field flash. But by varying the parameters of wavelengths and background illumination, a complete isolation of the rods and cone systems of the retina is obtained. This information is invaluable at times in early detection and diagnosis of retinal dysfunctions.


Fundus fluorescein angiography: a summary of theoretical concepts and clinical applications.

The basic concepts of fundus fluorescein angiography are presented along with historical and physiological background. Fundus photographs, as well as angiograms, are used to highlight the text.


The electroretinogram: concepts and clinical applications.

This paper describes the clinical applications of the electroretinogram (ERG). Its purpose is to familiarize the practitioner with the capabilities, and limitations, of the ERG. This discussion includes ERG theory, test procedures, and interpretation of results.


Visual implications of diagnostic advances in neurology.

Two major diagnostic advances in neurology which have visual implications are Computerized Axial Tomography (CAT Scan) and Evoked Potentials. This paper briefly discusses the basic concepts of the CAT Scan and of Evoked Potentials. In addition, it includes select clinical applicability of each.


Static vs dynamic visual field evaluation; with emphasis on the utility of the Friedmann Visual Field Analyzer.

This paper contrasts static and dynamic perimetry, highlighting the advantages and disadvantages of each. A type of static perimetry, the Friedmann Visual Field Analyzer, is discussed in some detail. A clinical study, comparing the sensitivity of the Friedmann Visual Field Analyzer and B&L Autoplot in detecting field defects in known glaucoma patients, is presented. The results of this study support the notion that static fields (utilizing the FVFA) are significantly more sensitive than dynamic fields (utilizing the B&L Autoplot) in detecting field defects in known glaucoma patients.


Variola minor in Braganca Paulista County, 1956: household aggregation of the disease and the influence of household size on the attack rate.

Household aggregation of cases, one possible characteristic of person-to-person transmitted disease, was formally tested in one epidemic of variola minor by using a pair statistic. A significant result was found for all households as well as for households grouped by the type of environment, or by the phase of the epidemic growth in time. Secondary attack rates, when related to household size (number of susceptibles) showed only a marginal trend in rural households but no trend in urban or semi-rural households.


Gastroenteritis in London and Jamaica: a clinical and bacteriological study.

The flora of both faeces and small bowel lumen was studied in children with gastroenteritis from London, England, and Kingston, Jamaica. Clinical and laboratory differences between these two groups are described. All bacterial groups in the faeces were greatly altered during gastroenteritis and this particularly affected anaerobic organisms. These changes generally reverted rapidly to normal after the illness. The small bowel flora was also altered during gastroenteritis; there was a tendency for a wider range of organisms including anaerobes to be isolated from the children in Jamacia than from those in England.


Salmonellosis in Indonesia: phage type distribution of Salmonella typhi.

The distribution of phage types was studied among 577 strains of Salmonella typhi from Indonesia. Chemotype, colicinogeny, and tetrathionate reductase activity were also studied for most of these strains. The current phage type formula for Java was determined to be: A, D2, D6, E1a, E2, M1, and 46, but two other large groups of strains were also found, I + IV and degraded Vi+ strains. Significant differences in S. typhi strain distributions were noted between two localities on Java with respect to phage type and tetrathionate reductase activity. Comparisons were made with past phage typing studies in Jakarta as well as with more recent studies in other parts of south-east Asia. Phage types A, D1, D2, and E1 persisted at a rather steady level in Jakarta for 28 years. Evidence was found for epidemiological links to European and Asian areas. Antibiotic resistance among these Indonesian S. typhi strains was rare.


Epidermal tissue as a primary site of replication of lymphocytic choriomeningitis virus in small experimental hosts.

When fresh urine from LCM tolerantly infected mice was applied to small areas of excoriated skin of guinea-pigs undiluted or diluted 10(-1), a high LCM infectivity developed in the local dermal tissue within 3 days and quickly spread to the lymphatic system. The skin at this site of infection became erythematous 10--12 days after infection and a few days later a rash was often seen in the hairless skin around the mammary teats. A viraemia was first detected at about 8 days after infection and persisted for at least 8 days, during which time a high infectivity titre in skin not only at the infection site but also distal to it suggested that there was a generalized active infection of the dermis. Infectivity in the tongue was simultaneously high and probably associated with erosions of the tongue tip seen a day or two later than the teat rash. In similar experiments in hamsters and rabbits, indications were again that lightly injured dermis was a primary site of virus replication. These observations should lead to the dermal route receiving greater attention as a potential route of infection of man when exposed to infectious excretions of reservoir hosts of arena-viruses.


The increasing isolation of Serratia species from clinical specimens.

A 14-month survey was undertaken in a diagnostic bacteriology laboratory to determine the incidence of Serratia spp. in routine clinical specimens. Gram-negative organisms with enterobacteria-like colonies were tested by a simple screening procedure. Fifty-eight strains of S. marcescens and two strains of S. liquefaciens were isolated from 59 patients. The strains were usually non-pigmented and exhibited multiple antibiotic resistance. Serotyping and determination of bacteriocine sensitivity patterns revealed that the majority of infections were sporadic, although episodes of cross-infection did occur. S. marcescens was considered to contribute significantly to morbidity and mortality in 53% of patients and appears to be of increasing importance in hospital-acquired infections.


The cultural and biochemical characters of Streptococcus milleri strains isolated from human sources.

A collection of 346 strains of Streptococcus milleri from a variety of human sources was examined culturally and biochemically, and for the presence of Lancefield group antigens. Most of the strains were non-haemolytic and ungroupable, but 25% were beta haemolytic and 19% were alpha haemolytic; 28% possessed a group antigen (A, 5%; C, 6%, F, 14%, G, 3%). These antigens were present in 69% of beta-haemolytic but in only 13% of alpha-haemolytic or non-haemolytic strains; beta haemolysis occurred in 82% of group-F strains, 43% of other groupable strains and 11% of ungroupable strains.The following reactions were given by > 80% of S. milleri strains: hydrolysis of arginine and aesculin, a positive Voges-Proskauer reaction, and acidification of trehalose, lactose, salicin and sucrose. A minority of strains showed enhancement of growth by CO(2), bile tolerance, NaCl tolerance, and ability to acidify other sugars, notably mannitol, raffinose and melibiose. Departures from the modal pattern of biochemical reactions showed a weak correlation with the type of haemolysis and the presence or absence of a group antigen but were not sufficiently systematic for clear-cut subdivisions to be recognized within the species.S. milleri therefore appeared to comprise a ;central' group of non-haemolytic strains that rarely formed a Lancefield-group antigen, in which the aesculin reaction was nearly always positive, lactose was usually acidified, and a considerable minority showed enhancement of growth by CO(2) and bile tolerance. Deviations from this pattern were of two main types. (1) ;Loss' of one or more of these reactions, which tended to be associated with beta-haemolysis and presence of a group antigen. In these respects, alpha-haemolytic strains tended to occupy an intermediate position. (2) ;Gain' of the ability to acidify additional sugars, notably raffinose and melibiose or mannitol; this occurred mainly among otherwise typical non-haemolytic strains that were rarely groupable.Only 12% of isolations from the bloodstream of patients suffering from systemic infections were beta haemolytic and only 18% possessed a group antigen, but a considerably greater proportion of those from visceral abscesses were beta haemolytic (28%). Among isolations from superficial lesions in some body sites there were considerably greater proportions of beta-haemolytic and groupable strains; thus, nearly one-half of those isolated from the abdomen other than the female genital tract were beta-haemolytic and over one-half were groupable. On the other hand, strains from the teeth and gums were nearly always non-haemolytic and ungroupable, and most vaginal isolations were of non-haemolytic strains with a wide sugar-fermentation pattern.


A garment for use in the operating theatre: the effect upon bacterial shedding.

In operating theatres the air is mainly contaminated with bacteria shed from the human skin. The emission of bacteria can be prevented by wearing clothing of impervious material, while normal cotton clothing does not decrease the shedding of bacteria. In this study shedding of viable bacteria from 20 test-persons wearing an operating theatre suit, composed of 65% polyester and 35% cotton (Diolen), was investigated in a test-chamber and compared with that when normal clothing was worn. The use of this operating-theatre suit resulted in a significant reduction (50--75%) in the number of bacteria-carrying particles in the air of the test-chamber and in an operating room when everyone present wore this suit. A combination of the suit with knee-high boots showed a further reduction in the dispersal of colony forming units. The dispersion from female subjects wearing an operating-theatre frock was significantly higher than when wearing an operating-theatre suit.


Comparison of the tuberculin Tine Test+ and the Mantoux.

Re-evaluation of recently published figures comparing the tuberculin Tine Test+ and the Mantoux showed that, contrary to original opinion, the Tine Test+ did perform satisfactorily, the reason for the apparent ineffectiveness lying in the original interpretation of the results.


Influenza A(H1N1) viruses of the 1977/78 outbreak: isolation and haemagglutination properties.

During the H1N1 outbreak of 1977/8, the virus was isolated in embryonated eggs from 59 out of 76 patients (78%) with the serologically confirmed infection. A similar isolation frequency has been achieved during a period of six H3N2 outbreaks since 1972/3. The H1N1 strains were isolated less frequently from late specimens (collected 4--6 days from the onset of illness) and more often only in the second passage compared with the H3N2 viruses. The new H1N1 strains resembled those prevalent in the 1950s with respect to their ability to agglutinate erythrocytes of certain laboratory animals and to be eluted from these, and thus differed from the H3N2 viruses.


Detergents compared with each other and with antiseptics as skin 'degerming' agents.

Three detergent preparations (bar soap, 'Hibiscrub' base and 'LIC 76'), TWO ANTISEPTic preparations (0.5% chlorhexidine in 95% ethyl alcohol and an alcohol jelly, 'Alcogel'), and one antiseptic-detergent solution (4% chlorhexidine gluconate in a detergent base, 'Hibiscrub') were compared for their effectiveness, on a single use, in reducing the yield of bacteria from the hands of volunteers. The antiseptic and antiseptic--detergent preparations were more effective than the detergents, with a mean reduction in yield of skin bacteria of 96.0% after use of alcoholic chlorhexidine and of 81.2% after use of Hibiscrub. One of the detergents, LIC 76, appeared more effective than the others, causing a mean reduction in the yield of skin bacteria of 41.5%, compared with reductions of 4.6% by the Hibiscrub detergent base and an increase of 3.2% with bar soap; unlike the other detergents, LIC 76 was found to have appreciable bacteristatic and bactericidal properties.


Early complement components in NZB/NZW mice. I. The first component.

Sera from periodic bleedings of B/W female mice were assayed for C1 hemolytic activity. It peaked at 3 to 4 months of age and declined to very low levels by 6 to 7 months. Activity remained low even in long-lived animals.


In vitro activation of complement by isolated human heart subcellular membranes.

Activation of human complement (C) occurred in vitro when mitochondrial membranes isolated from normal human heart tissue were incubated with normal human serum. This activation, as measured by C3 depletion, was not completely inhibited by blocking classical pathway activity in serum treated with EGTA, in C2-deficient serum, or in C1-depleted serum, nor in serum heated at 50 degrees C for 30 min to block the alternative pathway, but it could be prevented by blocking the classical and the alternative pathway simultaneously with EDTA, or by treating heated serum (50 degrees C. 30 min) with EGTA. Factor B was converted in normal serum as well as in EGTA-treated serum, but not in EDTA-treated serum. Mitochondrial membranes had no direct enzymatic or other activity that could inactivate functionally or highly purified C4 or C3, but the membranes could bind and activate C1 either in serum or in functionally pure C1 preparations. C4 also bound to the mitochondrial membranes only in the presence of C1. These data suggest that the activation of C by heart subcellular membranes involved both the classical and the alternative pathways, that the mitochondrial membrane preparations were capable of forming stabel complexes with C1 and C4, but not C3, and that the mitochondrial membrane preparations did not contain enzymes or have inherent properties that could directly cause C3 conversion.


Characterization of an antiserum specific for the Fabalpha fragment. Its use for detection of alpha-heavy chain disease protein by immunoselection.

An antiserum to the Fabalpha fragment of human IgA was prepared by injecting K-and lambda-type IgA1 myeloma protein Fab fragments in complete Freund's adjuvant into a goat. After appropriate absorptions, the antiserum reacted specifically with Fabalpha fragments, normal IgA, and K- and lambda-type IgA1 and IgA2 myeloma proteins. This antiserum contained predominantly antibodies to determinants shared by different IgA myeloma proteins and expressed on isolated alpha chains. These determinants were presumably located on the first constant domain of the alpha chain (Fd fragment). When incorporated into agarose, the anti-Fabalpha antiserum is a potentially valuable reagent for screening human sera for alpha-heavy chain disease proteins by an immunoelectrophoretic method of immunoselection.


Allogeneic placenta is a paternal strain antigen immunoabsorbent.

A partially purified 125I-labeled antibody directed against the murine major histocompatibility complex and injected i.v. pregnant females is selectively absorbed by the placenta when the fetus bears the appropriate targen antigen. The placenta specifically absorbs the antibody at both 13 days and 17 days of gestation, and the differential uptake is found as early as 2 hr after injection and as late as 12 hr. It is also seen in 13-day placentas that have had the fetus removed surgically, indicating that fetal circulation is not essential for the absorption to take place. Specificity controls eliminate the possibility that the differential uptake is due to Fc receptor binding or undefined effects of hybrid vigor. This increase in binding is not seen in the livers, spleens, kidneys, or lungs of the pregnant females. There is a slight decrease in radioactivity in the serum. These results indicate that the placenta is a paternal antigen-bearing immunoabsorbent that could serve as a barrier between the maternal immune response and the semi-allogeneic fetus.


Ascaris and eosinophil. II. Isolation and characterization of eosinophil chemotactic factor and neutrophil chemotactic factor of parasite in Ascaris antigen.

In the previous study, we found an eosinophil chemotactic factor of the parasite (ECF-P) in extracts from the body cavity fluid of the Ascaris species. In this study, the physiochemical characteristics of ECF-P were determined in order to elucidate the relationship between ECF-P and Ascaris antigens. ECF-P was active in both in vivo and in vitro chemotactic assay systems and was found to be heat labile, nondialyzable, and stable to lyophilization. The estimated m.w. was approximately 30,000 daltons. The isoelectric point of ECF-P was 8.5. It was quite different from the Ascaris antigen in the same extract, or various Ascaris allergens that had been reported previously by other researchers. Furthermore, ECF-P did not show any activity in eliciting a guinea pig passive cutaneous anaphylaxis (PCA) reaction. From these experimental results, we concluded that ECF-P is not identical to Ascaris antigens. In addition, we found that the neutrophil chemotactic factor of the parasite (NCF-P), with at least two components, is present in the Ascaris extract, and is also heat labile and resistant to dialysis and lyophilization, but separable from coexistent ECF-P by reason of their different physiochemical properties.


The identification of actin as a major lymphocyte component.

Lymphocytes derived from pig mesenteric lymph node and mouse spleen have been shown to contain a major protein conponent which a) co-migrates on one-dimensional (SDS-PAGE with purified pig muscle actin, and b) possesses the same isoelectric point as muscle actin on two-dimensional gel electrophoresis. Further, limited proteolytic digestion of the lymphocyte actin-comigrating band by S. aureus V8-protease revealed extensive primary sequence hemology with purified muscle actin. Based on the virtual identity of molecular size charge, and primary sequence, we conclude that the actin-comigrating band represents lymphocyte actin. The appearance of lymphocyte actin as a nonspecific "contaminant" of immune precipitates has also been discussed.


Activation of the classical complement pathway by nephritic factor bound to the alternative pathway C3/C5 convertase.

Nephritic Factor (NF), the potent alternative pathway activator, which is occasionally found in association with certain types of nephritis has recently been identified as an IgG class autoantibody specific for the C3 convertase (C3bB) of the alternative pathway. In these studies we have examined the possibility that the cell-bound NF-stabilized C3 convertase (EC3 bBNF) binds and activates the first component of the classical pathway of complement. EC3bBNF bound C1q, and the extent of binding was dependent upon the number of NF molecules bound per cell and decreased parallel to the dissociation and release of NF from the cells. Interaction of C1 with bound NF resulted in its activation as shown by the proteolytic conversion of proenzyme C1s to its activated form C1s. As was the case with C1q binding, C1 activation was dependent on the number of NF molecules bound per cell. Thus the NF-stabilized C3 convertase binds and activates C1.


High responder rabbits to SRBC, a familial incidence.

Three litters of rabbits from our closed colony when immunized with multiple i.v. injections of SRBC gave unusually high titers compared to other rabbits immunized in a similar fashion. Data suggest that this response is a familial occurrence. The antisera from the high responding rabbits also exhibited other unique properties in addition to the high agglutinin titers. The early bleedings had significantly higher amounts of IgG anti-SRBC antibodies but similar amounts of IgM anti-SRBC antibodies relative to normal rabbits immunized with SRBC. The IgG antibodies from the high responders were low avidity antibodies and may exhibit limited heterogeneity.


Spontaneous cytotoxicity. III. Inhibition of human monocyte mediated cytotoxicity by pokeweed mitogen.

We have shown that human mononuclear cells become spontaneously cytotoxic to a variety of erythrocyte targets after 7 days of in vitro culture. This phenomenon of spontaneous cytotoxicity occurs in low concentration of either autologous serum of FCS and is independent of any known stimulant. It does not require exogenous antibody, mitogen, or antigen. Once expressed, the cytotoxic cell has little, if any, specificity. Monocytes appear to be responsible for this observed in vitro cytotoxicity. We now present evidence that the development of spontaneous monocyte-mediated cytotoxicity in vitro is completely reversed by the addition of optimally mitogenic doses of PWM. PWM appears to reverse monocyte-mediated killing by stimulating a potent cell-mediated suppressor. This suppressor cell is radiosensitive and its action is blocked by inhibitors of protein synthesis. In contrast to other mitogen-driven suppressor models, PWM will inhibit spontaneous monocyte-mediated cytotoxicity even when added as late as 6 hr before the end of a 7-day culture.


Physical association of histocompatibility antigens and tumor-associated antigens on the surface of murine lymphoma cells.

Serologic and immunochemical techniques were used to characterize plasma membrane antigens of the murine lymphoma 6C3HED. Syngeneic C3H/HeJ anti-6C3HED antisera were prepared and were shown by cytotoxicity, absorption, and immunoprecipitation tests to be specifically reactive with the lymphoma cells. Alloantisera to H-2.23 were found to be unreactive with intact lymphoma cells by cytotoxicity and absorption assays. However, H-2.23 antigens were detectable in tumor cell lysates by inhibition of cytotoxicity and immunoprecipitation. SDS-PAGE electropherograms of immunoprecipitates of radiolabeled tumor cell extracts obtained with anti-6C3HED and anti-H2.23 antisera were essentially identical; proteins with Mr of approximately 70,000, 45,000, and 12,000 were apparent in both. An immuno-adsorbent of insolubilized anti-6C3HED removed H-2.23 antigens detectable in lymphoma extracts, but had no effect on H-2.23 antigens in normal splenocyte extracts. These data indicate that H-2.23 and TAA are physically associated on 6C3HED cells.


Mouse antibody-dependent eosinophil and macrophage adherence and damage to schistosomula of Schistosoma mansoni.

The effects of sera from S. mansoni-infected mice together with eosinophil- and macrophage-rich peritoneal exudate cells (PEC) on the schistosomula were examined in vitro. Adherence to and killing of the parasite by PEC is mediated through a complement-independent opsonizing IgG. Destruction of schistosomula was assessed morphologically and by evaluating their infectivity to normal mice. By using ferritin-labeled anti-mouse immunoglobulin, the antibody was localized on the surface of the schistosomula and at the cell-organism areas of contact. The serum antibody activity appears with the onset of egg production, does not correlate well with the duration of infection, and is relatively specific among schistosome species. Eosinophils and macrophages were demonstrated by electron microscopy to adhere firmly to the surface of the organisms; the latter cells were shown invading the schistosomula. Furthermore, depletion of eosinophils by prior incubation of PEC with monospecific anti-eosinophil serum blocked the early adherence phase, but a substantial part of PEC ability to kill the schistosomula was retained.


The expression of the allelic gene of murine C3 in fetal and neonatal mice.

A single co-dominant gene locus linked to H-2, tentatively designated C3-1 locus by us, controls allotypic variation of murine complement (C) C3 as previously described. We used this genetically determined variation of murine C3 as a marker in the probe for the synthesis of this C component in fetal and neonatal mice. Phenotypes of murine C3 were determined by a combined use of analytical isoelectric focusing and immunofixation and by antigenic analysis with alloantiserum directed to the gene product of one of the alleles at the C3-1 locus. Fetuses and neonates of inbred mice (BALB/c and NC strains), F1 hybrids, and backcross progeny express the same C3 phenotype as the one observed in adult mice of the corresponding C3 genotype. No evidence for the occurrence of "fetal C3" in fetal mice was obtained. Furthermore, allotypic differences of C3 between mother and fetus or neonates provide evidence that murine C3 is synthesized by fetus or neonate and is not transferred from mother transplacentally nor via colostrum. In summary, mice synthesize C3 according to the allelic structural gene(s) at the C3-1 locus even during their early developmental life.


Electrical measurements of complement-mediated membrane damage in cultured nerve and muscle cells.

Cultured neuroblastoma-glioma hybrid cells and primary mouse and rat muscle cells were studied by using intracellular microelectrodes to monitor membrane electrical potential and resistance changes during complement-mediated lysis. The cell membrane was TNP modified under mild conditions and subsequently coated with rabbit IgG anti-TNP, with no electrical changes observed. However, upon addition of guinea pig C the membrane potential dropped from approximately -50 mv to less than -12 mv within a few minutes, with parallel decreases in electrical resistance. Ten to 60 min after these electrical changes the cells became stainable with trypan blue. No electrical changes or trypan blue staining was observed in the absence of antibody or with heat-inactivated C. With more dilute C so that only a fraction of cells became trypan blue positive, all cells nevertheless showed the membrane electrical changes; surviving cells recovered their original membrane properties within 1 hr. Thus the C-mediated damage to the membrane measured electrically is not in itself sufficient to ensure the subsequent death of the cell. The early electrical changes observed appear to be comparable to increases in 86Rb efflux measured by others.


Lactoperoxidase catalyzed radioiodination of cell surface immunoglobulin: incorporated radioactivity may not reflect relative cell surface Ig density.

Rabbit and mouse splenic lymphocytes were radioiodinated by the lactoperoxidase technique, extracted with non-ionic detergent, immunoprecipitated with high titered rabbit anti-kappa antisera, and compared by SDS-PAGE. Mouse sIg peaks were reproducibly larger in size than rabbit sIg peaks (often greater than 10 times). Neither differences in incorporation of label into the rabbit cell surface, nor differences in average sIg density explain this result. Total TCA-precipitable radioactivity was similar in each species. Estimation of the relative amounts of sIg in the mouse and rabbit showed similar average sIg densities. Differences in detergent solubility, proteolytic lability, or antisera used also do not adequately account for this difference. Thus, these data indicate that radioactivity incorporated after lactoperoxidase catalyzed cell surface radioiodination may not reflect cell surface Ig density. Conclusions about cell surface density based upon relative incorporation of radioactivity should be confirmed by other approaches.


Mouse C4 (Ss): three-step purification of its C4c fragment and production of a monospecific antiserum.

We report here that a large fragment of mouse C4 (C4c) can be easily purified from serum with good recoveries, and used to produce a monospecific antiserum. The method of isolation of C4c is based on the observation that C4 is easily activated and fragmented in mouse serum. The fragments bind to the C4-binding protein (C4-bp), a newly described macromolecular component of the complement system. The high m.w. complexes were precipitated by dialysis of the serum against 0.1 M acetate buffer, pH 5.0, and purified by passage of the dissolved precipitate through a Sephadex G-200 column. The complexes were dissociated at high salt concentration, and the C4 fragments were isolated by passage of the mixture through a second Sephadex G-200 column. One of the C4 fragments (C4c) was used to immunize rabbits, and a monospecific antiserum was obtained, which showed a reaction of identity between C4 and Slp. Therefore the C4c fragment of C4 and the Slp protein are structurally related.


The use of staphylococcal protein A in the mixed agglutination test.

A mixed agglutination (MA) test employing staphylococcal protein A (SpA) instead of an antiglobulin reagent has been developed for the detection of IgG antibodies bound to cell-surface antigens. In the SpA MA test, a single indicator system may be used for the detection of IgG antibodies from several mammalian species. The sensitivity of this test compares favorably with that of the conventional MA test in the detection of most mammalian IgG. The main advantage of the conventional SpA MA test is its usefulness in the study of antigens on cells because of adherence of antibody-coated indicator erythrocytes.


Immunoassay for serum thyroxine monitored by chemiluminescence.

An immunoassay for thyroxine (T4) monitored by chemiluminescence was evaluated with clinical serums. A thyroxine-label conjugate (T4-L) and serum samples were applied sequentially to alkaline Sephadex G-25 columns which adsorbed the thyroxine species. Other serum components and potential interferents were washed from the column with barbital buffer. Subsequently addition of antibody initiated the binding reaction. After 1 h incubation, the antibody was eluted from the column with a barbital buffer wash. The bound T4-L in the eluate was oxidized in a chemiluminescent detection reaction. The peak light intensity, attained in about 1 sec, was related to T4 concentration by means of standards. The intra-assay precision of the chemiluminescence immunoassay was +/-5% (C.V.). Statistical comparison of T4 levels determined for 28 serums by this method and a reference assay was acceptable (y = 0.95x + 5.9, r = 0.98, Sy square root of y . 100 = 13.1%).


Effect of retinoids on the differentiation of chick embryo metatarsal skin explants.

Twelve retinoids were evaluated in organ culture for activity in modulating epithelial differentiation of metatarsal skin explants from 13-day chick embryos. The epithelium differentiated into a squamous, keratinizing epidermis; but, in the presence of active retinoids, keratinization was inhibited, and a mucous metaplasia developed. The methyl-keto and 1-methoxyethyl cyclopentenyl analogs of retinoic acid were about tenfold more effective than retinoic acid in altering epithelial differentiation. The dichlorophenyl analog exhibited about the same activity as retinoic acid. The following analogs were one-half to one-third as effective as retinoic acid in inhibiting keratinization: the chlorotrimethylphenyl analog of retinoic acid and the 13-cis, 10-fluoro analog of trimethylmethoxyphenyl methyl retinoate. The other 7 retinoids were essentially not active at the concentration tested (1.4--2.0 x 10(-5) M). The activity of synthetic retinoids in altering epithelial differentiation may be related to their ability to affect or treat epithelial lesions provided that modification of the retinoid molecule can enhance its activity and decrease toxicity.


Determining skin thickness with pulsed ultra sound.

A high frequency, high resolution ultrasonic echo technique is presented for determining the thickness of human skin. An experimental series is described in which the accuracy of this new technique is demonstrated by comparison with a radiological method of proven accuracy. The "ultrasonic biometric ruler" is shown to provide an accurate, simple, noninvasive method for measuring full-thickness human skin. In addition to the determination of skin thickness, it is demonstrated that the underlying subcutaneous fat and muscle can also be noninvasively "explored" with the possibility of identifying a variety of skin and underlying tissue lesions.


Experimental inoculation of dermatophytes on psoriatic skin.

Two dermatophytic agents, Trichophyton rubrum and Trichophyton mentagrophytes were used in the experimental inoculation of psoriatic lesions in 27 volunteer patients (13 males and 14 females with an age range from 17 to 72 yr). T. rubrum was inoculated in 14 patients and T. mentagrophytes (varietas granulare) in 13. For purposes of control in each patient the same agent was also inoculated on healthy skin. Results were noted once a week for a period of 1 mo. A positive response was found in 12 of the 14 patients inoculated with T. rubrum but only in 4 of the 13 inoculated with T. mentagrophytes. Healthy skin was found to have a positive response in only 5 cases, in 4 with T. rubrum, and in 1 with T. mentagrophytes. These results confirm the possibility of invasion of psoriatic lesions by dermatophytes and suggest that the coexistance of psoriasis and dermatophytic infection may be not always purely coincidental. The importance of the awareness of the presence of mycotic lesions and their possible spread with the application of steroid ointments under occlusive dressings in a psoriatic patient is stressed.


Penetration of chromium in intact human skin in vivo.

Graphite furnace atomic absorption spectroscopy was modified to permit analysis of solid material, thereby fully utilizing the high sensitivity of the method. Tangential sections of biopsies from skin tested epicutaneously with K2Cr2O7 were analyzed for the content of chromium 5, 24, and 72 hr after application of the test substance in various concentrations, either in petrolatum or in water. The concentration of chromium as a rule showed 2 maxima, one at the dermal-epidermal junction and the other in upper mid-dermis. The content of chromium was of the same order of magnitude at all 3 times after application. This indicates that a steady state is reached already after 5 hr and that penetration is continuing at 72 hr. The chromium content in normal skin was 0.4-2 ppm. K2Cr2O7 in petrolatum penetrated to a greater extent than in water. The relative penetration of chromium was lower for 0.5% concentration than for 0.05%. A complex of chromium and glycine was found to penetrate into the skin but to a lower extent than K2Cr2O7. It also elicited a positive epicutaneous test reaction in 1 out of 2 individuals hypersensitive to chromium.


Transfer of paraphenylenediamine delayed-type hypersensitivity: a comparative investigation in the guinea pig, using arteriovenous cross-transfusion and parabiosis.

Arteriovenous cross-transfusion and parabiosis were used to study transfer of paraphenylenediamine (PPDA) hypersensitivity in guinea pigs. Nine out of 10 transfusion experiments were successful, resulting in definite positive test reactions in the recipient animals, and the remaining experiment was evaluated as possibly positive. Six out of 10 parabiosis experiments were successful, a further 2 were negative and the remaining 2 were evaluated as possibly positive. The mean test reactions in the recipients were stronger in the transfusion experiments than in the parabiosis experiments (0.001 less than P less than 0.01 for 0.5% PPDA). In control experiments, where the "donors" were nonsensitized guinea pigs negative test reactions were obtained. It was concluded that the transfusion technique was the most reliable of the 2 methods, when studying transfer of PPDA hypersensitivity.


Identification of a novel cell type in peripheral lymphoid organs of mice. V. Purification of spleen dendritic cells, new surface markers, and maintenance in vitro.

Dendritic cells (DCs; 1) have been purified from mouse spleen in good yield. Spleen cell suspensions were floated on dense bovine plasma albumin (BPA) columns, and the low density fraction was adhered to glass (2). The adherent cells consisted of DCs and immature macrophages most of which eluted in a viable state from the culture dish after overnight incubation. The macrophages were then removed by selective rosetting with opsonized erythrocytes and recentrifugation on dense BPA. This protocol resulted in a purified DC fraction, containing 1--3 X 10(5) DCs/spleen, which was homogeneous and distinctive in its properties. All cells exhibited the phase contrast and transmission electron microscopy (EM) cytologic features that were previously described for freshly isolated adherent DCs. By scanning EM, most purified DCs exhibited a remarkable array of bulbous protrusions of varying length and shape, unlike any other lymphoid cell. All DCs expressed surface Ia and other major histocompatibility complex (MHC)-linked alloantigens. DCs, however, lacked surface Ig and T-cell antigens, and did not bind or interiorize opsonized erythrocytes. Purified DCs have been maintined in vitro for 3 days. Recovery of cultured purified cells was 70% or more of starting cell numbers. When [3H]uridine-tagged DCs were mixed with nonlabeled heterogeneous spleen cells, 70--80% of the labeled DCs were recovered as viable cells 2--3 days later. Purified DCs did not readhere to tissue culture surfaces and did not proliferate, even when cultured with mitogenic doses of concanavalin A and lipopolysaccharide. Finally, DCs did not change their cytologic or surface properties after 3 days of culture. These observations extend the evidence that DCs are a novel cell type and provide useful properties and techniques for their further study.


Role of marrow-derived monocytes and mesangial cells in removal of immune complexes from renal glomeruli.

Phagocytosis of intravenously administered immune complexes by cells in the mesangium was investigated. The model used was that of exchange marrow transplantation between Chediak-Higashi (CH) mice and syngeneic partners after X-irradiation. This model was chosen since marrow-derived macrophages could be differentiated from resident mesangial cells by the presence of the characteristic giant lysosomes in phagocytic cells of the CH mice. Injected immune complexes were cleared normally and localized in the glomerular mesangium in CH or C57BL/6J mice receiving either C57BL/6J or CH marrow. C57BL/6J mice with CH marrow injected with immune complexes prepared with reduced and alkylated antibodies accumulated many cells within the mesangium that contained both giant lysosomes and electron dense deposits. Deposits were not found in cells with subplasmalemmal microfilaments and perpheral dense bodies. Conversely, the cells in the mesangium of CH mice with C57BL/6J marrow that contained electron dense deposits were devoid of giant lysosomes. Based on these observations, we concluded that (a) marrow-derived monocytes contribute to mesangial hypercellularity after deposition of immune complexes and (b) phagocytosis of immune complexes localized in the glomerular mesangium was by marrow-derived monocytes rather than by mesangial cells.


RNA and protein synthesis in human peripheral blood polymorphonuclear leukocytes.

Polymorphonuclear leukocytes purified from human peripheral blood synthesized RNA and proteins when placed in cell culture. Autoradiography of the cultured cells revealed that a majority of mature PMNs were engaged in macromolecule synthesis, and an analysis of newly synthesized proteins by SDS-polyacrylamide gel electrophoresis showed that many different polypeptide chains were synthesized by these cells. The rate of [3H]uridine incorporation and the pattern of newly synthesized proteins were modulated by Con A and glucocorticoids. These results suggest that in spite of their short lifetime and a large performed enzymatic apparatus, mature PMNs retain a substantial capacity for RNA and protein synthesis; and, further, that modulation of macromolecule synthesis forms part of the mechanism by which PMNs respond to inflammatory and anti-flammatory stimuli.


Differential response of early erythropoietic and granulopoietic progenitors to dexamethasone and cortisone.

The sensitivity of erythropoietic (BFU-E) and granulopoietic (CFU-C) progenitor cells to dexamethasone and cortisone was studied in cultures of mouse bone marrow. Although the log dose-response relationships had a similar form, the BFU-E were much more sensitive than the CFU-C to either glucocorticoid. The dexamethasone concentration for 50% inhibition was 3 X 10)-9) M for BFU-E and 60 X 10(-9) M for CFU-C. The differential sensitivity to cortisone was even greater, with 60% inhibition of BFU-E and 18% inhibition of CFU-C at 0.1 microgram/ml. These findings suggest a specific rather than a general response to glucocorticoids and indicate that granulocyte-macrophage progenitors are less affected than early erythroid progenitors by physiologic concentrations of these hormones.


Persistence and pathogenicity of defective Friend spleen focus-forming virus. Decreased transplantability of hemopoietic cells as a marker for preleukemic change.

A latent form of persistent infection can be established in susceptible adult mice inoculated with a preparation of defective Friend spleen focus-forming virus (SFFV) purified free from standard leukemia-inducing helper virus (LLV-F). SFFV persistence was initially observed using an in vivo rescue technique in which SFFV could be directly rescued to form splenic foci of malignant erythropoiesis in mice. At approximately 30 d after virus inoculation however, SFFV could not be rescued after inoculation of LLV-F indicating that persistently infected (i.e., SFFV+) mice were either immume to exogenous helper virus or able to express SFFV-associated defective-interfering (DI) function(s). Persistent infection by SFFV was further documented using an in vitro rescue technique and ultimately resulted in the induction by SFFV of erythroleukemia in the absence of polycythemia or overt virus production. However, SFFV rescued by LLV-F from persistently infected normal and transformed hemopoietic cells was able to induce polycythemia in adult mice suggesting that this is a helper controlled property of the Friend virus complex. Transplantable SFFV-induced erythroleukemic cells could be retrieved from persistently infected yet histologically normal mice. The duration of SFFV persistence in normal spleen tissue suggests that the SFFV provirus resides in either a long-lived or pluripotent hemopoietic cell. Further, certain changes occurred, presumably in the membranes of persistently infected cells, which preceded the overt development of Friend leukemia and facilitated the definition of an SFFV preleukemic phase. Cell surface alterations were revealed using cell transfer techniques. Hemopoietic cells harboring a rescuable SFFV failed to proliferate when inoculated into lethally irradiated, syngeneic adult mice. In contrast, the transformed progeny of preleukemic cell populations and spleen cells transformed by FV complex (i.e., cells replicating both SFFV and LLV-F) were not rejected. This result suggests that histologically normal SFFV+ preleukemic cells express an antigen recognition site which is not present on overtly transformed cells and which may be a pertinent surveillance target for host anti-leukemogenic reactions.


C5b-9 dimer: isolation from complement lysed cells and ultrastructural identification with complement-dependent membrane lesions.

The membrane attack complex (MAC) of complement was extracted from the membranes of cells lysed by human complement and its properties were compared with those of the fluid phase complex SC5b-9. Upon sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunochemical analysis, the two isolated complexes had identical subunit compositions, except that the MAC lacked the S-protein. The sedimentation coefficient and molecular weight of the extracted and isolated MAC were, respectively, 33.5 S and 1.7 x 10(6) daltons, compared to 23 S and 1.0 x 10(6) dalton for SC5b-9. Because the molecular weight of the MAC is approximately two times greater than that of C5b-0 (800,000 daltons), the MAC is considered the dimer of C5b-9. Under specified conditions, the 33.5 S dimer could be converted to the 23 S monomer without dissociation of subunits. The MAC had the electron microscopic appearance and dimensions that are characteristic for the complement produced ultrastructural membrane lesions. SC5b-9 had a different ultrastructure that is dissimilar to the morphology of the lesions. The isolated MAC could be reincorporated into phospholipid bilayers and assumed on the surface of the resultant lipid vesicles the orientation and appearance of typical complement lesions.


Hyaluronidase-sensitive halos around adherent cells. Their role in blocking lymphocyte-mediated cytolysis.

A variety of adherent sarcoma, carcinoma and normal cells are surrounded in vitro by thick, transparent zones (approximately equal to 9 micron thick) that spleen cells and a variety of other cells and particles cannot penetrate. Seven lymphoblastoid cell lines did not possess such halos. The presence of these halos around adherent fibrosarcoma cells appeared to protect them from lymphocyte-mediated cytolysis. Hyaluronidase treatment, which destroyed the halo and allowed lymphocytes to approach the tumor cell membrane, enhanced the cytotoxic action of immune but not of normal spleen cells. These observations, in addition to highlighting a little-known feature of the cell surface, may also be of general relevance to the in vitro and in vivo killing of tumor cells by immune effector cells.


Regulation of IgE antibody production by serum molecules. III. Induction of suppressive activity by allogeneic lymphoid cell interactions and suppression of IgE synthesis by the allogeneic effect.

Antibody responses of the IgE class are, like other immunoglobulin classes, regulated by a finely-tuned network of complex cellular and molecular interactions (1). Previous studies conducted in our laboratory (2, 3) have provided new insights into the differences in control mechanisms that result in individuals manifesting either the high (allergic) or low (nonallergic) IgE responder phenotype. These experiments have shown that certain manipulations (i.e. low dose X-irradiation) convert normally low responder mice to high IgE responders, apparently by diminishing a suppressor T-cell mechanism which normally dampens, rather selectively, IgE antibody production in such individuals. Similar findings have been made by Watanabe et al. (4). Recently, we have been studying the types of manipulations that could reverse the high IgE responsive state back to a low one. These studies (2, 3, 5, 6) have demonstrated that the high IgE responses induced in low responder mice can be substantially diminished, and even abolished, by passively transfusing serum or ascetic fluid from donor mice previously inoculated with mycobacterial-containing complete Freund's adjuvant (CFA). Because the suppressive activity of CFA-immune serum or ascitic fluid is so highly selective for IgE antibody production, we have recently termed these serum substances suppressive factors of allergy (SFA) (2, 3). The present study was undertaken to determine whether alternative means, particularly those that avoid administration of CFA, could be devised for the induction of SFA. Herein, we report the effectiveness of allogeneic lymphoid cell interactions in inducing SFA, both in vivo and in vitro, as well as the potent suppressive effects of an in vivo allogeneic effect on irradiation enhanced IgE antibody production in low responder mice.


Distribution of lymphocytes identified by surface markers in murine strains with systemic lupus erythematosus-like syndromes.

The frequencies and absolute numbers of B and T cells in the lymphoid organs of five murine strains (NZB, (NZB X NZW)F1, BXSB, MRL/l, and MRL/n) with SLE-like syndromes were examined. We assessed the frequencies of cells bearing surface Ig, C3d and IgG Fc receptors, and theta-antigen. The sequential expression of Ig isotopes on developing B cells and the Ig isotypes expressed on adult B cells were ascertained. In addition, the Ly subsets and the expression of Ia antigens coded for by the I-J subregion of the mouse H-2 complex were examined. Compared to normal, older mice, New Zealand mice had low frequencies and absolute numbers of B cells, BXSB mice had a moderate B-cell proliferation, and MRL/l mice had normal absolute numbers of B cells but a reduced frequency concomitant with a massive T-cell proliferation. Old New Zealand mice and BXSB mice had reduced frequencies and absolute numbers of T cells compared to old controls. The developmental Ig-isotype diversity during the 1st wk of age was similar in normal mice and those with autoimmune manifestations. Mature B cells were present in lymphoid organs of New Zealand mice and BXSB mice as evidenced by the high frequency of C3d receptor-bearing cells and Ig-isotype expression (high ratio of IgM- to IgD-bearing cells) in adult spleen cells. Numbers of IgG Fc receptor-bearing cells were reduced in autoimmune mice with advanced age and disease. The proliferating T cells in MRL/l mice were found to be theta-antigen positive but Ly null. These theta+-, Ly null cells may have arisen from Ly123+ T cells. MRL/l and BXSB mice seemed normal in their content of T cells bearing Ia antigens coded for by the I-J subregion of H-2. Overall, mice with autoimmune manifestations appear to express perturbations in T and B cells with development of disease, and their patterns of change vary from one strain to another.


Study of sample collection and preparation methods for multi element analysis in liver tissue by proton induced X-ray emission (PIXE).

Large samples of tissue and biopsy specimen were taken from a single human liver, homogenized or paraffin-embedded, then analyzed by PIXE. Homogenates, including those of biopsies, proved to be suitable for PIXE-analysis of P, S, Ca, Mn, Fe, Cu, Zn, Se, Rb, Mo, Cd and, to a limited extent, Pb. Paraffin-embedding leads to a distinct elution of K, Rb, Cl and to a lesser elution of P, Zn, Se, Br and Mo. The contents of Ca and Sr, on the other hand, were higher in the paraffin-embedded sections. Sections of paraffin-embedded biopsies proved to be hardly suitable for PIXE-analysis.


Variation of trace element contents in a single human liver.

Multi element analysis of homogenised samples taken from three different areas of a single human liver was performed for 18 elements by proton induced X-ray emission (PIXE). Small (10--20%) but statistically significant variations between the element contents of these areas were found for Cl, K, Ca, Fe, Cu, Zn, Br, Rb and Mo. These differences are not parallel to each other. The variation of element content within one area of this single liver did not show any distinct correlation between different elements, nor did it significantly exceed the methodical errors of 3--10% except for lead. Thus we conclude from our measurements that the trace element analysis of small liver samples by PIXE is, with the exception of lead, fairly representative for the whole organ.


Immuno-chemical studies on the alkali-labile carbohydrate chains of human serum glycoproteins.

Human serum glycoproteins can be classified into those containing N-acetyl-D-galactosamine and into those lacking this hexosamine. The N-acetyl-D-galactosamine-containing serum glycoproteins have alkali-labile chains containing this hexosamine linked O-glycosidically to hydroxy amino acids. These alkali-labile chains can be demonstrated in neuraminic acid free serum glycoproteins by gas liquid chromatography and by using precipitating lectins from invertebrates and plants. They are represented by two chains, one containing only N-acetyl-D-galactosamine, the other with D-galactose linked (1--3) beta-glycosidically to this hexosamine forming a disaccharide. Serologically these two chains, which usually occur together on one molecule, can be characterized by their reaction with lectins from Helix pomatia (anti-A like) and from Agaricus bisporus and Arachis hypogaea (anti-TF specificity).


[Simultaneous determination of antiepileptic drugs in clinical laboratories: Problems of quality control and standardization (author's transl)].

Methods for determination of antiepileptic drugs in clinical laboratories are discussed with respect to precision, constant error control and economy. Experimental data for reproducibility and standardization are presented for gas chromatographic and photometric methods. Methods for choosing the most suitable procedure for any of the common analytical tasks that may arise in the clinical laboratory are recommended.


[Creatine kinase and creatine kinase isoenzyme MB: determination of normal values and values for myocardial infarct, using the new, optimized method with N-acetyl cysteine as the activator (author's transl)].

Preliminary results are reported for the determination of creatine kinase and its isoenzyme MB in healthy individuals and in patients with acute, transmural myocardial infarct, using N-acetyl cysteine as the activator of the enzyme. In healthy individuals, the upper normal limit for creatine kinase activity was 54 U/l; the decision level for the presence of creatine kinase MB activity 9 U/l. The limiting values were only slightly different from those for the GSH-activated creatine kinase, and the results with the old and new methods showed a linear relationship with a slope of 1.0. The differences in the activation of creatine kinase MM by GSH and by N-acetyl cysteine are discussed; the activities obtained for creatine kinase and creatine kinase-MB by the GSH-activation method cannot simply be recalculated to give the appropriate values of the N-acetyl cysteine activation method.