Development and application of the extended scale for dementia.

As part of an interdisciplinary study of organic dementia, a psychologic test for assessing the degree of dementia--the Extended Scale for Dementia--was developed through the expansion and rescoring of the original Mattis Dementia Scale. Statistical analyses of the 23 test items resulted in a scoring scheme which includes the "weighting" of items for scoring purposes. The test was successfully administered to 90 subjects from 6 hospitals in the London (Ontario) region. With use of the Extended Scale, it was possible to discriminate between dementia and non-dementia groups of psychogeriatric inpatients and to correlate the findings closely with those of another measure of the degree of dementia, viz, the London Psychogeriatric Rating Scale (Ment.). Dementia patients who were retested after 6-month and 12-month intervals showed a significant decline in scores. No significant scoring differences were noted between males and females or between Alzheimer dementia and multi-infarct dementia.


Counseling the isolated elderly.

Increasing interest is manifest in the development of alternative models of counseling for older persons. Irrespective of the particular model used, the counseling of aged persons who have emotional and psychologic problems may require a treatment plan for health, mental health, and social services, to resolve the problems and to assess the functional limitations. Health and mental health centers employing both professional and nonprofessional counselors need to determine the value of adding outreach components to their services, and agencies which already have outreach programs may need to determine their relevance and effectiveness. Effective outreach requires a high level of gerontologic knowledge and clinical skills with aged patients. Agencies and clinics can design and implement mechanisms for supporting the elderly as they make use of multiple sources of service. Mobilization of the family and other informal helping networks in the community, the establishment of better coordination mechanisms within clinic and service programs, and multiple-service integration will improve the efficiency and effectiveness of long-term community care and follow-up programs.


Cultural and sex differences in dietary patterns of the urban elderly.

A group of 182 elderly subjects (91 females, 91 males; 94 blacks, 88 whites) were interviewed with respect to background data, dietary habits, and health variables. Analysis of variance showed race and sex differences with respect to meal patterns and quality of diet. In addition, correlational analyses revealed associations between quality of diet and social class, education, health status, and the smoking of cigarettes. Associations were strongest for the black subjects.


Sensory deprivation and homeostasis.

Most afferent inputs are sensory stimulants, and physical activity is probably one of the most significant of all sensory stimulations. Gravity is a constant stimulus. These demands are also known as stress, and the stressor effect as the general adaptation syndrome (Selye). In this study, these demands are called sensory afferent stimuli, which are necessary triggers to start the hypothalamus-pituitary-adrenal mechanism and thus maintain the general mechanical, autonomic and metabolic functions of the body in the state of homeostasis. If the sensory stimulus is removed, there is no stressor effect and the cycle of response is eliminated. This results in a decreased hypothalamus-pituitary response and a decreased production of ACTH and corticoids. A decrease in the sensory input creates changes in the central nervous system and in the musculoskeletal system which are compatible with lack of the hypothalamus-pituitary responses. When a person becomes ill, physical inactivity as well as sensory and perceptual deprivation are at a maximum, and the absence of these stimuli interrupts the cycle of homeostasis. Physical activity is one of the most important stressors (sensory input) or triggering mechanisms for the cycle of homeostasis (hypothalamus-pituitary-adrenal cycle). Therefore, it is imperative to provide the patient with physical activities which ensure motion and take into account the effect of gravity. This is best accomplished by a program of physical, occupational and recreational therapy.


Incorporation of gerontology into medical education.

Despite the increasing importance of gerontology to medical practice, it has been difficult to incorporate it into the curriculum of most medical schools. The aim of this project was to explore the feasibility of introducing gerontology into medical education by utilizing the framework of an existing clinical program. A gerontologist used the structure and resources of a Psychiatry Consultation-Liaison Program for this purpose. Three groups of target trainees were identified: psychiatry residents on their Liaison rotation; junior medical students on their psychiatry clerkship; and medical-surgical house officers in the general hospital. The objectives were to impart didactic information and to counteract negative attitudes leading to disinterest in evaluating and treating the elderly. The results indicated a positive impact on all three trainee groups, and a diffusion of the gerontologic innovation from the initial locus of a clinical sub-unit into the medical center and the medical school.


Drug use in a geriatric population.

Drug usage was studied in a geriatric hypertension-screening program. A questionnaire was used to collect the required information from patients visiting the clinic during three successive years. The study included 1,711 patients, of whom 76.6 percent were regularly using a drug preparation. A consistent increase was noted in the average number of drug categories used with increasing age, from 1.6 in patients under 70 to 2.6 in patients over 84 years old. The most common drug categories involved were antihypertensive agents, cardiovascular drugs, vitamins, and internal analgesics.


Bronchoextrapleural fistula after extrapleural pneumonolysis.

This case illustrates the development of a bronchoextrapleural fistula 40 years after extrapleural pneumonolysis with placement of a paraffin pack. The various diagnostic possibilities in the presence of such a radiographic presentation are reviewed.


Erythrocyte sedimentation rate and disease in the elderly.

The erythrocyte sedimentation rate (ESR) is not always elevated in relation to disease. Of 188 elderly autopsy patients studied, 125 had nonmalignant disease and 63 had malignant disease. Of the patients with malignant disease, 13 percent had a clinical ESR of less than 10 mm/hr and 26 percent a rate of less than 20 mm/hr, the upper level of normal for the elderly. Of the patients with nonmalignant disease, 9.6 percent had an ESR of less than 10 mm/hr and 25.6 percent a rate of less than 20 mm/hr. Thus, in about a quarter of the patients, the ESR was below the top normal level for the elderly. Moreover, the ESR may be as high as 35--40 mm/hr in healthy aged persons. Therefore, the ESR is unreliable as a test for the presence of disease (malignant or nonmalignant) in the elderly.


Intraocular lenses and their follow-up in the optometric office.

A broad overview of intraocular lens implantation is presented for the optometrist. Included in this overview is a review of the most recent literature comparing complication rates for lens implants versus standard cataract surgery. A proportion of the paper addresses itself to the pre and post-operative optometric management of the pseudophakic patient. This paper is presented to give the optometrist a basic understanding of the expanding field of intraocular lens implantation as well as a reference list which will serve as a starting point for further review.


The influence of a national health policy on optometry and optometric education in Ontario.

Until recently Canada had little or no direction toward forming a National Health Policy. It was not until 1961 that a government agency served as an impetus in establishing a health care program. With this however, came the challenge for optometry to defend itself against the ophthalmological society in an effort to receive its fair share of recognition. Optometry met this challenge, both in the institutional and practical setting. As a result legislation was passed to establish very positive guidelines for the optometric community including the definition of optometry, peer review, standards of practice, continuing education, and the use of diagnostic drugs. Since then, Canada's Minister of Health issued a statement to elaborate on a number of health related issues. In this 1974 statement he emphasized the value of care should be comparable to that of cure. To meet this need a reemphasis on educational training would have to be met. Optometry has looked toward this policy to plan for its future educational objectives and methods of practice. Because these public policies have been formed in a positive way toward optometry the Canadian optometrist will continue to work toward the goal of being a better practitioner within the scope of this Canadian policy.


The current status of auotomobile running lights.

The results of a number of studies of the use of lights to make a vehicle more visible have shown that they are effective. Accident studies have shown a drop in accidents from 7 to 59%. Only one of the studies used properly designed and positioned presence and signal lights and yielded 59% fewer accidents. It is concluded that a 50% or greater reduction of accidents is probable when using a high mounted 360 degrees visibility signal and running light combination.


Multiple sclerosis: etiology and epidemiology.

Multiple sclerosis (MS) is a demyelinating, auto-immune disease which affect the central nervous system. The research seems to implicate a small or incomplete slow-growing virus as the causative agent. The measles virus has been suspected because of high titers of measles antibodies in MS patients' serum and cerebrospinal fluid. An immunological deficiency in MS patients may be linked to genetic factors such as HL-A tissue types and histocompatibility antigens. Genetics and socioeconomic conditions may also result in the distinct geographical pattern of MS distribution in the world.


Early identification of multiple sclerosis through ocularly manifested symptoms.

Eye care professionals can play an important role in the early detection of multiple sclerosis (MS) since the disease often has ocular manifestations such as optic neuritis. In conjunction with optic neuritis, symptoms such as ocular pain, acuity reduction, visual field loss, blurred or edematous discs, nystagmus, and diplopia may be present. Although MS still has no cure, early identification can aid in beginning the treatment of the symptoms.


Steady state VER measurement of visual system latency in normal and multiple sclerosis patients.

VERs were recorded from 30 normal and two multiple sclerosis (MS) subjects using a steady state checkerboard stimulus. The normal subjects produced a mean OU latency of 90.8 msec with a mean absolute interocular latency difference of 3.2 msec. Latencies from the MS patients were up to 13 standard deviations longer than the latencies from the normal subjects.


A clinical study: plastic lens warpage.

This clinical study of ophthalmic plastic lens warpage reviews prior work and confirms the presence of warpage. However, there were no patient symptoms related to the warpage, especially if within the ANSI Z80.1 standards.


Quality assurance and optometry.

About 120 years ago the first attempt to determine the quality of treatment was made. Today, in the face of increased consumer awareness, third party payments for optometric care, and the advent of National Health Insurance legislations optometry must prepare itself for quality assurance programs. Specific methods are described along with the steps necessary to accomplish the needs of the profession.


Toxocara canis.

Toxocara canis, commonly known as the dog ascrid, is a frequent parasite of the domestic dog. If the eggs of an adult worm are accidentally ingested by a human host, usually a child, a larval form of the worm develops that can survive for a limited period in various human tissues. The retina is one of these tissues and upon the death of this organism a granulomatous reaction occurs and a fibrotic mass is formed in the retina. This unusual ophthalmoscopic appearance is demonstrated by the presentation of three fundus photographs of patients with suspected Toxocara lesions. A knowledge of this parasite and its life cycle will aid in its recognition, treatment, and possible prevention.


A procedure for measuring the near-point fusional vergence reserves of young children.

This paper describes a modification of the standard method for measuring near-point fusional vergence reserves that reduces dependency upon patient's verbal responses by employing a fixation target that can be perceived accurately only when viewed binocularly. As such, the procedure is particularly useful with young children whose subjective responses often tend to be unreliable. Data from 34 preschool children are reported.


Visual imagery and written language.

The importance of visual imagery in optometric diagnosis and therapy is discussed, stressing its relationship to the auditory-verbal system and its effect on reading and spelling. Techniques are presented to aid in determining visual imagery capability as well as in aiding the patient to utilize this ability for analytical-verbal tasks.


Optical identification and traceability potentials.

This brief outline of optical identification potentials alerts law enforcement agencies to the early developments in the field. The present elective system of I.D. classification may become a mandatory program for universal traceability in the future.


The 1979 revision of Z80.1--the American National Standard recommendations for prescription ophthalmic lenses--the AOA position.

Since April, 1972, the American National Standard Requirements for First-Quality Prescription Ophthalmic Lenses, Z80.1-1972, has been in the process of revision. The American National Standards Institute procedures are explained as well as details of the new revised standard. In addition, an explanation of the AOA position on the revision is given. Finally, what this standard means to the practicing optometrists is discussed.


Z80.1 and the state of the art.

Ophthalmic standards such as ANSI Z80.1, 1979 (Recommendation for Prescription Ophthalmic Lenses) are developed by the joint effort of many disciplines from eye care practitioners to suppliers and manufacturers. The tolerances, agreed upon by consensus, are fundamentally controlled by the state of the art, that is upon the quality of product which can be reliably produced by competent personnel using standard methods of processing. Research has demonstrated that the Z80.1, 1972 standard had been set at a level above the state of the art for today's lenses. Some of the tolerances in the 1979 revision were therefore lowered so as to establish a more realistic guideline. The new standard, if diligently applied by both laboratories and dispensers, should lead, however, to an increase and not a reduction in product quality.


The optometrist's new role in the use of corrected curve lenses.

The new role is brought about by new laboratory practices in the use of stock semifinished and uncut lenses in filling orders and in the changes in standards for design requirements. A partial solution to this problem is proposed.


The Commission on Ophthalmic standards--the American Optometric Association's standards development program.

The American Optometric Association has been concerned with ophthalmic standards for over fifty years. This paper describes this involvement and how it has led to the creation of the Commission on Ophthalmic Standards. The commission's scope and function and the responsibilities of the Director of Standards Development are given. The progress of the commission's program has been temporarily delayed due to a proposed FTC rule on voluntary standards setting organizations.


Standards and technology.

Improvements in standards and technology go hand in hand. A careful study of standards as related to the state of the art always brings to light the need for improving technology. The study involved in revising the 1972 ANSI Z80.1 Standard for Ophthalmic Lenses has focused attention on several new and upcoming developments. These relate (1) to the predetermination of the edger setting, and the use of the perimeter of a lens, (2) the accurate control of centers, (3) the measurement of power and axis, and unwanted power in the periphery, (4) the use of computers in generating the shape of a lens and the location of the MRP, and (5) the standardization of shapes.


Tolerance to tolerances: some thoughts on the 1979 American National Standard recommendations for prescription ophthalmic lenses.

The 1979 revision of the ANSI Z80.1 Ophthalmic Lens Standards resulted in rather significant changes affecting three areas of ophthalmic lens requirements: 1) central refractive characteristics, 2) prismatic characterstics, and 3) factors relating to the magnification properties of lenses. Based primarily on what an optical laboratory could consistently produce, the ANSI Ophthalmic Lens Sub-committee decided to widen some of the tolerances over those expressed in previous revisions. This paper attempts to show how these new tolerances relate to certain aspects of patient sensitivity to errors induced by ophthalmic lenses conforming with the new revision.


The dotting lens.

A dotting lens is an auxiliary lens placed in front of the telescope of a focimeter. It images a dot on the center of the lens under test on the scale in the eyepiece. A hole in the dotting lens permits simultaneous meaurement of the sample.


The age of specialization and its future influence on optometry.

As overview of the future role of specialization in optometry is presented, and topics such as the necessity and desirability of optometric specialties, certification of specialists, referrals to specialists, training of specialists, and the impact of public opinion, governmental programs, expanded use of the optometric auxiliaries, and advances in technology on optometric specialization are examined.


The effect of praziquantel on Schistosoma mansoni.

3 x 10(-6)M praziquantel fails to completely paralyse miracidia and cercariae in a short time but they are not infective when maintained in the solution. 3 x 10(-5)M praziquantel prevents infected snails shedding cercariae but does not kill daughter sporocyts or developing cercariae. As the action of praziquantel on adult worms is not blocked by 10(-2)M mecamylamine, pempidine or carbachol, but is reduced by calcium depletion, it is suggested that praziquantel may act by permitting calcium influx to muscle cells causing them to contract.


Intestinal mast cells during experimental ancylostomiasis.

The alterations of mast cell counts induced in the gut of swiss albino mice by infection with Ancylostoma caninum larvae have been studied, both in single and multiple infections. It was found that mast cell rise was greater in females than males. In singly infected animals, mast cell increase was influenced by the infective dose of inoculated larvae. In case of repeatedly infected mice, mast cell counts were decreased because of their earlier degranulation.


Strigeoid trematodes of Malaysia with descriptions of a new genus and three new species.

Six species of strigeoid trematodes are reported from Malaysia. One new genus and 3 new species are described: Apatemon (Apatemon( jamesi sp. n (Strigeidae); cercaria Cotylurus sullivani sp. n. (Strigeidae); Neodiplostomum (Neodiplostomum) sp. (Diplostomatidae); Fibricola ramachandrani (Diplostomatidae); Pseudoscolopacitrema otteri gen. n. et sp. n. (Diplostomatidae); and cercaria Cyathocotyle malayi sp. n. (Cyathocotylidae). The life cycles of A. jamesi and C. malayi have also been investigated.


Dunnifilaria dilli sp. n. (Nematoda: Filarioidea) from Rattus koratensis in Thailand.

Dunnifilaria dilli, a new species of filariid from Rattus koratensis from Thailand is described. Adult worms were recovered from the lymphatic system. They differ from D. ramachandrani in their location in the host, the tail length, the shape of the spicules, and in the number of caudal papillae. Microfilariae are much smaller and unsheathed. Additional hosts include R. rattus and R. sabanus.


Accumulation of peroxidase in the cap rays of Acetabularia during the development of gametangia.

Accumulation of peroxidase was demonstrated by light and electron microscopy to occur in Acetabularia in certain regions of the cap rays in relation to the development of the gametangia (cysts). Peroxidase was found to be incorporated into special, cell wall-like obstructions that separate the cap rays from the stalk when the secondary nuclei have settled in the cap rays. It is assumed that peroxidase acts as an anti-microbial protectant of the gametangia.


Electronic sorting of granulocyte precursor cells from stimulated bone marrow.

A commerical cell sorter was used to obtain preparations of cells in various stages of granulocyte development from rabbit marrows stimulated by inflammatory response. Marrow cells were fractionated on density gradients of Ficoll/Hypaque and each fraction sorted using light scatter. Trial and error selection of appropriate gradient fractions and light scatter windows allowed sorting of early (blast cells, promyelocytes), intermediate (myelocytes, metamyelocytes) and late stage (band cells, polys) granulocytes with enhanced purity.


Epidemic mechanisms of type A influenza.

The antigenic varieties of influenza A virus isolated from 1968 to 1976 in a surveillance of a small, rather remote population were similar to those from England and Wales as a whole, despite frequent antigenic changes during the period. Household studies in the first two H3N2 influenza A epidemics found low attack rates within households, a high proportion (70%) of affected households with only one case of influenza, similar distributions of affected households in the two epidemics by the number of cases of influenza and similar distributions of the influenza cases by the day of their onset in the household outbreak. No serial interval could be demonstrated by cumulating household outbreaks. More than one minor variant was causing influenza contemporaneously in the same villages in several seasons, and different variants were on one occasion found on successive days in bedfellows. The regular occurrence of epidemics in winter was often accompanied by the disappearance of the epidemic variants and their replacement, after a virus-free interval, by new variants. These epidemiological findings seem best interpreted on the following tentative hypothesis. Influenza A sufferers do not transmit the virus during their illness; instead it rapidly becomes latent in their tissues so that they become symptomless carrier-hosts and develop specific immunity. Next season an extraneous seasonally mediated stimulus reactivates the latent virus residues so that the carrier-host becomes briefly infectious, though symptomless. Antigenic drift occurs because particles reconstituted to be identical with the progenitor virus cannot escape the specific immunity it has provoked in the carrier host. He can shed only mutants also determined by the progenitor virus. From the assortment of mutants shed by the carrier-host, his non-immune companions select that (those) which is best fitted to survive, and it rapidly causes influenzal illness. Epidemics consist largely or entirely of such persons sick with influenza caused by reactivated virus caught from symptomless carrier-hosts.


The relative resistance of f2 bacteriophage to inactivation by disinfectants.

f2 bacteriophage in the presence of fetal calf serum (at a final concentration of 10%) was exposed to six commonly used disinfectants for times of 10, 20 and 30 sec. At the end of exposure times skim milk neutralized the disinfectant activity and residual virus was assayed using the plaque technique. The 6 disinfectants considered were Javex, sodium hydroxide, ethanol, Wescodyne, One Stroke Ves-Phene and Sonacide. A 0.25% (w/v) solution of sodium hydroxide and 1/50 Javex (1200 parts/10(6) chlorine) were the most effective of the six disinfectants considered since 10(5) f2 bacteriophage were inactivated in 30 seconds in each instance. Since a 0.25% (w/v) solution of sodium hydroxide had a pH of 12.5 this made it too caustic to use as a disinfectant in many practical situations. It was concluded therefore that Javex at some dilution less than 1/50 (greater than 1200 parts/10(6) chlorine) was the most practical of the six disinfectants to use. Ethanol (95%, v/v) inactivated 10(3) f2 bacteriophage in 30 seconds while 1/20 Wescodyne and undiluted Sonacide inactivated 10(1)-virus particles. Ves-Phene at a dilution of 1/50 was a completely ineffective virucide during the 30 sec exposure. The resistance of f2 bacteriophage to inactivation by these six disinfectants was compared with that of echovirus 11 and coxsackievirus B5. In all instances except exposure to undiluted Sonacide, f2 was comparable in resistance to inactivation and in many cases had greater resistance.


The use of spore strips for monitoring the sterilization of bottled fluids.

A bacterial spore test has been developed which enables the efficacy of the sterilizing cycle recommended by the British Pharmaceutical Codex (1973) for bottled fluids to be accurately monitored. During a 14-month period this test detected faults in 3.3% of the sterilizing cycles, representing five distinct episodes of sterilization failure that passed unnoticed by the conventional controls of physical measurements and sterility testing. There were no failures of sterilization as detected by conventional techniques which were not indicated by the spore test.


The effect of long-term exposure to cigarette smoke on the height and specificity of the secondary immune response to influenza virus in a murine model system.

The effect of long-term exposure to cigarette smoke on the height and specificity of the secondary humoral immune response to influenza was investigated in a murine model system. It was shown that if mice were pre-immunized with a sub-lethal infection of influenza virus and then exposed to cigarette smoke daily for 36 weeks, they were able to mount a secondary immune response of normal height on subsequent challenge with the homologous virus strain. The response however, was less specific than that elicited in control mice, with high titres of cross-reacting antibody by haemagglutination-inhibition to the following strain in the same antigenic series. Recall of antibody to the previous strain in the antigenic series was not observed in either control or smoke-exposed animals. These results serve to correct an earlier discrepancy between the murine system and human studies in which the response to influenza infection in mice was depressed by prolonged exposure to cigarette smoke, whereas in man the response of smokers did not differ significantly from that of non-smokers. This apparent discrepancy had been caused by a lack of previous experience of influenza in the mice, which had therefore mounted a primary response, compared with the secondary response observed in the human studies.


The effect of air ionization on the air-borne transmission of experimental Newcastle disease virus infections in chickens.

The effect of artificial air-ionization on air-borne transmission of Newcastle disease virus (NDV) infection in chickens was studied in an isolated system consisting of two side-by-side cages with solid walls and a wire-gauze roof. During a 3-week observation period more than 90% of the uninoculated indicator chickens, housed in one of the cages, contracted the virus shed to the air by the NDV-inoculated, diseased birds in the neighbouring cage. This air-borne transmission of NDV was completely prevented by increasing the ion concentration in the test room by a constant negative corona discharge above the wire-gauze roof. On the other hand, spreading of the infection within a group of chickens housed in a single cage was not affected by air ionization. These and other results suggest that artificial air-ionization may protect animals from certain air-borne infections by interfering with microbial aerosol formation and/or by facilitating their decay.


IgA antibody response in acute rubella determined by solid-phase radioimmunoassay.

A solid-phase radioimmunoassay (RIA) for detecting rubella virus IgA serum antibodies was developed. Purified rubella virus grown in roller cultures of Vero cells was adsorbed onto polystyrene beads. The coated beads were then incubated with dilutions of serum, and rubella IgA antibodies which attached to the virus antigen on the solid-phase were subsequently detected with 125I-labelled anti-human-alpha antibodies. The specificity of the iodinated anti-human immunoglobulins was confirmed by RIA analysis of fractions obtained by chromatography of an early convalescent serum on an agarose column. A complete separation of IgM, IgA, and IgG was observed. A total of 144 serial serum specimens from 31 adult patients with an acute rubella infection were tested for rubella IgA antibodies, and the results were compared with the RIA IgG and IgM titres reported earlier from the same specimens. The RIA IgA response was detected in each of the 31 patients and the IgA antibodies appeared almost simultaneously with the IgG and IgM antibodies. The maximum titres, which were lower than the IgG and IgM titres, were reached in about 1 week after the onset of rash. In 6 patients out of 31 the IgA antibody response was transient and persisted approximately two months, while in the remaining 25 patients the IgA antibodies persisted throughout the study period of more than 5 months. The results obtained indicate that the presence of rubella IgA antibodies in serum is not an indication for a recent rubella infection.


Estimating the duration of latency and survival time of snails with schistosomiasis.

By means of techniques of analyses of survival data developed for cancer trials it is possible to study aspects of the natural history of the infection of schistosomiasis on the intermediate host of transmission, the snail. The simultaneous study of three response variables is largely based on a model of Lagakos (1976). When using this approach in the schistosomiasis setting it seems inappropriate to assume that one process, the duration of latency, follows an exponential distribution. Thus this stage is modified to follow a normal distribution and the derivatives required to obtain maximum-likelihood estimates and approximate variances of all parameters are provided. Simple graphical tools for assessing the validity of distributional assumptions in survival data are available from industrial research. The reader's attention is drawn to a paper by Nelson (1972). The relevance and application of these methods to the current problem are described in Section 4. In the event that the times to death of prepatent and patent snails do not follow exponential distributions as assumed in the primary model, a further modification is introduced to enable either or both to follow Weibull densities. Lastly it is possible to adapt both the primary model of Section Three and the modified model of Section Five to allow for the inclusion of auxiliary variables or covariates. Again the required derivatives to obtain maximum likelihood estimates and approximate variances are provided.


The alternative pathway C3/C5 convertase: chemical basis of factor B activation.

The structural basis of activation of the alternative pathway C3 convertase was explored. For this purpose a modified isolation procedure of the activating enzyme, Factor D, was elaborated. The procedure affords a 70,000-fold purification of the enzyme with a 20% yield. A simple assay was designed for the quantitation of both Factor D and Factor B activity. On the basis of activity measurements and amino acid analysis, Factor D concentration in plasma was estimated to be 1 microgram/ml. Highly purified Factor D was used to activate Factor B in the presence of C3b and Mg++. The resulting fragments, Ba and Bb, were characterized with respect to their circular dichroism spectra, amino acid compositions, reactive sulfhydryl groups, and partial amino- and carboxy-terminal sequences. The results indicate that the Ba fragment constitutes the amino-terminal region and the Bb fragment the carboxy-terminal region of Factor B. The bond in Factor B that is cleaved by Factor D is proposed to be an arginyl-lysine bond.


Latent lymphokines: isolation of guinea pig latent lymphocyte-derived chemotactic factor for monocytes. Its activation by trypsin and a soluble factor from macrophages.

Guinea pig lymphocytes when depleted of macrophages and stimulated by the T cell mitogen phytohemagglutinin produce a latent form of lymphocyte-derived chemotactic factor for monocytes (LDCF-M). Latent LDCF-M is also produced when immune lymphocytes are stimulated in vitro with specific antigen, horseradish peroxidase. Latent LDCF-M from both sources can be activated and converted to "classical" LDCF-M by trypsin and by a soluble factor obtained from sonicated macrophages. These observations suggest that macrophages may modulate lymphokine activities in vivo by releasing soluble factors that convert inactive latent lymphokines to biologically active substances.


The regulation of thymidine secretion by macrophages.

The secretion of thymidine by mononuclear phagocytes was correlated with the activity of the enzyme thymidine kinase (TK). Macrophages cultured in regular tissue culture medium released thymidine and did not express TK. However, when macrophages were incubated with medium conditioned by L cells, they expressed TK, incorporated 3H thymidine into trichloroacetic acid precipitable material, and ceased to secrete the nucleotide. Furthermore, replicating P388/D1 cells were induced to secrete thymidine by inhibiting TK with d-glucosamine. These results have demonstrated an inverse relationship between thymidine secretion and the expression of TK. They suggest that thymidine secretion by macrophages may be attributed to their lack of TK activity.


Mononuclear cells contain human transfer factor as assayed locally on the skin of dogs.

We prepared transfer factor from mononuclear (M) cells and polymorphonuclear (PMN) cells by sieving cell lysates through a P-2 Biogel column. By assaying column fractions by their ability to induce a local delayed-type reaction to antigen in canine skin, we determined that mononuclear cells most consistently contained skin activity. The elution patterns from P-2 Biogel of PMN or M-enriched preparations were less complex and much different from that seen with whole leukocytes.


Selective localization of radiolabeled immune lymphocytes into syngeneic tumors.

Newly formed long-lived small lymphocytes (LLSL) generated during the immunization of mice to tumor specific transplantation antigens (TSTA) of syngeneic MCA-induced sarcomas were labeled. We then studied the localization of these labeled cells into tumor implants. "Crisis-cross" experiments were performed in which MCA sarcomas with individually distinct TSTA were studied in parallel. A selective localization of LLSL into tumors to which the lymphocytes were immune was found when small tumor pieces were implanted into immune mice whose LLSL had been labeled. Selective localization was also detected upon adoptive transfer of immune LLSL to tumor-bearing mice, but only when these mice had, before transfer, received a sublethal dose of whole body irradiation.


Chemical cross-linking of Ia alloantigen alpha and beta chains with dimethyl 3,3'-dithiobispropionimidate.

We have examined the polypeptide chain composition of membrane-bound and detergent-solubilized Ia antigens using the chemical cross-linking reagent dimethyl 3,3'-dithiobispropionimidate (DTBP). Products of the I-E/C subregion of the major histocompatibility complex, which were solubilized from spleen cells with the detergent NP-40 and partially purified by affinity chromatography on lentil lectin-agarose, could be almost completely cross-linked by DTBP. Thus, the characteristic 33,000 m.v. (alpha) and 28,000 (beta) polypeptide chains seen on sodium dodecylsulfate polyacrylamide gels disappeared and a major new species of 60,000 m.w. appeared after cross-linking. When isolated and reduced with 2-mercaptoethanol, the 60,000 m.w. peak was found to be comprised to alpha and beta chains. Similar results were obtained when I-E/C, as well as I-A, alpha and beta chains were crosslinked on the cell surface. These data demonstrate that the alpha and beta chains of the Ia antigens exist primarily in the form of a dimer both in detergent solution and in situ.


Immune response in subacute sclerosing panencephalitis: reduced antibody response to the matrix protein of measles virus.

Immune precipitation was used to study the humoral immune response of patients with subacute sclerosing panencephalitis (SSPE). Patients with SSPE have a progressive infection of the CNS by measles or a measles variant despite high serum antibody levels to measles virus as measured by standard serologic techniques. However, when the antibody response to individual measles virus proteins was measured, we found a striking reduction in the ability of sera from patients with SSPE to precipitate the matrix (M) protein as compared to the precipitation of the M protein by sera from normal adults who had natural measles infection in childhood, or by convalescent sera obtained 3 to 5 weeks after a naturally occurring measles infection. The decreased antibody response to the M protein in sera from patients with SSPE occurred despite a vigorous antibody response to the other viral proteins, suggesting a selective defect in the production of antibody to a single viral protein. The reduced anti-M antibody in sera from patients with SSPE was demonstrated whether immune precipitation was performed with wild-type measles virus or SSPE virus proteins. These results suggest that in SSPE only small amounts of the M protein are produced. This result may help explain how measles virus persists in the central nervous system of patients with SSPE.


Genetic mapping of the locus controlling structural variations of murine C3 in the chromosome 17.

Backcross progeny, (NC X TF/GnLe)F1 X TF/GnLe, was tested for C3 allotype controlled by C3-1 and the expression of mutant gene tf, repeated loss and regrowth of hair. The recombination frequency between these two loci both located in chromosome 17 of the mouse was 24%. Taken together with our previous linkage data, C3-1 is now localized to a position 11 cm more distal than H-2 on chromosome 17.


Preparation of an antibody to mouse serum IgD.

Mice have recently been shown to have serum IgD. We have used affinity chromatography to partially purify mouse serum IgD, and have prepared a rabbit antiserum against this mouse Ig class. This antiserum, once adsorbed by IgD-depleted mouse serum bound to Sepharose, was isotype specific as determined by radioimmunoassay and fluorescence activated cell sorter analysis, and bound to the same splenic lymphocyte surface molecules as a hybridoma produced monoclonal anti-mouse delta antibody.


Studies on human lymphocytes in frozen tissue section: viability and spontaneous rosette formation with sheep erythrocytes.

Sixty to 70% viability of lymphocytes in frozen tissue section was obtained by using dimethylsulphoxide-containing medium as a cryoprotective agent and by conditioning time and temperature for freezing, making sections on a cryostat and thawing. The aviable and dead cells were differentiated by trypan blue exclusion test on section. Lymphocytes in tissue sections showed spontaneous rosette formation with sheep erythrocytes (E rosette) with specific localization for each lymphoid organ.


Immunofixation on cellulose acetate: an improved screening method for monoclonal immunoglobulins.

A rapid, simple and economical method is described for typing monoclonal immunoglobulins. It is a modification of the immunofixation electrophoresis method and cellulose acetate has been used as a supporing medium. It involves an initial electrophoretic separation followed by an antigen (Ag)-antibody (Ab) reaction 'in situ'. Eight samples can be typed on each 5.7 cm x 10.5 cm strip and only 20 microliter of commercial antiserum are required (about 2.5 microliter per sample). The method permits detection of monoclonal proteins at concentrations as low as 100 ng/microliter in only 60 min.


Solid-phase radioimmunoassay for the detection of antibodies to collagen.

A solid-phase radioimmunoassay has been developed using polystyrene tubes for the detection of class-specific antibodies to collagen. The optimal conditions for the adsorption of collagen onto the tubes, followed by incubation with the antisera and finally the radiolabelled antibody to the class-specific antibody are described. Studies carried out on antisera raised in rats and rabbits using this assay confirm its sensitivity and applicability.


Determination of avidity of anti-viral antibodies at 50% binding of antibody.

Binding studies with tobacco mosaic virus and specific IgG and Fab' fragments were done under conditions of 50% binding of antibody. When the log of the concentration of antigen sites required to bind 50% of the available antibody was plotted against the log of bound sites, a line was obtained which had a slope varying between 0 and 1. Instead of using the slope of this line as an index of avidity of the antiserum, it is suggested that the calculation K0 values over a range of reactant concentrations provides a better criterion for comparing the avidity of antiviral antibodies. The results of conventional binding tests were analyzed by means of Scatchard plots and were found to agree very closely with the data obtained at 50% binding of antibody.


Fluorescent labelling of proteins of lymphocyte plasma membranes.

Fluorescamine has been used for labelling proteins present on the surface of normal human peripheral blood lymphocytes. Under the conditions of study, 12 labelled proteins could be detected by SDS gel electrophoresis. This method may be of value in biochemical studies of lymphocyte membranes.


A micro macrophage migration inhibition test for the detection of cellular immunity in vitro.

A simple, economical, macrophage migration inhibition assay enabling multiple tests to be performed using microculture plates has been developed. The system confers many advantages over the standard in vitro assay of macrophage migration inhibition factor. It requires only 5 X 10(5) macrophages for each assay, small volumes (100 microliter) of test factors are consumed, 96 tests in each culture plate are easily assessed microscopically and the technique is easily and reproducibly executed. An adherent monolayer of activated fresh mouse peritoneal exudate cells (PEC) is established in each microculture well. A 'starting line' is scored across the monolayer by the application of a trimmed razor blade and the cells are dislodged and aspirated from one side of the line. Test factors from cultures of stimulated lymphocytes were added to the 'wounded' monolayer and the migration of macrophages from test and control cultures was measured after staining. The cells across the line were counted after incubation. Miration was shown to be time and temperature dependent, markedly stimulated by colchicine and appeared to associate with tuberculin antigen (PPD) skin testing and lymphocyte transformation in human subjects.


High volume antisera yields from rabbits using plasmapheresis.

A simple procedure for plasmapheresis of rabbits is described, permitting collection of approximately 75 ml of antisera to be obtained at weekly intervals. Evidence for maintenance of antibody titer throughout this interval is presented.


Isolation and characterization of hepatocytes and Kupffer cells.

Simplified isolation procedures are described for the parenchymal cell of the liver and the major non-parenchymal cell, the Kupffer cell. Hepatocytes are obtained in a purity of approximately 100%; a yield 10 X 10(6) cells/g liver tissue and the viability is greater than 85%. The recovery of Kupffer cells is 82%, viability 87% and purity 67%. Characterization of Kupffer cells is by the peroxidatic reaction, of hepatocytes by gluconeogenesis and also culture on collagen plates in a non-protein medium yielding albumin.


Relationship between actinic damage and chronologic aging in keratinocyte cultures of human skin.

The relationship of actinically-induced "premature aging" to chronological aging was studied in paired keratinocyte cultures obtained from the habitually sun-exposed (lateral) and nonexposed (medial) aspects of the arm of 5 male donors, aged 41 to 80 yr. In all cases, the number of cell generations in vitro was greater for cultures derived from sun-exposed skin, and this discrepancy increased with donor age and the severity of clinical aging changes. Hence, chronic sun exposure does accelerate aging in human skin by at least one previously established in vitro criterion: it decreased the lifespan of cultured keratinocytes. Plating efficiency was 11- to 32-fold higher for keratinocytes from chronically sun-exposed skin than nonexposed controls, perhaps reflecting the recognized carcinogenic potential of actinic radiation. Keratinocyte cultures appear to be as amenable to gerontologic studies as the already widely used human fibroblast cultures.


Histologic study of the regeneration of axillary hair after removal with subcutaneous tissue shaver.

It was observed that after subcutaneous tissue shaving for the radical therapy of hircismus and hyperhidrosis axillary hair often regrew. Histologic study of this phenomenon showed that hair bulb and most of the follicle up to a level near the sebaceous duct opening had been removed. Hair regrows from remnant outer root sheath, but only when sebaceous glands are preserved, that is when the upper portion of the follicular isthmus is intact. One or several solid epithelial pegs grow downward from the cut end of the trichilemma, and inner root sheath and new young hair are formed in its center. In hair peg stage, the lower tip of the hair follicle descends while new hair is growing in its center through the mitotic activity is growing in its center through the mitotic activity of hair germ cells and is prevented from pushing toward the skin surface by interlocking fusion between hair cuticula and sheath cuticula. Eventually, the epithelial cells wrap around a mass of mesenchymal cells and form a new bulb from which the terminal hair grows upward. The new matrix acquires a new complement of functioning melanocytes.


Distensibiltiy of the papaverine-induced passive vascular bed in dermis of generalized scleroderma.

Passive distensibility of of the vascular bed in cutaneous tissue of the hand was examined in 7 normal persons and 15 patients suffering from generalized scleroderma of the acrosclerosis type. Paralysis of vascular smooth muscle cells was obtained by locally injected papaverine. Increase in vascular transmural pressure was induced by lowering the limb. Blood flow was estimated by the local Xenon wash-out technique. There was no significant difference between the patients and the normals in relative blood flow during lowering, when injection volumes of 0.005 and 0.02 ml were used, indicating that distensibility of the vascular bed in cutaneous tissue in generalized scleroderma is not diminished. However, in generalized scleroderma an injection volume of 0.1 ml caused a significant decrease in relative blood flow during lowering. This phenomenon observed in the patients probably reflects changes in total tissue pressure opposing distension of the vascular bed. This suggests an altered pressure-volume relationship of the interstitial fluid spaces in cutaneous tissue of generalized scleroderma.


Local regulation of digital blood flow in generalized scleroderma.

A decreasing activity of the vascular bed upon changes in vascular transmural pressure and changes in arterial perfusion pressure head was observed from hand to fingers and from subcutis to cutis in patients suffering from generalized scleroderma. These findings are expressions of deteriorating vascular smooth muscle function. The reactive hyperemia response was decreased in subcutaneous as well as cutaneous tissue in patients fingers and in 4 of 7 patients the response was absent from the finger tip. The reactive hyperemia response of the patients resembled responses obtained in normal fingers during decrease in local perfusion blood pressure, during local cooling or after infiltration of norepinephrine around the digital arteries. This implied an increased digital artery resistance in generalizing scleroderma. A positive feed back loop between cooling, increase in digital artery resistance, increase in blood viscosity and a passive vascular bed is proposed as an important factor in Raynaud's phenomenon.


The acute effects of long-wave ultraviolet radiation on human skin.

The erythematogenic and melanogenic properties of polychromatic long-wave ultraviolet light (UV-A) has been re-examined. Redness appeared immediately after exposure and persisted for 24 hr with doses of about 50 Joules/cm2. With threshold erythemal doses, about 13 J/cm2, the redness faded after a few minutes. The response was not biphasic. Pigmentation also appeared immediately after exposure and faded rapidly with threshold doses of 4 J/cm2. With larger doses (18 J/cm2) immediate pigmentation gave way without fading to delayed pigmentation (true melanogenesis). Thus, the acute effects of UV-A, unlike other wavelengths within the UV-spectrum, are immediate and appear without latency. The responses are also most intense immediately after exposure.


Carcinoembryonic antigen: increased plasma levels in recessive epidermolysis bullosa.

Circulating plasma levels of the oncofetal antigen, carcinoembryonic antigen, were examined in 18 patients with various forms of epidermolysis bullosa. Carcinoembryonic antigen was markedly elevated in the plasma of 4 of 6 patients with recessive dystrophic epidermolysis bullosa and in 1 of 2 patients with recessive epidermolysis bullosa letalis. In contrast, patients with dominantly inherited forms of the disease, dominant epidermolysis bullosa simplex and dominant dystrophic epidermolysis bullosa, had normal levels of antigen. In the recessive patients, the plasma levels of carcinoembryonic antigen appeared to correlate with the severity of cutaneous involvement. Alternatively, it is possible that expression of carcinoembryonic antigen is genetically linked to certain forms of recessive epidermolysis bullosa or is part of a pleiotropic effect of the gene coding for the disease.


Histometric analysis of human skin in organ culture.

A histometric method is used for the study of human skin kept in organ culture in a defined medium for up to 10 days. The method provides quantitative, reproducible data on tissue survival, cell migration, and cellular differentiation (keratinization). With this method, the behavior of epidermal skin tissue can be effectively monitored during organ culture. As quantitative data are obtained, even subtle changes can be accurately demonstrated, and accumulated data may be subjected to statistical analysis. The various applications of this method are pointed out.


Assay of contact photosensitivity to musk ambrette in guinea pigs.

This study reports the induction of contact photodermatitis to musk ambrette, 2-methoxy-3,5-dinitro-4-methyl-t-butylbenzene, in guinea pigs. Photoallergic contact dermatitis was assayed using 2 alternative induction methods. Successful photosensitization was achieved only when the nuchal skin was stripped with scotch tape before application of musk ambrette and ultraviolet radiation. Induction methods utilizing nonstripped nuchal skin which induce photosensitivity to potent photoallergens were ineffective for musk ambrette. Phtotoxicity tests to musk ambrette at concentrations between 1 and 50% and a dose of 10.2 joules/cm2 from "Black Light" fluorescent tubes were all negative. Under identical irradiation conditions, anthracene at 0.9% and 8-methoxypsoralen at 1% were consistently positive. The mechanism of photosensitivity to musk ambrette appears to be photoallergic rather than phototoxic. The requirement for skin abrasion to induce photosensitization parallels the clinical reports of photosensitivity to musk ambrette in man.


Histogenesis of primary localized cutaneous amyloidosis: sequential change of epidermal keratinocytes to amyloid via filamentous degeneration.

Two cases of lichen amyloidosus and 8 cases of macular amyloidosis were examined by electron microscopy. Epidermal keratinocytes showed variable degrees of focal degeneration in the basal or lower Malpighian layer. The primary change was seen in cells which contain fibrillar (30 nm in thickness) cytoplasmic inclusion. The following developments seemed to lead to filamentous degeneration (colloid bodies): (1) aggregation of tonofilaments within the granular or fibrillar cytoplasm, (2) filamentous cells, which are composed of bundles of 7-nm thick filaments surrounded by cell membrane and desmosomes, and (3) filamentous masses composed of bundles or whorls of tightly packed 7-nm thick filaments in the intercellular spaces. At the dermo-epidermal junction, some of the filamentous masses were surrounded by the basal lamina of the epidermis and others were dropping into the dermis. Occasionally, loosened filaments (similar to amyloid filaments) were about to drop into the dermis. Early formation of amyloid islands consisted of electron-dense and electron-light parts. They were located directly beneath the epidermis. In the deeper postion of the papillary dermis and in the upper reticular dermis, the majority were electron-light masses. Electron dense parts were the densely packed 7-nm thick filaments, whereas electron-light parts were the typical straight amyloid filaments. Small tubular filaments were seen in common in the filamentous cells, filamentous masses, and amyloid islands. It is concluded that some of the amyloid substance in primary localized forms of cutaneous amyloidoses derive from the epidermal cells through filamentous degeneration.


Photoinactivation of skin fibroblasts by fractionated treatment with 8-methoxypsoralen and UVA.

Guinea pig skin fibroblasts treated with low doses of 8-methoxypsoralen (8-MOP) and long-wave ultraviolet light (UVA) showed a dose-dependent inhibition of 3H-Thymidine incorporation as determined by liquid scintillation counting. The minimum incubation time necessary to obtain constant inhibition rates was 60 min. By washing the drug was removed from the reactive sites within 30 min. Repeated light exposure at a constant concentration of 8-MOP caused a cumulative inhibition of DNA synthesis. Irradiation of 8-MOP-plus-UVA treated cells, from which the drug was removed, produced a small increase in photoinhibition. Split dose treatment at various time intervals (ranging from 1--48 hr) revealed inhibitory rates, which correspond to the total amount of UVA applied. No recovery effects were seen in cultures treated by single or multiple applications of 8-MOP-plus-UVA.


PUVA suppresses the proliferative stimulus produced by stripping on hairless mice.

Hairless mice were fed with 8-methoxypsoralen by gastric tube and exposed to UVA light to produce threshold phototoxic reactions. 3H-thymidine autoradiography revealed no significant difference of the epidermal labelling index as compared to that of nonirradiated animals (4.8 vs 6.2). Single PUVA exposures performed 2,8,14 and 24 hr after tape stripping lead to suppression of a wave of synchronized DNA synthesis present in nonirradiated control animals. These data demonstrate different reactions of stripped and unstripped epidermis to PUVA exposures and offer indirect evidence for the suppression of DNA synthesis in hyperproliferative skin disorders by PUVA in vivo.


New studies on the interaction between 8-methoxypsoralen and DNA in vitro.

Some aspects of the interactions between DNA and 8-methoxypsoralen (8-MOP) in its ground state (complex formation) or in its excited state (photobinding) have been investigated. 8-MOP shows a low affinity towards DNA in the complex formation; this fact minimizes the possible biological consequences deriving from this interaction, when it occurs in vivo. In covalent photobinding to DNA, 8-MOP forms mainly monofunctional adducts, and to a lesser extent bifunctional adducts, showing a behavior similar to that of other linearly condensed furocoumarins (psoralens); the ratio between mono- and bifunctional adducts was found to be 9:1. The covalent photobinding to DNA does not occur at random along the macromolecule, but preferentially at the level of specific receptor sites. The regions having an alternate sequence of A-T seem to be the best receptor sites for the formation of monoadducts while the regions containing an alternate sequence of A-T and C-G appeared to be the preferential sites for the cross-linkage formation.


Pulmonary and systemic defenses against challenge with Staphylococcus aureus in mice with pneumonia due to influenza A virus.

Pulmonary and systemic defenses against hematogenous challenge with 32P-labeled Staphylococcus aureus were measured 10 min, 8 hr, and 24 hr after intravenous injection of the bacteria in a mouse model of influenza virus pneumonia. Infection with influenza A virus did not alter bactericidal defenses in the liver and spleen, but pulmonary bactericidal activity measured 24 hr after infection was suppressed in virus-infected animals; 20% +/- 3% of the initially injected, viable bacteria were recovered from lungs of pneumonitic mice after 24 hr as compared with 9% +/- 1% from lungs of the uninfected mice. These data demonstrate that pulmonary infection with influenza virus does not alter antibacterial defenses of the liver and spleen but does suppress bactericidal activity in the lung.


Susceptibility of isolates of Bacteroides to the bactericidal activity of normal human serum.

Seventy-one strains of species from the Bacteroides fragilis group, including 46 isolates of B. fragilis, were tested for susceptibility to the bactericidal effect of serum from healthy subjects. Twenty-seven (38%) of the isolates were killed by serum. Isolates from feces were significantly more sensitive to serum than were isolates from patients with clinical infections. Killing of bacteria required heat-labile serum components and was an exponential function of serum concentration. Among the various species tested, B. fragilis was clearly the most resistant to bactericidal activity of serum. These observations may be important to the understanding of infections caused by the B. fragilis group, which contains the anaerobes of greatest clinical importance.


Inhibition by polyanions of adherence by Kanagawa-positive Vibrio parahaemolyticus: a physicochemical effect.

Rapid adherence of Kanagawa-positive Vibrio parahaemolyticus to human fetal intestinal cells has been demonstrated in monolayer cultures. In this study, polyanionic carbohydrates and glycoproteins of various biochemical compositions, structural configurations, and molecular weights were found to inhibit this adherence. Mono- and disaccharide components of inhibitory compounds did not themselves inhibit adherence. Levels of Ca++ in the test mixture were similar in both the presence and the absence of chondroitin sulfate, a potent inhibitor of adherence by V. parahaemolyticus. These results, which demonstrate a major effect of surface charge in this model of bacterial adherence, suggest that differences in surface charge may contribute to the varying degrees of adherence by different strains of V. parahaemolyticus. This striking effect of surface charge on adherence by V. parahaemolyticus underscores the importance of experimental conditions and of substances that affect the surface potential of the cell in the interpretation of findings in models of bacterial adherence.


Simultaneous administration of live, enteric-coated adenovirus types 4, 7 and 21 vaccines: safety and immunogenicity.

The safety of and the immune response to simultaneous administration of live, enteric-coated adenovirus type 4 (ADV-4), type 7 (ADV-7), and type 21 (ADV-21) vaccines were studied. Volunteers (476 men), randomly assigned to four study groups, received three vaccines (ADV-4, ADV-7, and ADV-21), two vaccines (ADV-4 and ADV-7), one vaccine (ADV-21), or no vaccine (placebo). Subjects were observed for three weeks, and no side effects due to vaccination occured. The percentages of susceptible subjects (those entering the study with a neutralizing antibody titer of less than 1:2 to each vaccine virus received) who seroconverted to ADV-4 were similar in both groups that received ADV-4 vaccine (78% of 77 subjects and 74% of 76). However, in the group that received three vaccines, only 62% of 77 subjects seroconverted to ADV-7, compared with 79% of 76 in the group that received two vaccines (P less than 0.05). Only 58% of 77 subjects in the three-vaccine group seroconverted to ADV-21, compared with 69% of 59 in the group that received one vaccine (P greater than 0.1).


Circulating staphylococcal antigen in humans and immune rabbits with endocarditis due to Staphylococcus aureus: inhibition of detection by preexisting antibodies.

A radioimmunoassay for staphylococcal antigen that had detected antigenemia in each of 12 nonimmune rabbits with staphylococcal endocarditis detected antigen in sera from one of nine humans and two of eight immune rabbits with Staphylococcus aureus endocarditis. Staphylococcal antigens could be detected at concentrations as low as 0.78 microgram/ml when diluted in normal rabbit serum, compared with 6.25 microgram/ml when diluted in normal human serum and greater than 25 microgram/ml when diluted in immune rabbit or human serum. Low titers of staphylococcal antibody were found in normal rabbit serum compared with immune rabbit and normal or immune human sera. Detection of staphylococcal antigen was inhibited when the antigen was diluted in the IgG and IgM fractions, but not in the albumin fraction, of normal human serum. This study demonstrated that antigenemia can be detected infrequently in patients and immune rabbits with staphylococcal endocarditis; staphylococcal antibodies inhibit detection of antigen, presumably through formation of immune complexes.


Studies of the kinetics of indium-111-labeled granulocytes.

Studies of the in vivo kinetics of granulocytes labeled in vitro with indium-111 were carried out in 10 normal subjects. The granulocyte suspension was prepared with a Ficoll-Hypaque density gradient and labeled with indium-111-oxine. No elution or reutilization of the radioisotope was demonstrable in vitro. The average intravascular recovery of indium-labeled granulocytes was 30% +/- 6, and the t 1/2 was 5.0 +/- 1.6 hr. Normal in vitro function of these cells was demonstrated by bacterial killing and chemotaxis assays. Because indium-111 decays by gamma emission, the fate of in vivo labeled granulocytes can be followed with scintigraphic techniques. Images obtained indicated normal uptake of activity in the liver and spleen. Effective in vivo function of indium-labeled granulocytes was demonstrated in four patients by the localization of radioactivity at sites of inflammation or abscess. Although the intravascular recovery with this method is lower than that reported for some other radioisotope methods, the comparison of abnormal findings with normal values by this technique is probably valid. Indium-labeled granulocytes should prove useful in the study of granulocyte collection, transfusion, histocompatibility, and storage.


Tetramethylbenzidine--a substitute for benzidine in hemoglobin analysis.

Plasma hemoglobin is routinely measured by some modification of the benzidine-hydrogen peroxide method, particularly at low concentrations. Because benzidine is carcinogenic, and thus impractical for clinical use under current restrictions, the identification of a noncarcinogenic benzidine derivative has become important. 3,3',5,5'-tetramethylbenzidine fulfills this criterion, and consequently a method is described in this paper which uses it for hemoglobin determinations in the range of 1 to 100 mg/dl. Absorption spectra show this method to be far less influenced by plasma proteins and other substances than the benzidine method, and moreover the sensitivity of TMB is at least seven times that of benzidine under comparable conditions.


Role of platelets in lysis of dilute plasma clots: requirement for metabolically active platelets.

The role of platelets in clot lysis has been investigated functionally with the use of dPRP clots formed at 4 degrees and shifted to 37 degrees. Clots handled in this manner lysed in 6 hr (+/- 1 hr), whereas clots formed at 4 degrees or at 37 degrees and held at those temperatures, or clots formed from dPPP did not lyse in less than 20 hr. dPRP clots having the shorter (6 hr) lysis time released 14C-5-HT at the time of the temperature shift. Preincubation of dPRP with antimycin A and 2-deoxy-D-glucose before addition of thrombin prolonged the clot lysis to 26 hr and inhibited release of 14C-5-HT at the time of the temperature shift. These studies demonstrate that metabolically active platelets are required to mediate the optimal clot lysis seen in the 4 degrees to 37 degrees system and that they continue to function (i.e., take up and release 5-hydroxytryptamine) after they have been incorporated into a clot. Thus the dPRP clot lysis system provides a model by which the timing and sequence of the interaction of metabolically active platelets with the fibrin framework of the formed clot can be studied.


Predisposing factors to thrombosis in systemic lupus erythematosus: possible relation to endothelial cell damage.

We studied 28 patients with SLE, five of whom had had thrombotic episodes. Platelet function and coagulation and fibrinolytic studies were performed to determine whether any of these factors may predispose to thrombosis in SLE. An inhibitor of blood coagulation was detected in 12 patients, and von Willebrand's syndrome was observed in two others. The most striking findings which could be correlated with thromboembolic phenomena were the increase in VII R:WF and the absence of plasminogen activator release after venous occlusion. Both proteins are synthesized by the endothelial cell, and the abnormalities observed are possibly related to damage of the vascular endothelium by immune complexes observed in SLE.


Virulent gentamicin-induced small colony variants of Staphylococcus aureus.

Stable nonhemolytic small colony variants were isolated in pure culture from nine of 30 Staphylococcus aureus clinical strains after incubation of log10 7.0 cfu for 48 hr in MH broth containing 1.0 microgram/ml gentamicin. The variants resembled Staphylococcus epidermidis on blood agar, but they were positive for tube coagulase and thermostable nuclease at 24 hr and fermented mannitol slowly. The infectivity and virulence of four variants were compared to four parent S. aureus and three S. epidermidis strains in a rabbit model of endocarditis. Log10 5.0 cfu of the variant S. aureus, parent S. aureus, or S. epidermidis strains were injected intravenously into rabbits with intracardiac catheters. Quantitative culture of vegetations demonstrated endocardial infection in 47 of 49 (96%) animals injected with S. aureus variants, 44 of 44 injected with S. aureus parent strains, and four of 21 (19%) S. epidermidis-injected animals. The mortality rate in untreated animals within 4 days was five of 49 (10%) for variant S. aureus, 33 of 44 (75%) for parent S. aureus, and 0 of 21 for S. epidermidis. Small colony variants of S. aureus may be mistaken for S. epidermidis, but the variants are significantly more infective than S. epidermidis and are more likely to cause endocarditis. Gentamicin-induced S. aureus small colony variants are as infective but less virulent than their parent S. aureus strains.


A proposed model for chemotactic deactivation: evidence for microtubule modulation of polymorphonuclear leukocyte chemotaxis.

Incubation of the CFs Gly-His-Glyc or CCF with PMNs in the absence of a gradient, resulted in a dose-dependent depression in chemotactic activity when, after washing, the cells were challenged with the CFs in a Boyden chamber. When the cells were preincubated with either CF and suitable concentrations of colchicine, the inhibition of chemotaxis that either of these agents induced when incubated with the cells alone was abolished. Deactivation reappeared when the optimal ratio between colchicine and CF was altered in either direction. Ultramicroscopic studies showed an increase in centriole-associated microtubules following incubation of cells with CFs. This increase was arrested by prior exposure of the cells to colchicine. Colchicine did not alter the specific binding of CCF to human neutrophils, and lumicolchicine had no effect on either chemotaxis or deactivation. Our data suggest that the control of PMN chemotaxis is predicted upon microtubule assembly evoked by cell interaction with a chemotactic gradient. Chemotaxis would be prevented by conditions that inappropriately organize responsive microtubules in either a polymerized or depolymerized configuration.


Effects of storage on platelet reactivity to arterial subendothelium during blood flow.

The usual in vitro methods for studies of platelets after storage do not measure physiological parameters such as capability for interaction with vessel wall subendothelium during blood flow conditions. For studies reported here we adapted a technique described by Baumgartner (Microvasc Res 5:167, 1973) to quantitate platelet interactions with subendothelium during blood flow and before and after storage at 4 degrees and 22 degrees and by a modified 4 degrees procedure called temperature cycling. The results indicated that platelets kept at 22 degrees for 72 hr interacted with subendothelium in quantities which were less than 10% that of fresh controls. However, storage at 4 degrees and by temperature cycling produced interactions equal to 71% and 42% of controls, respectively. The data also indicated that utilization of Baumgartner's technigue as described here will (1) provide an in vitro technique for identification of qualitative defects responsible for the delayed hemostatic effectiveness of 22 degrees-stored platelets and (2) permit an objective and quantitative analysis of prospective improvements in storage before in vivo function testing is necessary.


Reverse cross-coupling in the synthesis 3 alpha, 7 alpha-dihydroxy-5 beta-cholestanoic acid.

The present report describes the characterization of (24R and 24S)-27-nor-24-methyl-3 alpha, 7 alpha-dihydroxy-5 beta-cholestan-26-oic acids obtained in considerable amounts during the synthesis of (25RS)-3 alpha, 7 alpha-dihydroxy-5 beta-cholestan-26-oic acid by the electrolytic coupling of chenodeoxycholic acid and the half ester of methylsuccinic acid. The mixture of 24R and 24S diastereomers was resolved by analytical and preparative thin-layer chromatography and characterized by gas-liquid chromatography, proton magnetic resonance, and molecular rotation differences. For reference, the model compound, 27-nor-3 alpha, 7 alpha-dihydroxy-5 beta-cholestan-26-oic acid, was synthesized by electrolytic coupling of chenodeoxycholic acid and the half ester of succinic acid.


Gangliosides, cholesterol, and phospholipids were quantitated in the tissues of 11 human neural tumors and the cells of two gliomas cultured in vitro. All tumor tissues contained higher water concentrations but lower total lipid concentrations than either human grey or white matter. In general they contained less cholesterol, sphingomyelin, and serine glycerophospholipid but more choline glycerophospholipid than white matter. Concentrations of total ganglioside sialic acid were intermediate between grey and white matter. Compared with normal brain, all tumors had greater proportions of the structurally less complex gangliosides and smaller proportions of the more complex gangliosides. This was most marked in the rapidly growing tumors while the better differentiated astrocytomas contained the greatest proportions of complex gangliosides. The cells of the cultured tumors contained amounts of total lipid and total phospholipid similar to their parent tissues. However, the cultures had less cholesterol, sphingomyelin, and total ganglioside than their parent tissues. There were significant amounts of choline and ethanolamine plasmalogens in both cultures and parent tissues. The ganglioside patterns of both cultures were complex but they contained a greater proportion of structurally simpler gangliosides than their parent tissues.-Yates, A. J., D. K. Thompson, C. P. Boesel, C. Albrightson, and R. W. Hart. Lipid composition of human neural tumors.


Enzyme-substrate interaction in lipid monolayers. I. Experimental conditions and fundamental kinetics.

A systematic study has shown the importance of the different factors which are concerned with the action of lipase on a substrate (1,3-didecanoylglycerol). These consist of a) the process of adsorption of lipase to the surface, b) the necessity of limited stirring to reach equilibrium, and c) the persistence during the reaction process of the enzyme molecules adsorbed on the monolayer. On the basis of this preliminary investigation, a technique was established to analyze the mechanism of lipase action with defined quantities of enzyme and lipid segregated in the monolayer. Thus, the process of the reaction itself is separated from the adsorption process, and it is demonstrated that the quantity of substrate hydrolyzed per minute depends only on the quantity of initially adsorbed lipase and not on the quantity of substrate or on the surface concentration of the enzyme. An appropriate new definition of the rate is consequently adopted.


Enzyme-substrate interaction in lipid monolayers. II. Binding and activity of lipase in relation to enzyme and substrate concentration and to other factors.

With the limited stirring procedure used in the present work, substrate and enzyme together form a segregated and well-defined system on the surface. The lipase molecules responsible for the lipolysis are only those that are adsorbed on the glyceride monolayer. After a study of the stirring procedure, two series of systematic experiments were done: a) the bulk concentration of the enzyme was varied with different constant surface concentrations of the substrate, and b) the surface concentration of the substrate was varied with different constant bulk concentrations of the enzyme. The influence of the surface concentration of substrate on a) the rate of lipolysis, V,; b) the enzyme activity, a,; and c) the enzyme adsorption, Ze, were each determined by a different procedure. The values obtained verify the enzymic activity equation (a = V/Ze). The roles of other factors (Ca2+ ions, and pH) which govern the adsorption of the enzyme and its specific activity were also studied in preliminary experiments.


Metabolism of 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 26-tetrol and 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 25-tetrol into cholic acid in normal human subjects.

Side chain oxidation and cleavage of precursors in cholic acid synthesis is thought to involve initial hydroxylation at either position 25 or 26 of the side chain. Therefore, the conversion of 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 26-tetrol and 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 25-tetrol into cholic acid was studied in normal subjects after single intravenous injections of these labeled alcohols. Eighty-six percent and 82% of 5 beta-cholestane, 3 alpha, 7 alpha, 12 alpha, 26-tetrol was converted into cholic acid in two subjects, respectively. However, only 14 and 16% of the injected 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 25-tetrol was converted into cholic acid in two subjects, respectively. Thus, this study indicates that 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 25-tetrol is an inefficient substrate for cholic acid biosynthesis in man and that the major route of cholic acid synthesis probably involves the 26-hydroxylated intermediate.


The phase behavior of hydrated cholesterol.

The thermotropic phase behavior of cholesterol monohydrate in water was investigated by differential scanning calorimetry, polarizing light microscopy, and x-ray diffraction. In contrast to anhydrous cholesterol which undergoes a polymorphic crystalline transition at 39 degrees C and a crystalline to liquid transition at 151 degrees C, the closed system of cholesterol monohydrate and water exhibited three reversible endothermic transitions at 86, 123, and 157 degrees C. At 86 degrees C, cholesterol monohydrate loses its water of hydration, forming the high temperature polymorph of anhydrous cholesterol. At least 24 hours were required for re-hydration of cholesterol and the rate of hydration was dependent on the polymorphic crystalline form of anhydrous cholesterol. At 123 degrees C, anhydrous crystalline cholesterol in the presence of excess water undergoes a sharp transition to a birefringent liquid crystalline phase of smectic texture. The x-ray diffraction pattern obtained from this phase contained two sharp low-angle reflections at 37.4 and 18.7 A and a diffuse wide-angle reflection centered at 5.7 A, indicating a layered smectic type of liquid crystalline structure with each layer being two cholesterol molecules thick. The liquid crystalline phase is stable over the temperature range of 123 to 157 degrees C before melting to a liquid dispersed in water. The observation of a smectic liquid crystalline phase for hydrated cholesterol correlates with its high surface activity and helps to explain its ability to exist in high concentrations in biological membranes.


Modification of the fatty acid composition of cultured human fibroblasts.

The fatty acid composition of human skin fibroblasts grown in 10% dialyzed fetal calf serum can be modified considerably by adding supplemental fatty acids to the culture medium. The degree of modification was dependent on the concentration of added fatty acid over the range tested, 2.5 X 10(-5) to 1 X 10(-4) M. At the higher concentration, the extent of the modifications was as those which can be produced in nonhuman or malignant cell lines. Although the greatest changes were produced in the neutral lipid fraction, the cellular phospholipids also exhibited appreciable modifications. The phospholipids isolated from a microsomal fraction prepared from the cell homogenate exhibited similar changes in fatty acyl composition. These findings indicate that the human fibroblast can tolerate considerable variability in fatty acid composition, even in membrane phospholipids. The triglyceride content of the cells increased when they were grown in the presence of added fatty acids, but the phospholipid and cholesterol content remained unchanged. Growth was not affected by either oleic or linoleic acids, but it was reduced up to 50% when palmitic linolenic, or arachidonic acid was added in concentrations of 5 X 10(-5) M or above. Extensive modifications in phospholipid fatty acid composition also were produced in confluent monolayers of these fibroblasts. This suggest that some membrane lipid turnover occurs even when the cultures are not rapidly growing. Fatty acid modifications also were produced in the commercially available IMR-90 strain of human lung fibroblasts, suggesting that the ability to tolerate considerable differences in fatty acid composition is not a special property of the skin fibroblast line that was isolated locally.


A new method for the measurement of lipoprotein lipase in postheparin plasma using sodium dodecyl sulfate for the inactivation of hepatic triglyceride lipase.

Lipoprotein lipase (LPL) and hepatic triglyceride lipase (H-TGL) are lipolytic activities found in postheparin plasma. A simple and precise method for the direct determination of LPL in postheparin plasma is described. Pre-incubations of this plasma (45--60 min at 26 degrees C) with sodium dodecyl sulfate (35--50 mM) in 0.2 M Tris-HCl buffer, pH 8.2, results in the inactivation of H-TGL, while leaving LPL fully active. Direct determination of H-TGL is done in a separate aliquot of the same postheparin plasma sample using previously reported assay conditons that do not measure LPL. The sodium dodecyl sulfate-resistant lipolytic activity has the characteristics of LPL as judged by a) its activation by serum and by apolipoprotein C-II; b) its inactivation (over 90%) by 0.75 M NaCl; and c) its inactivation by a specific antiserum. No sodium dodecyl sulfate-resistant activity was found in postheparin plasma from a patient with LPL deficiency (primary type I hyperlipoproteinemia). An excellent correlation of values was obtained (r = 0.99) for 30 samples assayed after sodium dodecyl sulfate treatment and after immuno-inactivation of H-TGL. The intra-assay coefficient of variation was +/- 11% and 4% before and after normalization of values, respectively.


Modification of the Dittmer-Lester reagent for the detection of phospholipid derivatives on thin-layer chromatograms.

A simple modification of the Dittmer-Lester reagent is described that allow the detection of phospholipid derivatives at very low concentrations on silica gel and reversed-phase thin-layer plates. This modification, which involves the addition of acetic acid to the mixture, permits the observation of sharp blue spots on a white background. The specificity and sensitivity of the spray are discussed.


Electrocardiographic observations on the termination of supraventricular tachycardia by verapamil.

The effect of intravenous verapamil on the termination of supraventricular tachycardia (SVT) was studied by continuous electrocardiographic monitoring of 27 episodes of SVT. Progressive increase of the cycle length heralded conversion in eight episodes while cycle-length alternation preceded cessation of the arrhythmia in 13 episodes. In five patients the arrhythmia was either stopped or closely followed by a ventricular premature beat (VPB), followed by further VPBs in three. Runs of bizarre ventricular tachycardia followed initial sinus-beats in two patients. Sinus standstill, lasting 30 seconds, was observed in one patient. The first post SVT beats had an aberrant QRS configuration with a normal P-R interval in four cases and an aberrant QRS complex with a short P-R interval, resembling Wolff-Parkinson-White complexes, in a further seven patients. The possible mechanisms causing this variability of pre- and post-conversion period are discussed. It is suggested that some aspects of verapamil action may be explained by a parasympaticomimetic effect on the myocardium.


Conduction disturbances in patients with calcified mitral annulus diagnosed by echocardiography.

Clinical, radiologic and histo-pathological observations have suggested that conduction disturbances (CD) are frequent in patients with calcified mitral annulus (CMA). To determine the incidence of CD, 65 consecutive patients with CMA were studied. Sixty-five normal individuals, matched for age and sex served as a control group. CMA has been identified by M-mode echocardiography, cross-sectional echo and image amplifier fluoroscopy. CD were present in 23 patients with CMA as compared to seven patients in the control group (p less than 0.01). Three patients developed complete atrioventricular block (4.6%) during hospitalization requiring artificial pacemakers. We conclude that, 1) early detection of CMA in the elderly is useful in predicting a higher incidence of CD. 2) Complete heart block is more frequent in patients with CMA. 3) Frequent ECG follow-up in these patients has proven to be useful in avoiding serious clinical complications.


Electrocardiographic QRS changes induced by acute coronary ligation in the isolated rabbit heart.

The electrical fields associated with augmented R- and decreased S-wave amplitudes during acute, severe myocardial ischemia were studied in fifteen isolated rabbit heart preparations. Hearts were suspended in a spherical tank and perfused with oxygenated electrolyte solution. Electrocardiographic signals were recorded from electrodes on the tank's surface and processed by computerized methods. Fifteen minutes after suture ligation of the left anterior descending coronary artery, records from electrodes overlying the lesion demonstrated increased R-wave amplitude and ST-segment elevation. Fitting a single moving dipole to pre- and post-ligation potentials demonstrated that the ligation increased the dipolarity of the field and shifted the terminal QRS dipole to a position topographically related to the location of the ischemic lesion. The effects of injury were further assessed by study of fields computed by subtracting control from post-ligation data. This generated a dipolar field at the instant of maximal dipole moment whose strength directly correlated (r=0.74) with the area of non-perfused epicardium, as determined by post-ligation methylene blue infusion. These results suggest that the electrical fields generated during mid to late dipolarization by coronary ligation are qualitatively similar to those generated during repolarization, and may therefore be of similar clinical and/or experimental value.


Effects of stable and changing rates and premature ventricular beats on transient tachycardia-, pseudobradycardia-, and bradycardia-dependent bundle branch block alternans.

New circumstances under which bundle block (BBB) alternans may appear or disappear are described. 1) Tachycardia-dependent as well as bradycardia-dependent BBB alternans may begin after constant BBB is interrupted by a premature ventricular beat. Tachycardia- and bradycardia- dependence may be differentiated by the shape of the first beat after the pause. 2) When BBB alternans disappears during a constant ventricular rate, tachycardia-dependent BBB alternans changes to persistent normal or more normal intraventricular conduction, whereas bradycardia-dependent BBB alternans changes to a persistently greater degree of BBB. 3) BBB alternans appears to be tachycardia- or pseudobradycardia-dependent in relation to the cycle length and antegrade and retrograde refractory periods in the involved bundle branch. 4) BBB alternans may be recognized during persistent irregular ventricular action in atrial fibrillation. Here the recognition of BBB alternans depends upon the sequence of contours as well as upon the cycle lengths.


Orthogonal electrocardiographic study on progressive muscular dystrophy of the Duchenne type.

An attempt was made to investigate the incidence and significance of high frequency notches and slurs on the QRS complex in patients with progressive muscular dystrophy of the Duchenne type (PMD). The patients were classified into eight stages from the most mild, S(1), to the most severe, S(8), according to Swinyard-Deaver's criteria. Cases where the sum of high frequency notches in the combined leads exceeded none were generally limited to those more advanced than stage S(4). Also, the sum of the notch count tended to be higher in S(5) to S(6) than in the remaining groups and to be lower in the milder cases, S(1) to S(4), and most severe cases, S(7) and S(8). It should be emphasized that cases in groups S(7) and S(8) who had a history of congestive heart failure and/or developed congestive heart failure during the observation period, tended to show a smaller number of notches. High frequency slurs showed almost the same tendencies as the high frequency notches. It is thus anticipated that a significant increase or decrease in the number of notches and slurs on the QRS complex may be suggestive of more intense myocardial derangement.


Predicting coronary artery disease with treadmill stress testing: changes in R-wave amplitude compared with ST segment depression.

Coronary angiograms and treadmill stress tests were reviewed independently in 108 nonconsecutively selected cases. There were 16 patients (15%) with infarcts on ECG. Changes in R-wave amplitude and ST segments during exercise were evaluated to determine the sensitivity and specificity of each as a predictor of coronary artery disease (CAD). ST segment changes had a sensitivity of 49%, and a specificity of 74%. The sensitivity increased to 55% when infarcts were excluded. R-wave amplitude changes had a sensitivity of 68% and a specificity of 84%. The sensitivity increased to 78% when infarcts were excluded. An index formed by the sum of the change in R-wave amplitude and the magnitude of ST segment change yielded a sensitivity of 76% and specificity of 78%. The sensitivity increased to 84% when infarcts were excluded. There was no statistical difference between specificities for each criteria. Of those patients with an R-wave amplitude decrease, 69% had no coronary artery atherosclerosis, while 31% had significant lesions. Of those patients with no change or an increase in R-wave amplitude, 83% had coronary artery atherosclerosis, while 17% were normal. Of the 83% with coronary artery atherosclerosis, 81% had two and three vessel disease, while only 19% had single vessel disease. No change or an increase in R-wave amplitude during treadmill stress testing is a more reliable indicator of CAD in our laboratory than ST segment changes.


Body surface potential distributions in posterior ventricular pre-excitation.

Waveform of the QRS complex during ventricular pre-excitation is subject to the influence of both the site of pre-excitation and the time of pre-excitation relative to that of excitation via the normal AV path. This paper reports a case in which lead V1 of the electrocardiogram (ECG) could be altered from an R to an rS pattern by the administration of atropine sulfate. The provable mechanism was that of reduced conduction time in the normal AV path with altered time phase of normal excitation and pre-excitation. This mechanism was simulated in experiments on dogs and yielded similar findings. Body surface mapping in both the patient and the dogs provided evidence that pre-excitation could be recognized by that means with varied time phase of normal excitation and pre-excitation. It was demonstrated that the QRS complex of right sided precordial leads could be altered from an R to an rS pattern by altering the time phase of normal excitation and pre-excitation of the posterior ventricular wall. This alteration was related to the degree to which negative potentials on the anterior chest wall due to right ventricular breakthrough of normal activation developed in relation to the time of pre-excitation.


A scale for ECG rhythm analysis.

A simple scale, which allows the ECG reader to assess the regularity of the RR (or PP) interval and to detect the P wave train, has been presented.


Multiple mechanisms of tachycardias in a patient with the Wolff-Parkinson-White syndrome.

In a patient with the Wolff-Parkinson-White Syndrome we observed atrial fibrillation and three distinct paroxysmal re-entrant tachycardias. Intracardiac electrograms obtained during the tachycardias showed the mechanisms to be A-V nodal, accessory pathway and sinus node re-entry. When P wave morphology, R-P relationship and QRS configuration are considered, it is illustrated how these four tachyarrhythmias may be successfully diagnosed on the surface electrocardiogram. The therapeutic implications of multiple arrhythmias with different mechanisms in the Wolff-Parkinson-White Syndrome are discussed.


Ebstein's anomaly in association with anomalous nodoventricular conduction. Pre-operative and intra-operative electrophysiological studies.

A 13 year old girl with Ebstein's anomaly was investigated for refractory paroxysmal tachycardias and ventricular pre-excitation. Intracardiac electrophysiological studies demonstrated that ventricular pre-excitation was due to conduction in an anomalous nodo-ventricular pathway. Tachycardia occurred as a result of re-entry within the A-V node with pre-excitation during tachycardia due to conduction in the nodo-ventricular pathway. These tachycardias were controlled initially by medical therapy but because of increasing frequency of attacks, occasionally requiring D.C. conversion, further electrophysiological studies and epicardial mapping were undertaken. The epicardial surface of the right ventricle and right atrium were mapped during tachycardia. The results of the studies confirmed that a direct anomalous atrio-ventricular pathway was not present and that re-entrant tachycardia did not involve an accessory pathway of this type. A rapid atrial pacing system was implanted and paroxysmal tachycardias have been successfully controlled.


Plasma testosterone levels in intact and hemicastrated growing cockerels.

Changes in the levels of testosterone in plasma were measured by radioimmunoassay in blood samples taken at frequent intervals between 2 and 26 weeks of age from entire cockerels and cockerels hemicastrated before 2 weeks of age. In both groups the pattern of testosterone secretion could be divided into three clearly defined phases. In young birds, the levels of testosterone in plasma were low (0.3 ng/ml) but in the prepubertal period, at 11 weeks of age, they started to rise and continued to rise until 22 weeks of age when adult levels, which fluctuated between 2.5 and 3.5 ng/ml, were reached. In the immediate period after hemicastration, the concentration of testosterone decreased temporarily. From 11 weeks of age the levels of testosterone in the hemicastrated birds were approximately 75% of those in intact birds. These results are discussed in relation to the compensatory testicular hypertrophy which occurs in growing cockerels hemicastrated at an early age.


Inhibition of steroid 5 alpha-reductase in vivo: effect on suppression of luteinizing hormone and stimulation of accessory sex organs by testosterone in the orchidectomized rat.

An inhibitor of 5 alpha-reductase, the 17 beta-carboxylic acid derivative of testosterone (testosterone-17 beta CA), has been used to evaluate the importance of the 5 alpha-reduction of testosterone in its action on the suppression of LH secretion in male rats. The potential of testosterone-17 beta CA to inhibit the formation of 5 alpha-dihydrotestosterone (DHT) was first demonstrated in vitro. When homogenates of hypothalami or anterior pituitary glands were incubated with [3H]Testosterone in the presence of a 50-fold excess of testosterone-17 beta CA, the formation of labelled DHT was inhibited by more than 80%. Adult male rats that had been castrated for 1-2 months were fitted with chronic intravenous catheters and implanted with silicone elastomer sheets: one group received one sheet, 0.5-2.0 cm2 in size containing 1.6% testosterone, a second group received one 50 cm2 sheet containing 1.6% testosterone-17 beta CA and a third group received two sheets, one sheet 50 cm2 in size containing 1.6% testosterone-17 beta CA and the second ranging in size from 0.5 to 2.0 cm2 and containing 1.6% testosterone. Blood was withdrawn daily from each rat over a 4-5 day period after implantation of the steroids and the level of LH in the plasma was measured by radioimmunoassay. The seminal vesicles and the ventral prostate gland were removed at autopsy on day 4 or 5; the weights of these organs were shown to have increased progressively as the size of the implant of testosterone increased. In contrast, the level of LH in the plasma was suppressed to a comparable extent by implants of testosterone between 0.6 and 2 cm2, whereas a 0.5 cm2 implant of testosterone had no effect. Implants of testosterone-17 beta CA alone did not influence the weight of the accessory organs or the level of LH. When testosterone-17 beta CA and testosterone were implanted together, the growth-promoting effect of the latter on the accessory sex organs was significantly reduced. The effectiveness of testosterone in suppressing the level of LH in the plasma of these animals was not influenced by the presence of testosterone-17 beta CA and in certain instances the level was raised.


Production of erythropoietic colony-forming units and erythrocytes during chick embryo development: an attempt at modelization of chick embryo erythropoiesis.

The enumeration of erythropoietic colony-forming cells in vitro has allowed us to complete previous data on changes in the various erythroid cell populations during chick embryo-genesis. Erythrocytic colony-forming units in culture (CFU-cE) which are sensitive to avian erythropoietin appear in the blastoderm as soon as the 24th hour of development. They represent most likely precursors of the megalocytic erythropoiesis, and do not seem to derive from stem cells common with normocytic erythropoiesis. Data concerning vitelline normocytic erythropoiesis were analysed in a kinetic model based on stochastic change of the stem cells. From this model it appears that 17-20 cell divisions are required for differentiation of erythrocytes from stem cells.


Anomalous ipsilateral optic fibre projection in Xenopus induced by larval tectal ablation.

Unilateral left tectal ablation was carried out in Xenopus between stage 48 and 1 month after metamorphosis. Six to 12 weeks after metamorphosis the retinal projection from the right eye was examined with the use of [3H]proline autoradiography. The autoradiographs indicated that optic fibres whose tectal target was destroyed recrossed to the ipsilateral tectum and basal optic nucleus via the posterior and pretectal commissures. No anomalous recrossing occurred if the tectal ablation was carried out at stage 58 or later. The aberrant optic fibres were restricted to the rostrolateral and central parts of the ipsilateral tectum and they terminated in a discontinuous manner. It is concluded that available surfaces serving as contact guidance cues are needed to direct aberrant optic fibres to the ipsilateral tectum.