The incorporation of Ga-67 into the ferritin fraction of rabbit hepatocytes in vivo.

Twenty-four hours after the administration of Ga-67 citrate and Fe-59 citrate, rabbits were killed and their livers removed and homogenized. Labile proteins in the filtered liver homogenates were denatured; ferritin was then crystallized from the supernatants by cadmium sulfate. Sephadex G-200 gel filtration of the ferritin fractions was done to determine the distribution of molecular weights in the substances associated with Ga-67 and Fe-59. It was found that Ga-67 was incorporated into the crystallizable ferritin fraction of rabbit hepatocytes with approximately one-sixth the uptake of simultaneously administered Fe-59. Gel-filtration chromatography confirmed that both the Ga-67 and the Fe-59 of the crystallizable ferritin fraction were associated with substances of the appropriate molecular weight for ferritin.


Effects of dry and humid climates on exercise-induced asthma in children and preadolescents.

Among factors which possibly influence the responses of asthmatic children to exercise, climate has received little attention. This study was performed to determine whether the level of air humidity is a factor to be considered. Twenty asthmatic (extrinsic perennial) girls and boys, 6 to 14 yr of age, with unverified history of exercise-induced asthma (EIA) took part. They rested and exercised in a climatic chamber in dry (25% relative humidity) and humid (90%) sessions at 25 degrees to 26 degrees C. One to three weeks separated the sessions, the order of which was counterbalanced. No changes in pulmonary functions (FVC, FEV 1.0, MMEF, MBC) were found following a sitting period of 60 min in either climate. Five and ten minutes following the treadmill run, however, bronchoconstriction was distinctly more pronounced in the dry than in the humid climate. Exercise heart rate and the subjective rating of effort were not affected by climate. It was concluded that, under the above experimental conditions, EIA is more likely in dry air than in humid air, possibly due to heat loss at the airway mucosa caused by evaporation. High humidity of inspired air could be the reason why EIA is less prevalent in swimming, as compared with other modes of exercise.


Prevention of homocytotropic antibody formation and anaphylactic sensitization by prefeeding antigen.

Studies were conducted in rats to determine whether pre-feeding antigen can prevent the development of Bordetella pertussis-induced homocytotropic antibody (HCA) formation and anaphylactic sensitization to the fed antigen. DA rats fed ragweed for a minimum of 2 wk and Sprague-Dawley rats fed horse serum for at least 4 wk demonstrated specific unresponsiveness to anaphylactic sensitization as measured by their inability to form HCA and their normotensive response to intravenous challenge with antigen. This state was more easily attained when feeding of antigen was started at the age of 30 days than at 90 or 150 days, and resistance to anaphylactic sensitization could be maintained by continued feeding of antigen. However, if anaphylactically sensitized to the fed antigen. The results suggest that induction of anaphylactic sensitization can be prevented by pre-feeding antigen.


Pathways to the decisive extension of the human specific lifespan.

Three approaches to reversal or removal of gerogenic aggregations of macromolecules have shown promise. Of these the enzyme approach is the most gentle, and can be made specific. Aside from this, the lower the molecular weight of an enzyme, the better chance it will have to be immunologically tolerated as well as replicated synthetically in whole or in part. The chelating approach provides a powerful means for removing a single class of unwanted, random crosslinkages, i.e., those due to extraneous polyvalent metals such as lead, cadmium and aluminum. The free hydroxyl radical approach is the most penetrant and most versatile means for removing otherwise insoluble aggregates, but its very lack of specificity will demand great foresight in control and use. Together, these three methods, when properly applied, might bring some principal objectives of gerontology within closer range.


Stable chronic disease: a behavioral model.

A three-year study was undertaken in the general medical clinic of a private community hospital, to assess the health behavior, health status, and profile of function of stable chronic disease was developed and tested. It was shown that these patients used a disproportionate amount of health care services. Half of the group was treated by a nurse practitioner/physician team and half by a house officer/preceptor team. Patients in both groups behaved similarly. These patients: 1) made frequent demands for outpatient services but did not need more than average hospital care; 2) tended to have problems of socio-economic indigency; 3) were likely to have hypertension, obesity, arthritis, and functional disease; 4) were chiefly women; 5) required special visits 9 percent of the time, usually for exacerbations of illness or intercurrent health problems; 6) made greater demands if they had functional complaints as a primary or secondary health problem; and 7) viewed their health more positively and functioned at a higher level if they were over 65 years of age. It was also found that the nurse practitioner, working in consultation with a physician, was able to provide high-quality health care.


Speech and hearing problems in the geriatric patient.

Regardless of the physical and mental health of any elderly person, verbal communications tend to deteriorate with advancing years. A significant loss of hearing occurs in 30-50 percent of persons over age 65, and voice changes are virtually inevitable after the age of 60. These alterations in the organs of communication increase markedly in incidence and degree after age 70. Deafness is a symptom and not a disease. Presbycusis is the result of degenerative changes affecting the organ of Corti and its central connections. The effects of aging on the expressive system of human communications may take different forms, including changes in the pitch, intensity, and quality of the voice. Therapy consists of rehabilitation, substitution, and amplification for the preservation and maintenance of communications. For patients with presbycusis, rehabilitation may include speech reading, auditory training, speech improvement, hearing-aid instruction, and guidance in social adjustment. Patients with speech and voice impairment need re-education by a skilled speech pathologist. With the aid of communication specialists, the geriatric patient can attain a high level of satisfaction from the give-and-take of personal communications.


Pigmentation and musculoskeletal abnormalities in an aged state hospital population.

In a State hospital population, 686 patients (mean age, 65 years) were examined for patterned pigmentation on unexposed areas of the body. Of these patients, 285 (41.5 percent) met the criteria for pigmentation. There were no sex or age differences. Psychosis had been diagnosed in 96 percent of the pigmented group. All of the 239 patients examined orthopedically had vertebral malalignment greater than 15 degrees and often had other bony deformities. Nearly all of the pigmented group displayed dyskinetic activity. A possible mechanism for the production of the pigmentation-psychosis-skeletal deformity triad could be an alteration in the concentration of available melatonin. Continued research along these lines may lead to an effective means of pharmacologic treatment as well as better psychologic and physical care.


The role of geriatrics in a family practice residency program.

In an attempt to clarify the role of geriatrics in family practice, the program at St. Joseph's Hospital and Nursing Home of Yonkers is outlined. The objective of this Family Practice Program are: 1) to assist in identifying needs in a target population and in developing and providing services to meet these needs; 2) to train and educate the Family Practice resident in the multiple medical, social and psychiatric problems of late life; 3) to indoctrinate this resident in the unique and numerous needs of the institutional elderly and of the institution in which they reside; and 4) to develop new and comprehensive systems of health care that will permit the marginally ill elderly to remain in the community as long as possible.


Applied genetics for the practicing optometrist. Part II.

Part I included a discussion of the four modes of single gene inheritance and examples of ocular anomalies which follow these modes. Part II will conclude the discussion by presenting other transmission forms in conjunction with examples of ocular anomalies characteristic of each. These two articles are designed to serve only as a guide for familiarizing the practicing O.D. with the various types of inheritance patterns, and not as a comprehensive text on the genetics of ophthalmic diseases. The reader is referred to the reference list for in-depth information regarding specific disorders.


Four clinical techniques in cup/disc ratio estimation.

Four clinical techniques for estimating cup/disc ratio were compared. Small differences in magnitude of the cup/disc ratio were found with each technique studied. In addition, a mean cup/disc ratio of between 0.22 and 0.30 was found for the subjects studied. The clinical utility of four funduscopic examination methods is reviewed.


The mydriatic effect of tropicamide on light and dark irides.

Tropicamide 0.5% was topically applied to the eyes of thirteen subjects to study its effectiveness as a mydriatic. A comparison was made between the subjects with light colored irides and those with dark brown irides, and also between those who had worn contact lenses previous to dilation and those who had not. Results indicate that a minimum pupil size of 6 mm was achieved within 25 minutes after instillation and this dilation was independent of iris pigmentation. Typically, IOP was reduced during dilation with tropicamide.


Cataract: current concepts, signs, and symptomatology -- Part II.

This is the second portion of a two part series presented with the purpose of providing the optometrist with an overview of lens structure, cataract formation, classification, and ensuing clinical implications. Part II deals primarily with the clinical signs and symptomatology as encountered during the optometric examination. Each section of a general visual examination is presented in an attempt to demonstrate how cataracts of various types might influence test results. Some suggestions are presented for modification of optometric testing procedures along with some auxiliary procedures which may be utilized in evaluating the cataract patient.


A visual guidance program: evaluation report.

A group of 80 fourth-grade pupils were administered visual tests to identify poor eye synchronization. The tests were given at Ruby Thomas Elementary School, Las Vegas, Nevada during February 1971 by Drs. Fred Erlich and Harvey Ohriner. Pupils with unsatisfactory eye synchronization were divided into two groups, a treatment and non-treatment or control group. A second control group was randomly selected from pupils with satisfactory eye synchronization. Visual guidance exercises were given to the treatment group. Post test reading tests showed significant gains in reading for the treatment group. Control groups showed non-significant gains.


Maddox rod to determine axis in laser refraction.

The procedure described offers a technique for determination of cylinder axis which requires no additional equipment beyond the basic apparatus used in a conventional laser refraction. An application of the Maddox Rod allows the patient to subjectively select the proper cylinder axis so that conventional laser refraction may proceed more smoothly and accurately


Anaglyphic T.V. ping pong antisuppression trainer.

Orthoptic therapy often entails the repetitive usage of detailed and complicated training procedures. Amblyopia and antisuppression therapy in particular may require weeks or months of trainning. A multipurpose training device has been constructed from commercially available components and promises to maintain a high level of patient interest and cooperation throughout the training procedure.


The optometrists' view of site selection.

Numerous studies have attemted to describe and to explain the maldistribution of physicians, yet few studies have attempted to explain the distribution of other health care personnel. Knowledge of the factors involved in the locational decision of relatively facility-free optometric practitioners offers excellent research opportunities for social scientists and holds forth promise for state and regional optometric associations concerned with attracting qualified optometrists. This study focuses on the impact of homestate and age on the decision to locate an optometric practice both in a state and in a community. In a survey of North Carolina optometrists the impact of homesite was found to be a significant factor in the locational decision. In addition, optometrists under 35 years of age were found to exhibit different locational preferences from their older counterparts.


Unilateral Morgagnian cataract.

Morgagnian cataract is a hypermature lens in which the cortex liquifies and the dense nucleus sinks inferiorly. Aside from the total reduction of vision, serious sequelae may result. This report is of a unilateral Morgagnian cataract of unknown etiology for which a dilated pupil facilitated the differential diagnosis between cataract types.


Optometry in Israel--a clinical externship.

In recent years, it has become the policy of optometric institutions to expand the scope of the clinical education experience beyond the geographical locale of the parent facility. This article is a presentation of the background and description of one such clinical program organized by the New England College of Optometry with the cooperation of the Department of Ophthalmology Hadassah Hebrew Medical Center, Jerusalem, Israel.


Drugs prescribed for the elderly.

The elderly constitute an increasing fraction of the population. Their lengthy exposure to the hazards of life makes them more likely to be suffering from chronic conditions as well as the inevitable effects of age itself. Many are receiving or have received prolonged courses of treatment by one or more drugs. This report summarizes the common medications and the typical doses which were being administered to 833 patients of whom all but 79 were age 65 or over.


The role of the trainer in visual training.

The role of the trainer in assisting the patient to internalize changes in his visual process is discussed. Emphasis is placed on two aspects: (1) making the patient aware of the nature of the change in process desired, and (2) making the patient aware that the phenomena which he observes during vision training techniques, such as blurring and clearing, diplopia, and suppression of training targets, which he may perceive as occurring esternally reflections of his visual process and as such, are subject to his control.


Case report: familial cone dysfunction.

Four generations of a family with hereditary retinal degeneration underwent extensive optometric evaluation, including electrodiagnostic testing. The compiled data indicate a partial cone dysfunction transmitted as an autosomal dominant trait. The optometrist's role in the diagnosis of such conditions is discussed as well as his role in family counseling. The cone dysfunction under discussion is differentiated from other tapetoretinal degenerations.


Vision care in Nicaragua.

In recent years an increasing number of professionals have become interested in providing health care to the people in under-privileged countries. Today, optometrists are serving these people through various organizations. This article relates the experiences of three University of Houston College of Optometry fourth year students during their volunteer optometric service in Nicaragua in the summer of 1975. Most colleges and schools of optometry have similar programs; however, only Ohio State University and the University of Houston participate in the Amigos de las Americas program. Participation in these and other similar activities by the profession is necessary to maintain good public health and develop better public relations.


A multidiscipline approach in low vision rehabilitation.

The first low vision clinic was instituted at the Industrial Home for the Blind in 1953 in response to a growing demand that more effective use be made of the residual vision in blind persons. The technology was there. Eye practitioners certainly knew enough about vision problems to offer many of these people an opportunity to enter a new life. The problem was: how to put that knowledge to work through a sound service delivery system. By its twentieth anniversary in 1973 the clinic had seen almost 5,000 patients for low vision evaluation and had built a service which consistently provided substantial and useful improvement in vision through the use of optical aids to a majority of its patients. The low vision optometric staff works as an integral part of a rehabilitation team. That team consists of a number of professionals, including ophthalmologists, social workers, teachers, and a variety of rehabilitation instructors. All members work simultaneously to create a total program of rehabilitation for thepartially sighted. We feel this teamwork approach is of basic importance to effective rehabilitation.


Criteria for fibular sesamoidectomy in hallux abducto valgus correction.

Fibular sesamoidectomy is necessary in certain instances to achieve permanent correction of hallux abducto valgus deformities. The decision to remove the fibular sesamoid is usually based on the position of the tibial sesamoid (which is determined by clinical and radiographic examination). If the fibular sesamoid is in positions 4 to 7, it is difficult, if not impossible, to maintain the correction of the hallux abducto valgus; if the position is 1 through 3, the sesamoids can be moved medially and maintained by the crista which is still intact.


A new method for measuring the amount of correction to be obtained by using a crescentic osteotomy of the first metatarsal.

In performing a crescentic osteotomy of the first metatarsal, it is difficult to determine how far to move the first metatarsal to place it in the ideal position. In this paper, the author presents a new method of measurement which makes it possible for the surgeon to calculate easily the amount of correction necessary and to measure this correction at the time of surgery. His technique eliminates the delay of an x-ray in the operating room before closure.


Neurodermatitis: a skin condition that affects surgical wound healing.

Neurodermatitis, or lichen simplex chronicus, generally found in females between the ages of 25 and 55 years, is highly inflammatory and may become generalized. The patients are hypersensitive and are not good candidates for surgery as a Köbner-like phenomenon may occur at the site of the incision with consequent breakdown of the incision, wound dehiscence, and infection. If these patients are to respond normally to treatment, it is sometimes necessary for the surgeon to treat their tensions.


Synopsis of managing soft tissue podiatric surgical infections.

Every effort should be made to prevent the occurrence of postoperative foot infections. However, since they are not always avoidable, every surgeon must be prepared to manage these infections. Early diagnosis is essential in preventing overwhelming sepsis and minimizing disability, and immediate therapy is necessary to resolve the complication and prevent it from recurring.


Proximal and distal Aiken procedures.

The Aiken osteotomy is the procedure of choice in the surgical treatment of the transverse plane structural deformity of hallux abductus caused by a high distal articular set angle and/or obliquity or asymmetry of the distal phalanx. The ease of performance and the reliability of both the proximal and distal types of Aiken osteotomy have resulted in functional and cosmetic success in the treatment of hallux abductus in the past 50 years.


Differential diagnosis of the thrombophlebitis.

Thrombophlebitis is a disease which is increasing in incidence because of increased longevity and because the effect on the clotting factor by modern drugs is difficult to diagnose, and the rate of accuracy in clinical diagnosis seldom exceeds 50%. In this paper, the authors present a clinical review of thrombophlebitis and discuss clinical findings of other diseases which may mimic thrombophlebitis. It is their hope that their findings may help practitioners to diagnose the disease with a greater degree of accuracy.


Polymethylmethacrylate--a review.

Methylmethacrylate, an acrylic plastic used in dentistry, has recently been adapted as a cement in bone surgery. It has proved to be useful in implant surgery and total joint replacement, in malignant neoplastic fractures, and in the fixation of pins and screws in bone. However, problems such as recurring infections, thermal necrosis of tissue, depression of myocardial and hepatic function, and leakage of monomer may occur with the use of methylmethacrylate.


Interphalangeal joint hallux ossicle.

The interphalangeal joint hallux ossicles that podiatrists are concerned with are those which produce pain as a result of keratoma and bursal sac formation. The patient with interphalangeal ossicles usually has a well delineated, circumscribed, painful excrescence with surrounding soft tissue inflammation, underlying bursal sac formation, and atrophy of the facial planes inferior to the interphalangeal joint. Although an interphalangeal ossicle may be clinically palpable, radiographic examination will not always confirm the condition as many interphalangeal ossicles are fibrocartilaginous and therefore are radiolucent. Dr. Sharon discusses diagnosis and surgical approaches which may be used to correct the problem.


Juvenile aponeurotic fibroma.

Juvenile aponeurotic fibroma is an uncommon benign tumor which occurs mostly in the hands and wrists of young children. It has a tendency to recur, especially in younger individuals, and this, combined with a rather cellular microscopic appearance, has led to mistaken diagnoses of malignancy. No instances of metastasis have been reported. Conservative therapy, consisting of excisional biopsy without sacrifice of vital structures, is indicated.


Juvenile aponeurotic fibroma: a case report.

Juvenile aponeurotic fibroma is a rare lesion of the palmar surface of the hands and, occasionally, of the feet of children. It is benign tumor which is self-limited in growth, but which has a strong propensity to recur. Grossly and histologically it exhibits a tendency to invade all surrounding structures, but does not metastasize. Rapidity of growth seems to slow down with increasing age. It should be recognized and treated conservatively by repeated surgical removal.


Dermal angiomyoma of the upper extremity.

Twenty-four of a series of 85 dermal angiomyomata were located in the upper extremity distal to the elbow. Eleven occurred in the right arm and 13 appeared in the left. Women exceeded men by 13 to 1 in number. The greatest number occurred in the 40 to 70 year age group. One case recurred as a leiomyosarcoma. This report is intended to bring this lesion to the attention of the surgeon in the differential diagnosis of painless masses of the hand, wrist, or forearm.


Multiple chondrosarcomas in a hand: a case report.

In a patient with multiple enchondromas limited to the ring finger and the fifth ray of the left hand, four of the enchondromas underwent simultaneous malignant degeneration during a 7 year observation period. Roentgenograms showed the initial cortical expansion changing to cortical destruction and soft tissue invasion. Ray resection of the involved digits was performed with a good functional result.


Periosteal (juxtacortical) chondrosarcoma: report of a case occurring in the hand.

A rare case of periosteal (juxtacortical) chondrosarcoma of the hand is described. Inadequate excision was followed by recurrence which necessitated a transmetacarpal amputation to eradicate the malignant disease.


Bradypneic, a new mutation in mice causing slow breathing, runting, and early death.

A new recessive mutation, bradypneic (bd), causes variable weight retardation beginning as early as 3 to 5 days. Severely affected mice die within the first 2 weeks; less severely affected mice may survive to weaning or to adulthood and may be fertile. Bradypneic mice tested at 3 weeks or older breathed at half the normal rate, but breathing was deeper and O2 consumption per unit of body surface was normal. No obstruction was found in the nasal passages, pharynx, trachea, or bronchi. The lungs were somewhat emphysematous and, probably in consequence, the right atrium was enlarged. The only other pathological conditions found were dilation of some of the distal tubules of the kidney and large amounts of gas in the stomach and intestines. The investigation did not reveal the cause of the breathing defect, but it is possible that the breathing defect is responsible for the emphysema, intestinal gas, small size, and early death. Extensive linkage tests have not yet revealed the chromosomal location of bd.


A cytomorphologic approach to standardization of cell dispersal evaluation for gynecologic specimens.

Dispersal of cells in gynecologic specimens is a major concern in sample preparation for automated systems. A variety of dispersal techniques has been tested, with varying degrees of success reported by many laboratories. In order to have a more meaningful comparison of the results of different techniques and among investigators, a standardized evaluation procedure is desirable. The cytomorphologic element of evaluation of cell dispersal has been developed and tested in this laboratory and might be applied toward this purpose. This evaluation reveals the changes in relative numbers of single cells according to cell type and estimates cell loss as well as changes in cellular preservation. Groups of cells are tabulated according to whether they appear as definite tissue groups or as clumps of single cells. Because abnormal cells and cells from the transformation zone often occur in tissue groups, special emphasis is placed on dispersal analysis by cell type to reveal whether the diagnostically important cells and cells relating to specimen adequacy were optimally dispersed. Furthermore, the format of evaluation allows assessment of the applicability of dispersal procedures to preparation for both flow and slide-based systems of automated cytology.


Enrichment of the gynecologic cytologic specimen.

The principal objective of our group has been and continues to be the improvement of the biologic signal-to-noise ratio of the gynecologic cytologic specimen through: the dispersal of clusters and clumps, enrichment of the abnormal component of the dispersed specimen by the method of rate-zonal sedimentation under unit gravity, and monitoring the effects of these manipulations with biophysical non-destructive methods. This report is concerned only with presenting the current status of the effectiveness of our enrichment techniques both qualitatively and quantitatively. Zonal sedimentation creates a broad spatial spectrum of cells by virtue of their different velocities which are directly related to their areas. Certain fractions of this spectrum are extracted, concentrated and deposited on slides for subsequent manual differential analysis, evaluation and documentation. Qualitatively, fractions M1 and M2 show impressive removal of erythrocytes, leukocytes and debris. Quantitatively, enrichment factors vary from 0.1 (depletion) to as high as 26 yielding an average enrichment of about 6, a most rewarding improvement.


Ultrasonic disaggregation of cell clusters.

A technique of cluster disaggregation has been developed and applied to cervical cells in suspension. Preliminary results indicate that ultrasound is capable of cluster disaggregation under controlled energy conditions. Cytologic quality of the resulting preparation is adequate for cell analysis in flow systems.


Effect of ascorbic acid on intraocular pressure of normal and buphthalmic rabbits.

Compared with serum the aqueous humor of the eye is high in ascorbic acid. The administation of ascorbic acid appears to have a transient hypotonic effect on intraocular pressure in normal eyes. Its effect on glaucomatous eyes is not as well defined particularly in the case of the rabbit. In this communication we shown the ocular hypotensive effal (+/+) or have hereditary glaucoma (bu/bu). We also show that the transport of ascorbate from the serum to the aqueous is much slower in buphthalamic rabbits than in normal controls. Osmolarity is suggested as a possible hypotensive mechanism and a differential fluid transport rate between the blood and the aqueous for the different genotypes is suggested as a possible mechanism for the difference in duration.


Natural selection for an intermediate level of heterozygote activity in Japanese quail.

Latter's hypothesis of a molecular explanation of the advantage of heterozygotes is based on the intermediate character of such phenotypes. Under particular environmental conditions the efficiency of heterozygotes might clearly be superior to that of wild-type individuals. This hypothesis is illustrated by two examples of egg-white proteins in quail.


A procedure for dissociating Ayre scrape samples.

The dissociation of cervical cell suspensions after various chemical and enzymatic treatments was monitored by using the  Centrifugal Cytology rotor to produce glutaraldehyde-fixed dispersions on conventional microscope slides and subsequent Pap staining. A special program was written in RPG II to record and analyze the results of the dissociation experiments in terms of white blood cells and the true cervical cells ("other cells"), and the degree of dissociation and recovery of both classes of cells. Since accurate differential counts on the untreated Ayre scrapes were difficult, the samples were syringed gently to break up the large or adventitious clumps. Cumulated results from control preparations indicate that the white blood cells and "other cells" are composed respectively of 92 and 63% single cells. The cells were further dissociated by: dissolving the cervical mucin sequentially with dithiothreitol and iodoacetic acid; depolymerizing the nucleohistone gel with ribonuclease; solubilizing the desmosomes with EDTA; removing the remaining cellular agglutinins with Varidase; and finally mechanical dispersion by hypertonic shock. The optimum procedure for dissociation involves the use of ribonuclease, dithiothreitol, iodoacetic acid EDTA, Varidase and sucrose shock. The white blood cells are now monodisperse and 81% of the "other cells" are found as single cells. If nuclear separation by two diameters is considered sufficient 98% of the "other cells" are single. The slide preparations are now sufficiently good that a scanning system is feasible.


Two sex-linked mutations in Culex tarsalis.

Two sex-linked mutations are described for the Culex tarsalis mosquito. Bleached ocelli (bloc) affects the pigment in the larval and pupal ocelli, and fringe (fr) puts in disarray the costal and vein scales on the adult wing. Their importance is discussed in relation to the basic genetics for this species and the accumulation of multiple-marker stocks needed for the analysis of genetic systems that show potential for control.


Optimization of the binding of dissociated exfoliated cervico-vaginal cells to glass microscope slides.

In order to monitor the development of a cell dissociation technique, it was essential to utilize the Centrifugal Cytology rotor to produce glutaraldehyde-fixed even cellular dispersions. The Cytology rotor has been improved to insure rapid alignment with the centrifugal field during both acceleration and deceleration, and the fixative is now delivered to the surface of the slide. The dissociation of the cells results in a loss of their adhesion to glass slides. Three bonding agents were tested: (a) Poly-L-Lysine; (b) Mayer's albumin fixative; (c) positively charging the slides with a silicone coating. The results with 65% albumin-coated slides were clearly superior to the other two. The addition of a postfixation step of 95% ethanol/4% polyethylene glycol did not significantly affect the recovery of the cells, but did eliminate some unevenness in the Centrifugal Cytology preparations, flattened the cells and expedited the procedure.


Crypt cell isolation in the small intestine of the mouse.

A successful method has been developed for isolating viable single cell suspensions of intestinal crypt cells from the small bowel of the mouse. The lumen of the intestine was perfused with a 0.2% trypsin solution that dissociated the lining epithelial cells. Crypt cell isolation, which proves to be extremely critical, occurred under optimal mechanical and chemical configurations about 75 min after the beginning of the procedure.


Flow-microfluorometric analysis of nuclei isolated from various normal and malignant human epithelial tissues. A preliminary report.

In order to obviate some of the technical problems associated with preparation of monocellular cell suspensions required for flow fluorometry, isolation of nuclei from several types of benign and malignant human tissues was undertaken. Satisfactory preparations of nuclei were obtained from epithelia of the uterine cervix and colon and from lung tissue using the citric acid method. The sucrose method was effective with colonic epithelium only. Distribution of deoxyribonucleic acid content in these nuclei was measured based on green fluorescence of acridine orange and red fluorescence of propidium iodide in a Bio-Physics Cytofluorograph. Essentially diploid patterns of deoxyribonucleic acid distribution were observed for all benign samples regardless of tissue origin whereas the malignant samples gave histograms suggestive of abnormal deoxyribonucleic acid distribution. Preliminary observations on distribution of single-stranded nucleic acids using acridine orange red fluorescence showed marked differences between populations of benign and malignant nuclei. Isolated nuclei appear to be suitable for flow-through microfluorometric analysis and offer some significant advantages over intact cells.


Automated classification of blood cell neutrophils.

The classification of white blood cell neutrophils into band neutrophils (bands) and segmented neutrophils (segs) is a subproblem of the white blood cell differential count. This classification problem is not well defined for at least two reasons: (a) there are no unique quantitative definitions for bands and segs and (b) existing definitions use the shape of the nucleus as the only discriminating criterion. When cells are classified on a slide, decisions are made from the two-dimensional views of these three-dimensional cells. A problem arises because the exact shape of the nucleus becomes indeterminate when the nucleus overlaps so that the filament is hidden. To assess the importance of this problem, this paper quantitates the classification errors due to overlapped nuclei (ON). The results indicate that, using only neutrophils without ON, the classification accuracy is 89%. For neutrophils with ON, the classification accuracy is 65%. This suggests a classification strategy of first classifying neutrophils into three categories: (a) bands without ON, (b) segs without ON and (c) neutrophils with ON. Category III can then be further classified into segs and bands by other stretegies.


A volume-activated cell sorter.

A sorter that is activated by the resistance pulses of a Coulter orifice (volume detector) has been developed. The problem arising from the distortion of the volume signals produced by the electric droplet forming and charging voltages are avoided by special shielding and grounding of the volume detector. Experiments with beads and leukocytes are described. The maximal processing rate at this time is 2000 cells/sec, the purity of sorted fractions is better than 97%, and the viability of sorted cells is better than 80%.


The influence of particles on jet breakoff.

Experimental observations of the breakup of a liquid jet issuing from a vibrated nozzle support the view that particles passing through the orifice tend to create a surface disturbance which, depending on the phase relative to the nozzle vibration, may either add to or subtract from the disturbance created by the nozzle vibration. The result may be either a shortening or lengthening of the jet breakoff length, depending on phase. The implications for sorting large cells with a fluorescence-activated cell sorter are discussed.


Proof without prejudice: use of the Kolmogorov-Smirnov test for the analysis of histograms from flow systems and other sources.

In this paper the Kolmogorov-Smirnov statistical test for the analysis of histograms is presented. The test is discussed for both the two-sample case (comparing fn1(X) to fn2 (X)) and the one-sample case (comparing fn1 (X) to f(X)). Presentation of the specific algorithmic steps involved is done through development of an example where the data are from an experiment discussed elsewhere in this issue. It is shown that the two histograms examined come from two different parent populations at the 99.9% confidence level.


An improved electronic pantograph.

A device was developed to permit simultaneous observations of a microscopic field, under any light optic conditions, and permanent recording of the relative X-Y coordinates of the center of the field. A modification is described that allows gray scale values to be assigned to the plots. The system can be readily adapted to digital computer data entry.


[Spine density of lamina V pyramidal cells in the visual cortex of laboratory rats after lengthy dark exposure].

1. The spine density was investigated at the apical dendrites of lamina-V-pyramidal cells in the visual cortex of rats following dark rearing for 250 days. 2. There are no significant differences in the average spine density between rats reared in darkness and normal conditions. 3. The results are discussed in view of the significant differences found after dark rearing for 50 days. It is assumed that these present results are caused by a delay in spine development in younger animals, which is compensated in the adult stage.


[Comparative-quantitative investigations of brains of wild and laboratory rats].

By means of serial sections at equal intervals through the brains of 8 Norway rats and 8 laboratory rats the fresh volume sizes of the optic regions lateral geniculate nucleus (pars dorsalis), striate area, superior colliculus and tractus opticus were determined. Furthermore the sizes of neocortical white matter and grey matter were measured in the same way. On the basis of the intraspecific allometric relation between brain size and body size differences in size of the structures between wild and domestic animals could be established. Especially the subcortical terminal regions of visual input and the striate area showed size reductions of different intensity from Norway to laboratory rats, whereas the optic tract underwent increase. The volume of grey matter showed decrease too and for the white matter no size differences between both groups came out as a result. These changes in size of nervous tissue from Norway rats to laboratory rats were partly discussed as due to the special changes of environment through domestication but in this special case also as due to the effect caused by the albinism of the investigated laboratory rats.


[Brain organization of Amia, Lepisosteus and Polypterus: comparative morphology and quantitative analysys].

The quantitative study of the brain of Amia, Lepisosteus and Polypterus leads to results which are compared to those previously got on the Sturgeon and three species of Tleosts (the Rainbow Trout, the Carp and the Ballan wrasse). The volumetric analysis has been applied at first to the whole brain (encephalization indices) and later to the main subdivision of the brain: telencephalon, diencepahlon... (relation indices and relative volumes). Some result are discussed in regard to the various opinions expressed about the phylogenetic affinities of the studied species. According to the brain organisation, the Bichir (Polyterus) seems to be at a relatively high level of evolution, and no relationship can be expected with the Chondrosteans. Amia and Lepisosteus show a common general brain pattern, but some peculiarities allow to consider Amia as more primitive than Lepisosteus--although the osteological characteristics and the scale structure lead to put together the first one and the Teleosts.


[Enzyme histochemical investigations of the nucleus nervi oculomotorii of Salmo irideus (Gibbons 1855) during biomorphosis. I. Transmitter enzymes].

1. The activities of the transmitter enzymes acetylcholine esterase (AChE), monoamine oxidase (MAO) and gamma-aminobutyrate transaminase (GABA-TR) were investigated in the nucleus nervi oculomotorii of Salmo irideus (Teleostei) during biomorphosis. 2. AChE as well as MAO-activity could be demonstrated already before the time of hatching, whereas GABA-TR occured from the 10th day post hatching. 3. In earliest stages the AChE activity was found in the neuronal pericarya of the nucleus n. III, but later on a strong increase occured in vestibular presynaptic terminals establishing synaptic contacts with the lateral surface of the large pericarya, especially in the medial part of the dorsal subnucleus and the ventral subnucleus. In adults the enzyme activity was localized in vestibular nerve fiber terminals exclusively due to the loss of enzyme activity in the pericarya. 4. The activity of MAO was found surrounding the neurons of the nucleus nervi oculomotorii with stronger deposits of reaction product at the medial surface that is not occupied by cholinergic terminals. 5. Besides non reacting neurons in all parts of the nucleus some cells in the transition zone between the medial and lateral area of the dorsal subnucleus, exhibited high cytoplasmatic GABA-TR activity. 6. The different ways of chemical transmission between nerve terminals and the neurons of the nucleus nervi oculomotorii are discussed.


The telencephala of Division I of the teleost fishes.

A comparative neuroanatomical investigation of the telencephala of Division I of teleost fishes (Greenwood, Rosen, Weitzman, and Myers, 1966) has demonstrated that homologies with other vertebrates can be made on the basis of the relationships of commonly recognized fiber bundles such as the olfactory tracts, forebrain bundles and commissures as well as those of cell types in the nuclear regions. A description and illustrations of the telecephalon of a typical representative brain (American eel, Anguilla rostrata) of Division I are presented with comparisons with the other elopomorphs and with the clupeomorphs. Although the brains may differ in gross appearance, all have certain unique microscopic configurations. They lack lateral ventricles and have a dorsal membrane as do all actinopterygeans. The brains of these fishes characteristically have (1) apposed sessile (or attenuated penduncluar) olfactory bulbs (2) which lack the typical vertebrate laminar pattern, (3) the preoptic region is unusually large although (4) the recess may not extend rostral to the anterior commissure, (5) the striatal and amygdalar areas are large and especially well differentiated, and (6) a nucleus glomerulosus is lacking. Other fishes differ from those in Division I in one or more of these anatomical features. These criteria may be used to differentiate teleosts of Division I from those of other Divisions.


The amino acid pattern of the british diet.

The amounts of the essential amino acids in the average British household food supply have been calculated and compared with those recommended by FAO/WHO. There were large excesses of all essential amino acids, with threonine being the most limiting. When similar calculations were made ten years ago, the sulphur-containing amino acids were the most limiting. The change results not so much from the greater amounts of animal protein which are now eaten as from changes in the reference pattern of amino acids.


Breast feeding practice of the hillside people of Thailand and its effect on the growth of their offspring.

Based on measurement of weight, height, head circumference, arm muscle and fat area, it seems that hilltribe infants in the first year of life can achieve a maximum growth potential similar to the general international standard of reference when they are on a total breast feeding regime. Theoretical calculations suggest that the undernourished hilltribe mothers may have a high capacity to lactate, especially in the first-year postpartum. Thus the practice of total breast feeding should be preserved and encouraged in every possible way. Also the infant should receive dietary supplements as he approaches the end of the first year.


Studies on the cytotoxic activity of human lymphocytes. II. Interactions between IgG and Fc receptors leading to inhibition of K cell function.

The inhibition of human K cell-mediated cytolysis by a variety of immunoglobulin preparations was studied. It was found that inhibition was not a simple function of IgG concentration but was dependent on the mode of immunoglobulin presentation. The relative inhibitory capacity of IgG preparations on a weight basis was as follows: cell associated Ig larger than or equal to immobilized immune complexes greater than insoluble immune complexes greater than soluble immune complexes greater than heat-aggregated Ig greater than monomeric Ig. These results imply that the inhibitory capacity of immunoglobulin is determined by ligand (Fc) multivalency. It was found that although interactions between immunoglobulin and Fc receptors occur at low temperatures, these interactions lead neither to lysis nor to K cell inactivation. A temperature-sensitive post-binding event was required both for lysis to occur and for K cells to be inactivated. These findings lead us to propose that an IgG-induced redistribution ("capping") of Fc receptors on the K cell surface may be required for cytolysis and for effector -ell inactivation.


Metabolic requirements for hormone-induced resistance to antibody-complement mediated killing of tumor cells.

Line-1 guinea pig hepatoma cells are susceptible to killing by anti-Forssman IgM antibody plus guinea pig complement (GPC). When these tumor cells are incubated with insulin, epinephrine, hydrocortisone, or prednisolone, the cells show a marked reduction in their susceptibility to antibody-C-killing. If the ability of the cells to synthesize DNA, RNA, and protein is impaired by pretreatment with metabolic inhibitors, x-irradiation, or culture in nutrient-deficient media, the hormones are no longer effective in rendering the cells resistant to killing. If only DNA synthesis is impaired, but not RNA and protein synthesis, the hormones are effective. The inability of cells inhibited in their macromolecular synthesis to be rendered resistant to killing after hormone treatment is not due to an inability of the cells to bind hormone.


Immune responses of outbred guinea pigs to the sequential polymer poly(L-Tyr-L-Glu-L-Ala-Gly): association of response to strain 13 histocompatibility locus.

Outbred Hartley guinea pigs were immunized with the sequential polymer poly(L-Tyr-L-Glu-L-Ala-Gly) (T-G-A-Gly)n, the random copolymer poly(Glu50,Tyr50) (GT), and a low dose of dinitrophenylated guinea pig albumin (DNP-GPA). Only the guinea pigs that carry strain 13 histocompatibility antigen responded to these three antigens as measured by the antigen-binding assay and delayed skin reaction. The immune response (Ir) genes controlling responses to (T-G-A-Gly)n, GT, or DNP-GPA, were found to be distinctive genes. Nonresponder inbred strain 2 guinea pigs were able to produce anti(T-G-A-Gly)n antibodies when immunized with (T-G-A-Gly)n and methylated bovine serum albumin (MBSA) complex, although there was no delayed skin reaction and in vitro cultured lymphocytes did not respond. The in vitro response to (T-G-A-Gly)n of cultured lymphocytes from responder animals was inhibited by strain 2 antistrain 13 alloantisera but not the strain 13 anti-strain 2 alloantisera.


Humoral immunostimulation. VIII. Increased incorporation of phosphate and turnover of phosphatidylinositol in cells treated with antibody.

Uptake of phosphate (32P) in L cells was modulated by reaction with anti-L cell antibody. A biphasic response was noted with high, cytotoxic concentrations inhibiting 32P uptake and low, cytostimulatory concentrations stimulating 32P uptake. Stimulation of 32P uptake was dependent upon multivalent binding as immune IgG and F(ab')2 were effective, but univalent Fab' was ineffective in enhancing 32P uptake into cells. Antibody stimulation of 32P uptake appeared to be an energy independent process and to take place by activation of 32P membrane transport with an increased Vmax (19.3 pmoles/min to 25.3 pmoles/min), but the same Km (0.22 mM). Isolation and measurement of cellular (primarily membrane) phospholipids demonstrated a dramatic increase of (2-fold) specific radioactivity in phosphatidylinositol. Early turnover of phosphatidylinositol may be an important signal for tumor cells to grow at an enhanced rate when exposed to cytostimulatory concentrations of antibody.


Synthesis and secretion of a mitogenic protein by macrophages: description of a superinduction phenomenon.

We have examined the biosynthesis and fate of the mitogenic protein (MP)2 made by macrophages. This mitogenic protein was assayed by its stimulation of DNA synthesis of thymocytes. Macrophages freshly harvested from the peritoneal cavity did not contain MP in the cell. The MP, however, rapidly appeared in the cell, reaching maximum amount by 1 to 2 hr, maintaining its concentration up to about 6 hr, and then disappearing. The disappearance of MP did not correlate with its secretion into the medium. The elaboration of MP stimulated by contact with the dish was inhibited by cycloheximide. Exposure of the cells to inhibitors of protein synthesis after 1 to 2 hr of culture at a time when the MP was already made produced a striking increase in secretion. The MP inside the cell did not appear to decay in the presence of cycloheximide. This result thus represents another example of superinduction. The intracellular fate of MP appears to be under the regulation of a control protein.


Antibody-dependent killing of erythrocyte and tumor targets by macrophage-related cell lines: enhancement by PPD and LPS.

Five murine macrophage or monocyte-related tumor cell lines in culture were compared for antibody-dependent lysis (ADL) of a tumor target (T lymphoma EL4) and lysis and phagocytosis of sheep erythrocytes (RBC). Some lines were effective only with RBC targets, while others were capable of lysing these and tumor targets. Both murine alloantisera to H-2 and Thy 1.2 antigens on the target cells and rabbit anti-mouse spleen and anti-mouse thymus sera directed target lysis, while normal mouse or rabbit sera were inactive. Preincubation of macrophage line RAW264 with lipopolysaccharide (LPS) or purified protein derivative from Mycobacteria (PPD) for 2 days resulted in an average of 76% or 86% increase, respectively, in ADL of RBC, although there was no stimulation in RBC phagocytosis or in nonspecific or antibody-dependent lysis of tumor targets. These results indicate that macrophage cell types are capable of antibody-dependent tumor lysis, that macrophages are probably heterogeneous in effector cell activities, and that they can be stimulated to increased ADL in the complete absence of other cell types.


Physicochemical characterization of a vascular permeability factor produced by con A-stimulated human lymphocytes.

Stimulation of human lymphocytes with Con A resulted in the liberation of a soluble lymphokine that increased the permeability of guinea pig skin capillaries. This factor, termed vascular permeability factor (VPF), was characterized by physicochemical methods. Upon gel filtration chromatography on Sephadex G-100, it eluted in a narrow peak with an apparent molecular weight of 12,000 daltons. The sedimentation rate of VPF, estimated by sucrose density gradient ultracentrifugation, was 1.8 S. Polyacrylamide gel electrophoresis showed the biologic activity to migrate in the beta region; the factor displayed a pI of 6.4 upon isoelectric focusing. These characteristics allow VPF to be distinguished from other human lymphokines.


Purification of C3b inactivator and demonstration of its two polypeptide chain structure.

C3b inactivator (C3bINA) was isolated from plasma by sequential ammonium sulfate gradient solubilization, anion exchange chromatography, gel filtration, and repeat ammonium sulfate gradient solubilization. The final product was pure as assessed by alkaline disc gel electrophoresis, isoelectric focusing, and SDS polyacrylamide gel electrophoresis, and elicited a monospecific rabbit antiserum. The normal serum concentration of C3bINA was found to be 53 +/- 9 microgram/ml (mean +/- S.D.). Heterogeneity of purified C3bINA was apparent on alkaline disc gel electrophoresis and isoelectric focusing, but not with SDS polyacrylamide gel electrophoresis and thus is attributed to forms of C3bINA that differ in charge rather than in size. SDS polyacrylamide gel electrophoresis of unreduced, alkylated C3bINA yielded a single stained band with an apparent m.w. of 93,000, whereas the reduced protein demonstrated two bands of 55,000 and 42,000 m.w., thereby establishing a composition of two disulfide-linked polypeptide chains for C3bINA.


The rate of membrane turnover by the hairy cells of leukemic reticuloendotheliosis: a kinetic study of a receptor for IgM.

The kinetics of turnover of a surface receptor for IgM were studied in six cases of hairy cell leukemia and four cases of chronic lymphocytic leukemia and the results were used as an indicator of membrane turnover in the pathologic cells of these two chronic leukemias. Rosette formation, shedding, and reexpression with EA (IgM) by hairy cells, a process which was shown to require protein synthesis, followed a cyclical pattern with a modal peak-to-peak reexpression time of 6 hr. Membrane turnover time in the cases of chronic lymphocytic leukemia was 12 hr or more. This work indicates that a reduced membrane turnover rate is not an essential feature of chronic leukemias. The more general application of this type of approach to the study of membrane turnover in normal cells and their pathologic counterparts are discussed.


Antibody-dependent, complement-mediated homologous and cross-cytolysis of togavirus-infected cells.

The appearance of viral induced antigen(s) on Sindbis and Semliki Forest (SF) virus-infected chick embryo cells is detected with homologous mouse anti-viral antibody-dependent, complement-mediated cytolysis in a 51Cr release assay. Such antigen(s) is detected within 3 to 4 hr after infection whereas viral cytopathologic effects do not occur until 10 to 12 hr. In addition, significant heterologous cytolysis (anti-Sindbis on SF virus-infected cells, or, anti-SFV on Sindbis-infected cells) could be observed with serum samples tested. These sera were assayed for neutralizing and hemaggluatination-inhibition activities. Heterologous cytolysis cannot be correlated with hemagglutination-inhibition antibody titers nor is cross-neutralizing antibody detected. The possibility is discussed that another previously undefined antigen on infected cell membranes may be involved in heterologous cytolysis.


The role of antigen-antibody complexes in mediating immunologic unresponsiveness in the chicken.

A significant suppression of an adoptive secondary response to either BSA or sheep red blood cells (SRBC) resulted when chicken spleen cells were incubated with BSA-antibody (Ab) complexes formed in antigen excess (molar ratio Ag1Ab1.1). Serum from 14-day old birds rendered unresponsive at hatching with large doses of BSA also produced suppression. In addition the in vitro response to SRBC was suppressed by the above treatment. Complexes formed at equivalence either had no effect on the subsequent responses of primed cells or produced an enhancement. The fact that both the homologous and heterologous responses were inhibited could indicate that both were suppressed via a common pathway.


Plasma cells and epithelial immunoglobulins in the mouse mammary gland during pregnancy and lactation.

Mammary glands of virgin, pregnant, lactating, and post-weaning CAF1 mice were studied by immunofluorescence for epithelial immunoglobulins and stromal plasma cells. In normal virgins, both females and males, only occasional plasma cells and but scanty intraepithelial immunoglobulins were present. During pregnancy and the early days of lactation, an increase in the number of plasma cells occurs, and by 1 week of lactation, there is a marked increase in the number, and most are synthesizing IgA. This increment parallels the development and proliferation of the glandular epithelium, in anatomical relation to which the plasma cells are observed. The intraepithelial content of IgA is also maximal when the glandular epithelium is most developed. These findings are consistent with a local production of the IgA in milk. Weaning, or deliberate interruption of suckling for more than 10 days results in a sheep decrease in the number of IgA plasma cells and an involution of the epithelium, whose content of immunoglobulin also declines. The changes in the secretory IgA system which occur locally during lactation are thought to be hormone-dependent.


Complement activation in semisolid media: insolubilization of alternative pathway convertases in agar gels with C3 nephritic factor-containing sera.

It was previously shown that when normal human serum and purified properdin or serum containing nephritic factor were allowed to diffuse toward each other in agar, a stainable precipitin line formed only in the presence of an intact alternative pathway. In the present study we have shown that when guinea pig erythrocytes were incorporated in agar and normal human serum was allowed to diffuse toward C3 nephritic factor-containing serum, a line of hemolysis appeared that coincided with the stainable line. The line of hemolysis only formed in the presence of C3, factor B and Mg++, as does the properdin or C3 nephritic factor-induced stainable line. When C2-deficient serum was incorporated in agar with guinea pig erythrocytes and serum samples containing nephritic factor were applied in wells, rings of lysis developed, the areas of which correlated significantly with nephritic factor activity. When partially purified nephritic factor (contaminated only with IgG) or serum containing nephritic factor was subjected to electrophoresis in agar-guinea pig erythrocyte gels and overlaid with normal human serum, bands of lysis developed that required factor B and C3. Lysis of guinea pig erythrocytes in this system appears to be due to insolubilization of alternative pathway convertases with activation of the membrane attack unit C5-C9.


Enhancement of C56-initiated lysis by cell-bound C3 fragments: evidence for a mechanism independent of the prior binding of C56 to C3b.

Cell-bound C3b can reversibly bind C56, the activated complex of the fifth (C5) and sixth (C6) components of complement, and in this way potentiate C56-initiated lysis by favoring the formation of C567 at the cell surface. We report here another way in which cell-bound C3 fragments can enhance C56-initiated lysis, which involves C567 generated in the fluid phase rather than at the cell surface. Evidence for the involvement of fluid phase C567 was obtained by use of dextran sulfate, which is known to inhibit the hemolysis of E mediated by fluid phase C567. Dextran sulfate strongly inhibited the formation of C567 sites on cells bearing C4b and C3b (EAC4b3b) as well as on unmodified E when C56 and C7 were added simultaneously to the cells. By contrast, dextran sulfate had virtually no effect on the reaction sequence involving the prior binding of C56 to C3b and subsequent formation of C567 at the cell surface. Treatment of EAC4b3b with either anti-C3 Fab' fragments or the C3b inactivator reduced but did not eliminate the enhancement of hemolysis, raising the possibilities that a C3 fragment(s) other than C3b also can enhance C56-initiated lysis and/or that the enhancement is indirect without a requirement for an interaction between C567 and the cell-bound C3 fragment itself.


Allotype suppression at the CHn locus of IgM in heterozygous rabbits with concomitant suppression at the VHa locus of all Ig classes.

Doubly heterozygous offspring, malen83alpha1/femalen82alpha2 or malen81alpha1/femalen82alpha2, of doubly homozygous dams, n82alpha2/n82alpha2, were injected neonatally with anti-n83 or anti-n81 antibody directed to the paternal n locus allotype. In each injected rabbit, the paternal n83 or n81 allotypes were suppressed and increased amounts of the maternal n82 allotype accounted for almost all of the serum IgM. This is the first report of allotype suppression in the rabbit mediated by antibody directed toward the CH region of any Ig class. A concomitant decrease in the paternal a1 Ig allotype and a compensatory increase in the maternal a2 allotype were also found. This finding supports the concept that IgM-bearing cells are the precursors of IgG-secreting cells.


Initiation of bone resorption by the classical and alternative C pathways and its mediation by prostaglandins.

Activation of C by immunoglobulins reactive with cell surface antigens stimulated synthesis of prostaglandin E and resultant release of 45Ca from organ cultures of fetal rat bones labeled in utero. Absorption of the C source (rabbit serum) with rat spleen cells or dilution of C abolished this activity which was, however, restored by addition of immunoglobulin preparations from sera of rabbits immunized with either rat erythrocytes or sonicated fetal rat bones. The active reconstitution factors were present in the 50% (NH4)2SO4 cut of the antisera and were eluted from Sephadex G-200 in both the 19S and 7S fractions and from DEAE cellulose in the IgG-containing fraction. The C-dependent resorption of bone by the F(ab')2 fragments of the IgG antibody implicated a role for the alternative C pathway in this event. Complete inhibition of this biologic effect by indomethacin and detection of enhanced levels of prostaglandin E in the media of cultures containing antibody and C demonstrated a role for prostaglandins as mediators in the destruction of bone initiated by immune activation of C.


Echinococcus granulosus: identification of subunits of the major antigens.

This paper describes a procedure for obtaining the two "major" Echinococcus granulosa antigens (antigens 4 and 5) from sheep hydatid fluid. Purified antigen 4 (m.w. 400,000 and over) and antigens 5 (m.w. 150,000) were separated in order to identify molecular subunits of each. SDS 12.5% polyacrylamide gel electrophoresis of antigen 4 gave a single band for which a m.w. of 67,000 was calculated. When 5% 2-mercaptoethanol was added to the antigen 4 preparation, SDS 12.5% polyacrylamide gel electrophoresis gave two bands (m.w. 47,000 and 20,000). SDS 12.5% polyacrylamide electrophoresis of antigen 5, with or without 2-mercaptoethanol, gave at least three bands with m.w. ranging from 10,500 to 20,000. Low m.w. subunits of both antigens were eluted in a single peak from a preparatory SDS 7.5% acrylamide gel electrophoresis; these subunits were found to be immunogenic and antigenic.


Spontaneous cell-mediated cytotoxicity by human peripheral blood lymphocytes in vitro.

With isolated subpopulations of human peripheral blood mononuclear cells, spontaneous cell-mediated cytotoxicity (SCMC) and antibody-dependent cellular cytotoxicity (ADCC) were demonstrated to be functions of a population of surface immunoglobulin (sIg)-negative, Fc receptor-positive lymphocytes. Organ distribution studies revealed that SCMC and ADCC effector cell activity occurred in parallel, being present in spleen and absent in tonsils. Treatment of effector cells with trypsin significantly inhibited SCMC but not ADCC and this inhibition was not reversed by short-term incubation in autologous plasma. These results demonstrate that SCMC and ADCC involve different cytotoxicity mechanisms and are consistent with two subsets of effector cells with separate functional capabilities or a single effector cell mediating both forms of cytotoxicity through separate mechanisms.


Characterization of antigen-binding receptors in vitro. I. An equilibrium method for measuring antigen binding in murine cells.

This study reports a novel method for studying the binding of soluble antigen (bovine serum albumin, BSA) to surface receptors on lymphoid cell populations under equilibrium conditions in the presence of 10% normal rabbit serum or 20 mg/ml of ovalbumin. Virtually no nonspecific uptake was demonstrated to nonlymphoid tissues. Detectable quantities of BSA could be found on both nonimmune and immune lymphoid populations. The binding of BSA was demonstrated to be antigen specific and to be proportional to the numbers of binding cells. The quantity of antigen bound was proportional to the free antigen exposed to the population and saturation could be achieved with 1 to 2 microgram of antigen/1.2 ml of culture, per 10 X 10(7) cells. The kinetics of antigen binding was very rapid and occurred within 10 min at 4, 27, and 37 degrees C. The binding was independent of the viability of cells. Binding was antigen specific and could be partially blocked by cross-reacting serum albumins, but not by non-cross-reacting albumins. Binding was independent of cytophylic antibody concentrations in the serum.


Lytic activity of C5-9 complexes for erythrocytes from the species other than sheep: C9 rather than C8-dependent variation in lytic activity.

With the reactive lysis system, a form of hemolysis mediated solely by the late-acting complement components (C56, C7, C8, and C9), guinea pig C9 (C9gp) was found to be very inefficient in inducing the lysis of guinea pig and mouse erythrocytes bearing human C5-8. By contrast, C9gp could efficiently induce the lysis of sheep and goat erythrocytes bearing human C5-8. By contrast, C9gp could efficiently induce the lysis of sheep and goat erythrocytes bearing human C5-8. Human C9 was efficient in the lysis of erythrocytes from the species mentioned above. Further study showed: 1) the observed inefficiency in the lysis of guinea pig erythrocytes was not due to incompatibility between human C7 or C8 and C9gp; 2) C9gp could efficiently bind to guinea pig erythrocytes bearing human C5-8 but was inactive in the subsequent lytic process. The present finding emphasizes a role for C9 in complement-mediated membrane damage which may not be a simple effector function of C8 action.


Serologic identification of hemopoietic progenitor cell antigens common to mouse and man.

An immunologic approach to the identification of human hemopoietic stem cell antigens has yielded two types of antisera. Both recognize antigens on hemopoietic progenitors which are identical or cross-reactive with determinants on sperm, brain, and, across the species barrier, with murine hemopoietic stem cells (CFU-S). The first antiserum was the result of an autoimmune reaction in a post-vasectomy patient. The second was produced in rabbits after injection of human brain cells.


Some immunochemical properties of Le(a)-and Le(b) -active substances in human urine.

Le(a)- and Le(b)-active substances present in normal human urine were isolated by sequential membrane filtration, column chromatography with DEAE-Sephadex (A-25), and gel filtration of Bio-Gel (P-6). Le(a)-active substance with the highest activity was obtained in the P-1 fraction (0-8 mg/1) from urine of a non-secretor type by the final gel filtration. Le(b)-active substance with the highest activity was also obtained in the P-1 fraction (0-2 mg/1) from urine of a secretor type. The P-1 of the non-secretor inhibited Le(a)-anti-Le(a) haemagglutination minimally at 4 microgram/ml, and the P-1 of the secretor inhibited Le(b)-anti-Le(b) haemagglutination at 1 microgram/ml. Both P-1 fractions possessing Le(a) or Le(b) activity were characteristic in having a much higher focuse content than other isolated fractions, and seemed to be complex carbohydrates with molecular weights of the order of 4-5 x 10(3).


The defect of Mk erythrocytes as revealed by sodium dodecylsulphate--polyacrylamide gel electrophoresis.

The membrane glycoprotein defect of Mk red cells was studied using discontinuous sodium dodecylsulphate--polyacrylamide gel electrophoresis, carbohydrate analytical procedures and lectins. The results suggest that Mk erythrocyte membranes contain only about half of the normal amount of the MN and Ss sialoglycoproteins, both molecules being not qualitatively different from normal. The major membrane protein exhibits an increased molecular weight in the above red cells. Some data on Mg and Miltenberger class five cells are presented for comparison. The results on Mk erythrocytes support the operon concept of the MNSs blood group system proposed previously.


Measurement of skin thickness: a comparison of two in vivo techniques with a conventional histometric method.

Two in vivo techniques which are rapid, inexpensive, and reproducible have been investigated. The first is a standardized radiological (xerographic) technique which we have shown is capable of detecting small degrees of dermal atrophy after the application of topical corticosteroid preparations for only one month. The second technique employs the Harpenden Skinfold Caliper used in an unconventional manner so as to exclude subcutaneous fat. We have shown that this too is capable of detecting dermal atrophy from the application of topical corticosteroids and that there is a strong correlation between the two techniques (r = 0.82, p less than 0.001). Histometric techniques, on the other hand, give inaccurate and erroneous results for dermal thickness.


Viscoelastic properties of intact human skin: instrumentation, hydration effects, and the contribution of the stratum corneum.

Past attempts to relate mechanical properties of the stratum corneum to its state of hydration or disease condition have been hampered by the lack of adequate in vivo methodology. With the technique described herein, viscoelastic properties of the skin surface can be measured in vivo or on excised skin in vitro. This method is sensitive enough to detect changes in the properties of the stratum corneum induced by topically applied agents or by mechanical disruption of it. The viscoelastic data suggest that the outermost layers of the stratum corneum may become dry in vivo despite their proximity to the underlying wet tissue, and that the instrument is sensitive enough to measure the changes that occur. It also appears that the in vivo behavior of the stratum corneum is similar to that of isolated stratum corneum membranes which become brittle when they are dried and supple when they are hydrated.


Thermoanalytical probes for the analysis of physical properties of stratum corneum.

Thermomechanical analysis (TMA) measurements were used to characterize the physical properties of normal and treated stratum corneum. TMA measures changes in the overall mechanical and dimensional properties of the material as a result of temperature-induced transformations such as lipid melting, protein conformational changes, and glass transitions. TMA indicates several softening transitions in normal human stratum corneum which occur at about 50, 150, 260 degrees C. Differential scanning calorimetry and thermomechanical analysis studies show that the 50 degrees C softening is related to lipid melting as well as protein side chain motion while the other observed transitions appear to result from a change in state of the protein organization and decomposition, respectively. These particular parameters are sensitive to changes in composition and macromolecular structure of the stratum corneum brought about by disease and chemical alteration. Shifts in the temperature of the second softening transition are observed between normal stratum corneum and stratum corneum treated with a retinoic acid product, occlusive therapy, petrolatum, propylene glycol-ethanol, and normal saline. These shifts imply a possible alteration of the higher-level organization of the keratin in the stratum corneum.


Stratum corneum studies with photoacoustic spectroscopy.

Conventional spectroscopic studies on opaque membranes are difficult to pursue because of excessive light scattering and complications arising from the specimen's surface. Recently, a new spectroscopic technique, photoacoustic spectroscopy, unlike conventional optical spectroscopy, has been demonstrated to be an informative technique amenable to spectroscopic studies of solids and membrane-like samples. We have applied photoacoustic spectroscopy to the study of hydration and maturation of newborn rat stratum corneum, and have obtained clean spectra in the 220 to 450 nm region indicative of a change in thermal diffusivity with increased hydration, and biochemical changes associated with the initial maturation period.


Measurement of intracorneal cohesion in man using in vivo techniques.

Measurement of intracorneal cohesion in vivo should provide quantitative information concerning the process of esquamation. In this investigation three techniques have been employed to measure the internal binding forces within the stratum corneum in vivo. The first technique, cohesography, directly measures the force required to remove stratum corneum of partial thickness from a known area of skin. Sex and site differences were detected using this method. A second technique employs a surfometer to measure the surface contours of the internal face of skin surface biopsies. Quantitative assessment of surfometer tracings reflects both internal structure of stratum corneum and its cohesive property. Positive correlations were found using these two techniques on the forearms of 16 normal subjects. A scrub technique has also been used which can deliver a controllable and measurable stimulus to the skin surface to release corneocytes. Counting the number of corneocytes released provides a further measure of internal cohesion of the stratum corneum but the results did not correlate with the other two techniques. It is suggested that the use of all three techniques might provide a profile of the cohesive property of the stratum corneum.


Frictional properties of skin.

To gain a more fundamental understanding of the frictional properties of skin, various parameters (hydration, oils, surfactants, etc) affecting skin friction were evaluated using a rotational friction technique. The rotational device utilized a motor-driven wheel. The wheel and drive shaft were connected to the motor through a flexible rubber coupling in such a way that friction encountered by the wheel caused a lateral displacement of the drive shaft which was monitored by a photocell and recorded on a strip-chart recorder.


Laser Doppler measurement of cutaneous blood flow.

This work describes an instrument for the noninvasive measurement of cutaneous blood flow velocity. The system utilizes the Doppler shift of laser light backscattered from moving red blood cells in the cutaneous microcirculation, the shift being obtained by an optical heterodyning technique. Comparison is made between this technique and the 133xenon clearance technique in measuring cutaneous flow in the forearms of normal volunteers. Variations in flow were obtained by inducing different degrees of solar erythema with an ultraviolet sunlamp. A Y on X linear regression yielded a regression coefficient = 0.89 (p less than 0.001, n = 16) between the two methods. The laser Doppler method appear to represent a practical technique for clinical evaluation of cutaneous blood flow in any skin surface.


Accounting for natural tension in the mechanical testing of human skin.

A low-pressure suction device has previously been used to obtain quantitative information on the mechanical properties of human skin. Reducing the raw data from this technique was complicated by the fact that the skin is already biaxially loaded in the natural state and a rational basis on which to compare results obtained from different subjects was not available. Using a strain gauged pretension device, a procedure for determining the natural state tension and extension fields in the skin has been developed. The natural tension was then relaxed in the direction of testing and the suction device used to determine the two-dimensional skin tension-extension ratio response of the skin at a constant testing rate. The data from a number of subjects have been used to develop a new multidimensional stress-strain theory in terms of two material constants that are related to basic material characteristics of the dermis and are uniquely determinable regardless of the natural state stress field. Tests performed on the upper backs of 23 healthy adult males were used to investigate the variations in the material constants with age and sun-exposure habits. Age variations were found to corroborate earlier studies, and variations with sun exposure habits were related to known dermal collagen and elastin changes due to ultraviolet radiation. The apparent sensitivity of the testing procedure to physiologic state variables holds out the hope that mechanical properties characterization will be a useful tool in the evaluation of the severity of certain pathologic states and the effect of therapy.


A ballistometer for the study of the plasto-elastic properties of skin.

The ballistrometer is based on the "drop impact" of a body onto a stationary surface. A collision in one dimension is provoked by allowing a hard body to drop from a given height onto the skin surface to be tested. After the collision, the impacting body undergoes a variable number of rebounds decreasing in amplitude. By measuring the height of the rebounds, the amount of energy returned by the tissue is calculated in terms of coefficient of restitution e. The equipment, consisting of a hammer unit, a feeder-amplifier, and a plotter, is described. Ballistometry has been carried out on skin areas of 46 normal subjects ranging in age from 8 to 80 years, as well as on pathologic and cadaveric skin. The results show a progressive decrease of e with advancing age, as well as differences related to various skin regions. The tests performed on pathologic skin showed a lowered e in epidermal hyperplasia, hyperkeratosis, sclerosis, and dermal infiltration. An increase of e was observed when a high water content was present in the skin.


The time-dependent mechanical properties of skin.

The mechanical properties of the skin were investigated by applying a torsional deformation to a circumscribed area. Results of the tests indicate that the skin is nonlinear and time dependent. The dynamic response shows that the phase angle is insensitive to frequency below 1 Hz; the peak torque amplitude increases slowly above 0.004 Hz. A self-consistent description is presented utilizing continuous relaxation spectra. The technique describes the stress relaxation data and predicts the form of the dynamic response.


A method of in vivo measurement of the elastic properties of skin in radiotherapy patients.

A portable apparatus which measures the force-displacement or elastic characteristics of skin in vivo has been constructed. A displacement transducer between two arms measures the extension of the skin as the arms are driven apart by means of a lead screw, and the force exerted on the skin is measured by means of two electrical resistance strain gauges attached at a reduced section on each of the arms. The apparatus was designed for compactness and lightness to enable it to be hand held. Illustrative results obtained on patients during and after courses of radiotherapy are presented.


Relation of electrical properties of skin to structure and physiologic state.

Interpretation of cutaneous electrical measurements in terms of structure or function requires special techniques for identifying the separate contributions of the various elements. Resistance measures have different implications depending upon whether the subject is sweating and whether the electrode preparation is wet or dry. Microelectrode measurement indicates that the stratum corneum and other epidermal layers represent a significant pathway for ion conductance. Impedance measurement allows estimation of skin capacitance and of the thickness of the capacitative element. When applied to data on excised stratum corneum, this analysis indicates the presence of a relatively impermeable layer less than 2 microns thick, a conclusion subject to some doubt because of uncertainty over the dielectric constant of wet keratin. The use of impedance measurement with closely spaced electrodes gives an indication of the hydration of the superficial horny layer and also demonstrates reabsorption of sweat from this region. Potential measurement at the surface reflects the relative internal resistance of two parallel sources, the sweat glands and an "epidermal generator."


Thermal properties of human skin related to nondestructive measurement of epidermal thickness.

The thermal and physical properties of skin are reviewed briefly in the light of their relationship to skin functions and their influence on temperature measurement and related methodology. Thermal conductivity and epidermal thickness have a direct bearing on the majority of skin functions as these significantly affect heating rates, thermal pain thresholds, and blister formation, as will be seen from the experimental data. The accumulated data on both temperature and pain threshold are used to estimate epidermal thickness in the intact individual. The procedure to accomplish this end utilizes the measured thermal pain threshold, surface temperature, exposure time, and incident energy on a standardized skin site (volar surface of the forearm) to obtain conductivity values. These values are then used in a two-layer system heat flow equation to determine epidermal thickness in other skin sites (fingers) referred to the standard area. Systematic exposures to various materials at high temperatures in contact-burn studies provide data for checking the reliability of this procedure by alternative computations and comparison with predicted tissue temperatures derived from earlier work. Certainly, blister formation and physical measurement of the excised epidermis would provide direct verification of the accuracy of the procedure but these measures have not been undertaken. It is quite possible that over a period of time verification data could be accumulated incidentally in the clinic where skin grafting procedures are carried out. For this purpose only relatively simple measurements of skin temperature and pain threshold, as described herein, would be required to be performed on skin graft donors to provide epidermal thickness measurements for comparison with direct measurements of excised skin immediately after removal.


Measurement of the thermal properties of human skin. A review.

The effective use of techniques to detect and interpret surface and subsurface temperature patterns depends upon the thermal and electromagnetic properties of the tissues under examination. A review is made of the existent data concerned with the thermal characteristics of skin. The thermal conductivity, k, and thermal inertia, kpc, as measured as a function of local temperature are presented. The radiation properties of the skin are presented as a function of wavelength. It is concluded that the information currently available about the thermal properties and their normal variation with body site, skin color, and age is quite limited.


Nonspecific protection of mice against influenza virus infection by local or systemic immunization with Bacille Calmette-Guérin.

Bacille Calmette-Guérin (BCG) has been used effectively to protect nonspecifically against bacterial infections and neoplasms, probably by enhancement of cell-mediated immunity. It has been suggested that cell-mediated immunity plays a role in the host defense against certain viral infections. In recent in vitro studies, macrophages from animals sensitized by BCG were more effective in lowering the titer of influenza virus than were macrophages from control animals. The purpose of this study was to investigate the in vivo effectiveness of nonspecific immune stimulation with BCG on influenza virus infection in mice. Immunization with BCG resulted in significant protection of mice. Also, the local (nasal) route of immunization was more effective than the systemic (intraperitoneal) route against the intranasal inoculum of virus, a finding which suggests that important role of local immunity, i.e., either earlier stimulation of secretory antibody or nonspecific cell-mediated immunity. The time course of the resistance ot infection suggests that interferon was not the protective mechanism.


Encephalitis in ferrets caused by a nonproductive strain of measles virus (D.R.) isolated from patient with subacute sclerosing panencephalitis.

A nonproductive, syncytiogenic strain (D.R.) of measles virus, isolated from a patient with subacute sclerosing panencephalitis (SSPE), was inoculated intracerebrally into ferrets in an attempt to induce subacute encephalitis. Inoculation of freeze-thawed syncytia before immunization was the least effective procedure, and inoculation of live syncytia after immunization with measles virus vaccine was the most effective procedure, for induction of subacute or persistent subclinical encephalitis in the animals. After the latter procedure three of five ferrets developed subacute or subclinical encephalitis, whereas ferrets inoculated with live syncytia without prior immunization consistently contracted acute fatal encephalitis in one to two weeks. The subacute encephalitis in ferrets was characterized by high titers of antibody to measles virus in serum. At the time of sacrifice 1.25, 4.5, or 8.0 months after inoculation, brains of the ferrets showed histologic lesions similar to those characteristic of SSPE, and nonproductive syncytiogenic measles virus was recovered from the brains of two of the animals. All three ferrets had greatly increased concentrations of gamma-globulin in their brains and high levels of neutralizing and hemagglutination-inhibiting antibodies to measles virus. Only one of these animals developed clinical signs 1.25 months after inoculation.


Temperature-sensitive mutants of influenza A virus. XIV. Production and evaluation of influenza A/Georgia/74-ts-1[E] recombinant viruses in human adults.

The two temperature-sensitive (ts) lesions present in influenza A/Hong Kong/68-ts-1[E] (H3N2 68) virus were transferred via genetic reassortment to influenza A/Georgia/74 (H3N2 74) wild-type virus. A recombinant clone possessing both ts lesions and the shutoff temperature of 38 C of the Hong Kong/68 ts donor and the two surface antigens of the Georgia/74 wild-type virus was administered to 32 seronegative adult volunteers. Thirty-one volunteers were infected, of whom only five experienced mild afebrile upper respiratory tract illness. The wild-type recipient virus was a cloned population that induced illness in five of six infected volunteers. Therfore, the attenuation exhibited by the Georgia/74-ts-1[E] virus could reasonably be assumed to be due to the acquisition of the two ts-1[E] lesions by the Georgia/74 wild-type virus. The serum and nasal wash antibody responses of the ts-1[E] vaccinees were equivalent to those of the volunteers who received wild-type virus. The two ts lesions present in the Hong Kong/68-ts-1[E] virus have now been transferred three times to a wild-type virus bearing a new hemagglutinin, and in each instance the new ts recombination exhibited a similar, satisfactory level of attenuation and antigenicity for adults. It seems likely that the transfer of the ts-1[E] lesions to any new influenza virus will regularly result in attenuation of a recombinat virus possessing the new surface antigens.


Quantitative bacteriology of the vaginal flora.

Quantitative bacteriology was performed on vaginal secretions from healthy adult women. The analysis included a single sample from 17 college students and 35 samples from five volunteers collected at intervals of three to five days throughout the menstrual cycle. Mean concentrations in all 52 specimens were 10(8.1) aerobic bacteria/g and 10(9.1) anaerobic bacteria/g. The rank of predominant organisms, according to rates of recovery in concentrations of greater than 10(5) colony-forming units/g, was anaerobic and facultative Lacrobacillus species, Peptococcus species, Bacteroides species, Staphylococcus epidermidis, Corynebacterium species, Peotostreptococcus species, and Eubacterium species. Sequential samples collected throughout the menstrual cycle showed relatively consistent mean levels of anaerobes and a significant decrease in concentrations of aerobes in premenstrual specimens compared with those in the specimens collected in the week following onset of menses. Analysis of sequential specimens from each of the five individuals showed considerable variation in species recovered. These data indicate that the vaginal flora in healthy adult women is a dynamic ecosystem in which anaerobes are usually the numerically dominant bacteria.


Sensitivity of influenza A/New Jersey/8/76 (Hsw1N1) virus to amantadine-HCl.

The appearance of a swine type of influenza A among recruits in Fort Dix, New Jersey, in 1976 prompted an evaluation of the sensitivity of this virus to amantadine - HCl. Well-tolerated levels of the drug effectively blocked replication of the influenza A/New Jersey/8/76 (Hsw1N1) virus isolate in three laboratory models: monkey kidney cells, embryonated eggs, and intranasally infected mice. On a comparative basis this virus would appear to be more sensitive to amantadine - HCl than human influenza A strains, including the recent Victoria (1975) strain. All strains of influenza A virus continue to be sensitive to amantadine - HCl.


Immune response of the intestinal mucosa to cholera toxoid.

The intestinal immune response to cholera toxoid was studied in dogs and rats. Oral or intraperitoneal priming followed by duodenal boosting with toxoid reulted in antitoxin-containing plasma cells (ACC) in jejunal lamina propria of rats. Priming and boosting by the intraperitoneal route alone induced almost no jejunal response. Lamina propria ACC were derived largely from migrating immunoblasts, which appeared earlier among thoracic duct lymphocytes. Protection of dogs after repeated subcutaneous immunization with toxoid was mediated largely, or entirely, by serum-derived antibody. The sequence of subcutaneous priming and repeated oral boosting induced longer lasting protection mediated almost entirely by the local intestinal immune mechanism. The results suggest that cholera immunization should be specifically designed to stimulate the intestinal immune mechanism and that the subcutaneous oral immunizing sequence may be an efficient way to do this.


Protective efficacy of an inactivated Mycoplasma pneumoniae vaccine.

The protective efficacy of a formalin-inactivated Mycoplasma pneumoniae vaccine was evaluated in a double-blind fashion in 7,861 Marine Corps recruits at Parris Island, South Carolina. Vaccine, administered in a 1-ml dose by a jet-injection device, was glass-grown and contained 264 microgram of protein nitrogen/ml. Phosphate-buffered saline with formalin was injected as a control. Systemic reactions to injection were similar in both groups, but the percentage of vaccinees with erythema (51%) and induration (52%) at 24 hr was significantly greater than the percentage of controls (2%) with these reaction (P less than 0.001). Twenty-one (0.5%) of 3,930 vaccinees and 43 (1.1%) of the 3.931 placebo recipients were hospitalized with pneumonia (chi2=7.61; P less than 0.01). Ten of 21 vaccinees and seven of 43 controls with pneumonia had a positive pharyngeal culture for M. pneumoniae (chi2=1.69; P =0.20), and fourfold rises  in titer of serum antibody were noted in five of 14 vaccinees and in 15 of 28 placebo recipients with pneumonia (chi2=7.90; P less 0.0005). Therefore, vaccine efficacy for M. pneumoniae-specific pneumonia was 42% as determined by cultures and 67% by serologic tests. The vaccine showed no protective efficacy for M. pneumoniae-specific bronchitis or for M. pneumoniae pharyngeal carriage in recrutis in training.


Changes in adenine nucleotide and mitochondrial metabolism of the kidney of burned rats and their relation to insulin.

In rats with third-degree burns, the blood glucose level increased remarkably, with a concomitant suppression of insulin secretion from the pancreas after an oral glucose load. The energy charge (ATP + 1/2 ADP/ATP + ADP + AMP) levels of the kidney decreased to 0.659 as compared with 0.858 of controls at 8 hr after the burn (p less than 0.001). The phosphorylative activity of the kidney mitochondria fell to one third of controls at 8 hr after the burn (p less than 0.001), and that of heart mitochondria decreased to approximately 70% (p less than 0.005); the fall in liver and brain was less remarkable. The decrease in mitochondrial phosphorylative activity was accompanied by a reduction in the respiratory control ratio, P/O ratio, and state 3 respiration. The concentrations of cytochrome a(+a3) in the kidney mitochondria decreased to 69.9% of controls at 8 hr after the burn (p less than 0.001), those of cytochrome b to 82.6%, and those of cytochrome c + c1 to 75.3% (p less than 0.001). The decreased energy charge and oxidative phosphorylation of the kidney in burned rats were remarkably restored by subcutaneous administration of insulin. It is suggested that a reduction in insulin secretion from the pancreas may play an important role in initiating an impairment of adenine nucleotide and mitochondrial metabolism of the kidney.


Pathogenesis of anemia in rats with Walker 256 carcinosarcoma.

The purpose of this study was to further clarify the pathophysiology of anemia in malignancy. To accomplish this end a total of 210 normal or splenectomized rats with or without the solid form of Walker 256 carcinosarcoma was studied. In vivo studies demonstrated that in stage I cancer (tumor weight less than 10% of body weight) a slightly shortened red cell survival resulted in a mild degree of anemia. With increasing tumor size, 51Cr red cells mass decreased further, in spite of extramedullary erythropoiesis and a slightly increased incorporation transferrin-bound iron into red cells. Splenectomized rats with stage II cancer developed a more profound degree of anemia associated with a significantly decreased incorporation of 59Fe into red cells. Marrow cell culture studies demonstrated that heme synthesis in response to erythropoietin in stage I cancer was not significantly different from normal, but in rats with stage II cancer (tumor weight greater than 10% of body weight) heme synthesis in response to erythropoietin was markedly decreased. In vitro studies demonstrated that plasma erythropoietin levels were appropriately increased in most rats with transplanted malignancy. These studies indicate that bone marrow heme synthesis in response to erythropoietin is impaired in rats with the anemia of advanced malignancy.


Hepatic lesions and hemolysis following administration of 3alpha, 7alpha, 12alpha-trihydroxy-5beta-cholestan-26-oyl taurine to rats.

Patients with a metabolic block in the conversion of THCA into cholic acid develop cirrhosis and hemolysis, and die of hepatic failure. In these patients, THCA is largely conjugated to taurine (tauro-THCA) and excreted instead of being converted into cholic acid. In the present study, the effects of tauro-THCA on hemolysis, bile flow, and hepatic morphology were evaluated in bile fistula rats. All rats infused with tauro-THCA at rates of 0.25, 0.50 or 0.75 micronmol/min developed hemolysis with hemoglobinuria. A direct toxic effect of tauro-THCA on washed human red blood cell membranes was demonstrated at a concentration of 8 X 10(-4) M. Liver biopsy sections from rats infused for a 2 hr period with tauro-THCA were examined by electron microscopy and showed dilation of the rough endoplasmic reticulum and distortion of mitochondrial membranes. Cholestasis was not induced, since tauro-THCA actually caused a greater choleretic response for a given rate of bile salt excretion than did taurocholate. This study raises the possibility that the clinical liver disease seen in patients with a metabolic block in the conversion of THCA into cholic acid may be caused by tauro-THCA.


Differences in response to sodium administration in normotensive white and black subjects.

In order to examine possible differences in PRA, PA, and renal electrolyte excretion in normotensive black and white subjects, we studied 68 age-matched subjects: 19 white men, 19 black men, 15 white women, and 15 black women. PRA was measured before and after the infusion of 2 L of normal saline to evaluate suppression of the renin-aldosterone system. The excretion of sodium, potassium, and creatinine in response to the salt load was monitored for 24 hr. PRA and PA measurements were repeated after 2 hr of ambulation on the morning following salt loading. Dietary sodium was then restricted and oral furosemide (120 mg) was administered in order to assess PRA and PA responsiveness to sodium and volume depletion. White and black subjects had similar PRA and PA responses to normal saline and furosemide administration. Over a 24 hr period, black subjects excreted significantly less sodium and potassium following the salt load than did white subjects. The PRA values of black subjects 24 hr following the salt load were significantly suppressed as well. These observations suggest that there may be racial differences in renal sodium excretion reflected by decreased urinary sodium excretion and a relative suppression of PRA following a salt load in black persons.