Diffusing capacity, membrane diffusing capacity, capillary blood volume, pulmonary tissue volume, and cardiac output measured by a rebreathing technique. A rebreathing method for estimating diffusing capacity, membrane diffusing capacity, pulmonary capillary blood volume, pulmonary capillary blood flow, and pulmonary tissue volume consists of rebreathing into a bag for 15 sec while acetylene, (18O)-carbon monoxide, oxygen, and helium are continuously sampled by a mass spectrometer. Because the masses of carbon monoxide and nitrogen are nearly identical at 28, it was necessary to use a stable isotope, C18O, to distinguish this gas with the mass spectrometer. Comparison of the pulmonary capillary blood flow by the rebreathing technique with the simultaneously obtained indicator dilution measurement in anesthetized dogs revealed good agreement. Estimations of pulmonary tissue volume appeared to be quite reproducible and consistent; the values tended to be somewhat smaller and less variable among normal subjects than reported by other investigators. After subtraction of capillary blood volume, tissue volume was 311 plus or minus 73 ml at rest and increased significantly to 352 plus or minus 61 ml at 75 watts of exercise. Pulmonary tissue volume in dogs using the rebreathing method averaged 9.2 ml per kg of body weight, a mean comparable to previously reported estimates using the ether plethysmographic method. The slope of pulmonary capillary blood flow (cardiac output in normal subjects) as a function of oxygen consumption during exercise in normal subjects of 0.0060 times oxygen consumption in milliliter per min was identical to published values. The rebreathing technique provides a rapid, reliable, noninvasive method for estimating pulmonary hemodynamic parameters.


Liver granulomas containing copper in vineyard sprayer's lung. A new etiology of hepatic granulomatosis. In a previous publication the writers described a new pulmonary disease in rural workers who sprayed vineyards with Bordeaux Mixture, a copper sulfate solution neutralized with hydrated lime for the prevention of mildew. The study of the liver of these patients at autopsy and percutaneous biopsy revealed histiocytic and noncaseating granulomas containing inclusions of copper, identified by histochemical techniques. The occupational exposure to copper sulfate, the characteristic pulmonary lesions of vineyard sprayer's lung, and the presence of copper in the liver of these patients define this new variety of hepatic granulomatosis.


Autoanalytic (colormetric) determinations of SGOT in isoniazid recipients are reliable. A study was made to determine whether the ingestion of isoniazid interfered with the colormetric reaction used in most autoanalyzers to measure serum glutamic oxaloacetic transaminase (SGOT). Colormetric (autoanalytic) and noncolormetric (enzymatic) measurements of SGOT were made on a venous blood sample from each of 100 hospital employees who were taking isoniazid to prevent tuberculosis. The 2 techniques of measuring SGOT yielded comparable results. It was concluded that autoanalytic (colormetric) measurements of SGOT in isoniazid recipients are reliable.


Cysticercosis: an unusual cause of papilledema. A case of cerebral cysticercosis with papilledema is presented in a 44-year-old Mexican woman who progressed to optic atrophy despite a ventriculoatrial shunt. The diagnosis was made on testing the cerebral spinal fluid for the indirect hemagglutination response to cysticercosis. The problems of the pork tape-worm as well as several references from the literature are discussed pertaining to ocular and cerebral involvement. Although this disease is not common in the United States we should think of it in cases of papilledema in patients from endemic areas and in those who have traveled in those areas and eaten pork not properly cooked or frozen.


Simplified refracting technique in keratoconus. A simple but effective technique to refract keratoconus patients is presented. The theoretical objection to these methods are discussed. In addition, a formula to calculate lenticular astigmatism is presented.


Further use of polyglycolic acid (Dexon) sutures in cataract and other intraocular surgery. Polyglycolic acid sutures were used in 239 cases of intraocular surgery (164,6-0 size, 75, 7-0 size). Most of these (226) were cataract extractions. The results were excellent, with minimal complications. The 6-0 suture was somewhat stiff and associated with some tissue drag. The 7-0 size overcame these deficiencies and was considered the closest approach to date to the ideal absorbable suture. In addition, these appears to be an inhibition to bacterial penetration by PGA.


Retinal telangiectasis: delayed response to photocoagulation. A case suggestive of early Coats' disease treated cautiously with xenon arc photocoagulation is presented. Although adequate reaction was observed, the abnormal vessels persisted through the initial follow-up period. However, 9 months after treatment the vessels were found to be obliterated.


Evaluation of a new tropical steroid, fluorometholone, in the postoperative course of the cataract operation. This is a study of the effect of steroids on ocular tension, in the postoperative course, in a group of 23 patients operated upon simultaneously for cataract in both eyes. In the right eye fluorometholone and in the left eye dexamethasone drops were instilled 3 times a day. Tensions which were somewhat higher were appreciated after the third week of the postoperative course in the eyes treated with dexamethasone.


The influence of tropicamide on intraocular pressure. Elevation of intraocular pressure by many cycloplegic drugs has been reported to occur in eyes in which the filtration angle remained open during mydriasis. Tropicamide 1% is also capable of this phenomenon in patients with open angle glaucoma who are being medicated with pilocarpine. However, in normal eyes as well as those with untreated open angle glaucoma, this cycloplegic has a relatively small effect on intraocular pressure. It is postulated that this difference between the subject groups may be due to competitive inhibition of the miotic at the receptor site in those treated with pilocarpine. In 3 patients in whom the intraocular tension elevation was found to be quite marked, heavy pigments showers in the anterior chamber occurred which suggested an outflow obstructive phenomenon similar to that seen in "Pigmentary Glaucoma."


The importance and accuracy of the water drinking test and tonography. The water drinking test was performed on 122 glaucoma suspect patients using the standardized Schiotz tonometer and Goldmann applanation tonometer. Additionally, the tonography was performed on 12 glaucoma suspect patients before the water test and on 69 glaucoma suspect patients after the water test. Our results showed that the most accurate test is the applanation water test. However, the accuracy of tonography was also increased if the tonography was performed after the water drinking test. The evaluation of the water drinking test only by Schiotz tonometer was not satisfactory. By using the applanation tonometry the sensitivity of the water test could be improved by 70%.


Amaurosis congenita (Leber).

Amaurosis congenita (Leber). A case of Leber's amaurosis congenita is presented. The patient was bilaterally and congenitally blind. She had nonrecordable ERG responses and a striking fundus appearance with multiple irregularly shaped yellowish white flecks deep in the retina, peripheraly distributed over an area extending from behind the equator to the ora serrata. No flecks were present in narrow bands on both sides of the retinal vessels and in the whole nasal portion of the fundus. Fine powdery granules of pigment were diffusely spread in the areas that were devoid of flecks. The macula and the optic disc showed no gross abnormality. Although this fundus appearance resembles that of the flecked retina syndrome and Oguchi's disease, retinal function in this case is more profoundly affected than in either of the two.


Long-standing anterior dislocation of the crystalline lens. We believe this is the first reported case of an anteriorly dislocated lens that produced no pain or discomfort for at least 4 months, ie, March to July 1972. Even after July, the symptoms were not severe enough for this man to seek medical attention until September 1972. The lens may have been dislocated as early as age 5 since, according to his history, surgery was suggested at age 7 for the right eye. A pupillary block may have developed in March 1972, causing an intraocular pressure rise which eventually led to enucleation in September 1972. The microscopic evidence of fibrovascular adhesions of the lens to the iris and cornea documents the long-standing nature of this condition.


Chiasmatic arachnoiditis and empty sella: report and discussion of a case. The case is presented of a 5-year-old boy with progressive visual loss and physical findings suggestive of pituitary dysfunction. A craniotomy revealed opticochiasmatic arachnoiditis and empty sella. The concomitance of these conditions has not been reported. Both entities are associated with arachnoid cysts or cyst-like swellings of the arachnoid, according to the literature. It is proposed that a primary arachnoid cyst may have displaced the pituitary in the sella resulting in secondary inflammatory responses which affected the chiasm. An alternate, but similar explanation, would involve a perichiasmatic arachnoiditis of unknown cause resulting in cystic invasion of the sella. Rupture of the presumed cyst is believed to have occurred late in the preoperative course, and to have been responsible for aggravation of the symptoms.


A follow-up of blind diabetic patients.

A follow-up of blind diabetic patients. This report concerns the subsequent fate of 95 blind diabetics who were using a Seeing Eye  guide dog in 1964. In 1973, 9 years later, 40 were known to be dead, 31 were known to be alive, and no data were available on 24. The average interval from onset of severe blindness to death in 37 of the 40 who had died is 9.4 years. The mean age at death was 43.2 years for the group. This compares favorably to our original study which showed an average survival from onset to blindness to death of 5.8 years, and an average age at death of 35.9 years. Because follow-up data are incomplete for the 1964-1973 interval, the true figures may be poorer than the ones reported now. In any case, it is apparent that the life expectancy of the blind diabetic is still poor after the onset of blindness from severe retinopathy.


Microspectrophotometric estimates of non-histone proteins in cell nuclei displaying differing degrees of chromatin condensation. Nuclei isolated from mouse thymus, kidney, and liver were fixed in ethanol-acetic acetic acid; treated with dilute acid to extract histones; stained by three protein end-group procedures; and measured by scanning, integrating microspectrophotometry. Measurements were also made of nuclei isolated from the same organs and stained by the Feulgen procedure for DNA. Protein end-group procedures included pH 2.8 Biebrich scarlet (for basic groups), mercury orange (for sulfhydryl groups), and mercury orange after thioglycolate reduction (for the sum of sulfhydryl and disulfide groups). With the exception of the comparison between Feulgen-stained 2C liver and kidney nuclei, the integrated extinction values obtained for nuclei of a given organ differed significantly from the measurements of nuclei from other organs, regardless of the staining procedure. Furthermore, the integrated extinction values for 2C nuclei were highest in larger, more vesicular nuclei (from liver and kidney) and lowest in condensed thymocyte nuclei, except in the case of measurements of the disulfide content of the nuclei. In this instance, the values of integrated extinction were highest in condensed thymocyte nuclei, intermediate in kidney nuclei, and lowest in 2C liver nuclei. When 2C, 4C, and 8C liver nuclei were compared, the integrated extinction values of 4C nuclei were found to be approximately twice those of 2C nuclei whose disulfide and Feulgen values were, respectively, higher and lower than expected. The greater disulfide values and reduced Feulgen values obtained in thymocyte and 8C liver nuclei might be related to a greater degree of chromatin condensation in these nuclei, and therefore, to a reduction or selective restriction of their RNA transcriptional capacities.


Mandibular movements of the albino rat during feeding. Jaw movements of albino rats during biting and mastication of relatively hard food were recorded by means of conventional and X-ray cinematography. Mandibular kinetics have been analysed in the context of passive mechanical limits imposed by jaw morphology, particularly of the joints, and by the food itself. Movements have been described in terms of degrees of gape, condylar translation and horizontal rotation of the rami about the symphysis. During biting the condyle remains in the anterior two-thirds of the fossa, moves forward as the jaw opens and the converse. The rami usually spread well apart; the lower incisors are usually approximated. Incised food particles are transported toward the molars by means of coordinated jaw and tongue movements. The prominent palatal rugae of the diastemal region abet this process. In the power stroke of mastication, the mandible shifts forward as the lower toothrows move a little inward; the condyles occupy the posterior two-thirds of the fossa. All movements seen were bilaterally symmetrical. Simultaneous chewing occurred on both sides. It is suggested that the lingual components in the primarily anterior power stroke enhance grinding efficiency. A movable symphysis appears to be of critical importance in facilitating this type of mastication.


Ultrastructural studies of regenerating spines of the sea urchin Strongylocentrotus purpuratus. I. Cell types without spherules. The fine structure of regenerating tips of spines of the sea urchin Strongylocentrotus purpuratus was investigated. Each conical tip consisted of an inner dermis, which deposits and contains the calcite skeleton, and an external layer of epidermis. Although cell types termed spherulecytes containing large, intracellular membrane bound spherules were also present in spine tissues, only epidermal and dermal cell types lacking such spherules are described in this paper. The epidermis was composed largely of free cells representing several functional types. Over the apical portion of the tip these cells occurred in groups, while proximally they were distributed within longitudinal grooves present along the periphery of the spine from the base to the tip. The terminal portions of apical processes extending from some of the epidermal cells formed a thin, contiguous outer layer consisting of small individual islands of cytoplasm bearing microvilli. Adjacent islands were connected around the periphery by a junctional complex extending roughly 200 A in depth in which the opposing plasma membranes were separated by a narrow gap about 145 A in width bridged by amorphous material. Other epidermal cells were closely associated with the basal lamina, which was 900 A in thickness and delineated the dermoepidermal junction; some of these cells appeared to synthesize the lamina, while others may be sensory nerve cells. The dermis at the spine tip also consisted of several functional types of free cells; the most interesting of these was the calcoblast, which deposits the skeleton. Calcoblasts extended a thin, cytoplasmic skeletal sheath which surrounded the tips and adjacent proximal portions of each of the longitudinally oriented microspines comprising the regenerating skeleton, and distally, formed a conical extracellular channel ahead of the mineralizing tip. The intimate relationship between calcoblasts and the growing mineral surface strongly suggests that these cells directly control both the kinetics of mineral deposition and morphogenesis of the skeleton. Other cell types in the dermis were precalcoblasts and phagocytes. Precalcoblasts may function as fibroblasts and are possible precursors of calcoblasts. Closely associated with the basal lamina at the dermoepidermal junction were extracellular unbanded anchoring fi0rils 150 A to 200 A51 in diameter. Scattered proximally among dermal cells were other extracellular fibrils, presumably collagenous, about 300 A in diameter wit


Some anatomical aspects of the cardiovascular system of Antarctic seals and their possible functional significance in diving. The hearts and ascending aortae of 11 Weddell seals, Leptonychotes weddelli, three adult Crabeater seals, Lobodon carcinophagus, two adult Ross seals, Ommatophoca rossi, and one adult Leopard seal, Hydrurga leptonyx, were examined for comparison with terrestrial forms. The Weddell seal specimens were from animals ranging in age from midterm in fetal development to mature adults. All specimens were collected in 1971, 1972, and 1973, from McMurdo Sound and the Ross Sea, Antarctica. The phocid hearts were characteristically broader and flatter than those of other carnivore families and they tended toward bifid apices. The heart form indices (height/circumference) averaged 31.5 compared to 39.0 for felids. The right ventricular chambers of the Antarctic seals were found to average longer in Weddells and narrower in all, than those reported for four other carnivore families. An elastic enlargement was present in the ascending aortae of all seals. The largest diameter of the aortic bulb averaged 25.5 mm more than the base of the aorta in the adult Weddell seals which represented an increase of 72.5% over the base. It is suggested that the general heart form, and especially the ascending aortae, are anatomical adaptations to diving. The compressed heart makes possible unimpaired function when the chest is compressed during deep dives. The aortic bulb maintains mean arterial blood pressure and perfusion of the brain and cardiac tissue during diving bradycardia.


Upper digestive tract burns caused by Denalan denture cleanser powder. Denture cleansers are ubiquitous compounds frequently found in the household. Severe oral cavity burns were clinically observed in a two-year-old female who accidentally ingested a denture cleanser powder, Denalan. Experimental studies were carried out to investigate the caustic, chemical and histopathological properties of this compound. Denalan was found to be a powerful alkali agent which caused severe upper digestive tract burns.


Application of inverse filtering for detecting laryngeal pathology. Inverse filtering analysis was performed on acoustic speech samples of the vowel sound "ah" produced by ten selected patients and ten normal adults. The inverse filter output signal is the residue obtained by automatically extracting the effects of the supraglottal structure from the acoustic speech signal. For normal speakers, the residue consists of a series of rather sharp periodic spikes having low amplitude noise between periods. For pathological voices, the residue consists of lower amplitude, less sharp spikes having higher amplitude noise between periods. Comparisons between the acoustic speech waveforms and the residue signals illustrate the superiority of the residue signal for detecting irregularity, and thus suggests the feasibility of applying inverse filtering as an aid to laryngeal diagnostics.


Demoplastic fibroma of the mandible. A case report. Case of a 4-year-old boy with a desmoplastic fibroma of the mandible. Desmoplastic fibromas are benign fibrous neoplasms which are easily confused with other fibrous tumors. Although benign, they usually display aggressive local tissue extension and frequently recur when treated conservatively. At the present time, wide, local excision appears to be the treatment of choice.


Sensorineural impairment in unilateral otosclerosis. Audiometric findings were evaluated in a group of patients with clinical unilateral otosclerosis. A comparison was made between bone conduction threshold in the better hearing ear and the ear with the mixed hearing loss. Over 50% of the patients had a significant elevation in the bone conduction thresholds in the ear with clinical otosclerosis as compared to the uninvolved ear. A pair of temporal bones from a patient with unilateral otosclerosis and unilateral mixed hearing loss is presented.


Air bypass voice prosthesis for vocal rehabilitation of laryngectomees. An air bypass voice prosthesis and the refinement of a surgical technique has been developed. It involves a one-stage, low retrograde cervical esophagostomy that is functionally and cosmetically feasible since it permits hidden use of the device, while eliminating fistula tract stenosis and aspiration. Most importantly, it provides a natural speech mechanism without any training period. Patients can regain immediate speech following installation of a voice prosthesis with a minimum of maintenance required. Ultimately, they have demonstrated greater self-reliance and developed a high degree of self-esteem which has enabled them to resume a normal way of life.


Prophylaxis of predisposed otitis externa. Evaluation, in the guinea pig ear, of fourteen water repellent or therapeutic compounds has resulted in the recommended use of 360 Medical Fluid as a prophylactic agent for those persons with a high risk of otitis externa. This compound was found to adhere well, be easy to apply and was nonirritating. Use of this material prevented adverse bacterial growth in ear canals exposed to water for up to seven days. The significance of maintaining the normal Gram positive bacterial flora in the external ear canal and the role of cerumen in maintaining a healthy meatal surface is discussed. A system for monitoring the health of the ear canal through measurement of the Gram positive/Gram negative bacterial ratio is suggested.


Some aspects of fetal laryngeal development. Some aspects of morphological and physiological development of the fetal larynx are presented. Histochemical confirmation and determinations of fetal glandular activity are needed. Anomalous development of the larynx is still conjecture, but congenital cystic formation with glandular development appears possible after the fifth month of human fetal life.


The nature and crystal growth of otoconia in the rat. Several types of otoconia are present in the macular regions of young rats. These include multifaceted, transitional and rounded body forms, some variant otoconia and a few rhombohedrons. The adult form has typically rounded by nonsmooth body surfaces and pointed ends with three planar faces. The multifaceted and transitional otoconia fracture and etch more readily than do the adult type. The differences in properties of the otoconia are considered in the light of known facts concerning inorganic crystal nucleating and growth. This integrated approach indicates that many otoconia originate by seeding of multiple subunits on an organic substrate and develop by the mechanism of parallel growth. The basic structural unit is the rhombohedron. By analogy to inorganic crystals of calcite, it would seem that the typical otoconium grows on the end faces but growth on the side faces is suppressed by some unknown chemical factor. Some otoconia are exceptions, evidently seeding and growing in the pure rhombohedral form. Decalcification of cleaved otoconia shows that organic material is incorporated during growth. The observations are interpreted to indicate that organic substance influences growth and achievement of the adult otoconial form.


Recording auditory-nerve potentials as an office procedure. Recording auditory-nerve potentials from human subjects is already a routine procedure in the laboratory. In order to bring such recording capabilities into the office of practicing otologists, a number of difficulties had to be overcome. First, a small signal averager was built and incorporated into a stimulus generating and response recording system. The entire system was made portable and self-sufficient. The effects of electrical interference and background acoustic noise were shown to be tolerable. After studies of how responses vary with electrode location, electrodes were designed to be placed on the ear canal so that no invasive procedures were necessary. Methods were found to simplify the procedure so that recordings can be made in a matter of minutes by one person working alone.


Remote extracochlear versus intracochlear recordings in the guinea pig. A comparison was made of the whole-nerve action potential recorded from a wire electrode placed on the tympanic membrane (TM) (remote extracochlear recording) and than recorded from intracochlear electrodes in the first and third turn scala tympani of the guinea pig. Cochlear microphonics and summating potentials were also studied. The expected decreased sensitivity at the TM site was observed in all responses, although the microphonic (CM) was diminished more that the action potential (AP) as compared to the responses monitored via the ST1 electrode. Nevertheless, the TM-recorded cochlear potentials largely mimicked the ST1-recorded potentials in a qualitative manner, i.e., similar configurations of the CM pseudo-threshold functions. It was consistently observed that the N2 of the AP is proportionally larger (relative to N1) in the TM recording than in the ST1 recording. These phenomena were attributed to the complex spread of the bioelectrical potentials in the nonhomogeneous volume conductor formed by the tissues of the temporal bone. It was suggested that the observed change in the N1/N2 ratio at the different sites of recording is due to the different contributions of second-order neuronal discharges to the compound AP. In effect, the TM electrode "sees" proportionally more activity from second-order fibers than does the ST1 electrode, and the ST3 electrode, less.


Occult breast carcinomas detected by xeroradiography: clinical considerations. One hundred and eighty-five breast cancers were demonstrated by xeroradiography in 6,238 patients. Sixty-two cancers were occult, and constituted one-third of the demonstrable breast cancers. Some of the patients with clinically nonpalpable carcinomas had high risk factors known to be associated with the development of breast cancer. The diagnosis was usually established and confirmed by wide excisional biopsy of the area containing the suspicious lesion. In some instances the lesion was localized preoperatively by xeroradiography using a technique of skin markings. Specimen xeroradiography was utilized if there were calcifications seen on the preoperative xeroradiograph. There was a significantly decreased incidence of axillary node metastases in the occult cases when compared to the dominant mass cases.


Tympanometric pattern classification in relation to middle ear effusions. Tympanometric evaluation using an otoadmittance meter and X-Y plotter was performed on 129 ears of 70 children with history of recurrent acute otitis media, or evidence otoscopically of persistent middle ear effusion, or both. Myringotomy, performed immediately following the tympanometric procedure, confirmed the presence or absence of effusion. Following myringotomy, tympanometric patterns, as shown by susceptance and conductance tracings at 220 and 660 Hz, were identified and middle ear pressures and otoadmittance peak values were determined. These findings were compared and criteria were developed which best determined the presence or absence of effusion. The results revealed the following: 1) High negative middle ear pressure is not necessarily a reliable indicator of middle ear effusion. 2) Tympanometry can be used reliably as an indicator of effusion. A combination of pattern classification and susceptance criteria enabled correct prediction of effusion in 93% of these children. One pattern at B660 was found to be pathognomonic of effusion. 3) In general, otoadmittance at 660 Hz appears to be a better indicator of effusion than 220 Hz.


Sudden infant death syndrome: plasma vitamin E levels and dietary factors. Vitamin E and selenium deficiency have previously been suggested to be responsible for the Sudden Infant Death Syndrome (SIDS). New experimental data reveal that this is not the case since vitamin E as well as plasma selenium levels of SID infants are approximately equal to those of normal controls. Although breast feeding was believed to have a protective effect against SIDS, a statistical study of groups of SID- and control infants in San Diego County indicate no such correlation. Totally or partially breast-fed SID infants actually died at an earlier age than those fed by formula only (p=0.02). Compared to matched normal controls, SID infants appear to have received a less varied diet with a lower incidence of extradietary vitamin supplementation (p=0.02). There is also a somewhat greater prevalence of mothers smoking during pregnancy in the SID group (one-tail p=0.05).


Nonacoustic stimulation of the middle ear muscle reflex. The purpose of these experiments was to determine the incidence of the middle ear reflex in response to several nonacoustic (tactile and air jet) stimuli among subjects with normal hearing who had an acoustic reflex and selected patients with severe hearing loss. The results demonstrate that the incidence of response to tactile stimulation increases as the facial area stimulated approaches the auricle. The response to an air jet stimulus directed toward the eye is high; however, the clinical utility of the air jet may be limited because it often results in a startle reaction and head movement, and the response appears to fatigue easily. In normal listeners the response to auricular air jet stimulation probably results from both acoustic and tactile stimulation. The presence of a reflex to tactile stimulation, together with normal tympanometry constitutes strong evidence of a normal middle ear; but the absence of a reflex to acoustic or tactile stimuli still leads to an ambiguous determination of potential stapedial muscle function.


Significance of a solitary lung shadow in patients with breast cancer. From 1949 through 1972 at Memorial Sloan-Kettering Cancer Institute, 72 breast cancer patients developed a synchronous or metachronous solitary lung shadow. Forty-three had separate primary lung cancers; 23 had breast carcinoma metastases and six had benign lung lesions. Of these, 47% were asymptomatic and the lesions were discovered by routine chest roentgenograms. Among 49 patients who underwent pulmonary procedures, secondary involvement of regional lymphatics was noted in 22, nine of which were daughter metastases secondary to the breast deposit. In such a setting, a radical lobectomy (or pneumonectomy) would seem the operation of choice. Four patients who had primary lung cancer and five patients with pulmonary breast metastases survived five years after the thoracic procedure. Prolonged survival in these patients bore no relationship to age, sex, state of axillary lymph nodes or length of interlude between the breast and the lung cancer. Early diagnosis, the extent of the cancer and adequate excision seem to influence end results. When other primary cancers were present, in addition to those of breast and lung, these did not seem to diminish the chance for survival provided they were treated as if they existed alone. The need for strict periodic and continual followup examinations, not only in breast, but also for all cancer patients is essential for early detection of metastases or new primaries. Chest roentgenograms are an integral part of such a plan. The ambiguity of a solitary lung shadow (or at the most two or three opacities) with a known primary breast cancer requires clarification without delay as to its histologic nature so that proper treatment can be instituted and an accurate prognosis given.


Rapid microanalysis of cholesterol in bile and serum by gas chromatography. Bile and serum cholesterol concentrations were analyzed by the same saponification, extraction and gas chromatographic procedure, utilizing stigmasterol as an internal standard and trifluoroacetate derivatization. The method was rapid and highly sensitive, requiring only 20 to 50 mul of sample. Comparable levels of precision were achieved by parallel analysis of a series of bile and serum samples by the present method and the Abell colorimetric method; a high degree of correlation was found between these two methods.


Plasma cerebrosides in stroke and multiple sclerosis. This investigation was conducted to determine whether or not plasma galactosyl ceramides were elevated in patients with stroke and multiple sclerosis, and to determine glycosyl ceramide concentrations in older, normal subjects. It was hypothesized that central nervous system destruction, like that which occurs in stroke or in the demyelination characteristic of multiple sclerosis, might be reflected by changes in plasma glycosyl ceramides, specifically by an increased percentage of galactosyl ceramide. Glycosyl ceramides were analyzed in duplicate 10 ml aliquots of plasma from each of seven patients with stroke, five patients with multiple sclerosis and five control subjects age-matched with stroke patients. Mean percentages of galactosyl ceramide for both controls (11.06 percent) and multiple sclerosis (11.40 percent) were strikingly similar. The percent of galactosyl ceramides for stroke was slightly elevated (14.32 percent) but there were no significant differences at the p=greater than 0.05 level.


What is clinical smoke poisoning?

What is clinical smoke poisoning? In this 13-year study, 51 patients were admitted with the primary diagnosis of "smoke poisoning" "carbon monoxide (CO) poisoning" or "respiratory burns." Forty patients (78%) had diagnosis of smoke poisoning with minor or no skin burns. The study indicated that clinical diagnosis of CO poisoning cannot be made reliably without carboxyhemoglobin (COHg) determination and that smoke poisoning patients often had CO poisoning. Seventeen of 19 smoke poisoning patients (89%) had CO poisoning above COHb levels of 15% saturation. Carbon monoxide was successfully removed from the blood by improving alveolar ventilation and oxygen concentration. However, there were 2 smoke poisoning deaths as the result of gaseous chemical injury. There was a correlation coefficient of 0.87 between initial COHg levels and patients' hospital days primarily determined by patients' pulmonary complications. Since CO is non-irritating, COHb levels may be used as an additional indicator of suspected pulmonary injury by noxious combustion gases.


Characterization of elevated fibrin split products following thermal injury. These studies establish that the staphylococcal clumping test is superior to the tanned red cell hemagglutination inhibition immunoassay for monitoring fibrin split product concentration in burn sera. It is strongly suggested that the principal circulating degradation product is a complex of soluble fibrin monomer with fragment D. Finally, there does not appear to be any effect on measured fibrin split product concentration in sera of burn patients receiving prophylactic heparin or aspirin.


Automated bichromatic analysis of serum ceruloplasmin. An automated micromethod is described for measurement of serum ceruloplasmin by assay of its p-phenylenediamine oxidase activity using the Abbott bichoromatic analyzer. Ten mul of serum are mixed with 250 mul of p-phenylenediamine (PPD) substrate (9.2 mmole per liter) in acetate buffer (0.1 mole per liter, pH 5.45). Spectrophotometric measurements of the rate of formation of the purple oxidation porduct of PPD are performed after a 10 min delay for thermal equilibration at 37 degrees and for avoidance of the lag-phase of the enzymatic reaction. The coefficients of variation of replicate analyses of normal serum by this technique are 1.1 percent (within-the-run) and 3.3 percent (day-to-day). Measurements of ceruloplasmin concentrations in serums from 75 patients by this automated method provided close correlation with measurements by a manual reference procedure (correlation coefficient=0.973). The mean concentration of ceruloplasmin in serums from 64 healthy men was 29 mg per dl (central 95th percentile limits=22 to 40 mg per dl).


Coagulation dynamics following thermal injury: effect of heparin and protamine sulfate. A burned rat model was developed to examine post-burn alterations in coagulation. Fibrin split product concentration, as measured by the staphylococcal clumping test, was noted to rise significantly within the first 24 hours following injury. Prophylactic in vivo systemic anticoagulation with heparin was ineffective in modifying this response. However, systemic administration of protamine sulfate prevented post-burn elevation of fibrin split products. In vitro fibrin split product concentration in burn sera following the addition of heparin and protamine sulfate, was also analyzed. The results of these experiments elucidated the biochemical effects of protamine sulfate on circulating fibrin degradation products in the rat burn model.


Villous tumors of the duodenum.

Villous tumors of the duodenum. Villous tumors of the duodenum are rare, but treatment may be problematic because of their association with invasive adenocarcinoma. Two cases of villous tumor of the duodenum are described and 39 other reported cases are reviewed. Presenting symptoms were bleeding 27%; obstruction 24%; jaundice 22% and vague dyspepsia 20%. Diagnosis may be made by radiographic barium contrast evaluation of the duodenum, especially with the addition of air contrast hypotonic studies and by fibro-optic endoscopy. Twenty-seven per cent of villous tumors of the duodenum are associated with adenocarcinoma. Invasive tumor is more common in patients over 50 years old (35%), in tumors of the third and fourth portions of the duodenum (44%) and in tumors over 4 cm in diameter (30%). Local excision is the treatment of choice for benign lesions. Pancreatico-duodenectomy is recommended for tumors which include invasive carcinoma in patients without distal metastases.


Experimental and clinical application of microvascular surgery. The object of these studies was the development of a technique whereby a vessel of 1 mm or smaller could be easily anastomosed. In total replantation of rabbit ears, only 3 ears survived replantation in 80 attempts in the first series. In the second series of 80, 35 ears survived completely, 3 ears partially survived and 8 additional ears survived the postoperative period, but the rabbits died within 2 weeks due to diarrhea. Thirty-five ears failed and venous obstruction was the primary cause. Free transplantation of a great toe and a second toe to recreate a thumb is also described. The operative procedure of the great toe to thumb transplantation does not appear to be difficult. Six months after operation, the function of the newly created thumb was good. The second toe to thumb transfer was more difficult technically compared to the great toe to thumb transfer due to the much smaller size of the digital vessels of the second toe. However, removal of the second toe causes less disadvantage than the removal of the great toe. Nine months postoperatively, the function of the newly created thumb was satisfactory. Digital replantation is described with 36 cases with 65 complete finger amputations. Thirty-two with 45 fingers survived the reattachment. Repair of one artery per digit is sufficient, although two probably are better. A minimum of 2 veins per digit must be repaired. Prompt re-exploration is imperative if postoperative circulatory impairment occurs. In this group, arterial obstruction was more frequent than venous obstruction as the cause of replantation failure. The function of the replanted fingers is satisfactory in the majority of the cases.


Femoroperoneal bypass: evaluation of potential for revascularization of the severely ischemic lower extremity. In 79 patients in whom distal small vessel bypass with autogenous vein was used for revascularization because of gangrene, gangrenous ulceration or rest pain, 14 had femoroperoneal bypasses. Femorotibial or femoroperoneal bypasses were performed in those patients in whom no popliteal runoff was present on pre-operative arteriogram. Femoroperoneal bypass was performed in preference to primary amputation in each case. Nine of 14 (64.3%) of femoroperoneal bypasses were functional whereas 57 of 79 (72.2%) of total distal bypasses to small vessels were functional. Salvage of severely ischemic lower extremities was achieved in 5 of 14 (35.7%) patients after femoroperoneal bypass and in 46 of 65 (70.8%) patients after bypass to anterior tibial or posterior tibial arteries. Graft patency without limb salvage occurred in 4 of 9 (44.4%) patients with patent femoroperoneal bypasses and in only 2 of 48 (4.2%) of patients with femorotibial bypass. Although limb salvage rate is considerably less with femoroperoneal than femorotibial or femoropopliteal bypass, attempted limb revascularization by peroneal bypasses is preferable to primary amputation in patients with rest pain, gangrenous ulceration or gangrene.


Effect of antrectomy and subsequent vagotomy on the serum gastrin response to food in dogs. The effect of food on serum gastrin and gastric acid secretion has been studied in dogs with denervated pouches before and after antrectomy and subsequent vagotomy. A Billroth I anastomosis was used in one group of dogs and a Billroth II in the other. Serum gastrin was measured by radioimmunoassay. In both groups of dogs antrectomy significantly depressed mean basal levels of serum gastrin and abolished the rise in serum gastrin in response to a meat meal. Meal-induced pouch acid secretion was considerably lowered by antrectomy after either Billroth I or Billroth II anastomosis. Vagotomy after antrectomy increased basal levels of gastrin, but did not restore the serum gastrin response to a meat meal in either group of dogs. It is suggested that biologically active forms of gastrin are released from the antrum in response to a meal. Biologically inactive basal levels of gastrin apparently originate from extra-antral sources. The post-vagotomy increase in basal (static) gastrin suggests vagal control of the metabolism of static, extra-antral gastrin.


Determinants of cardiac output during experimental therapeutic hemodilution. In 20 dogs, blood loss of 20 ml/kg during thoracotomy was immediately replaced with 40 ml/kg of 2.5% dextran-40 or dextran-70 in saline. An immediate rise in blood volume, associated with a decrease in hematocrit and blood viscosity, produced a marked rise in cardiac output. This effect was slightly greater with dextran-40. During the first four hours after infusion of dextran-40, both blood volume and cardiac output fell below control levels. Blood volume and cardiac output remained at or near control levels during this period in the group receiving dextran-70. In both groups, blood viscosity and hematocrit remained low throughout the study, but cardiac output increased only in response to an increase in blood volume. Twenty-four hours after infusion of either dextran-40 or dextran-70, hematocrit had decreased further, presumably due to postoperative blood loss while blood volume and cardiac output remained at or near control levels. During isovolemic hemodilution in this experimental setting, cardiac output responded primarily to changes in blood volume rather than blood viscosity. Therefore, decreased oxygen availability during hemodilution was compensated only by a rise in oxygen extraction.


Efficacy of intravenous clindamycin and methicillin in gram-positive soft tissue infections. In a comparative study on a general surgical service, intravenous clindamycin phosphate or methicillin was used to treat a variety of soft tissue infections due to gram-positive organisms, chiefly staphylococci. The infections were rated according to severity, responsible organisms, and site of the infection. Excellent or good clinical and bacteriologic responses were obtained with both clindamycin and methicillin as adjuncts to basic surgical therapy in these soft tissue infections. The adverse effects of each drug were detailed, and were comparable. Clindamycin phosphate is a satisfactory substitute for methicillin in soft tissue infections secondary to gram-positive organisms.


The ability of an elemental diet to support nutrition and adaptation in the short gut syndrome. 85% enterectomy or jejunal biopsy was randomly carried out on 80 male Sprague-Dawley rats, 300-350 gm. After recovery, rats in each group were randomly fed either elemental diet (ED) or rat chow. All rats were sacrificed 8 weeks postoperatively. Biopsied rats gained more weight with ED than chow (22% vs. 18%; p less than .01). Resection produced greater weight loss than biopsy (p less than .01) and slower gain (p less than .01). The failure to gain was most marked in ED rats, significantly less than in the chow group (p less than .05) by week two. A fall in hemoglobin, protein and albumin was seen: neither the fall nor difference between the groups was significant (p greater than .05). Enterectomized chow rats ate 22% more than their controls (p greater than .05). Increase in bowel length after resection, equally divided between jejunum and ileum, was 21% with ED and 26% with chow. Gut diameter after resection increased 104% and 98% with chow and ED, respectively. Villus height increased 65% in the jejunum of both enterectomized groups and 85% and 78% with chow and ED, respectively, in the ileum. Changes in gut length, diameter and villus height are significant in each group (p less than .01) but not between groups (p greater than .05).


The differentiation of primary hyperparathyroidism from the hypercalcemia of malignancy. The presence of hypercalcemia in patients with known cancers may be due to the cancers themselves, or to co-existing primary hyperparathyroidism. The differentiation of primary hyperparathyroidism from the hypercalcemia of malignancy is important since the relief of distressing symptoms and prevention of hypercalcemic crises and renal failure can be accomplished relatively easily by parathyroid surgery in the former condition, and only with difficulty, at times, with fluids and drugs in the latter condition. The histories of three recent patients are presented, which demonstrate the difficulties inherent in the differentiation of these conditions. These patients were ultimately found at operation to have primary hyperparathyroidism in addition to malignancies of the cervix, adrenal gland and kidney. In our experience the following have been helpful in establishing a diagnosis; history of hypercalcemia prior to development of cancer, the type of cancer itself, the effect of cancer therapy on the hypercalcemia, and selective venous sampling with radioimmunoassay for parathyroid hormone.


Effect of agitation on platelet aggregation and microaggregate formation in banked blood. Blood stored at 4 C in ACD or CPD solution develops microaggregates composed primarily of fibrin and platelets. This debris has been implicated in the pathogenesis of posttraumatic pulmonary insufficiency in man. Recent work indicates that gentle agitation of the blood during storage appears to decrease debris formation. These studies were undertaken to establish more clearly the effect of agitation on debris formation. Blood was drawn in CPD from healthy young males, non-aspirin ingesting donors and stored at 4 C. One-half of the bags were gently and continuously agitated for 21 days and the other half remained stationary. At the end of the storage period, platelet counts and screen filtration pressures were measured. Agitated blood showed significantly less debris formation and significantly higher platelet counts. Gentle agitation was shown to be an effective method for preventing debris formation in banked blood.


Arterial injury and massive blood loss: a case report of management of pelvic gunshot injury with femoro-subscrotal-femoral bypass and 116 units of blood. A case of massive shotgun injury to the left thigh and hip is reported. The patient received 116 units of blood, and a femoro-subscrotal-femoral vein graft was employed to save the left leg. A Teflon wool blood transfusion filter, used from the beginning of therapy, was believed to have been a major factor in preventing significant pulmonary complications.


Mouse myeloma. A model for studies of cell kinetics. The mouse plasma cell tumor Adj PC-5 grows slowly due to a large loss of cells from the growth fraction into nonprolifeative, end-stage cells. All tumor cells with the capacity to form a colony appear to be in cell cycle. Marked tumor specificity of several alkylating agents could not be explaned by differences in the porliferative state of myeloma and normal marrow cells. The sensitivity of different mouse myelomas to an alkylating agent varies considerably. The factors determining whether a mouse myeloma is sensitive to an alkylating agent are probably related to structure of the agent and intrinsic properties of the cell, rather than to the agent's mechanism of action.


In vitro and in vivo alterations of long-term murine plasmacytoma cultures. Mouse myeloma cell line (SLU-5) that has undergone lengthy propagation in vitro and become nononcogenic was compared with an earlier oncogenic passage of cells. Immunogenicity of cells cultured for various periods of time was compared with that of the original tumor (MOPC-21). The nononcogenic cells were most immunogenic. Cells that were nononcogenic in normal mice produced tumors in nonlethally irradiated mice. Clones isolated from oncogenic and nononcogenic populations varied widely with respect to ability to produce tumors in mice and to specific globulin production.


Monitoring myelomatosis.

Monitoring myelomatosis. There is evidence for the value of the following tests in the diagnosis, monitoring, and prognosis of myelomatosis: (1) serum paraprotein measurements, (2) urine paraprotein (including Bence Jones) measurements, (3) serum ablumin, (4) serum urea, (5) proteins in the urine other than those in 2, and (6) hemoglobin level. During treatment, increased rate of rise in 1 or 2, disproportionate increase in 2, emergence of related paraprotein, loss of 1 or 2 with reticulosarcomatous change, and monocytic leukemia suggest that more malignant subclones can emerge from the original myeloma clone, possibly due to drugs acting on DNA.


Clinical classification of plasma cell myeloma. Clinical and electrophoretic data were evaluated in 334 consecutive patients with myeloma or monoclonal peaks on serum or urine electrophoresis. Of the 242 patients with myeloma, 7% had localized plasmacytomas with absent or low level monoclonal peaks on electrophoresis and received only radiotherapy to focal disease areas. Chemotherapy was also withheld from eight other patients in an indolent clinical phase of multiple myeloma. Disease progression was apparent in about one third of the patients with localized and indolent myeloma within 12 months. Forty-three patients had idiopathic peaks on serum electrophoresis; more than 90% were of the IgG type with levels less than 3.0 gm/100 ml. Serial elecrtophoreses, immunoglobulin quantitations, and skeletal radiographs are recommended for the evaluation of patients with idiopathic peaks, the classification of early phases of myeloma, and the confirmation of tumor mass change.


Remission maintenance therapy for multiple myeloma. The effects of various regimens of melphalan combination chemotherapy were evaluated in 508 patients with multiple myeloma. No value was confirmed from the addition of procarbazine or vincristine sulfate to melphalan-prednisone combinations. Ninety-six patients who responded to treatment were allocated at random to one of three maintenance regimens, namely intermittent courses of carmustine with prednisone, continued courses of melphalan with prednisone, or no chemotherapy. There were no differences in the frequency of relapse, the remission duration, or the survival time among these maintenace groups. The frequencies of pneumonia and herpes zoster were higher in patients receiving continued chemotherapy. Continued melphalan-prednisone chemotherapy after the first year is of no major value to responding patients with multiple cyeloma. Attempts to reduce tumor mass maximally with a change in therapy are justified.


Treatment of myeloma. Comparison of melphalan, chlorambucil, and azathioprine. A randomized study compared the response of patients with multiple myeloma to chlorambucil, melphalan, and azathioprine. All patients also received a combination of prednisone and fluoxymesterone. Seventy-three of 86 patients entered on the study could have evaluations. The results indicate that melphatan produced more responses than either azathioprine or chlorambucil, but responses to both of these agents were observed. No difference was noted between survival curves for patients with no poor-risk factors as compared to those having at least one poor-risk factor. The only poor-risk factor affecting survival in this group of patients was the blood urea nitrogen level.


Sequential therapy compared with combination therapy in multiple myeloma. A timed sequential chemotherapeutic regimen for multiple myeloma was desinged, based on plasma cell kinetics. A randomized study comparing this regimen with the combination of intermittent melphalan and prednisone was started after an adequate pilot study. Studies with tritiated thymidine labeling and mitotic indexes were performed on patients treated with sequential therapy. Of 13 patients treated with the combination therapy, the responses were as follows: two, objective improvement; nine, subjective improvement; and two, no responses. Of nine patients treated with sequential therapy, the responses in six patients who could have evaluations were as follows: one, objective improvement; two, subjective improvement; one, no response; one, with progressive disease; and one, cardiac death. The two studies showed differences in pertubation of cell kinetics of myeloma that may be related to exponential growth and immunoblobulin types.


Present status of immunoglobulin research. Antibodies have been known as important proteins, and, in recent years, understanding of their composition, synthesis, and genetic control has progressed rapidly. These aspects, as well as developments in the areas of structure, including function correlations, membrane immunoglobulin as recognition units, and antibody as regulator molecule, are reviewed. The flow of information development in recent years serves as a basis for projections into coming years, especially in the areas of more aggressive use of antibodies in therapy.


Chemical typing of immunoglobulins and their subtypes. An analytical method, described herein, is based on differences in the primary structure of immunoglobulins and their subclasses. This method is independent of the supply of specific antisera, is simple and inexpensive to perform, and is fully reproducible. The distribution of subtypes of IgG and IgA resembled published results from immunological studies. Finding of differences in the distribution of subclasses of IgG proteins in patients with light-chain disease or in the absence of M-protein is of special interest.


Correlation of abnormal immunoglobulin with clinical features of myeloma. Physical and chemical characteristics of the abnormal immunoglobulin as well as the immunoglobulin type are important determinants of the clinical features and prognosis in myeloma. A review of 270 cases of myeloma studied by the Acute Leukemia Group B showed that survival was adversely affected by azotemia. The presence of Bence Jones proteinuria adversely influenced survival even in patients who were not initially azotemic. The presence of lambda-type Bence Jones proteinuria resulted in greatly shortened survival in contrast to kappa-type Bence Jones proteinuria in the absence, or presence, of a serum IgG M-protein. The adverse effect of lambda-type Bence Jones protein could not be demonstrated in patients with IgA myeloma. Patients with IgA myeloma were found to have a noticeably shorter survival than those with IgG myeloma.


Mu-chain disease.

Mu-chain disease. The clinical and laboratory features of the seven known patients with heavy chain disease showed that all but one had chronic lymphocytic leukemia. Two common and striking features are the presence of kappa-type Bence Jones proteins and vacuolated plasma cells in the marrow.


IgD multiple myeloma. Review of 133 cases. One hundred thirty-three patients had IgD myeloma. The IgD comprises 0.8% of M-components in general and 2.1% of myelomas in particular. Males predominate and 65% of the patients are younger than 60 years at the diagnosis. More than half of the patients have lymphadenopathy, hepatomegaly, or splenomegaly. Extraosseous spread and amyloidosis are frequent. Severe anemia and azotemia are common. Total serum protein and IgD M-component levels are usually not high. LAMBDA-type light chains are found in 90% of IgD M-components. Bence Jones proteinimia is frequent and Bence Jones proteinuria appears in almost all patients. Mean survival is 13.7 months from diagnosis. The IgD is different from IgG and IgA myeloma, indicating that the clinical picture and course of multiple cyeloma may be related to the class and type of M-component.


Psychodynamic changes in untreated neurotic patients. II. Apparently genuine improvements. This work is part of a study of 45 neurotic patients who were seen for consultation at the Tavistock Clinic but who never received treatment, and who, by the time they were asked to come for follow-up, had never been interviewed by a psychiatrist more than twice in their whole lives. In a previous report, we described 13 patients who were at least "improved" on purely symptomatic criteria but whom we did not regard as improved on dynamic criteria. Here, we describe 11 patients who were judged to be improved on dynamic criteria. These patients are of extraordinary interest, providing not only direct evidence of therapeutic mechanisms in everyday lifeān answer to a question posed by Strupp and Bergin in 1969but also, quite unexpectedly, evidence about the therapeutic effects of single interviews.


Psychoanalysis, "focal psychotherapy," and the nature of the therapeutic influence. It has traditionally been asserted that the nature of the therapeutic influence in psychoanalysis is qualitatively different from that in "psychotherapy." This thesis is considered untendable. Analysis of a case history by Balint, Ornstein, and Balint shows evidence that therapeutic change can be conceptualized more parsimoniously along lines other than those traditionally preferred. The therapist establishes himself as a good parent or authority figure vis-a-vis the patient, and within that context mediates important lessons in nonneurotic constructive living. Given strong motivation to seek change, the patient is won over to a point of view different from the one that has, in essential respects, guided his life in the past, and he has to make these teachings his own. Analytic therapy is an education for optimal personal freedom in the context of social living.


Growth hormone responses to hypoglycemia in postmenopausal depressed women. Human growth hormone (HGH) responses to insulin-induced hypoglycemia were measured in ten postmenopausal women suffering from primary unipolar depressive illness, and in ten age-matched normal postmenopausal women. The mean maximal HGH response in the depressed patients was 4.6 plus or minus 4.4 ng/ml, and in the normals 13.3 plus or minus 9.8 ng/ml (P less than .05). All of the normal subjects had clinically adequate HGH responses, in contrast to only four of the depressed patients (P less than .01). The blood glucose responses were virtually the same in the two groups. Since brain catecholamines play a major role in mediating HGH responses to hypoglycemia, the findings are consistent with the hypothesis of diminished functional catecholaminergic activity in the depressed patients.


Familial associations with EEG variants in manic-depressive disease. Electroencephalographic (EEG) studies of 60 patients with bipolar manic-depressive disease disclosed an incidence of small sharp spikes plus a few other variations in 47% of the sample. In women these EEG features were significantly associated with a history of mental illness in the patient's mother or the maternal side of the family and an absence of mental disorder in the fathers. The reverse was true of women probands without these EEG characteristics. In the men small sharp spikes did not relate to parental psychopathology but half of the sisters of men with these EEG characteristics were found to be mentally ill. On the basis of these observations and previous work, we hypothesize that the small sharp spike EEG pattern might be an inherited characteristic related in some way to the familal transmission of manic-depressive disease.


Creativity and psychosis. An examination of conceptual style. The performance of creative writers on the Goldstein-Sheerer Object-Sorting Test was compared with that of admitted manic and schizophrenic patients. Writers and manics tended to show more behavioral and conceptual overinclusion, but the writers showed substantially more richness and the manics more idiosyncratic thinking. Schizophrenics tended to be underinclusive rather than overinclusive and showed less richness and bizarreness than the writers and manics. These data imply that the conceptual style of writers may resemble mania more than schizophrenia and that, if overinclusiveness is an index of thought disorder, manics may have a more florid thought disorder than schizophrenics.


Addicted and nonaddicted drug users. A comparison of drug usage patterns. Adetailed analysis of drug usage patterns of 101 multidrug-using soldiers disclosed little differentiation between individuals with a history of heroin addiction (N=68) from demographically similar multidrug-using (but nonaddicted) peers (N=33). Prior to the onset of herion addiction, relatively few differences in drug usage patterns emerged between the two groups, and what differences there were indicated more extensive drug use and a more rapid progression of drug use among the nonaddicted users. A "stepping-stone hypothesis" of heroin addiction is refuted by these data. Differences in drug use emerge after the intiation of heroin. Within a few months after first heroin use, those who did not become addicted returned to their preheroin experimentation levels of drug taking while the addicted group maintained an escalating pattern of opiate use.


Utilization of medical services by transferred employees. Differential effect of life change on health. Some of the health consequences of being transferred to a new job and a new community were examined. The medical records of transferred employees for one year following transfer were compared to the records of a matched control group of more permanently located employees. Because of the authors' clinical observation that an employee's job description was related to the medical response to transfer, the professional and nonprofessional (production and clerical) employees were examined as separate groups. The results demonstrated a greater utilization of health services by transferred nonprofessional, but not by transferred professional employees. The results are discussed in the light of current research in life changes and their relationship to illness.


Dynamic content analysis.

Dynamic content analysis. An experiment was performed to demonstrate that methods of linguistic content analysis suitable for the study of intra-interview processes are both possible and practicable. Six hundred thirty-eight randomly selected psychiatrist were sent rating protocols of machine-mediated interview transcripts (transcripts of interviews conducted by means of remotely located teletypes) and were asked to rate each patient communication on one or more of 13 diagnosis-related rating dimensions. Rank correlation statistics on the raters' levels of agreement were found to be significantly high in 11 of the 13 dimensions. In contrast to the commonly held opinion that psychiatrist lack consistency in their application of diagnostic terms, this study suggests that psychiatrists generally agree in their usage of these terms, provided that the behavioral samples being evaluated are appropriately delimited.


Ocular involvement in disseminated intravascular coagulopathy. Seven cases illustrate the ocular pathology of disseminated intravascular coagulopathy in its several stages, and correlate these changes with the clinical manifestations. The prime abnormalities consisted of detachments of the retina, vacuolar disruption of the pigment epithelium, choroidal hemorrhage, and, most importantly, thrombotic occlusion of the choriocapillaris and of the juxtaposed vessels in the submacular and peripapillary regions. The changes were characteristically symmetrical in the two eyes.


Geographic atrophy of the retinal pigment epithelium. A manifestation of senile macular degeneration. Eleven patients with geographic atrophy of the retinal pigment epithelium secondary to senil macular degeneration are differentiated from central areolar choroidal sclerosis. Although these lesions appear morphologically similar to central areolar choroidal sclerosis, they differ strikingly in age of onset, family history, association with senile disciform macular degeneration, and pathologic features.


Flecked retina. Appearance secondary to oxalate crystals from methoxyflurane anesthesia. Calcium oxalate crystals were demonstrated in the retinal pigment epithelium (RPE) of a 66-year old white man with changes in the fundus oculi consistent with the flecked retina ayndrome. The patient had a history of rheumatoid arthritis, mild hypertension, and mild renal insufficiency presumed due to his hypertension. He underwent prolonged abdominal surgery under methoxyflurane anesthesia, following which he developed acute irreversible renal failure. Calcium oxalate crystals were demonstrated postoperatively in a kidney biopsy specimen. He subsequently was maintained on renal dialysis with decreasing renal function. Several weeks before his death fundus examination revealed a picture suggestive of a flecked retina syndrome. At autopsy, widespread oxalosis was found including crystals in the RPE and in some areas in the neural retina and cillary epithelium.


Varix of the iris.

Varix of the iris. A varix of the iris as a primary vascular lesion without evidence of reactive vascular proliferation secondary to obstructed outflow of a systemic vascular lesion occurred.  No similar cases have been found in the current literature.


Peripheral iridectomy with scleral cautery for glaucoma. Peripheral iridectomy with cautery (Scheie operation) was carried out on 113 eyes with glaucoma. The results were so favorable and the complications so few that we believe this operation still has an important place in the surgical management of the glaucomas and is noteworthy in the light of the trend to more precise surgery of the canal of Schlemm and the trabeculae.


Pilocarpine toxicity. Effects on the rabbit corneal endothelium. The effects of pilocarpine on the healthy rabbit corneal endothelium were studied by in vitro perfusion, photography, and electron microscopy. The toxicity of endothelial cells to pilocarpine was doserelated. Photography showed shrinkage of cells exposed to pilocarpine. The most striking ultrastructure changes were margination of nuclear heterochromatin and cytoplasmic vacuolation.


Effects of drug vehicles on ocular contact time. Radioactive sodium pentechnetate Tc 99m was added to commonly used ophthalmic drug vehicles of saline, 1% methyicellulose, 1.4% polyvinyl alcohol, and a 6:4 mixture of white petrolatum-mineral oil ointment. These drug vehicles containing 99m Tc were placed on both rabbit and human eyes and the rate of disappearance of the radioactivity determined. The ocular retention of 99m Tc by the eye was much longer in the nonblinking eye regardless of the vehicle used. The rate of loss of 99m Tc from the eye was fastest with saline and slowest with the ointment, with methylcellulose and polyvinyl alcohol in between. The longest contact time of the drug vehicle with the eye is afforded by the use of the ointment vehicle and the covering of both eyes.


Teratogenicities of ophthalmic drugs. I. Antiviral ophthalmic drugs. Nonradioactive idoxuridine (IDU,5-iodo-2'-deoxyuridine), while not teratogenic to rats, does produce fetal maliformations in rabbits when administered topically to the eye in doses similar to those used clinically, 0.1% four times a day for 12 days. These maliformations include exophthalmos and clubbing of the forelegs. By contrast, trifluorothymidine (F3TdR), another highly effective antiherpetic agent currently under investigation but not available for general use, was found not to be teratogenic to rabbits, even when given in concentrations tenfold greater than the doses used to produce idoxuridine teratogenicity.


Teratogenicities of ophthalmic drugs. II. Teratogenicities and tissue accumulation of thimerosal. Under the conditions of this study, systemically or topically applied thimerosal was found to have no teratogenic effect even when given in concentrations approaching the 50% lethal dose of these compounds. A comparison of topical and subcutaneous administration of thimerosal to rabbits shows that a substantial concentration of mercury was present in blood and tissues of the treated animals and their offspring. Thimerosal was found to cross the blood-brain and placenta barriers.


Human subretinal fluid. Its cellular and subcellular components. A light and electron microscopical study of subretinal fluid removed at surgery for rhegmatogenous retinal detachment showed a heterogeneity of cell types originating from the neural retina, pigment epithelium, and wandering cells of the blood. Subcellular organelles sedimenting in the centrifuged pellets could be identified as to cell of origin, health of the parent cell, and time since cell rupture. Acellular specimens were typical of angiomatous detachments or long-standing rhegmatogenous detachments. Because various mechanisms of cellular pathophysiology contribute to the destructive and reparative processes during retina detachment, the cytologic analysis of subretinal fluid becomes a useful tool in understanding the biological processes that affect visual recovery or impairment after successful retinal detachment surgery.


Visual evoked responses in the assessment of field defects in glaucoma. A technique for producing steady state visual evoked responses (VERs) to pattern reversal stimulation of retinal areas corresponding to discrete field quadrants is described. Patients examined included 10 normal individuals, 11 patients with ocular hypertension, and 21 with glaucomatous field defects (unilateral except in two cases with bilateral defects). The VERs relating to normal homonymous field quadrants were in phase in all three groups, excluding patients with cataracts. The VERs corresponding to quadrants with small field defects showed a phase shift compared with responses from normal homonymous quadrants in all cases, but were not detectable if the defect filled more than three quarters of the quadrant tested in most cases. The method offers a means for the objective assessment of visual fields in patients with unilateral optic nerve lesions, provided that central visual acuity is 20/80 or better


Pituitary carcinoma with liver metastases and Cushing syndrome. Report of a case. A 38-year-old woman complained of headache, visual impairment, and diabetes insipidus for 18 months prior to rapid onset of Cushing syndrome and amenorrhea. An autopsy disclosed a pituitary carchinoma invading the right cavernous sinus and two liver metastases. The tumor cells were chromophobic and agranular, with scarce polymorphism and rare mitoses. A review of the literature showed that the association of Cushing syndrome to pituitary carcinomas increases in direct proportion to the occurence of metastases. Such association was noted in almost half of the reported cases of pituitary carcinomas with extracranial dissemination.


Ultrastructure of articular cartilage in pyogenic arthritis. The articular cartilage in pyogenic arthritis, induced in rabbit by intra-articular injection of Staphylococcus aureus, was studied with the electron microscope. The cartilage showed erosion of the surface and degeneration and necrosis of the chondrocytes starting from 24 hours onward, following the intra-articular injection of bacteria. The surface erosion and chrondrocyte necrosis progressed rapidly. The changes are believed to be due mainly to liberation of lysosomal enzymes from the synovial cells and polymorphonuclear leukocytes.


Recovery of pancreas from mild puromycin-induced injury. A histologic and ultrastructure study in rats. Pancreatic acinar cells undergo degeneration or necrosis following injection of puromycin intraperitoneally in rats. The purpose of this study was to characterize recovery following injection of four hourly doses of puromycin, 40mg/kg of body weight, examining the pancreas histologically and by electron microscopy. The number of dividing acinar cells increased following injury. By 12 to 24 hours following treatment, electron microscopy showed numerous autophagic vacuoles and intracisternal granules in the cells. By 48 hours, these were largely cleared from surviving cells although the intracisternal  granules persisted in isolated acinar cells as long as 144 hours. At 24 hours, there was debris in the acinar lumens and interstitial space. We conclude that some acinar cells injured by puromycin may survive and be restored to normal structure; that surviving acinar cells can extrude autophagic vacuoles; and that necrotic acinar cells are replaced by regeneration following puromycin-induced injury in rats.


Rhinosporidium seeberi. An ultrastructural study of its endosporulation phase and trophocyte phase. The endosporulation phase of Rhinosporidium seeberi is the part of the life cycle most vulnerable to drug therapy. The laminated body formed from the precleavage sporangium has double-membraned tubular and vesicular structures within concentric layers. Similar tubular structures are also found in the developing sporoblast. The development of trophocytes involves three distinct phases. The early trophocyte retains many of the characteristics of the sporoblast as well as endoplasmic reticulum and other radiating filamentous structures in the cell wall. The itermediate tropocyte is larger (20 mug to 30mug m) and contains lipid bodies and microbodies, whereas the mature trophocyte is smaller (7mug m to9mug m) and microbodies are sparse or absent.


Myocardial injury from transthoracic defibrillator countershock. Myocardial damage resulting from transthoracic administration of direct current with cardiac defilbrillators present in clinical use was studied in 66 dogs. Electrodes were applied to the thoracic skin. All direct current discharges were delivered via a commercially available defilbrillator. Most of the animals received ten consecutive discharges with a dial setting of 400 watt-seconds. Animals were killed from 3 to 14 days after receiving the discharges. Myocarial necrosis was produced in most of the animals. The lesions were characterized by sharply localized areas of muscle necrosis that progressed to fibrous scars. Mineralization of damaged muscle and florid proliferation of large mononuclear cells were striking features of the lesions.


Selective filtration in the isolation of independent clones of Streptomyces. A single procedure for an efficient discrimination among the different size components of heterogeneous populations of Streptomyces spore suspensions is described. Conidia connected by substances of hydrophobic nature make obtaining all the spores in independent units difficult. For the isolation of pure clones, single spore suspensions were prepared by selective filtration through uniform and controlled pore size membranes. Single spore-enriched suspensions may be a useful tool for increasing the yield of auxotrophs, ensuring independent mutation during mutagenic treatments. This procedure can be extended to other sporulating species.


Premonitory sign of heart block in acute posterior myocardial infarction. The appearance of the ARS complex in leads V3R and V4R was analysed in a series of 94 patients with acute posterior myocardial infarction. The cases of posterior myocardial infarction with direct signs of injury (ST segment elevation with a rise of 0.5 mm or more of point F and/or QS pattern) in leads V3R and/or V4R were complicated three times as often by atrioventricular block as those in which such signs were absent (66% and 22%, respectively; P smaller than 0.001). When one of these signs was present in leads V3R and/or V4R, the disorder of conduction was "severe" (complete atrioventricular block or sinotrial block with pauses) in half the cases and "unstable" (bradycardia below 50 beats/min; ventricular pause with or without syncope; widening of QRS complex; ventricular hyperexcitability) in one-third, justifying the introduction of a stimulating catheter. Such disorders were found, respectively, only 1 in 7 (14%), and less than 1 in 10 (8%) when these signs were absent (P smaller than 0.001). The association of ST segment elevation and QS pattern was rarer (15 cases) than the isolated finding of either sign. It was found in the most severe disorders of atrioventricular conduction. The changes observed in leads V3R and/or V4R before the appearance of atrioventricular block enable one to predict which patients with posterior myocardial infarction are the most likely to develop atrioventricular block. These electrocardiographic features seem to indicate septal involvement.


Pulmonary vascular changes associated with isolated mitral stenosis in India. Pulmonary vascular changes were studied in 100 cases of isolated mitral stenosis; these included 90 patients in whom lung biopsies were obtained at valvotomy and 10 patients who came to necropsy. Medial thickness of the pulmonary arteries was measured in each case and in 12 cases was correlated with the haemodynamic data. Most patients were young, 78 being 30 years of age or less and 42 under 20 years or less. Males predominated 2:1. All patients with mitral stenosis showed varying degrees of vascular and other associated parenchymal changes. The most conspicuous were those observed in the muscular branches of the pulmonary artery in which the media was thickened in all cases, moderately in 44 and considerably in 28 cases. Dilation lesions representing grade 4 lesions of hypertensive pulmonary vascular disease (Heath and Edwards, 1958), hitherto not described in mitral stenosis, were observed in 4 cases. The intima was found to be frequently abnormal, showing oedema, fibrosis, and, more importantly, variable degrees of muscularization, often suggesting the incipient formation of a second media. Arteries and arterioles were often occluded by thrombi in various stages of organization, and the freshly formed channels tended to acquire a muscular lining. Arterioles were muscularized in all cases, and in many there was a pronounced intimal proliferation. Other changes included medial hypertrophy in the veins and and occasional muscularization and dilatation of the lymphatics. A notable feature was hypertrophy of the musculature of the bronchiolo-alvelar system seen in a majority of cases. The alveolar walls showed variable degrees of thickening and fibrosis, intimal proliferation of alveolar capillaries, and "epithelialization" of alveoli. Haemosiderosis was present in 70 cases. On the whole the more severe changes were observed more often in the younger subjects, further supporting the observation that rheumatic mitral stenosis in India commonly affects the juvenile age groups and is characterized by association with severe pulmonary hypertension. Medial hypertrophy was proportional to the level of pulmonary artery pressure.


Pulmonary blood flow distribution after banding of pulmonary artery. Radioisotope lung scanning was used to investigate the distribution of pulmonary blood flow after banding of the pulmonary artery in children with a left-to-right shunt and pulmonary hypertension. An abnormal distribution of blood flow in the lung on the side of the operation approach was observed in all patients in the first three weeks following surgery. Abnormalities were still observed in 17 of 21 children 10 months to more than 8 years after the banding operation. There was no significant relation between the occurrence of these abnormalities and time after surgery. Diminished flow to the zones of the right lung was observed less frequently. The incidence of abnormalities in flow distribution was also high preoperatively. Respiratory complications in infants with large left-to-right shunts were considered to be responsible for most of the abnormal blood flow distributions observed. Radioactive lung scanning was found to be a valuable diagnostic method in the early and late postoperative period in infants and small children. It was more sensitive than the other techniques used in revealing deviation of blood flow from one lung in those cases with shifting of the applied band.


Echoventriculographic detection, localization, and quantification of left ventricular asynergy in acute myocardial infarction. A correlative echo- and electrocardiographic study. "Echoventriculography", an echocardiographic method specially developed to scan the regional function of the left ventricle, is introduced for studying left ventricular wall motion alteration in patients with acute myocardial infarction. Purposeful probe directions, a 2:1 magnification, and careful adjustment of the gain and reject levels allowed a direct echocardiographic scanning of practically the entire left ventricle. Technically acceptable echoventriculograms were obtained from the upper and lower halves of the septal, anterior, lateral, and postero-inferior left ventricle segments in all observations on 30 consecutive patients with acute myocardial infarction. Various degrees of regional left ventricular asynergy were present in 100 per cent of the patients with acute myocardial infarction. In contrast, synergic ventricular segmental wall motion was observed in 40 healthy subjects. Pronounced asynergy was already detectable within 12 hours from onset of the symptoms of acute myocardial infarction. Echoventriculography detected acute left ventricular asynergy as well in the anteroseptal or lateral as in the posteroinferior locations. The anterior and/or septal infarction (13 of the 30 patientsy always showed a paradoxical systolic motion of the, generally large, infarcted areas. The amplitude of abnormal outward motion was up to 5 mm. In the posteroinferior infarctions (17 patients) akinetic or hypokinetic modes prevailed. The contractile function of the uninvolved segments could be measured at the same time. Hypercontractile left ventricular wall motion was common in these healthy areas in acute myocardial infarction. These findings provide useful insight into the various components of the overall left ventricular pump function in acute myocardial infarction. The validity of the echoventriculographic evaluations of the segmental left ventricular function subsets was further confirmed in 2 patients undergoing left ventricular cineangiographic studies and in 2 by necropsy. The site of the asynergic left ventricular wall motion abnormalities correlated excellently with electrocardiographic prediction of the site of acute myocardial infarction. The echoventriculographic analysis proved to be more accurate in detecting asynergy than was the electrocardiogram. This new echoventriculographic method may become a useful tool for serial noninvasive alalysis of left ventricular performance, in detecting both the asynergic areas and the reserve function of the normal regions in acute myocardial infarction.


Familial pulmonary stenosis with underdeveloped or normal right ventricle. A family with 3 cases of pulmonary stenosis with underdeveloped or normal right ventricle is described. In the family there have also been some spontaneous abortions and many infant deaths, and it seems possible that these were also related to congenital cardiac abnormalities. Recognized syndromes with pulmonary stenosis and reports on familial occurrence of this malformation are reviewed. The cases reported in this paper differ from these previously described syndromes. The possibility of sex-influenced monogenic inheritance is discussed.


Extreme pulmonary hypertension caused by mitral valve disease. Natural history and results of surgery. Five hundred and eighty six patients with mitral valve disease were studied with cardiac catheterization between 1961 and 1972; 48 (8.2%) had extreme pulmonary hypertension (resting systolic pulmonary artery pressure of 80 mmHg or above and pulmonary vascular resistance of 10 units or greater) and of these patients, 27 underwent cardiac surgery. The operative mortality for mitral valvotomy was 11 per cent and for mitral valve replacement 56 per cent. The overall mortality was 31 per cent. The risks of operation were increased in those with a long history of cardiac symptoms, those over 50 years of age, and in the presence of associated aortic valve disease. The mean survival for those patients not having operation was only 2.4 plus or minus 0.5 years. The mean follow-up period for those surviving operation has been 5.8 plus or minus 0.6 years, and symptomatic improvement has been good.


Myocardial lactate metabolism during isometric hand grip test. Comparison with pacing tachycardia. Twenty-five patients with chest pain were studied by left ventriculography and coronary arteriography. Myocardial metabolic studies were done during control state, pacing tachycardia, isometric hand grip at 30 per cent of maximum force, and combined hand grip plus pacing tachycardia. Nine patients had myocardial lactate abnormality (group 1) as evidenced by myocardial lactate production or decreased extraction (less than 10%). Though tension time index and triple product (left ventricular ejection time times HR times systolic pressure) as determinants of myocardial oxygen consumption were highest during combined hand grip plus pacing tachycardia, myocardial lactate abnormalities were most frequent during pacing tachycardia. The present study indicates that isometric hand grip even if performed during pacing tachycardia is not a sensitive test for detection of myocardial lactate abnormalities. The rising level of arterial lactate during isometric hand grip is the most likely mechanism of positive myocardial arteriovenous lactate difference.


The pattern of intramural veins of the left ventricle of the human heart. A detailed injection, microradiographic, and histological necropsy study of 18 hearts showed that the pattern of myocardial veins differed from the arterial pattern. In "normal" hearts, large drainage veins began in the subendocardial zone and coursed fairly directly towards the epicardium, maintaining a comparatively even calibre throughout. Smaller, but similar, drainage veins also begain in the middle of the myocardial wall. In the outer myocardium small groups of these vessels converged to form a single vein which then entered, almost at right angles, the pericardial veins. In general, small veins within the myocardium entered directly into the large drainage veins without any extensive intermediate sized venous network. In "abnormal" hearts with left ventricular hypertrophy the myocardial veins appeared basically normal but were more widely separated and consequently overall less dense on microradiography. Interruption of main drainage veins and loss of normal pattern were seen in areas of myocardial fibrosis associated with coronary artery disease. In a heart with severe generalized coronary artery atheroma the normal pattern in the inner half of the wall was replaced by a "plexus" of small veins in which the majority of vessels coursed circumferentially. The possible significance of both the normal and abnormal vein pattern is discussed.


Classification of ventricular pre-excitation. Vectorcardiographic study. In a study of 45 cases of ventricular pre-excitation, 19 were classified as type A and 20 as type B according to Rosenbaum's criteria, which depend on the polarity of the major deflections in the right praecordial leads and not, as is commonly thought, on the direction of the delta vector. Six cases that could not be classified as type A or type B were termed intermediate. Vectorcardiograms were recorded from 29, and these showed a wide but continuous range of values for both the delta and the main QRS vectors in all three planes. Any classification based on these features must, therefore, depend on arbitrary quantitative data. Three patients in this series had associated right bundle-branch block. A review of the published reports on the association of pre-excitation and bundle-branch block failed to provide a rational basis for the classification of pre-excitation. It is emphasized that Rosenbaum's classification is empirical and its validity is questioned.


Pericardial defect presenting as acute pericarditis. A case of herniation of the left atrial appendage, through a defect in the pericardium, is described in a 16-year-old boy who was below the third percentile for height and weight. The clinical presentation was an acute pericarditis. Widespread inversion of T waves on the electrocardiogram and collapse of the lower lobe of the left lung with pleural effusion developed. Thoracotomy showed strangulation of the atrial appendage which was excised, and the pericardial defect repaired. After operation there was a striking improvement in the electrocardiogram and a rapid gain in weight. Syndromes caused by pericardial defects are reviewed.


Proteolysis of paramyosin from Mercenaria mercenaria and properties of its most stable segment. The helical muscle protein paramyosin appears to consist of three segments of approximately equal size that differ in stability to guanidine hydrochloride and heat. The N-terminal segment is most stable and the C-terminal segment is least stable. These differences in stability serve as a basis for design of proteolytic digestions to specifically remove segments of low and intermediate stability. Thus, at room temperature only the C-terminal region was susceptible to digestion by pepsin or trypsin. Proteolytic removal of the latter region resulted in the accumulation of the remaining two-thirds of the paramyosin molecule as a segment (PPC-1) of 140,000 daltons that was still in a stable helical conformation. Proceeding to more rigorous conditions, papain digestion of either paramyosin or PPC-1 in 4 M guanidine-HCl that would be expected to destabilize all but the N-terminal segment did result in cleavage of all except that region. The N-terminal region accumulated as a helical segment of 74,000 daltons (PPC-2) if digestion was limited to 1.5 hr or a smaller segment of 58,000 daltons (PPC-3) if digestion continued for 24 hr. Stability of the three PPC segments to guanidine-HCl and heat was measured by change in fluorescence of tyrosyl residues upon loss of the helical conformation. The stability of the segments corresponded well with the stability of those regions in the paramyosin molecule from which the segments were believed to have come. Amino acid composition of the PPC segments and of paramyosin were all very similar, and prediction of relative stability of these helical proteins from inspection of gross amino acid composition does not appear promising.


Complex formation between naphthothiopheneethanolamines and deoxyribonucleic acids. Reduced viscosities, viscometric titrations, fluorescence, and flow dichroism measurements indicate that the aromatic ring system of naphthothiopheneethanolamines (NA) is intercalated on binding to DNA. No marked change in binding was found on substituting a dibutylamino group for a piperidyl ring on the side chain. Ionic strength studies, however, did indicate that the positive charge on the side chain is quite important for binding. Both the binding equilibrium constant and the number of binding sites, as determined from a Scatchard plot of spectrophotometric results, decreased on increasing the sodium chloride concentration. The increase in Tm for several DNA samples, caused by adding NA to DNA at a fixed ratio, showed a marked dependence on the percentage of AT base pairs. The conclusion drawn from these data is that the DNA-NA complex involves high specificity of the NA for AT base pairs, intercalation of the aromatic ring system, and binding of the NA side chain in the DNA minor groove with strong interaction between the positively charged side chain and the DNA phosphate groups.


Thermodynamic properties of superhelical DNAs. Binding isotherms of ethidium to the superhelical DNA from phage PM2, and to PM2 DNA containing -1 single-chain scission per molecule, have been determined at six temperatures from 2.5 to 50 degrees, in 3M CsCl-0.01 M Na3EDTA. Spectrophotometric measurements in both the visible and ultraviolet (uv) regions were used to obtain the binding isotherms. The isotherm at 20 degrees was also obtained by determining the free ethidium concentrations in equilibrium with DNA-ethidium complexes in boundary sedimentation experiments. A simple thermodynamic analysis shows that for a superhelical DNA with v bound ethidium per nucleotide, the change in free energy per unit change in the number of superhelical turns T, dGT/dT, is directly related the ratio of the free ethidium concentrations, cf,v and cf,v, in equilibrium with the ethidium complexes of the superhelical and the nicked DNA, respectively, at the same values of v. The relationship is dGT/dT = (360/E)RTln (cf,v/cf,v), where e is the unwinding angle of the DNA helix per bound ethidium molecule. Experimentally, it was found that ln (cf,v/cf,v) = a1(v - vc), where a1 is a constant at a given temperature and vc is the value of v at which the originally superhelical DNA is completely relaxed, i.e., containing no superhelical turns. The equation relating dGT/dT and v, upon transformation and integration, gives finite difference GT,v=o = -a1NRTvc2/2, where finite difference GT,v=0 is the free energy of superhelix formation of the superhelical DNA in the absence of ethidium and N is the number of nucleotides per DNA molecule. This equation is independent of the unwinding angle e. The values of a1 are 11.2 plus or minus 0.9, 11.2 plus or minus 0.7, 11.2 plus or minus 0.6, 10.7 plus or minus 0.8, 10.0 pl.us or minus 1.0, and 9.8 plus or minus 1.4 at 2.5, 10, 20, 30, 40, and 50 degrees, respectively. Measurements with lambda b2b5c DNA, and monomeric and trimeric lambda dv DNA, indicate that the constant a1 is insensitive to the molecular length of DNA. The present results are compared with the previous results of Bauer and Vinograd (Bauer, W., and Vinograd, M. (1970), J. Mol. Biol. 47, 419), obtained by a statistical mechanical analysis of the ethidium binding isotherms (determined by density gradient centrifugation measurements) to two forms of simian virus 40 DNA at 25 degrees. The effects of superhelical turns on a number of processes, such as the binding of a ligand which unwinds or winds the DNA helix, the denaturation of a DNA segment, and the formation of hair-pinned structures from base sequences with a twofold rotational symmetry, are discussed from the thermodynamic point of view, based on the present results.


Nuclear magnetic resonance studies of 2'- and 3'-ribonucleotide structures in solution. A systematic 220-MHz proton nuclear magnetic resonance (nmr) study has been made of all common purine and pyrimidine 2'(3')-ribonucleotides in D2O solutions at 20 plus or minus 2 degrees. Spectra for the entire series were measured under similar conditions of concentration, temperature, and ionic strength, thereby facilitating intercomparisons of spectral properties. Spectral assignments were accomplished with the aid of selected 31P-1H decoupling experiments and accurate values of nmr parameters were derived by simulation-iteration procedures. A detailed analysis of the coupling constants and chemical shifts permitted a determination of conformational properties for ribose rings, exocyclic carbinol and phosphate groups, and the orientation of base-ribose rings. Following procedures described elsewhere, an evaluation was made of ribose ring pseudorotational parameters for each 2'(3')-nucleotide. The results show that both the degree of pucker and pseudorotational angle vary only slightly throughout the entire series of molecules, and lie within the ranges found in the crystalline state. Furthermore, the ribose rings are in rapid equilibrium between N type (C(3')-ENDO, C(2')-exo) and S type (C(2')-endo, C(3')-exo) conformers, N forms and is formed by S, with an S type conformer favored in purine 2'(3')-ribonucleotides (-60:40) while pyrimidines exhibit approximately equal compositions. Thus, the phosphate location on the ring has less of an effect on ring properties than the nature of the base ring. An analysis is also reported of rotamer equilibria about C(4')-C(5'), C(2')-O(2'), and C(3')-O(3') bonds. For the former the nmr coupling constant data are consistent with a predominant gg rotamer (-70%) with gt and tg rotamers populated to the extent of -20 and -10%, respectively. No correlation of the type seen for 5'-nucleotides appears to exist between C(4')-C(5') gg population and ribose ring equilibrium composition. For 2'-nucleotides the 31P-H(2') coupling data indicate a preferred C(3')g, C(1')t conformer about C(2')-O(2') in agreement with 13C nmr results. A less definitive rotamer analysis follows from observed J31P,H(3') values, but when these results are combined with relevant chemical shift data for deoxynucleosides and nucleotides the evidence strongly points to essentially free rotation and approximately equal rotamer populations about C(3')-O(3'). Chemical shift differences between purine and pyrimidine 2'(3')-ribonucleotides are qualitatively accounted for by "in-plane" purine diamagnetic anisotropy effects. Also, the greater magnitude for purine deshieldings in 2'(3')-nucleotides relative to 5'-nucleotides is explained by a more favored syn:anti ratio in the former in line with recent nucle