Mortality and its causes among Moslems, Druze and Christians in Israel. The mortality rates of Moslems, Druze and Christians in Israel have decreased strikingly since the early 1950s, mainly because of a decline in mortality among children and younger women. There has been an increase in mortality among middle-aged and elderly men. Mortality from infective diseases had declined, but these remain important causes of death, particularly among Moslems and Druze. Mortality from external causes has increased in recent years. The mortality rate from ischemic heart disease has risen sharply; it is especially high among Christian men. Mortality due to congenital anomalies is relatively high among Moslems and Druze.


Infant morbidity and mortality.

Infant morbidity and mortality. The steady decline in the infant mortality rate among Jews showed a temporary, distressing reversal during the years immediately following the establishment of the State of Israel, but thereafter the rate again declined and the reduction was maintained. The rate fell to 13.9/1,000 live births in 1977. During recent years, there has been a significant parallel fall in the number of deaths during the first six days of life, reaching 8.0/1,000 live births in 1977. Hospital admissions, as an indicator of morbidity, now show less children with infantile diarrhea, formerly a major cause of hospitalization. Although Israel has reached reasonably satisfactory standards of infant health, there remains much room for improvement.


Infective diseases in Israel. Changing patterns over 30 years. Geography, climate, socioeconomic conditions and public health action are factors that have influenced the incidence and pattern of infective disease in Israel. During the British Mandate, the epidemiologic picture was dominated by tropical and subtropical diseases. The problems related to infective diseases and their control during the 30 years of Israel's statehood, especially those connected with the mass immigration of the 1950s, are examined in detail. Diphtheria, poliomyelitis and whooping cough were quickly brought under control by means of immunization programs, and measles, later and more slowly. Tuberculosis constituted a serious burden in the early years of the State but has been successfully subdued through the combined effects of BCG vaccination, screening, case finding, systematic chemotherapy and improved socioeconimic conditions. In contrast, diarrheal diseases--while eliminated as a major cause of infant mortality--are still highly prevalent in the general population. Some specific local infective disease problems of topical scientific interest are briefly discussed.


Cancer patterns in Israel: selected aspects. Data on cancer incidence in Israel have been collected by the Israel Cancer Registry since 1960. Overall incidence patterns are similar to those observed in other "Westernized" countries. In the total Jewish population, lung cancer accounts for one in seven cancers in males and breast cancer, for one in four cancers in females. The main time trends are: a decrease in stomach cancer, a halt in the increase of male lung cancer and a continuing rise for female lung cancer, female breast cancer, cancer of the colon and rectum, and malignant melanoma. Cancer of the uterine cervix may be on the rise in Israel-born women. The relatively high incidence of cancer of the esophagus in immigrants from Iran and Yemen and of cancer of the nasopharynx in immigrants from North Africa probably reflect incidence patterns in their countries of origin.


Breast cancer in Israel: incidence and mortality, 1960-75. Between 1960 and 1975, the incidence of breast cancer in Israel rose 20%. A steady rise in incidence was seen for women aged 60 or more years, whereas for women younger than 60 years, the incidence pattern was less regular, showing a definite rise only in the period 1970-75. Mortality from breast cancer also rose among women aged 60 or more years, but much more slowly than incidence. Among women younger than 60 years, breast cancer mortality was essentially unchanged. When mortality rates from a disease rise more slowly than incidence rates, it can be inferred that a greater proportion of new cases are being cured or are experiencing longer survival. It would appear that this is the case with breast cancer in Israel, and this trend, probably the result of earlier diagnosis, is encouraging.


Ischemic heart disease in Israel: changes over 30 years. As in most of the industralized nations of the world, ischemic heart disease is the major cause of death in the adult population of Israel. The first two and a half decades since the founding of the State in 1948 were typified by a constant rise in morbidity and mortality from this disease in all ethnic groups in this heterogeneous population. Major changes in health-related behavior in the immigrant groups resulted in a rise in the disease rates that ranked this country among those with the highest incidence rates in the world. Since 1972-74 there is evidence of a fall in mortality in all ethnic groups. There is less conclusive evidence of a possible fall in morbidity as reflected by hospital admissions. These reductions are too recent for their full significance and the possible factors involved to be clear.


Epidemiologic aspects of rheumatic fever and rheumatic heart disease in Israel. Rheuamtic fever and rheumatic heart disease are still important public health problems in Israel. Rheumatic fever is a seasonal disease, more common during the cold months of the year. A high incidence of rheumatic fever followed the immigration waves of the 1950s, in contrast with a decline in new cases over the last 10 years among both Jews and non-Jews. In the same period (1950s to 1975), a decline in the numbers of hospitalized patients was also reported. After 1970 mortality rates of rheumatic fever dropped to zero. The prevalence of rheumatic heart disease has declined in children, as shown in school studies, but in adults, a different trend is seen. The number of hospitalized patients has increased, in parallel with the increasing population, to more than 3,000 hospitalized patients per year. While a plateau curve is seen in the mortality rates of males of all ethnic origins studied, among females there is a slight decline in mortality. The discrepancy between the decline in occurrence of rheumatic fever and rheumatic heart disease in children and the increase in prevalence of rheumatic heart disease in adults is explained by the massive immigration from countries with a high prevalence of rheumatic heart disease.


On the shape of the hospital industry long run average cost curve. Empirical studies of the hospital industry have produced conflicting results with respect to the shape of the industry's long run average cost (LRAC) curve. Some of the studies have found a classical U-shaped curve. Others have produced results indicating that the LRAC curve is much closer to being L-shaped. Some theoretical support exists for both sets of findings. While classical theory predicts that the LRAC curve will be U-shaped, Alchian has presented theoretical arguments explaining why such curves would be L-shaped. This paper reconciles the results of these studies. The basis for the reconciliation is recognition of the failure of individual hospitals to produce all their individual product lines at efficient volumes. Such inefficient production is feasible and perhaps common, given the incentive structure which exists under current cost reimbursement systems. The implication of this paper is that large hospitals may have a greater potential for scale economies than has previously been recognized.


Cost of elective surgery and utilization of ancillary services in teaching hospitals. Measures of surgical utilization studied are the number of elective tests performed preoperatively and the total cost per case. The unit of analysis is a matched pair of patients who underwent the same elective procedure, one a Veterans Administration patient, and the other a municipal or voluntary hospital patient. Federal ownership of the hospital ahd the strongest impact on tests and cost per case. On average, costs for the VA patients were 52 percent more per case. The foreign medical graduate variable had a large positive (inflationary) effect on the number of tests, but a slight downward influence in the cost regressions. The fraction of surgeons with faculty appointments had a strong negative (curtailing) impact on elective testing, but an upward influence on cost per case. Additional variables such as age, average laboratory turnaround time, and fraction of the medical school's students doing their surgical clerkship at the hospital ahd a slight upward influence on utilization. The three policy issues raised in the study involve changing the hospital reimbursement incentives, targeting continuing education programs to categories of staff that need it most, and redistributing faculty and students.


[Necrotizing enterocolitis in the neonatal period. Revue of 19 cases proved by anatomopathologic examination]. Nineteen newborn infants with necrotizing enterocolitis, proved by autopsy or surgical examination, are presented. In this study the absence of blood in the stools and of pneumatosis intestinalis did not exclude the diagnosis of necrotizing enterocolitis. The role of umbilical arterial and venous catheters and of exchange transfusion in the pathogenesis of the disease was less than has been previously reported. There was no evidence for emboli of thrombosis in the mesenteric vessels. On the other hand, the probable importance of nonocclusive mesenteric arterial insufficiency is supported by several observations, including the high incidence of antecedent hypoxaemic episodes, the time of onset of the subsequent gastrointestinal pathology and the localization of the lesions. An infectious component was demonstrated, but our results do not allow to state precisely the time of its occurrence.


Relationship between clinical conditions, radiographic findings and pulmonary functions in patients with cystic fibrosis. Clinical and radiological findings, quantified by special scores, in 36 patients with cystic fibrosis are compared with lung function measurements, including the ratio of RV/TLC and FEV/VC and the arterial blood gases at rest and during exercise. Although respiratory function tests were found to correlate well with both the pulmonary finding score and the chest radiographic score, distinction of three severity groups was possible only by a vector calculation of the blood gases in PaO2-PaCO2-diagram, at rest and during exercise. Thus, measurements of pO2 and pCO2 at rest and on exercise appear to be a helpful adjuvant to routine spirometry for the individual appraisal of the degree of lung involvement, performance and care level.


Complement in cystic fibrosis.

Complement in cystic fibrosis. Quantitative and functional assessments were made of both the classical and alternative pathways of complement activation in sera from 23 patients with cystic fibrosis. The classical pathway functioned similarly in patients and controls as measured by CH50 titre. Alternative pathway function, initiated in patient sera by incubation with inulin, was equal to that of controls as determined by cleavage of Factor B and C3, and by the consumption of terminal components. Factor B, however, was more readily activated in patient than in control sera. This rapid alteration of Factor B did not lead to accelerated or more extensive activation of the terminal complement components via the alternative pathway when assessed by C3 cleavage and the consumption of terminal components. Thus, a complement deficiency was not found. The importance of the easily activated Factor B is undefined.


Testicular volume and function in adolescence after breech delivery. Testicular volume and pubertal development were investigated in 108 boys born by breech delivery aged 14 1/2 to 19 1/2 years. Mean testicular volume was slightly but not significantly lower than in a control group of the same age. Pubertal development was normal. One boy had unilateral and two boys bilateral testicular atrophy (one with azoospermia and one with oligospermia). In a control group of 135 boys delivered from vertex position ther were three cases with unilateral and none with bilateral testicular atrophy. In both groups all cases with unilateral testicular atrophy could be explained by postnatal testicular injury, whereas the two cases of bilateral atrophy in the group of breech delivered boys remained unexplained. They may have originated from local trauma during breech delivery. It is concluded that testicular atrophy as a complication of breech delivery is seen with a frequency of only 2% or less.


[Multidisciplinary treatment of rhabdomyosarcoma in childhood]. From 1974 to 1978, 21 children with rhabdomyosarcoma were treated in a combined interdisciplinary protocol. All children received a polychemotherapy for 18 months. Radiotherapy and surgery were used according to the stage and localization of the tumor. The overall two-year survival rate is 86 +/- 9%, the tumor free survival rate is 78 +/- 11%. 7 patients have no evidence of disease after the end of treatment.


Pseudo-vitamin D deficiency rickets: report of a case. A 19-month-old boy with repeated convulsions from the age of 7 months was found to have pseudo-vitamin D deficiency rickets. The convulsions were attributed to hypocalcaemia which is one of the main features of this type of rickets. High doses of vitamin D produced clinical, biochemical and radiographic cure.


Congenital defect in intracellular cobalamin metabolism resulting in homocysteinuria and methylmalonic aciduria. I. Case report and histopathology. The case is described of an infant who suffered from progressive, severe dystrophy, hemolytic and megaloblastic anemia, hematuria, proteinuria and slight uremia. He died at 4 months of age following two acute episodes of heart failure. Abnormally increased excretion of methylmalonate and homocystine was detected by our screening program for metabolic disorders. Amino acid analyses showed that the plasma and urine levels of methionine were very low whereas those of cystathionine were raised. Vitamin B12 deficiency, malabsorption or abnormal cobalamin transport were excluded by a normal serum total cobalamin and normal transcobalamins. These findings suggested a congenital error of cobalamin metabolism. Treatment with vitamin B12 resulted in a biochemical though not a clinical response. Postmortem examination revealed severe vascular lesions with changes in the kidney characteristic of thrombotic microangiopathy supporting a diagnosis of hemolytic-uremic syndrome. It is assumed that the elevated plasma homocysteine induced the vascular lesions by causing detachment of endothelium.


Human alveolar macrophages spontaneous reduction of BSPT salt. Alveolar macrophages of non smoking and smoking human adults reduce BSPT salt spontaneously. The staining obtained is located on three cell membrane systems: the endoplasmic reticulum, the Golgi apparatus and the nuclear envelope. Methylene blue MB inhibits BSPT reduction. The smokers alvelolar macrophages have less positivity than those of the non smokers. The endogenous cell substrate revealed in this work is the initial common pathway of two different oxidative chains bounded to the microsome. One acts with cytochrome P 450 for chemical detoxification by hydroxylation, the other one acts with cytochrome B5 for lipid oxidation or peroxidation and both may be connected with the cell bactericidal system.


Acridine orange staining of the mammalian fibroblast cell coat. Acridine orange selectively binds to glycos- and galactosaminoglycan (GAG) compounds in the presence of Na+ in low concentrations. We have worked out a cytochemical method, which is suitable for the specific demonstration of surface GAG components. The method consists of a glutaraldehyde prefixation, an acridine orange block-staining for 48 h and an OsO4 postfixation for some hours. The specificity of the staining was verified with the help of fluorimetric--in vitro--measurements and enzymatic digestions, respectively.


[Comparative endoscopic and histological findings in chronic sinusitis (author's transl)]. Comparative studies between endoscopic and histological findings in 93 patients with chronic sinusitis showed that the diagnosis based upon endoscopic mucosal appearances does not compare necessarily with the histological findings. Therefore the endoscopic findings are questionable indications for surgery without histological confirmation.


[Temporary total deafness in disseminated encephalomyelitis (author's transl)]. Total deafness, which recovered, in an 18 year old woman with disseminated encephalomyelitis is reported. The deafness involved the left ear, but vestibular abnormalities were demonstrated bilaterally. After some weeks both auditory and vestibular responses returned to normal but nearly 2 years later a temporary left ear hearing loss of 30 dB and a spontaneous nystagmus occurred.


[Extent and frequency of noise-induced hearing loss in miners depending on the intensity and duration of noise exposure]. The audiograms of 218 coalminers were studied and it was found that 37.7% had hearing losses attributable to noise. Based upon sound level measurements and duration of exposure the risk of hearing loss in mining can be also predicted and its statistical probability and its risk measured.


[Anesthesia and adenectomy under visual control with bipolar electrocoagulation]. To make complete removal of the adenoid tissues possible, it is necessary to perform surgery under optical control. The technique for the operation is described, utilizing newly-designed instruments and bipolar electrocoagulation for control of bleeding.


[A trilobed rotation cheek flap for closure of facial defects (author's transl)]. Rotation flaps of the cheek as described by Imre and by Esser are well known. In some situations we use a "tri-lobed cheek flap" to close defects of the cheek, external nose or temple. This flap is described.


An atlas of microscopic dissection following the attic approach to the cat's middle ear. A brief illustrated atlas demonstrates the progressive relationships of the anatomical structures of the cat's middle ear as one dissects these from the attic approach. There was an attempt to demonstrate the nerves of the lateral semicircular canal and the utricle but without preserving the footplate of the stapes to afford an unobstructed view of the nerves. For the first time, the anatomical features of the cat's middle are are brought together in a progressively illustrated and single source.


A multivariate strategy for prediction of psychoacoustic performance from electroacoustic characteristics of hearing aids. Sentence length psychoacoustic material was processed through 16 hearing aids and presented to 220 listeners. Twenty-nine indices of electroacoustic performance were abstracted from a testing program under laboratory conditions. Through a multivariate statistical strategy, psychoacoustic achievement was shown to be positively correlated to and could be predicted by a knowledge of white noise gain, bandwidth below and above 1 kHz, frequency response regularity, and transiet distortion (multiple R = 0.77; F = 2.04; df = 5,10). It was concluded that a multivariate statistical tactic, when used with discretion, can be a useful and powerful tool to predict psychoacoustic performance from a knowledge of hearing aid electroacoustic characteristics.


Cardiovascular response to acoustic stimuli in one-, two-, and three-month-old infants. Fifteen normal infants were seen at the ages of 1, 2, and 3 months. Cardiac responses to auditory stimuli of various frequencies and intensities were evaluated to determine the effects of age of the infant, frequency of the stimulus, and intensity of the stimulus. Heart rate was measured by a fingertip pulse counter and recorded on a polygraph. While the heart rate was being monitored, the infant was presented with a series of tone bursts of 1.5-sec duration with an interstimulus interval of 13.5 sec. Frequencies and intensities of stimuli were found to have no effects on the response. The cardiac rate response was influenced more by the ages of the infants than by stimulus parameters. Variability of the data for two-month-old children seemed to indicate a transition occurring at this age. However, the major influence upon the response was found to be the prestimulus heart rate.


Brainstem electrical responses from selected tone pip stimuli. Brainstem-electrical responses were obtained from 10 normal hearing adult subjects using frequency specific tone pips as stimuli. The four frequency specific tone pips (500, 1000, 2000, and 4000 Hz) were diamond shaped with a 2.5-msec rise/fall time. Each tone pip was presented at four intensity levels (70, 50, 30, and 10 dB hearing threshold level), and graphic recordings were made for each frequency at the specific intensity levels. Frequency specific Wave V intensity-latency functions were plotted, and these results were compared to tone pip data obtained in previous studies. In addition, suggested test procedures for obtaining tone pip brainstem-evoked responses under diagnostic conditions are discussed.


Acoustic reflex thresholds using conventional and tracking methods. Relationship between acoustic reflex thresholds obtained using conventional and tracking procedures was investigated. In the tracking method, the intensity of the stimulus from a self-recording audiometer is adjusted by the experimenter to bracket the acoustic reflex threshold as visually observed on the meter of an acoustic impedance measuring device. Acoustic reflex thresholds were obtained to pure tones of 500, 1000, and 4000 Hz. High correlations and no significant differences were obtained between the two threshold methods. It was concluded that the tracking method which allows easier control of stimulus parameters and response recording can provide acoustic reflex threshold data similar to those obtained using a conventional method.


Spread and detection of psoroptic scabies of cattle in the United States. Psoroptic scabies of cattle causes substantial economic losses. An eradication program begun in 1905 virtually eradicated the disease by 1950. Complacency was followed by a resurgence of the disease, particularly in the 1970s, when more than 1,000 outbreaks were reported. Program effectiveness was reduced when the disease was often not detected or reported for several months after herds became infested. Further spread was enhanced by infested cattle moving undetected through market channels.


Influence of high pass filtering on the intelligibility of amplitude-compressed speech. Monosyllabic triplet word intelligibility scores were obtained from normal-hearing and hearing-impaired, loudness-recruiting subjects under two experimental conditions: (1) high-pass (1200 Hz)-filtered, linear amplification, and (2) high-pass (1200 Hz)-filtered, compression amplification using input-to-output ratios of 5:1 and 20:1. Test materials were administered at increased sensation levels of 0, 10, 20, and 30 dB. In general, speech intelligibility was slightly enhanced for normal and hearing-impaired listeners, but only at lower sensation levels. Moreover, the improvement was observed only under the filtered, compression amplification condition for both groups. No important differences were observed between the two compression ratios used. This compression advantage may or may not be observed in clinical hearing aid evaluations.


A discussion of some temporal characteristics of electroacoustic impedance bridges. The purpose of this study was to describe an acoustic method for measuring temporal characteristics of electroacoustic impedance bridges, and to present data on the temporal responses of three widely used electroacoustic instruments. The potential effects of these instrument parameters on measured characteristics of the acoustic reflex are discussed.


A comparison of the effect on consonant discrimination of combining low- and high-frequency passbands in normal, congenital, and adventitious hearing-impaired subjects. To investigate consonant information in low frequencies, a low-frequency band which contains negligible consonant information in isolation was added to a high-frequency band. There was a significant increase from the high-frequency band-alone score for normal-hearing subjects when the bands were presented at favorable low-frequency/high-frequency bands sensation levels either to the same ear or opposite ears. For congenitally and adventitiously hearing-impaired subjects, there was a significant increase only when the bands were presented to opposite ears. A clinical application of these results is split-band amplification whereby low frequencies were added to one aid and removed from the other aid. Removal of low frequencies from one aid reduces internal noise, attenuates unwanted background noise, reduces cochlear masking, and thus optimally amplifies that part of the speech spectrum which conveys most of the consonant information. The low-frequency response aid adds additional information, including pitch, rhythm, and intonation.


Prognosis after surgical excision of fibrosarcomas in cats. Forty-four cats from which fibrosarcomas had been excised were studied for a minimum of 3 years, of until death. Two factors, tumor site and mitotic index, were found to be of prognostic significance, but tumor size, duration of growth, and histologic appearance were not. Following removal of fibrosarcomas from the pinna or flank in 6 cats, none died as a result of the tumor but 24 of 35 (70%) cats with fibrosarcoma in the skin of the head, back, or limbs were euthanatized because of local recurrence, usually within 9 months of surgery. The median survival time of cats with a tumor of mitotic index 6 or more in the head, back, or limbs was 16 weeks, compared with 128 weeks when the mitotic index was less than 6. Eleven percent of cats developed pulmonary metastases.


Diabetes insipidus in a cow.

Diabetes insipidus in a cow. Diabetes insipidus was diagnosed in a 6-year-old Holstein-Friesian cow with a history of recurrent fever, ketosis, lymphadenopathy, and inappetence. The diagnosis was based on the clinical findings, response to exogenous vasopressin, and lack of urine concentration in a water deprivation test. The disease gradually regressed over a period of 1 year and did not recur.


Anemia and osteopetrosis in a dog.

Anemia and osteopetrosis in a dog. A 1-year-old, male Australian Shepherd Dog with consanguineous parents was discovered to have severe nonregenerative anemia associated with osteopetrosis. Diagnosis of the bone abnormality was established by skeletal radiography and microscopic examination of a rib biopsy specimen. The anemia was attributed to failure to develop normal marrow cavities combined with failure of extramedullary erythropoiesis. Although blood transfusions sustained the dog's life for 15 months, the dog died of a hemolytic transfusion reaction.


Histopathologic evidence for parvovirus infection in dogs. Two dogs kenneled at a commercial establishment died of a disease manifested as severe hemorrhagic diarrhea. Histopathologic findings in both dogs bore a striking resemblance to those typical of feline infectious enteritis. Viral particles resembling members of the Parvoviridae family were observed in the contents of the small intestine of one of the dogs. The disease was similar to the recently recognized canine gastroenteritis associated with a parvovirus-like agent.


Crystalline corneal opacities in the Siberian Husky. Bilaterally symmetric opacities were detected in the corneal stroma of 78 (14%) of 560 Siberian Huskies, aged 7 months to 12 years, examined in ophthalmology screening clinics. The opacities were round or horizontally oval and consisted of a diffuse gray homogeneous haze in the anterior stroma or an array of fine polychromatic crystals in the posterior stroma, or both. The corneas were not inflamed. The frequency of occurrence and density of the opacities increased with age. Several affected dogs were closely related, but a specific inheritance pattern could not be established. Light and electron microscopy disclosed clusters of extracellular, thin, needle-shaped, crystalline clefts. Histochemical stains on frozen sections identified neutral fats, phospholipids, and cholesterol as components of the crystals.


Hereditary spinal muscular atrophy in Brittany Spaniels: clinical manifestations. A progressive neurologic disease was identified in a family of closely related Britanny Spaniel dogs. The disease developed in dogs less than 1 year old and characteristically resulted in paraspinal and proximal pelvic limb muscular atrophy. Neurologic examination, electrodiagnostic testing, and histologic studies indicated that the disease results from motor horn cell degeneration. A colony of affected dogs was established, and breeding studies incriminated an inherited basis for the disease.


Central corneal endothelial cell counts in children. Our data indicate that corneal endothelial cell density in a normal eye remains essentially unchanged from ages five to 20. Changes in cell density and morphology during the first five years of life need further investigation. In this study, the mean endothelial cell count of a normal population of children was 2696 cells/mm2. Children undergoing cataract aspiration had a mean cell loss from the normal of 7.2%. The cell loss rates for push-pull aspiration-irrigation or phacoemulsification cataract extraction were not statistically different. The implantation of an intraocular lens was associated with a 33.3% endothelial cell loss when compared to the control or normal population of eyes. A mean loss of 38.0% was noted when the IOL eyes were compared to their normal fellow eyes.


Sterile uveitis and intraocular lens implantation. Cases of sterile hypopyon following intraocular lens implantation, accumulated over a 4 1/2-year period, were subjected to critical clinical review and analysis. In approximately two-thirds of the cases, the anterior chamber cleared within three to four weeks without significant sequelae. The remaining cases followed a chronic or recurrent course. Recurrent anterior segment reaction appears to be a definite indication for early implant removal. The final visual results in this group of cases were less than satisfactory, suggesting that the appearance of sterile hypopyon after implant surgery is an ominous clinical sign which may prelude secondary membrane formation, chronic cystoid maculopathy, or even phthisis. The etiology of sterile hypopyon remains vague.


A closed technique for repositioning dislocated iris plane lenses. Of 450 iris plane IOL's implanted over a 3 1/2-year period, 20 (5%) dislocated. Four of these lenses dislocated into the vitreous and were either floated or surgically reoriented into the anterior chamber to give final visual acuities of 20/30 or better. Sixteen lenses dislocated into the anterior chamber and were repositioned using a closed bimanual technique. After lens repositioning, eighteen eyes achieved 20/30 or better vision. One eye's vision was 20/200 and another case had finger-counting vision.


The Shearing-style intraocular lens and the posterior chamber. We observed the Shearing-style intraocular lens in human cadaver eyes and in dog eyes. In human cadaver eyes the lens loops rest in the ciliary body sulcus without exerting undue pressure; the only major problem occurred when the posterior capsule was torn and a loop found its way into the pars ciliaris of the ciliary body or into the vitreous base. Seven Shearing-style lenses were implanted in four dogs: six-month preliminary histopathology showed four of these lenses to be well tolerated with abundant synechiae formation around the loops. Two eyes exhibited loop pressure necrosis due to slight lens displacement.


Comparative endothelial biomicroscopy.

Comparative endothelial biomicroscopy. Calibrating comparative endothelial biomicroscopy with a corneal endothelial microscope (Heyer-Schulte Medical Optics), we have obtained clinically useful estimates of endothelial cell density. This simple and inexpensive technique allows rapid examination and evaluation of a large number of endothelial cells across various corneal diameters. It is, however, a clinical rather than investigative procedure and is not intended to replace the quantitative features of endothelial specular photomicrography for scientific studies.


The Knolle-Pearce irrigating loop.

The Knolle-Pearce irrigating loop. Use of an irrigating lens loop which maximizes fluid flow eliminates the need for direct pressure or counter-pressure as the lens nucleus is removed from the anterior segment and also protects the internal structure of the globe from surgical trauma.


An investigation of allied health faculty attitudes toward faculty development concepts. Allied health faculty attitudes toward faculty development issues were determined by using a semantic differential technique. The faculty development issues included: (1) faculty evaluation by students, (2) faculty evaluation by peers, (3) faculty evaluation by chairperson and/or program director, (4) continuing education and (5) faculty tenure. Subjects (n=128) included students and graduates of a graduate program in allied health education and administration (program affiliates) with a master's degree in education. Other subjects included faculty located within geographic proximity to the graduate program who had received a master's degree other than in education as their most advanced degree (nonprogram affiliates). Differences in allied health faculty attitudes toward the faculty development issues were attributed to the degree to which faculty had been socialized into the educational system. Four factors emerged from factor analytic procedures on which subject attitudes were compared: the merit, organization, style and rigor of faculty development activities. Significant differences were found among program affiliates but no between program affiliates and nonprogram affiliates.


Teacher evaluation in the health professions. This paper reviews current concepts and practices of teacher evaluation as it exists in health professions educational programs. Problems associated with teacher evaluation and the necessity for well designed evaluation systems are explored. Recommendations are made for the development of objective teacher evaluation procedures.


Outcomes of teacher preparation in allied health sciences. Formal preparation of allied health instructors for responsibilities of teaching and educational leadership has been encouraged through substantial federal grants, institutional funds and university programming. A review of the literature reveals very little regarding formal measurements of outcomes in terms of career progress of the graduates their professional activities and their level of satisfaction with the educational content and preparation as a teacher. This study focuses on the achievements and opinions of 106 graduates of a master's degree teacher preparation program. Graduates were found to be employed in positions commensurate with their preparation, and rewarded by their employers with career advancement, salary increases and promotions. The graduates believed that they had gained the skills needed. They also identified the strengths and weaknesses of the program. Implications of the findings are discussed.


Analyzing and developing effective instructional behaviors for allied health educators. This article discusses interaction alaysis, a procedure for objectively recording and analyzing the verbal interchange between instructors and students, which can aid instructors in setting goals for improving their instruction. The following questions are addressed: Why should instructors study their own behaviors? What conditions are necessary for studying one's teaching behaviors? What is interaction analysis? How does one implement interaction analysis? How does one interpret the results of interaction analysis? What does research indicate about interaction analysis? The authors mention the well-known Flanders System of Interaction Analysis, and outline the use of a simpler system, the System for Coding Interaction with Multiple Phases (SCIMP), developed by Townsend.


Allied health careers special resources and services program: increasing the probabilities of success for 'high risk' students in allied health career programs. This article describes a successful developmental program specifically designed for academically "high risk" students entering a two-year community college career program in allied health. The program consisted of providing an intensive three-week instructional program to students before they entered the allied health career program, and subsequently providing an ongoing support system of tutoring, counseling and career development activities. Participants attained higher levels of academic performance and retention than nonparticipants.


Controlled biosynthesis of neoviridogriseins, new homologues of viridogrisein. III. Production, structures and biological properties of neoviridogriseins I and III. The high response of our isolate of Streptomyces griseoviridus was exploited to provoke the synthesis of new viridogrisein homologues by adding various amino acids to the culture medium in an attempt to replace the alanine, sarcosine, leucine and phenylsarcosine moieties of virdogrisein. Among the amino acids added, L- and DL-alpha-amino-n-butyric acid and L-methionine gave new TLC spots which we named neoviridogriseins I and III and neoviridogrisein VII, respectively. The structures of neoviridogriseins I and III were elucidated: In both compounds, the alanine moiety of viridogrisein is replaced by a L-alpha-amino-n-butyric acid residue; furthermore, in neoviridogrisein I, the allo-hydroxy-D-proline is replaced by D-proline.


Comparative in vitro activity of LY 127935 (6059-S), seven cephalosporins, three aminoglycosides, carbenicillin, and ticarcillin. LY 127935 (6059-S), a new semi-synthetic beta-lactam antibiotic was tested simultaneously with 6 cephalosporins, 3 aminoglycosides, carbenicillin and ticarcillin against 398 clinical isolates of Gram-negative bacilli and Gram-positive cocci. Many of the organisms were selected for study because of known resistance to one or more of the clinically available antibiotics tested. Escherichia coli, Klebsiella, Serratia and Providencia were susceptible to LY 127935. Some resistant strains of Enterobacter, Proteus, Pseudomonas aeruginosa and Acinetobacter were also resistant to LY 127935, but many of the strains resistant to other antibiotics were susceptible to LY 127935. The activity of LY 127935 against Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus viridans and Streptococcus bovis was similar to that of cephalexin and cephradine. LY 127935 was not active against methicillin-resistant S. aureus nor enterococcus.


Identification of the active metabolites of the isoxazolylpenicillins by means of mass-spectrometry. Electron-impact mass-spectrometry of the methyl esters of the isoxazolylpenicillins and of their active metabolites showed the latter to be formed from their parent compounds by hydroxylation of the 5-methyl group.


Production of 9-beta-D-arabinofuranosyladenine by a new species of Streptomyces and its herbicidal activity. 9-beta-D-Arabinofuranosyladenine (ara-A) was produced by a new species of Streptomyces designated as S. herbaceus. Ara-A was found to have potent herbicidal activity against Echinochloa crus-galli, Digitaria adscendens and Chenopodium ficifolium by the treatment with ara-A before germination of these plants. However, Oryza sativa had strong resistance to ara-A.


Synthesis and antibacterial activity of 1-N-(1,3-dihydroxy-2-propyl)kanamycin B (UK-31,214). 1-N-(1,3-Dihydroxy-2-propyl)kanamycin B was prepared and its in vitro activity against aminoglycoside-sensitive and aminoglycoside-resistant organisms was compared with that of kanamycin B and gentamicin. This kanamycin B derivative (code No. UK-31,214) demonstrated potent activity in all of these tests and gave good protection in experimental infections in mice.


Studies on bacterial cell wall inhibitors. VI. Screening method for the specific inhibitors of peptidoglycan synthesis. A screening method was established for selecting new specific inhibitors of bacterial cell wall peptidoglycan synthesis. In the primary test, culture broths of soil isolates were selected based on relative microbial activity. A culture, to be retained, must be active against Bacillus subtilis and lack activities against Acholeplasma laidawii. In the secondary test, inhibitors of bacterial cell wall synthesis were identified by their ability to prevent the incorporation of meso-[3H]diaminopimelic acid but not to prevent the incorporation of L-[4C]leucine into the acid-insoluble macromolecular fraction of growing cells of Bacillus sp. ATCC 21206 (Dpm-). As the tertiary test, inhibitors with molecular weights under 1,000 were selected by passage through a Diaflo UM-2 membrane. By this screening procedure, six known antibiotics and one new one were picked out from ten thousand soil isolates.


Studies on bacterial cell wall inhibitors. VII. Azureomycins A and B, new antibiotics produced by Pseudonocardia azurea nov. sp. Taxonomy of the producing organism, isolation, characterization and biological properties. Two new basic water-soluble antibiotics, azureomycins A and B, were isolated from the culture broth of an actinomycete, strain AM-3696, designated as Pseudonocardia azurea nov. sp. The antibiotics exhibit moderate antimicrobial activities against Gram-positive bacteria including penicillin-resistant Staphylococcus, Mycobacterium and Clostridium. They inhibit the synthesis of bacterial cell wall peptidoglycan.


Studies on antibiotics BN-227 and BN-227-F, new antibiotics. I. Taxonomy, isolation and characterization. The two new antibiotics, BN-227 and BN-227-F, were isolated from the fermentation broth of Pseudomonas sp. BN-227. BN-227 has a molecular formula C7H9NO3, and melts at 115 degrees C. BN-227-F has a molecular formula C21H24N3O9Fe, and melts at 156 degrees C. BN-227-F is a chelate compound consisting of three similar ligands (antibiotic BN-227) and ferric ion. The two antibiotics have antimicrobial activity against Gram-positive and Gram-negative bacteria.


G1549, a new cyclic hydroxamic acid antibiotic, isolated from culture broth of Pseudomonas alcaligenes. Antibiotic G1549, isolated from culture broth of Pseudomonas alcaligenes, is a new cyclic hydroxamic acid with a 1-hydroxy-2(1H)-pyridinone structure that complexes with metals. The structure of G1549 is suggested to be 1-hydroxy-5-methoxy-6-methyl-2(1H)-pyridinone. In vitro, G1549 and its copper and ferric complexes show moderate activity against Gram-positive bacteria, fungi and Trichomonas vaginalis. Topical application of G1549 and its copper and ferric complexes protect guinea pigs against cutaneous infection with Microsporum canis. The compounds, however, have some systemic toxicity in mice.


Gunacin, a new quinone antibiotic from Ustilago sp. In a screening program for antibiotics which were antagonized by cysteine, a strain, which was characterized as Ustilago sp., was found to produce a new quinone antibiotic, gunacin. The molecular weight M+ = 348.084 determined by mass spectroscopy, corresponds to a molecular formula of C17H16O8. Further spectroscopic data prove that gunacin is a new antibiotic. The antibiotic possesses a good inhibitory effect against mycoplasmas and Gram-positive bacteria including multi-resistant strains. It also possesses a weak activity against Gram-negative bacteria with the exception of Proteus vulgaris, which is more strongly inhibited. The main activity against fungi is found against Trichophyton mentagrophytes. Gunacin shows an inhibition of the DNA synthesis in vivo, is antagonized by mercapto compounds and possesses an acute toxicity of LD50 = 16 mg/kg i.p. and LD50 = 12 mg/kg i.v. in mice. Against HeLa-cell the antibiotic shows an ED50 = 12.11 microgram/ml. Thirty five microgram/ml of gunacin induces 1,063 interferon units.


Antibiotics from basidiomycetes. IX. Oudemansin, an antifungal antibiotic from Oudemansiella mucida (Schrader ex Fr.) Hoehnel (Agaricales). From mycelial cultures of Oudemansiella mucida a crystalline optically active antibiotic, oudemansin (2), has been isolated; its structure is closely related to strobilurin A (1). The relative configuration of oudemansin have been determined by X-ray analysis. The antibiotic exhibits strong antifungal properties and inhibits respiration in fungi, cells of the ascitic form of EHRLICH carcinoma, and rat liver mitochondria.


Studies on the biosynthesis of clavulanic acid. II. Chemical degradations of 14C-labelled clavulanic acid. Two chemical degradations of clavulanic acid are described which are useful for locating label in 14C-clavulanate. In the first, the beta-hydroxyethylidene side chain of p-bromobenzyl clavulanate is removed by ozonolysis to give p-bromobenzyl (2R, 5R)-3,7-dioxo-4-oxa-1-azabicyclo [3.2.0] heptane-2-carboxylate. The second involves the reaction of p-bromobenzyl clavulanate with dibenzylamine in methanol, to isolate the three beta-lactam carbons as methyl trans-3-(N-N-dibenzyl)amino acrylate. These techniques were used to degrade clavulanic acid derived from fermentations fed with 2-14C-acetate or universally 14C-labelled glycerol. The amount of label retained in the degradation products was in agreement with the distribution of 13C in clavulanic acid derived from 2-13C-acetate, or 1,3-13C2-glycerol, as observed by 13C-NMR.


Syntheses of sporaricin analogues, 2-deoxy-4-N-glycyl-6-O-(alpha-nebrosaminyl)fortamine and its 3-de-O-methyl compound. 2-Deoxy-4-N-glycyl-6-O-(alpha-nebrosaminyl)fortamine (21) and 3-de-O-methyl-2-deoxy-4-N-glycyl-6-O-(alpha-nebrosaminyl)fortamine (27) were prepared starting from lividamine. The syntheses include four key steps, that is, transformation of 2-deoxystreptamine moiety of lividamine to 4-N,3-O-didemethyl-2-deoxyfortamine, selective 4-N-methylation of the new aminocyclitol moiety, selective attachment of a glycyl residue to the methylamino group at C-4 and selective amination at C-6'.


Orally active esters of cephalosporin antibiotics. II. Synthesis and biological properties of the acetoxymethyl ester of cefamandole. The synthesis of the acetoxymethyl (AOM) ester of cefamandole (CM) is described. The sparingly soluble ester is shown to be well absorbed orally by mice, but only when administered in solution in a partially non-aqueous vehicle, 50% propylene glycol. Neither the ester in aqueous suspension nor the sodium salt of CM in solution is well absorbed orally. The rate of oral absorption of the ester from solution is very rapid as shown by the early peak time and shape of the plasma level curve. Oral bioavailability from solution is at least 60% and is apparently limited only by hydrolysis or precipitation of a variable portion of the ester dose in the intestinal lumen prior to absorption.


Immunological studies on sporamycin-treated animals. Sporamycin showed a remarkable tumor regressive activity against sarcoma-180 with a single 5 mg/kg dose of intravenous administration. This antitumor effect on tumor and host animals was examined immunologically. As the results: (1) When sarcoma-180 tumor cells were used as an antigen macrophage migration inhibition reaction by spleen cells derived from the tumor-bearing mice treated with sporamycin was positive at day 7 approximately 14 after the medication and was negative thereafter. (2) The delayed hypersensitivity tested by the foot-pad reaction was positive in tumor-bearing mice treated with sporamycin, and no decrease of foot pad reaction was observed, whereas this reaction decreased remarkably in non-treated tumor-bearing mice. (3) Sarcoma-180 tumor cells were mixed with spleen cells derived from sporamycin-treated mice, and were inoculated into normal dd mice. The growth of tumor cells was inhibited markedly, but no inhibition of tumor growth was observed in case of spleen cells derived from non-treated tumor bearing mice. (4) Combined treatment of sporamycin with PS-K, an immunopotentiator, showed a remarkable synergistic effect.


Studies on Actinomycetales producing antibiotics only on agar culture. I. Screening, taxonomy and morphology-productivity relationship of Streptomyces halstedii, strain SF-1993. Soil Actinomycetales that produce antibiotics during growth on agar but not in submerged culture were searched for and about 25 strains were obtained. One of these, Streptomyces halstedii, strain SF-1993, which produces an antibiotic designated SF-1993 was studied taxonomically and morphologically. The antibiotic-productivity of strain SF-1993 was correlated with mycelial morphology. The vegetative mycelium was filamentous in antibiotic-producing agar cultures, but fragmented in non-producing submerged cultures. By maintaining submerged cells filamentous, production of antibiotic in the submerged fermentations was accomplished. Filamentation was maintained by the use of diluted media or non-fragmented mutant substrains.


Studies on Actinomycetales producing antibiotics only on agar culture. II. Isolation, structure and biological properties of N-carbamoyl-D-glucosamine (substance SF-1993). N-Carbamoyl-D-glucosamine was isolated from the culture of Streptomyces halstedii SF-1993. It showed weak antimicrobial activity against some Gram-negative bacteria and some fungi.


Structure of nanaomycin E, a new nanaomycin. A new component, nanaomycin E, has been isolated from the culture broth of Streptomyces rosa var, notoensis, which had been found to produce nanaomycins A, B, C and D. Nanaomycin E was an epoxy derivative of nanaomycin A and was converted into nanaomycin A and 4a-epi-nanaomycin B by treatment with sodium hydrosulfite in an acidic aqueous solution. 4a-epi-Nanaomycin B was quantiatively converted into nanaomycin A under alkaline conditions.


Enzymatic conversion of aclacinomycin A to Y by a specific oxidoreductase in Streptomyces. A specific oxidoreductase converting aclacinomycin A to a new analog, aclacinomycin Y, was purified to apparent homogeneity from the culture filtrate of aclacinomycin-producing microorganisms. The isolated enzyme was a weakly acidic protein (isoelectric point, 5.9) with a molecular weight of about 72,000. The enzymatic reaction requires molecular oxygen and has a pH optimum at 5.5. The enzyme catalyzed an oxidation of the terminal sugar, L-cinerulose, of the trisaccharide moiety of aclacinomycin A to L-aculose (2,3,6-trideoxyhex-2-enopyranos-4-ulose) with removal of two electrons. Studies of substrate specificity revealed that the enzyme is an oxidoreductase capable of modifying anthracyclic triglycosides by oxidizing their terminal sugars.


Pharmacokinetic study of a new semisynthetic cephalosporin (AL-226) in rabbits. The pharmacokinetics of AL-226, a new semi-synthetic cephalosporin, were studied after i.v. and i.m. administrations to rabbits at doses of 10, 20, 30 and 40 mg/kg. The average values of the pharmacokinetic parameters after bolus i.v. administration of the antibiotic expressed by an open two-compartment kinetic model were: alpha = 0.131 min.-1, beta = 0.0341 min-1, K12 = 0.0287 min.-1, K21 = 0.0552 min.-1, K13 = 0.081 min.-1 a linear relationship between C0 and dose was found. The urinary excretion constants after i.v. administration were: 0.0266 min.-1, 0.0294 min-1, 0.0289 min.-1, 0.0306 min.-1, for doses of 10, 20, 30 and 40 mg/kg, respectively. The pharmacokinetic parameters after i.v. administration were used to determine the absorption constant: 0.0525 min.-1, 0.057 min.-1, 0.0596 min.-1, 0.0511 min.-1, after i.m. administration for doses of 10, 20, 30 and 40 mg/kg, respectively.


Metabolism of tritium- and carbon-14-labeled tiamulin in dogs, rats, and pigs. The metabolism of tiamulin hydrogen fumarate, labeled with 3H, 14C, or both, was studied in dogs, rats, and weanling pigs. After a dose of radiolabeled tiamulin, all three species excreted more radioactivity in feces (via bile) than in urine. Dogs absorbed 86% of a single oral dose of tiamulin-3H, and the disposition of the compound was similar after a single or multiple dosage regimen. The ratio of antimicrobial activity to total radioactivity in dog plasma was only about 0.25, and was still less in dog urine. After dosing with tiamulin-14C, rats and pigs excreted at least 1% of the dose as 14CO2 in expired air. In dual-labeled studies, pigs excreted less total 14C than 3H and had greater residues of 14C than 3H in edible tissues, blood, and plasma. After the administration of tiamulin-14C to pigs, radioactivity was incorporated into liver glycogen, indicating metabolic cleavage of the side chain of tiamulin. Tiamulin-3H is the isotopically-labeled compound of choice for studying metabolism and tissue residues in animals.


Interaction of aminoglycoside antibiotics with phospholipid liposomes studies by microelectrophoresis. Microelectrophoresis was found to be a rapid method for studying the interaction between aminoglycoside antibiotics and liposomes prepared from acid phospholipids. The ability to cause charge reversal of liposomes prepared from phosphatidyl inositol was ranked in the order neomycin greater than gentamicin greater than tobramycin greater than amikacin greater than kanamycin greater than ribostamycin greater than streptomycin greater than dihydrostreptomycin greater than Ca2+. Similar results were obtained with liposomes prepared from a mixture (8: 2) of phosphatidyl choline and phosphatidic acid, but no effect was detectable with neutral liposomes made from phosphatidyl choline only. The results support the view that the attraction between positively charged nitrogen groups on the antibiotics and the negatively charged groups of acidic phospholipids are predominantly responsible for the interaction. Extension of the studies to ionic strengths and calcium concentrations similar to those found in vivo showed a reduction, but not elimination, of the observed effects.


Method for obtaining ovine uterine secretions from unilaterally pregnant ewes. Development of the ovine conceptus was confined to the uterine horn ipsilateral to the corpus luteum (CL) by placing a ligature around that uterine horn at a point near the uterine body on day 5 of pregnancy. On day 140 of gestation, seven of 10 ewes were still pregnant and from 21 to 815 ml of uterine fluid (488 +/- 94 ml, X +/- SEM) were collected from the nongravid uterine horn. Total recoverable protein (X +/- SEM) was 13.4 +/- 3.4 grams. Polyacrylamide gel electrophoresis of the reduced proteins in presence of sodium dodecyl sulfate indicated that protein composition of uterine fluid was distinct from that of colostrum, serum, amniotic fluid, and allantoic fluid, and revealed the presence of two major polypeptides with molecular weights of about 57,000 and 58,500, respectively, plus numerous other minor components. Gel filtration on columns of Sephadex G-200 and Sepharose CL-6B suggested that these polypeptides formed a series of aggregates of high molecular weight when kept under nonreducing conditions. Glucose (.18 +/- .03 mg/ml), but not fructose, was present in uterine fluid. In addition, high levels of prostaglandin F (451.4 +/- 83.3 ng/ml) were present.


High pressure liquid chromatographic determination of sodium saccharin, sodium benzoate, and caffeine in soda beverages: collaborative study. A collaborative study for the determination of sodium saccharin, sodium benzoate, and caffeine in 3 types of soda beverage, cola, grape, and lemon-lime, has been completed using reverse phase high pressure liquid chromatography with a muC18 column and acetic acid mobile phase. Recoveries for sodium saccharin were 98.6, 98.0, and 99.1%; for sodium benzoate, 100.6, 102.6, and 100.6%; and for caffeine, 100.8, 101.4, and 101.1%, respectively. The method has been adopted as official first action.


Gas-liquid chromatographic-mass spectrometric determination of alpha- and beta-naphthylamines in FD&C Red No. 2 (amaranth). A method is described for the simultaneous quantitation of trace amounts of alpha- (alpha-NA) and beta-naphthylamines (beta-NA) with detectability in the 0.1 ppb range and sensitivity of 50 picomoles in certified food grade amaranth (FD&C Red No. 2; C.I. Food Red 9; CI 16185). The amaranth sample is extracted with benzene, and the evoporated residue is derivatized with perfluorooctanoic anhydride. The resulting derivatives are separated by gas-liquid chromatography and identified and quantitated by mass spectrometric monitoring of the m/e at 539.04. The method was used for quantitation of alpha-NA and beta-NA in randomly chosen samples of amaranth. Of 11 samples from different manufacturers, 5 were free of the beta-isomer; the remaining samples contained up to 1.2 ppb beta-NA. The concentration of alpha-NA ranged from no detectable amount to 970 ppb; the majority of the samples contained less than 7 ppb.


Analysis of fat-soluble vitamins. XXII. High performance liquid chromatographic determination of vitamin D in concentrates. Collaborative study. A collaborative study was carried out which compared the official chemical method (43.B14-43.B24), the HPLC method according to Hofsass et al. including maleic anhydride treatment, and the HLPC procedure according to De Vries et al. for vitamin D concentrates. A total of 396 samples were distributed to 33 collaborators for analysis. Five laboratories performed both the chemical and the HPLC methods. Five laboratories performed the Hofsass method and 16 laboratories performed the De Vries method. The results for the chemical method agreed with the theoretical values for the samples, and the standard deviation was comparable to that obtained in previous AOAC collaborative studies. Collaborative results for the Hofsass method were low. In addition, incorrect use of a fixed conversion factor (1/0.586) and necessity of a double chromatogrpahic system on a non-treated and a treated vitamin D sample reduce the effectiveness of the method. There were no adverse reactions to the DE Vries HPLC method. It is recommended that the method be adopted official first action as an alternative procedure for determining vitamin D in concentrates, excluding powders containing irradiated 7-dehydrocholesterol.


Semiautomated method for riboflavin in food products: collaborative study. A collaborative study was conducted comparing a semiautomated riboflavin method with a manual riboflavin method for 10 food products. Six laboratories provided results from the semiautomated method and 16 laboratories used the manual technique. The semiautomated method was more repeatable within a laboratory than was the manual method. The semiautomated method results compared favorably with the manual method for all 10 products.


Analysis of foods for iodine: interlaboratory study. An accurate, reproducible method for less than or equal to 1 ppm iodine in foods is required for nutritional labeling. In order to ascertain the current status of iodine analysis in foods, 7 samples, representing different food classes, were analyzed by 8 laboratories. Six laboratories used their modifications of the Ce-As-I catalytic method preceded by alkaline dry ashing. Two laboratories used neutron activation analysis (NAA), with differing radiochemical separations. The study showed wide discrepancy in analytical results. Mean relative standard deviation for all laboratories was 77.9% between laboratories; 19.1% within-laboratories. Laboratories using NAA had only slightly better precision than did laboratories using the chemical method. The lowest level reported on the entire group of samples ranged among laboratories from 0.0089 to 0.65 ppm. Figures reported by a laboratory are, in general, consistently high or consistently low. The only differences in methodology which may possibly correlate with level of iodine obtained are the use of NAA technique and use of manual, rather than automated, colorimetry.


Atomic absorption determination of tin in foods: collaborative study. Samples of green beans, applesauce, and a fruit juice were fortified with tin at 3 levels. Collaborators were asked to digest the samples, using HNO3-H2SO4, add methanol to enhance the absorption signal, and aspirate directly, using a nitrous oxide-acetylene flame. Results were received from 8 laboratories including 4 from Europe. However, only 6 laboratories used the prescribed methodology. All results were considered acceptable. The method has been adopted as interim official first action.


Lead, fluoride, and other elements in bonemeal supplements. The Pb, Cd, F, Al, Cr, Cu, Fe, Mn, Mo, Ni, Ti, and Zn content of 20 commercial bonemeal supplements was determined. Samples were mineralized with nitric and perchloric acids prior to determination of all elements except F, for which a diffusion method was used. Pb and Cd were determined by differential pulse anodic stripping voltammetry, F was measured using an ion selective electrode, and all other elements were determined by inductively coupled argon plasma spectroscopy. The mean recoveries of Pb and F were 97 and 99%, respectively. The concentration range of Pb was 1.5-8.7 microgram/g. Cd was quantitated in only one sample at a level of 2.5 microgram/g; all other samples were estimated to contain less than 0.05 microgram Cd/g. The concentration of F ranged from 261 to 921 microgram/g.


Atomic absorption spectroscopic determination of molybdenum in plant tissue and blood plasma. A method is presented for determining molybdenum in plant tissue and blood plasma by flame and flameless atomic absorption spectroscopy (AAS). The samples are digested with HNO3-HCIO4, and the metal is separated by chelation, extracted into methyl isobutyl ketone, and analyzed by AAS. Recoveries of molybdenum from fortified samples ranged from 92.0 to 98.9%. The limits of detection on the flame and graphite furnace were 10 and 2 ng/mL, respectively.


Microdiffusion and fluoride-specific electrode determination of fluoride in foods. A simple and accurate method was developed for routine determination of fluoride in foods. Hydrogen fluoride is diffused 20 hr at 50 degrees C from fresh or freeze-dried samples (0.1 g dry wt) in polystyrene petri dishes containing 2 mL 40% HCIO4 and 0.3 g Ag2SO4, and is absorbed on the lids, previously spotted with 0.1 mL 0.5M NaOH. The absorbent layer is dissolved in 2 mL buffer solution, and the fluoride is measured potentiometrically. The method was verified by analysis of NBS Standard Reference Materials; recovery from 28 spiked infant foods (average = 99%, range = 75-135%); and comparison of results with colorimetry results for the same diffusates, after modification to handle 1 g samples. Relative standard deviations varied from 4 to 20% day to day. Detection limits were below 0.05 microgram/g dry weight.


Minicolumn detection methods for aflatoxin in yellow corn: collaborative study. The CPC modified method, the Holaday modified method, and the combination of the 2 procedures have been compared. The CPC modified method involves more cleanup steps but has a more sensitive column. The Holaday modified procedure has fewer cleanup steps, but the column is more difficult to interpret. The combination CPC-Holaday, which has proven to be the most satisfactory, combines the speed and simplicity of the Holaday extraction and the sensitivity of the Velasco minicolumn used in the CPC method. Levels of 10 ng/g were detected by 89% of the collaborating laboratories using the combination, Holaday-Velasco, method. The combination method has been adopted as official first action.


Production of aflatoxin in cocoa beans.

Production of aflatoxin in cocoa beans. Aflatoxin was produced in both non-autoclaved and autoclaved Ivory Coast cocoa beans inoculated with Aspergillus parasiticus NRRL 2999 under optimum laboratory growth conditions. Total aflatoxin levels ranged from 213 to 5597 ng/g substrate. Aflatoxin was quantitated by using high pressure liquid chromatography (HPLC). Raw, non-autoclaved cocoa beans, also inoculated with aspergilli, produced 6359 ng aflatoxin/g substrate. Variation in aflatoxin production between bean varieties was observed. Total aflatoxin levels of 10,446 and 23,076 ng/g substrate were obtained on Ivory Coast beans inoculated with A. parasiticus NRRL 2999 and NRRL 3240, respectively. Aflatoxin production on Trinidad and Malaysian beans was 28 and 65 ng aflatoxin/g substrate. These data support previously reported low level natural aflatoxin contamination in cocoa.


Extraction, cleanup, and quantitative determination of aflatoxins B1 ANd M1 in beef liver. A method for the determination of aflatoxin B1 in eggs was applicable for aflatoxin B1 in liver, but ineffective for aflatoxin M1 in liver because of poor recovery of added aflatoxin and interferences in thin layer chromatography. The method was modified by the addition of citric acid to the extracting solvent and ammonium sulfate to the extract solution for removing protein. The elution system for silica gel column cleanup was also changed by substituting methanol for acetone, and adding a step for confirmation of aflatoxin M1 identity. The method has been used successfully for survey and research on aflatoxin residues in animal tissues.


High pressure liquid chromatographic determination of chloramine-T in food. A high pressure liquid chromatographic (HPLC) method for determining chloramine-T (N-chloro-N-sodium-p-toluenesulfonadmie) in foods such as ice cream, minced meat, and shrimp is described. Deproteinized samples are treated with sulfite to convert chloramine-T to p-toluenesulfonamide (p-TSA) and extracted, and HPLC analysis is performed on concentrated extracts. Sample extracts are chromatographed on a reverse phase 10 micrometers Lichrosorb RP-18 column with acetonitrile-water (190), and quantitated by an ultraviolet detector (220 nm) and digital integrator. The relationship between recorded peak area and concentration was linear to 0.600 micrograms p-TSA/microL. The detection limit was 2.5 ng pTSA/microL, corresponding to a chloramine-T concentration of 0.8 mg/kg in samples. Recoveries of added chloramine-T were 88% for ice cream, 73% for minced meat, and 51% for shrimp. Precision data indicate a relative standard deviation of 1.54 and 2.14% for the complete analysis of ice cream with levels of 15 and 63 mg chloramine-T/kg (n = 5 determinations), respectively. The HPLC method was applied to chloramine-T determinations in 62 ice cream, 25 minced meat, and 25 shrimp samples.


Microdetermination of naphthionic acid in serum and amniotic fluid. A simple, rapid, sensitive, and reproducible microdetermination of naphthionic acid (NA) in serum and amniotic fluid is described. The detection limit of the method is 1 ng NA in 20 microL serum, and 3 ng NA in 50 microL amniotic fluid. The concentration of nonderivatized NA was measured by fluorescence spectrophotometry (excitation 328 nm, emission 420 nm) of the supernate, after precipitation of proteins with absolute ethanol and heating for 30 min at 75 degrees C. Standard deviations of determinations for 2, 10, and 50 ng NA in 20 microL serum were 10.1, 7.59, and 7.64%, respectively. An analyst can perform about 100 determinations daily; results are available within 2 hr of sampling. A modification of the procedure to permit quantitation of NA in urine is also described.


Liquid chromatographic determination of methscopolamine bromide in various veterinary formulations. A reverse phase paired ion liquid chromatographic procedure is described for determining methscopolamine bromide (Pamine) in the presence of neomycin sulfate in 5 veterinary formulations (Biosol M). Methscopolamine bromide is separated from neomycin sulfate and other formulation excipients by extraction into ethanol. The sample preparations are then concentrated and dissolved in water before chromatography. Recovery of methscopolamine bromide added to 5 placebo formulations ranged from 99.7 to 100.7%, with relative standard deviations of less than 2%. Specificity of the method with regard to potential degradation products is demonstrated.


Colorimetric determination of hydralazine with 9-chloroacridine. A spectrophotometric assay for hydralazine hydrochloride based on the interaction of the drug with 9-chloroacridine has been developed. The interaction shows an absorption maximum at 460 nm and is affected by temperature, heating time, and quantity of acridine reagent used. Color development is maximum when the drug is heated in the presence of a 30-fold molar excess of the acridine in a 50 +/- 1 degree C water bath for 1 hr. The method detects hydralazine hydrochloride in the 10(-5)--10(-6)M range with sensitivity to 0.2 micrograms/mL and 3--4% accuracy. Typical calibration data obtained from linear regression analysis of absorbance at various drug concentrations show r = 0.9997 (n = 6). Hydralazine can be determined in dosage forms that also contain varying quantities of reserpine and hydrochlorothiazide. The 9-chloroacridine method is as sensitive as other spectrophotometric procedures for hydralazine but also more accurate and precise, and involves fewer manipulative steps.


Determination of edrophonium chloride injection USP in the presence of preservatives. The determination of Edrophonium Chloride Injection involves a modification of a procedure for phenylephrine and offers an alternative to the compendial assay. Edrophonium chloride forms an ion-pair with di-(2-ethylhexyl) phosphoric acid which can be extracted with an immiscible solvent such as ether. When this procedure is coupled with partition chromatography, the drug can be isolated from preservatives and quantitatively determined by its ultraviolet absorbance. Recovery data on synthetic mixtures ranged from 99.8 to 101.9% and were in good agreement with the compendial method.


Computerized gas-liquid chromatographic method for content uniformity analysis of dicyclomine hydrochloride capsules and tablets. An automated, computerized method is presented for the content uniformity determination of dicyclomine hydrochloride capsules and tablets, using up to 4 automatic sampler-equipped gas chromatographs interfaced with a minicomputer. A 3% OV-17 column is used with anthracene as an internal standard. Five sample injections are bracketed by standard mixtures containing about 90 and 110% of the labeled dicyclomine hydrochloride. Data are taken on-line simultaneously from each gas chromatograph and a computer-generated report is produced. Calculations use a BASIC program with linear fit of the 90 and 110% standard mixture. Individual tablet or capsule results are printed in milligrams and per cent declared, including summary calculations of average, high, low, standard deviation, and coefficient of variation. The GLC results are comparable (within 1%) to those obtained using the USP procedure.


Identification of analogs of the hallucinogen cyclohexamine. The hallucinogenic drug 1-phenylcyclohexylamine and its N-methyl-, N-ethyl-, N-propyl-,N-isopropyl-, and N-(beta-phenylethyl)-derivatives are identified by spectroscopic techniques. Reference mass and infrarred spectra and gas-liquid and thin layer chromatographic data are provided and discussed.


Spectrophotometric determination of piperazine and its salts in pharmaceuticals. A spectrophotometric determination of piperazine and some of its salts is described. The method depends on the UV measurement of the N-nitroso derivatives formed by the interaction of piperazine with nitrous acid. The chromophore is developed by heating the reaction mixture at 80 degrees C for 15 min, at pH 2.3--2.6. Beer's law is obeyed in the range 1--15 micrograms/mL.


Gas-liquid chromatographics determination of Bayer 73 in fish, aquatic invertebrates, mud, and water. A gas-liquid chromatographic (GLC) method is described for determining residues of Bayer 73 (2-aminoethanol salt of 2',5-dichloro-4'-nitrosalicylanilide) in fish muscle, aquatic invertebrates, mud, and water by analyzing for 2-chloro-4-nitroaniline (CNA), a hydrolysis product of Bayer 73. Bayer 73 residues are extracted from fish muscle tissue, invertebrates, and mud with acetone-formic acid (98+2), and partitioned from water samples with chloroform. After sample cleanup by solvent and acid-base partitioning, the concentrated extract is hydrolyzed with 2N NaOH and H2O2 for 10 min at 95 degrees C. The CNA is then partitioned into hexane-ethyl ether (7+3) and determined by electron capture GLC. Average recoveries were 88% for fish, 82% for invertebrates, 82% for mud, and 98% for water at 3 or more fortification levels.


Gas-liquid chromatographic determination of thiourea in citrus peels. A gas-liquid chromatographic (GLC) method was developed for the detection and determination of thiourea in citrus peels. After the peel is extracted with ethyl ether, the ether extract is adsorbed on sodium sulfate together with water. Thiourea is recoverd from both the sodium sulfate and the peel residue with ethyl acetate-acetone(2+1). The extracted mixture is cleaned on an alumina column, the eluate is concentrated under vacuum, and thiourea is extracted from the concentrate with sodium carbonate solution. GLC was carried out on the prepared benzoyl derivative of thiourea. The average recoveries of thiourea from lemon peel were 85.3, 93.1, and 97.6% at the fortification levels of 1, 10, and 100 ppm, respectively. The detection limit was low as 0.08 ppm.


Cleanup of biologigal samples for determining p,p'-DDT and its metabolites. A method is described for extracting tissue samples with hexane-acetone and hydrolyzing the fat and protein with aqueous sodium hydroxide. p,p'-DDT and its metabolites are measured on a gas chromatograph with an electron capture detector after cleanup on an alumina column. The new cleanup procedure gives 98% recoveries for p,p'-DDT and its metabolites added to fat in vitro, compared with 84% recovery by a modified commonly used method. Comparison by Student's t-test of the new method with the modified method on 32 cattle biopsy, 46 birds, and 20 fish samples for in vivo incurred p,pi-DDT residues showed that the results by the new method were significantly higher at the 95% confidence level. Alkali-hydrolyzed extract did not contaminate the detector. The new procedure avoids the use of dimethylformamide and shortens the time of analyses. A correlation (r=0.68) was observed between the fat-soluble protein concentration of the n-hexane extract and the p,p'-DDT residues.


Gas-liquid chromatographic determination of bromide ion in lettuce: interlaboratory studies. The gas-liquid chromatographic method for the determination of bromide ion, as originally described for grain by Heuser and Scudamore, was adapted for use on lettuce, and later on products of other vegetable or of animal origin. After optimization of the ashing and derivatization conditions, 2 interlaboratory studies were carried out in The Netherlands. In the first, a preparatory study, 6 laboratories investigated freeze-dried lettuce; in the second definitive study, 8 laboratories investigated both freeze-dried and fresh lettuce. Four lots of lettuce with bromide contents ranging from about 5 to about 200 mg/kg were investigated. Average bromide contents (mg/kg) and between-laboratory coefficients of variation were 5.6 (11.0%), 13.3 (18.4%), 29.9 (8.3%), and 209 (12.8%). The results obtained are normal for the type of work and the conditions of the study. The method will be recommended for regulatory use in The Netherlands and in international organizations.


Colorimetric determination of caffeic acid in plant materials. A new colorimetric method is described for determining caffeic acid content in plant materials. Caffeic acid is separated by thin layer chromatography from the alcoholic extract, and color is developed using 0.5% aqueous thiosemicarbazide solution under alkaline conditions. The absorbance is read at 475 nm. Lambert-Beer's law is obeyed in the concentration range 0.37-17.5 micrograms caffeic acid/mL. The method is reproducible and has been applied to the estimation of caffeic acid in carrot roots.


Stability of three forms of vitamin B6 to laboratory light conditions. Pyridoxine.HCl, pyridoxal.HCl, and pyridoxamine.2HCl solutions were exposed to several laboratory light treatments, and the resulting vitamin retentions were determined by the AOAC microbiological method. The 5 treatments compared were total darkness, regular laboratory light, low actinic glass protection, yellow incandescent light, and golden fluorescent light. All treatments were imposed for 8 and 15 hr, and with the vitamin solutions at both a low and a high pH. Regular laboratory light was the most destructive to the vitamins, with greater destruction at higher pH and longer exposure time in all cases. Pyridoxine retentions ranged from 97 (pH 4.5, 8 hr) to 66% (pH 7, 15 hr); pyridoxal from 97 (pH 4.5, 8 hr) to 55% (pH 6, 15 hr); and pyridoxamine from 81 (pH 4.5, 8 hr) to 47% (pH 8, 15 hr). Retentions in low actinic glassware or in clear glassware under yellow or golden fluorescent light were essentially com,lete, ranging from 94 to 106% over all treatments and all 3 forms. Results showed that either of the 2 subdued light conditions, yellow or golden fluorescent light, is suitable in vitamin B6 assays and that low actinic glassware is suitable for storing sample solutions.


Use of background correction to improve the accuracy of the selective reduction method for determining methyl mercury in fish. Background correction is used with a previously published flameless atomic absorption method for the concurrent determination of inorganic and methyl mercury. The use of background correction simplifies the procedure, but, more importantly, eliminates errors in mercury readings due to broad band absorption by other volatile compounds produced during the determination.


Sample preparation of some shelled treenuts and peanuts in a vertical cutter-mixer for mycotoxin analysis. A procedure has been devised for more rapid preparation of large samples of nutmeats (10-80 lb) for mycotoxin analysis without the use of grinding aids, by combining the grinding and mixing steps into one operation. Ease of cleaning equipment facilitates the preparation of several samples in a short time. The procedure was tested using shelled raw and roasted peanuts, and raw almonds, walnuts, and pecans. The reduction of particle size to pass a 20 mesh screen was attained using a 25 qt Hobart vertical cutter-mixer (VCM) equipped with standard serrated blades and a 40 qt VCM using either a standard serrated blade or a smooth-edge blade modified with sharp-edge notches to increase its cutting ability. Portions of ground composite were removed at various time intervals and particle sizes were measured. Original and check analyses of 12 naturally contaminated samples over a 3 year period indicate that this procedure is practical and reproducible.


Comparison of two extraction procedures for recovering nitrite from cured meat products. A study was undertaken to determine the extent of loss of nitrite during the analytical workup of cured meats for nitrite analysis. Two extraction techniques were compared: (A) a relatively long (2 hr) and hot (80 degrees C) procedure carried out at the natural pH of the sample, and (B) the method of Sen and Donaldson that specifies an alkaline solution and much milder conditions (20 min at 50 degrees C at pH 8) for extraction. The data obtained suggest that considerable losses of nitrite can occur with the former procedure, especially in acidic samples. The overall percentage recoveries of added nitrite by the 2 methods ranged between 20 and 112, and 80 and 114, respectively. Average percentage recoveries for the acidic (pH less than 5) samples only were 47.2 for Method A as compared with 100.2 for Method B.


Long-term storage of microorganisms used in antimicrobial effectiveness tests. Liquid nitrogen storage of organisms designated in the U.S. Pharmacopeia (XIX) for the assessment of antimicrobial effectiveness is described. By use of simple apparatus and procedures, 85-95% viabilities of pre-frozen cells are attained immediately upon freezing. Viability levels have remained constant over a 3-year period and the use of such inocula has considerably improved the reliability of the test. The superiority of liquid nitrogen storage for microbial test inocular over other types of storage is discussed.


Identification of nosiheptide in feeds and detection of residues in animal tissues. Nosiheptic is determined in fermentation broths of Streptomyces actuosus either by a microbiological method using Staphylococcus aureus or, more easily, by an automated colorimetric method. The results obtained with both methods correspond well for concentrations greater than 100 microgram/mL with a standard deviation of 1-3%. For determination of nosiheptide as a feed additive, the microbiological assay is made more specific by pretreatment with petroleum ether and 1N HCl. Standard deviation is less than 4%, and the assay is sensitive to 1 ppm. Nosiheptide is identified in feed containing other frequently used antibiotics by thin layer chromatography with bioautography; sensitivity is 1 ppm. The absence of traces of nosiheptide in tissues of treated swine and broiler is confirmed by microbiological diffusion, sensitive to 0.025 ppm.


Modified method for carbadox in feeds: collaborative study. The official first action method for carbadox in swine feed, 42.C01-42.C04, was modified in 2 respects. First, the samples were leached overnight at room temperature instead of boiled for 1 hr. This change avoided problems with overheating and excessive evaporation. Second, the dilution scheme for samples spiked with carbadox standard solution was changed to give absorbance values that were within the optimum working range of all types of spectrophotometers. The modified procedure was collaboratively studied by 21 laboratories. The repeatability standard deviation (sigma0) and reproducibility standard deviation (sigmax) were sigma0 = 0.00029% and sigmax = 0.00056% (8.9% of grand mean) for feeds containing 0.00617% carbadox; and sigma = 0.0012% and sigmax = 0.0019% (9.3% of grand mean) for feeds containing 0.0198% carbadox. The between-laboratory variance ratio was significant for feeds containing 0.0198% carbadox. The mean per cent of intent values for feeds containing 0.00617% carbadox and 0.0198% carbadox were 102% and 104%, respectively. In general, the statistical results were comparable to those previously obtained for the official first action method. Consequently, the modified procedure is not recommended as a replacement for the official first action method.


Bacillus stearothermophilus disk assay for detection of residual penicillins in milk: collaborative study. A collaborative study was performed on a Bacillus stearothermophilus paper disk method designed to detect residual levels of 4 antibiotic drugs in whole market milk. This method is a modification of an earlier procedure developed for the International Dairy Federation. Whole milk samples spiked at low levels with ampicillin, cephapirin, cloxacillin, and penicillin G were sent frozen to 11 collaborating laboratories with instructions to assay them promptly according to the method provided. Five of the laboratories reported inconclusive results due to technical difficulties encountered with the method. The 6 remaining laboratories all detected levels of 0.005-0.008 microgram or unit/mL for penicillin G, ampicillin, and cephapirin and 0.05-0.08 microgram/mL for cloxacillin. The most commonly used official methods, the Sarcina lutea (Micrococcus luteus) cylinder plate method and the Bacillus subtilis paper disk method, can detect levels of 0.01 and 0.05 unit penicillin G/mL, respectively. The B. stearothermophilus method is rapid, simple to perform, and more sensitive than present official methods. The method has been adopted as official first action for the detection of penicillins in milk.


Isolation and characterization of chromatin subfractions from rat liver. Rat-liver chromatin was digested with micrococcal nuclease at low ionic strength in the presence of a low concentration of CaCl2. The nuclease digest was successfully separated into three fractions, P1, P2, and P3, by gel filtration on a column of Sepharose 2B. P1 fraction was shown to be a mixture of long fragments of partially digested chromatin by the sedimentation profile or by electrophoresis of DNA. P2 fraction contained four histones H2A, H2B, H3, and H4 in almost equal amounts, together with nonhistone protein of low molecular weight. The DNA was composed of three or four fragments less than 300 base pairs long. From the Kav value of the P2 fraction, the average size was estimated to be about 240 base pairs. On analytical ultracentrifugation, this fraction exhibited a monophasic boundary and a sedimentation value of 13.7S. P3 fraction contained nonhistone proteins which showed a molecular weight larger than that of H1 histone. The size of DNA was estimated to be less than 50 base pairs from the Kav value. Based on these results, the P2 fraction was concluded to consist of nucleosome monomer enriched in nonhistone proteins. The P3 fraction is presumably the nuclease-sensitive or internucleosome portion, which contains small amounts of nonhistone proteins.


Identification of suberimidate cross-linking sites of four histone sequences in H1-depleted chromatin. Histone arrangement in nucleosome core. The arrangement of 8 histones in the nucleosome core has been investigated by identifying the sites of 4 histone sequences cross-linked with a bifunctional amino-group reagent, dimethyl suberimidate, selected from among 4 diimidoesters of various linker lengths examined. H1-depleted calf thymus chromatin was allowed to react with 14C-labeled suberimidate at pH 8.5 and 0 degrees C. The cross-linked chromatin was then digested exhaustively with trypsin. Almost all the histone fragments were released from the chromatin with 0.25 M HCl and chromatographed on several columns and on paper. Cross-linked peptides were detected by analyzing the content of radioactive suberimidoylbislysine after acid hydrolysis. The chromatographic procedure developed here showed that the whole histone fragments contained 29 mol% of the total linked reagent as suberimidoylbisylsine. The 5 finally purified cross-linked peptides were identified from the total and N-terminal amino acids of each pair of peptides separated by two-dimensional cellulose thin layer chromatography after cutting the linker by ammonolysis. Thus, intramolecular cross-linking was found between Lys-5 and Lys-9 of H2A, and Lys-34 and Lys-85 of H2B, while intermolecular cross-linking was found between Lys-24 (or 27) of H2B and Lys-74 of H2A, Lys-85 of H2B and Lys-91 of H4, and Lys-120 of H2B and Lys-115 of H3 and/or Lys-77 of H4. Most of these lysine residues are located in the DNA-binding segments of the 4 histone sequences identified previously [Kato, Y. & Iwai, K, (1977) J. Biochem. 81, 621--630]. All the 5 or 6 cross-links can be located in a heterotypic tetramer consisting of one molecule each of H2A, H2B, H3, and H4, and a model of the histone arrangement in the tetramer is proposed. Two such tetramers may compose to the histone octamer in the nucleosome core.


Isolation and identification of the reaction product of alpha-hydroxy-gamma-carboxymuconic epsilon-semialdehyde dehydrogenase. The reaction product of enzymic dehydrogenation of alpha-hydroxy-gamma-carboxymuconic epsilon-semialdehyde (HCMS) was isolated. The analytical data (elemental analyses, IR spectra, mass spectra, proton NMR spectra, and UV spectra) showed that the product was not alpha-hydroxy-gamma-carboxymuconic acid (HCMA), the expected primary product of HCMS dehydrogenation, but a lactone of HCMA. The structure of the lactone was tentatively determined as alpha-pyrone-4,6-dicarboxylic acid. HCMS was converted stoichiometrically to the lactone by the purified NAD(P)-linked HCMS dehydrogenase. The lactone was actively metabolized by a cell-free extract prepared from Pseudomonas ochraceae grown on phthalic acid, suggesting that it may be a metabolic intermediate in the bacterial metabolism of protocatechuic acid. Furthermore, a method for the chemical synthesis of the lactose of HCMA is presented and some of its chemical and biochemical properties are described.


Occurrence of GTP cyclohydrolase I in Bacillus stearothermophilus. A GTP cyclohydrolase which catalyzes the removal of carbon 8 of GTP as formic acid to yield a single pteridine compound occurs in an obligate thermophile Bacillus stearothermophilus ATCC 8005. The enzyme was purified 5.5-fold. Its molecular weight and Stoke's radius were estimated as 105,000 and 45.3 A, respectively. The Km for GTP was 0.98 microM. The temperature and pH optima for activity were 60-65 degrees C and 8.0-8.4, respectively. No divalent cation was required for the reaction. The pteridine product was 3'-triphosphate of 2-amino-4-hydroxy-6-(D-erythro-1',2',3'-trihydroxypropyl)-7,8-dihydropteridine (dihydroneopterin triphosphate), identified by isolating its immediate derivative, 2',3'-cyclic phosphate of 2-amino-4-hydroxy-6-(D-erythro-1',2',3'-trihydroxypropyl)pteridine (neopterin cyclic phosphate). The radioactive product from [8-14C]GTP agreed with 14C-formate. Molar ratio of formate release to pteridine formation was 1.0.


The effects of Na+ and Mg2+ on the thermal denaturation of native and acetylated spinach ferredoxins. The effects of metal ions on the thermal denaturation and Mg2+ binding of native spinach ferredoxin and its acetylated derivative were investigated. The denaturation of ferredoxin in a metal-free solution at 40 degrees C was quickly prevented by the addition of Mg2+ or Na+ at appropriate concentrations. The metal concentrations required for 50% protection from thermal denaturation were 1.54 x 10(-4) M Mg2+ or 8.0 x 10(-3) M Na+ for native ferredoxin and 1.05 x 10(-3) M Mg2+ or 6.0 x 10(-2) M Na+ for acetylated ferredoxin. It was also found that native ferredoxin in the presence of over 20 mM Mg2+ was almost completely protected from thermal denaturation at 40 degrees C. The D-form which has been observed in acetylated ferredoxin by Masaki et al. (1977) (J. Biochem. 81, 1-9) was confirmed to be present in native ferredoxin at high temperature (49 degrees C) and is suggested to be an important form in the denaturation processes of the ferredoxin molecule.


Isolation and properties of spiramycin I 3-hydroxyl acylase from Streptomyces and ambofaciens. An enzyme preparation able to acylate the hydroxyl group at C-3 of the lactone ring of spiramycin was obtained from the spiramycin-producing strain, Streptomyces ambofaciens ISP-5053. The enzyme was purified about 33-fold from the crude extract by means of ammonium sulfate fractionation, diethylaminoethyl (DEAE) cellulose batchwise elution and DEAE cellulose column chromatography. The optimum pH for the enzyme activity was 8.5. The enzyme was activated by Ca2++, Mg2+, and Mn2+ in this order, but was inhibited by various SH reagents. Spiramycin I was the best substrate for the enzyme. The enzyme showed no preference between acetyl-CoA and propionyl-CoA.


Structure of Streptomyces erythraeus lysozyme at 6 A resolution. A 6 A resolution electron density map has been calculated for a bacterial lysozyme produced by Streptomyces erythraeus. This lysozyme differs from the vertebrate lysozyme in its size, amino acid composition, and specificity. The structure was determined by the method of isomorphous replacement. Three heavy atom derivatives were obtained by soaking crystals of the lysozyme in HgCl2, K2PtCl4, and UO2(NO3)26H2O. The resulting electron density map clearly shows the molecular boundary. The molecule is ellipsoidal in shape with average dimensions 50 A X 35 A X 35 A. High resolution analysis and sequence analysis of the molecule are in progress.


Proteinase inhibitors from a mimosoideae legume, Albizzia julibrissin. Homologues of soybean trypsin inhibitor (Kunitz). Four proteinase inhibitors, A-II, A-III, B-I, and B-II, were isolated from seeds of Albizzia julibrissin (silk tree) of the subfamily Mimosoideae, which is often regarded as the most primitive group of the Leguminosae plants. They were all of the high-molecular weight type (21,600 for A-II and A-III, and 19,000 for B-I and B-II), and composed of two polypeptide chains, linked together by a disulfide bond. A-II (A-III) inhibited bovine trypsin and alpha-chymotrypsin probably at an identical site. B-I (BII) inactivated bovine alpha-chymotrypsin and porcine elastase. Sequence analyses of A-II and B-II revealed a considerable homology with soybean trypsin inhibitor (Kunitz) but suggested the presence of an about 20-amino acid insertion in the A-II molecule.


Interaction of studies of a 100,000 dalton chymotryptic fragment of rabbit skeletal M-line protein with the S2 subfragment of myosin. Controlled chymotryptic digestion of the 165,000 dalton component of the M-line of rabbit skeletal muscle, followed by Biogel P-150 chromatography of the digest, has led to the isolation of a homogeneous 100,000 dalton species. This fragment was found, by both sedimentation equilibrium and gel filtration chromatography, to interact with heavy meromyosin subfragment 2 (HMM-S2). The persistence of this fragment, after chymotryptic digestion, to bind HMM-S2, along with the known insensitivity of the M-line to proteolysis, suggests a structural role for the parent 165,000 dalton component along the lines of the M-filament, as suggested by the Knappeis and Carlsen model for M-line structure.


The linkage between the four-step binding of oxygen and the binding of heterotropic anionic ligands in hemoglobin. The linkage between the four-step binding of oxygen and the binding of heterotropic anionic ligands in hemoglobin was investigated by accurately measuring and analyzing the oxygen equilibrium curves of human adult hemoglobin in the presence and absence of various concentrations of one or two of the following materials: chloride (Cl-), 2,3-diphosphoglycerate (DPG), and inositol hexaphosphate (IHP). Each equilibrium curve was analyzed according to the Adair equation to evaluate the four-step oxygen equilibrium constants (Adair constants) and the median oxygen pressure. The binding constants of the anions for the molecular species of hemoglobin carrying j oxygen molecules, Hb(O2)j(j=0,1,...,4), were evaluated from the dependences of the Adair constants and the median oxygen pressure on the anion concentration by introducing a model which takes the competitive binding of Cl- and DPG or IHP into account. Assumptions made in the model are: (a) the hemoglobin molecule has two oxygen-linked binding sites for Cl- which are equivalent and independent and (b) no Cl- can be bound to hemoglobin to which DPG or IHP is already bound and vice versa. Thus, we could obtain values for the intrinsic binding constants of Cl- and DPG, i.e., the constants in the absence of other competitive anions. For IHP, only the binding constants and apparent binding constants for Hb and Hb(O2)2 were obtained. Values of the Cl- binding constants and apparent binding constants for DPG and IHP, i.e., the binding constants in the presence of Cl- for Hb and Hb(O2)4, were in reasonable agreement with literature values. From the binding constants we calculated anion binding curves for Hb(O2)j(J=0,1,...,4), the number of anions bound to Hb(O2)J, And the relationship between fractional anion saturation of hemoglobin and fractional oxygen saturation. The numbers of released anions are not uniform with respect to oxygenation step. This non-uniformity is the reason for the changes in the shape of the oxygen equilibrium curve with anion concentration changes and for the non-uniform dependences of the Adair constants on anion concentration, and also results in non-linear relations between anion saturation and oxygen saturation. The anion binding constants and various binding properties of the anions derived from those constants are consistent with those observed by other investigators using different techniques, indicating that the present model describes the oxygen-linked competitive anion binding well.


A fluorescence stopped-flow study on troponin labeled with N-ethyl maleimide and N-(p-(2-benzimidazolyl)phenyl) maleimide. The kinetics of the conformational change of the troponin-C (TN-C) subunit in N-(p-(2-benzimidazolyl)phenyl) maleimide (BIPM)-N-ethyl maleimide (NEM)-labeled troponin induced by calcium binding or removal were studied with the fluorescence stopped-flow method. The kinetic process of the conformational change was biphasic, the rate constants of the two phases were determined as a function of the free calcium ion concentration of the protein solution. The kinetic behaviour of the conformational change of TN-C in BIPM-NEM-labeled troponin was explained by a simple molecular kinetic mechanism: (Formula: see text) This molecular kinetic mechanism is different from that of the isolated TN-C which we found in the previous work (1). That is, formation of a complex of TN-C with troponin-I (TN-I) and troponin-T (TN-T) modifies the molecular kinetic mechanism of the conformational change of TN-C.


Ultrastructural morphology of three-dimensional colonies of cells derived from a hepatocellular carcinoma. Cultured hepatocellular carcinoma cells were studied during anchorage-independent growth in semi solid medium (Methocel). The regular occurrence of mitotic figures both at the surface and within the colonies precludes the possibility of such colonies being formed by re-aggregation. The estimated population doubling time in the three-dimensional (3-D) colonies is consistent with those two-dimensional of (2-D) colonies. Structures resembling bile canaliculi were observed between the closely opposed membranes from the well packed adjacent cells. Cell surface and ultrastructural features of the colonies and individual cells are presented and comparisons made with 2-D growth of normal and malignant liver cells in vitro. The formation of 3-D colonies may not only be an assay for transformed cells but also for predicting the type of tumors produced by re-innoculation of the in vitro transformed cells.


Comparison of a water-soluble and a water-insoluble podophyllotoxin derivative in murine neoplasms. Comparison of a water-soluble (27-487) with a water-insoluble (VP-16-213) podophyllotoxin derivative in seven murine neoplasms revealed slight therapeutic superiortiy of VP-16-213 but also slightly greater toxicity when compared with 27-487. A strikingly good response of the Taper liver tumor to both compounds was observed, providing further incentive to the use of podophyllotoxin derivatives in human hepatomas.