Kitten ganglion cells: dendritic field size at 3 weeks of age and correlation with receptive field size. A Golgi study of beta (brisk-X type) ganglion cells has been done to compare ganglion cells in the retina of 3-week-old and adult cats. An anatomical basis for the large receptive field centers found in the immature kitten retina was sought. Kitten beta-type ganglion cells have significantly smaller dendritic spreads than adult beta cells; the dendrites of the kitten cells must still grow to reach their final adult size. Therefore a synaptic basis for the large receptive field size of the immature cells is suggested.


Paramyxovirdae.

Paramyxovirdae. Iit is proposed that the family paramyxovirdae shall comprise three genera: paramyxovirus, Morbillivirus, and Pneumovirus.


Hepatitis B vaccine: efficacy in high-risk settings, a two-year study. A formalin-treated hepatitis B vaccine in the form of purified hepatitis B surface antigen (HBsAg) was prepared from asymptomatic human HBsAg carriers. Its safety and potency were tested in 5 chimpanzees. The vaccine was administered to 264-individuals. The results of the first 173 immunizations--46 hemodialysis patients and 127 staff members--are presented. Potency was ascertained and efficacy assessed by the development of humoral immune responses to HBsAg (anti-HBs antibody) and seroepidemiologic studies of vaccinated and nonvaccinated subjects. The results, 2 years after immunization, suggest that the vaccine was protective against hepatitis B infection in high-risk hemodialysis settings. Preliminary studies with an inactivated hepatitis B vaccine similarly prepared, but with aluminum hydroxide as adjuvant, indicate that a such a preparation induces a more rapid and stronger anti-HBs response.


Effect of supraoptimal temperatures upon tobacco mosaic virus RNA replicase. An examination of the effect of supraoptimal temperatures upon tobacco mosaic virus (TMV) RNA replicase demonstrated that the enzyme activity rapidly decreased at 40 degrees whether incubated in vitro after extraction or in leaves at that temperature prior to extraction and assay. The loss of replicase activity paralled the decline in capacity for in vivo incorporation of 32P into TMV RNA at 25 degrees following different periods of incubation at 40 degrews. The optimal temperature for replicase activity in vitro 35 degrees, and greater activity occurred at 40 than at 25 degrees. After about 15 min incubation at 40 degrees, in vivo incorporation of 32P into double-stranded TMV RNA became temperature-sensitive. This function could not be reproduced with the in vitro replicase system.


Hepatitis B core particles with endogenous DNA polymerase activity from chimpanzee liver. Hepatitis B core antigen (HBcAg) particles, approximately 27-28 nm in diameter and rho = 1.30-1.35 g/cm3, were purified from the liver of a chimpanzee experimentally infected with hepatitis B virus (HBV) while under cyclophosphamide treatment. The purified HBcAg particles incorporated radioactive deoxythymidine triphosphate. The product was precipitable by trichloroacetic acid and sensitive to DNase, but resistant to digestion by RNase. The reaction required four deoxyribonucleosise triphosphates- dATP, dCTP, dGTP and dTTP. Exogenous template did not enhance the reaction. From these findings, it was suggested that HBcAg particles purified from the HBV-infected chimpanzee liver contained DNA polymerase and endogenous DNA.


An insect cell line persistently infected with a baculovirus-like particle. A persistent infection by a baculovirus-like particle was found in the established lepidopteran (Heliothis zea) cell line, IMC-HZ-1. The virus caused CPE in less than 1% of the IMC-HZ-1 cells, as measured by phase-contrast and electron microscopy. Transmission tests showed that four lepidopteran cell lines were susceptible to the persistent virus (designated as IMC-HZ-I-NOV). In inoculated TN-368 cell cultures, 90--100% infection was achieved. The ultrastructure and development of IMC-HZ-I-NOV in cell cultures were similar to known baculoviruses, and it is probable that this persistent virus is a member of the family Baculoviridae. Two lepidopterous species (Estigmene acrea and H. zea) inoculated with IMC-HZ-1-NOV by intrahemocoelic injection and/or per os feeding of larvae were not susceptible.


Patterns of primary thyroid failure.

Patterns of primary thyroid failure. Serum thyroxine(T4), triiodothyronine (T3) and thyroid-stimulating hormone (TSH) levels were measured in 170 patients with suspected hypothyroidism. Only patients with elevated serum TSH levels indicative of some degree of primary thyroid failure were included in the study. They appeared to represent four distinct classes of thyroid failure: class 1 (19.5%) had normal T4 and normal T3; class 2 (8%) had low T4 and elevated T3; class 3 (37%) had low T4 and normal T3; and class 4 (35.5%) had low T4 and low T3. TSH levels were increased in all classes. Of 10 patients who were followed without treatment and evaluated after three and six months, six showed increasing clinical and chemical evidence of hypothyroidism and were reclassified, two did not progress and two patients improved. Our study showed that serum T3 levels may be normal or even elevated in hypothyroidism and, therefore, are not a useful indicator. The data also suggested that serum T4 is the main determinant of the metabolic state of the patient, since class 3 patients, the largest single group, had normal serum T3 levels despite striking clinical evidence of hypothyroidism. Thus, a normal serum T3 in the absence of a normal serum T4 is not generally sufficient for maintenance of a euthyroid state. Finally, our study showed that in primary thyroid failure there is better reciprocal correlation between the levels of serum TSH and T4 than between those of serum TSH and T3.


Low levels of serum calcium, phosphorus and plasma 25-hydroxy vitamin D in cirrhosis of the liver. In a retrospective analysis of 78 patients with liver cirrhosis, we found low serum levels of calcium and phosphorus. The low calcium levels showed a better correlation with high activity of aspartate aminotransferase than with low levels of albumin. In addition, there was a relationship between low calcium and low phosphorus levels. Therefore, factors other than, and in addition to, hypoalbuminemia seem to be responsible for the low calcium and phosphorus levels in cirrhosis patients. Although low levels of serum 25-hydroxyvitamin D were found in 23 of our patients, there was no indication that hypovitaminosis D was causative factor in the hypocalcemia and hypophosphatemia.


1alpha-hydroxycholecalciferol: a promising therapeutic approach for renal osteodystrophy. The effectiveness of 1alpha-hydroxycholecalciferol [1alpha-OH)D3] in healing uremic bone disease was investigated in 10 uremic patients on maintenance dialysis. Seven patients received 1alpha-(OH)D3, 2 microgram/day, for six months, and three patients served as a control group. Bone density was determined from X-rays of the second phalanx of the second digit. The apparent intestinal calcium absorption improved on treatment with 1alpha-(OH)D3, serum calcium levels and bone density significantly increased, and a significant decrease in serum alkaline phosphatase activity was observed. In one patient, reversal of uremic myopathy occurred after 17 days of 1alpha-(OH)D3 therapy.


Effect of an egg-rich diet on plasma lipids and proteins in severely burned patients. Eight severely burned young patients were maintained on an oral hyperalimentation regimen of about 7,000 calories--including 314 proteins, 600 g carbohydrates and 336 g fats, containing greater than 8 g cholesterol--based mainly on a daily ingestion of 35 eggs. Serum cholesterol and protein, and plasma lipoprotein levels were measured during period of 30 days on the diet. The mean serum cholesterol level prior to the egg-rich diet was low [108 +/- 42 (SD) mg/dl] and it remained within the normal range during the study. The initial mean serum total protein level of 4.1 +/- 0.5 (SD) rose to 6.5 +/- 0.7 g/dl by the 19th day and stayed high throughout the remainder of the study. Plasma lipoprotein levels, assessed by nephelometry and reported as concentrations of large, medium and small particles, were not increased. It is assumed that the high demand for energy during recovery from burns prevented abnormal rises in plasma lipid levels on fat-rich diet.


Absence of formation of cerebrospinal fluid in the spinal subarachnoid space of the cat. It is widely believed that formation of cerebrospinal fluid (CSF) takes place within the cerebral ventricles--80 to 85% being produced by the choroid plexus and the remaining 15 to 20% being secreted by the lining ependyma. It has also been suggested that CSF is formed within the subarachnoid space. To investigate this possibility. CSF formation was measured in the isolated subarachnoid space of the cat. In 16 animals that underwent 26 steady-state perfusions, the rate of formation of CSF was found to be only 0.00005 +/- 0.00056 (SE) ml/min. It is therefore concluded that little or no CSF is formed in the spinal subarachnoid space of the cat.


Scanning electron microscopy studies on the origin and structure of matrix vesicles in epiphyseal cartilage from young rats. Epiphyseal plates of young rats were examined by scanning electron microscopy to study the distribution, morphology and origin or the matrix vesicles (calcifying globules) and the results were compared with the findings on transmission electron microscopy. The matrix vesicles, 0.1 micrometer in diameter, were found to cover the intercolumnar septa and to increase gradually in number from the zone of proliferation to the zone of provisional calcification. In the intercolumnar septa of hypertrophic and calcifying cartilage, the vesicles aggregated to form calcospherites, i.e. mineralized globules about 1 micrometer in diameter. Chondrocytes showed numerous cell processes that were longer and more abundant in the more mature cells. In the zone of maturation many processes had enlarged tips with small globular projections ("bulges"), about 0.1 micrometer in diameter, which resembled the matrix vesicles covering chondrocytic lacunae. In the zones of hypertrophy and provisional calcification, many of these bulges aggregated to form larger, spherical structures. It is suggested that matrix vesicles in cartilage originate by budding from cell processess, and that in the zone of provisional calcification they aggregate to form the calcospherites, which are the loci of initial calcification of cartilage.


Significance of hyperprolactinemia in 70 women with amenorrhea. The clinical, radiological and hormonal findings in 70 amenorrheic patients are described. Hyperprolactinemia was found in 18 patients (25.7%) who were divided into three groups: nine patients with functional secondary amenorrhea, six patients with radiological evidence of pituitary tumor, and three patients with primary amenorrhea. One of the latter group had primary hypothyroidism. Plasma gonadotropin concentrations were normal or low in most patients, with the exception of five women with ovarian failure in whom the levels were significantly raised. Galactorrhea was diagnosed in 23 patients (32.8%), and in 16 of them plasma prolactin levels were elevated. Recent experience has shown that reduction of prolactin levels to normal by removal of prolactin-secreting tumors or by treatment with bromocriptine results in the restoration of normal menstrual cycles and fertility. The hormonal and radiological identification and characterization of hyperprolactinemic amenorrhea are therefore of prime importance.


Vaginal adenosis in a 48-year-old woman. Vaginal adenosis was diagnosed in a 48-year-old woman who had been on estradiol therapy in the past. The possible role of hormonal factors in the development of vaginal adenosis in older women is discussed.


Total hip replacement in a patient on hemodialysis. A 52-year-old woman, on intermittent hemodialysis because of chronic renal failure, underwent total hip replacement for severe osteoarthritic changes. The operation was performed under epidural anesthesia, the postoperative course was uneventful, and the patient was completely relieved of pain and limitation of movement. Although total hip replacement is a major surgical procedure, it is suggested that the operation can, when considered necessary, by successfully carried out in a patient on hemodialysis, provided that careful attention is paid to the pre- and postoperative care.


[Generalized acute pustulosis. An unusual presentation of leukocytoclastic vasculitis]. After an acute course of pharyngitis and bronchitis treated with Bactrim (trimethoprim and sulfamethoxazol), a 52 year old woman suddenly developed a widespread, sterile pustular eruption with predilection of the distal parts of the extremities. The histological and electron microscopical examinations showed intraepidermal pustules with underlying distinct leukocytoclastic vasculitis. Direct immunofluorescence revealed the prescence of intra- and perivascular deposits of C3 in the upper part of the dermis. Drug- and/or infection allergy is suggested to be responsible for the etiopathogenetic mechanism of leukocytoclastic vasculitis.


[Porphyria cutanea tarda--a multifactorial hereditary disease? A working hypothesis]. The influence of genetic and hepatotoxic factors in the development of porphyria cutanea tarda was studied in 51 patients with this disease. According to their data and findings in literature, the following hypothesis was made: porphyria cutanea tarda is assumed to be a disease following the rules of the polygenetical heredity. The appearance of the disease seems to be influenced by genetic rules, and hepatotoxic factors are playing a role in the manifestation of the disease.


[The value of C3-complement and ceruloplasmin determinations for the diagnosis of infections of the seminiferous tract]. 25 percent of 776 andrologic patients showed C3c-complement and coeruloplasmin--determined by radial immundiffusion--in their ejaculate. No correlation could be found between these globulines and possible bacterial infections of the genital tract.


[Neurotrophic heel ulcers due to spondylolisthesis]. A case of neurotrophic ulcers on both heels is reported, which is due to congenital spondylolisthesis. Spondylolisthesis as the etiology of the neurotrophic ulcer has not yet been described.


[Subungual melanoma with linear nail pigmentation]. A 53 year old male patient presented with a dark-brown linear pigmentation of the thumb nail due to a subungual melanoma mainly of the superficial spreading melanoma-type. Melanin-induced nail pigmentations are recommended to be treated by a resection in local anaemia of the dorsal two thirds of the terminal phalanx. This procedure combines sufficient radical surgery with a good functional and acceptable cosmetic result.


Quality assurance practice in rural and urban hospital clinical laboratories. This survey examined quality assurance practice in matched pairs of rural and urban hospital clinical laboratories. Questions directed at personnel qualifications, laboratory management, internal quality control mechanisms, and proficiency testing enrollment revealed no major differences between the rural-urban pairs. Deficiencies observed were either generic or size-related. Most laboratories were directed by physicians; however, almost half spent 5 or less hours per week in the laboratory. All respondents employed at least one individual professionally certified at the "technologist" level. Virtually all respondents insisted they employ a laboratory management system for ensuring quality performance, although 20% did not document their practice. Internal quality control practice varied widely. Many respondents recognized shortcomings related to volume and budget, and requested specific training courses for technical personnel. At least 20% of the laboratories were not enrolled in any recognized proficiency testing program, an integral part of the total quality assurance process.


An improved polystyrene polymeric XAD-2 resin column extraction of 5beta-cholestan-3beta-ol from polluted water. Appropriate methods and criteria for judging the degree of sewage contamination of water and its suitability for drinking or recreational uses are essential to safeguard the public health. 5beta-cholestan-3beta-ol seems to satisfy many, if not all, of the criteria required of a good indicator of fecal pollution. It was shown that this fecal sterol was strongly adsorbed to the polystyrene polymeric XAD-2 adsorbents at low pH, resulting in a 100% retention. The adsorbed sterols could be easily removed from the columns with acetone adjusted to pH 8.5-9 with concentrated NH4OH. It has also been demonstrated that large volumes of both fresh and sea water samples can be extracted by the "closed" column method in a relatively short time. The sensitivity of the column exceeded that of the conventional liquid-liquid partitioning procedure. With some modifications, the column extraction process can be incorporated in a fully- or semi-automated analytical procedure.


The comparison of viral antibody titers of acid-precipitated and non-precipitated mouse ascitic fluids. Mouse immune ascitic fluid has become a primary source of antibody for diagnostic, reference, and research work in many virus laboratories. The inherent disadvantage of ascitic fluid is that it repeatedly forms clots and subsequently loses volume. The acid-precipitation method of Chiewsilp and McCown eliminates the clot formation and does not appreciably alter the antibody titers for several arboviruses, varicella, rabies and influenza viruses.


[Operative therapy for chronic otitis media and middle ear cholesteatoma with preservation of the posterior auditory canal wall ("intact wall technique") (author's transl)]. The late results of two different procedures for middle ear surgery are compared 2-10 years following surgery. A total of 478 ears were operated, of which 367 had cholesteatomas and 111 chronic granulating otitis without cholesteatoma. a) In conservative radical operations, tympanoplasties, meatoplasties with fascia and obliteration with muscle (298 ears) gave satisfactory and stable results: 8% required re-operation, 2.8% had residual cholesteatoma, 0.9% had recurrent cholesteatoma, 15% had perforations, and 90% had dry operative cavities. b) In modified "Intact Wall Technique" with small cortical mastoidectomies and epitympanic control openings, 7% had residual cholesteatomas, 2% recurrent cholesteatomas, and 11% required re-operation. The following conclusions were made: The "Intact Wall Technique" is a safe method for the surgical management of granulating otitis media. However, the procedure is not considered to be safe when disease involves cholesteatoma because of the significant frequency of recurrent disease or the development of retractions which can then form cholesteatoma. Under such circumstances, combination of the "Intact Wall Technique" with obliteration may give more stable results.


[The clinical presentation and management of otogenic intracranial disease (author's transl)]. Since the introduction of antibiotics in the treatment of ear infection the number of otogenic intracranial complications has decreased. Furthermore the clinical presentation of these complications has likewise since changed. The traditional symptoms and signs as mentioned in the literature are replaced by an indefinite inflammatory course, which is finally succeeded by rapid progress of the disease despite the use of antibiotics. Experiences with 22 patients are reported. Temporal lobe brain abscesses occurred in 11 patients, meningitis occurred in 3 others and lateral sinus thrombophlebitis occurred in 8, in 2 of whom meningitis also occurred. Five patients died as a result of the intracranial complication. After establishing the diagnosis the recommended treatment is the immediate surgical clearance of the primary aural abscess, together with if possible drainage of the intracranial abscess or evacuation of the lateral sinus disease under appropriate antibiotic cover. Whether the patient with intracranial disease is subsequently handled in the ORL or Neurosurgical wards depends upon ward facilities and interdepartmental liaison and cooperation.


[Reflections on the technique of inserting the esophagoscope (author's transl)]. Due to the muscular attachments of the hyoid bone to the posterior part of the base of the skull, and the larynx to the sternum, the larynx will be pressed posteriorly against the vertebral column if the head is kept in a reclined position. This causes a greater resistance when trying to open the hypopharynx with the esophagoscope. If, however, the head is held forward, this resistance will be considerably reduced.


[Laryngeal and pharyngeal endoscopy and the disinfection of the endoscope in routine practice (author's transl)]. The usefulness in routine ENT practice of a laryngo-epipharyngoscope with a Hopkins lateral telescope is reported. The chemical disinfection of this endoscope is bacteriologically proven to be effective, and is discussed.


[Osteoradionecrosis resulting from cobalt-60 therapy for head and neck tumors (author's transl)]. From 1966--1974, 628 patients were irradiated with Cobalt-60 as treatment for tumors of the head and neck. Tumor doses ranged from 6200 R to 8000 R. An osteoradionecrosis was discovered in 17 patients, and was mostly localized in the mandible. Most of the patients developed the necrosis during the first two years following completion of their radiation, although necrosis was also found up to 9 years following therapy. Prophylactic treatment of teeth should be completed before beginning radiation therapy, but such does not exclude the risk of necrosis.


[The longterm results of osteoplastic frontal sinus surgery for recurrent disease (author's transl)]. The osteoplastic frontal sinus operation permits the complete removal of sinus mucosa and, with blockage of the frontonasal duct, the reliable obliteration of the sinus cavity. Two patients, who had had several surgical procedures for recurrent frontal sinusitis, became symptom free after this procedure. After an interval of 5 to 6 years the cosmetic result remained satisfactory and ossified obliteration of the sinus was demonstrable radiologically.


[Post-laryngectomy vocal rehabilitation with a tracheophypopharyngeal shunt (author's transl)]. Oesophageal speech is the accepted method of post-laryngectomy speech rehabilitation, but still has a high failure rate. Among the different surgical procedures that create a tracheo-oesophageal fistula the method of Komorn based on a concept of Conley appears to have the greatest theoretical advantages. In this procedure a fistula is created with its inferior opening into the oesophagus and its superior opening into the trachea. Komorn's technique has been utilized in a modified form in 12 patients and in 6, because of wound problems, useful speech was not achieved. Five patients have achieved excellent speech without difficulty.


Pure tone delayed auditory feedback: effect of instructions. This study compared the effect of instructions to the subject upon key tapping performance with pure-tone delayed auditory feedback. One set of instructions stressed the importance of maintaining tapping rate whereas a second set emphasized consistency of the pattern. A third set stressed rate and pattern equality. The results lead to the recommendation that instructions stress equally both rate and pattern consistency.


Elicitation of the acoustic reflex with synthetically produced consonant and vowel stimuli. Acoustic reflexes were elicited from seven normal hearing subjects with synthesized isolated consonant and vowel speech stimuli. Stimulus intensity and duration were controlled so that equivalent amplitudes and durations from 100 to 300 msec were produced. The recorded stimulus items were presented over an earphone to the subject's right ear, while susceptance changes caused by bilateral acoustic reflex activity were monitored in the subject's left ear. Analysis of variance revealed no significant difference between reflex threshold values for isolated synthetic consonant and vowel stimuli. Main effects of stimulus duration and subjects and the two-way interaction of phoneme type by subject were found to be significant.


Negative needle deflection of the acoustic reflex in otosclerotics. The preoperative audiometric and electroacoustic impedance test results of 24 surgically confirmed otosclerotics are reviewed with special attention given to the direction of needle deflection during acoustic reflex testing. Information on the direction of needle deflection during acoustic reflex testing was gathered at 5 or 10 dB above reflex threshold. In 12 of the 24 patients acoustic reflexes were present, and in each case the response included negative needle deflections. The presence or absence of the acoustic reflex seemed to be related to the reported duration of the hearing loss.


Changes in ventilatory pattern induced by intravenous anesthetic agents in human subjects. The tidal volume-inspiratory duration relationship was studied during air breathing and rebreathing in conscious and anesthetized human subjects using three different intravenous agents. The results observed have been compared with similar experiments carried out in cats. Its has been shown that anesthesia provokes an increase in breathing rate associated with a decrease in tidal volume in human subjects; an opposite effect on breathing rate was observed in cats. Thus, the tidal volume-inspiratory duration relationship, although very similar in the conscious cat or human subject, is very different under anesthesia. The results were quite consistent in a given species whatever the nature of the drug used. It is suggested that modifications of the breathing rate by anesthesia, related to animal species, are caused by central effects of the drug. These effects are probably mediated by different actions on inputs to the inspiratory "off-switch" mechanisms in the two species.


Effects of physical training and lean body mass of plasma amino acids in man. Postabsorptive plasma amino acid and insulin concentrations were determined in subjects with hyperplastic obesity and in nonobese controls before and after a 6-wk period of physical training. After the training period the plasma concentrations of insulin and leucine decreased and the concentration of alanine increased in the obese subjects. No changes were noticed in the controls. The obese subjects had elevated plasma levels of valine, isoleucine, leucine, tyrosine, and phenylalanine before as well as after physical training. The concentrations of these amino acids were correlated to the plasma insulin level and to lean body mass before but only to lean body mass after physical training. It is suggested that the lean body mass, whick is higher in hyperplastic obesity, contributes to the elevated concentrations of amino acids, and it is unlikely that the insulin decreases in the obese subjects after physical training is mediated through an effect of amino acids on insulin secretion.


Thresholds of aural overload, word discrimination in noise, and the "fragile ear". This study examined the relationship between two previously proposed measures of the fragile ear, the aural overload test and word discrimination score in noise. Ten normal-hearing young adults served as subjects. Aural overload thresholds were determined for fundamental frequencies of 500, 1000, and 2000 Hz. Word intelligibility in noise was assessed with tests using an open response-set (Northwestern University Auditory Test No. 6) and closed response-set (Modified Rhyme Test) at a variety of primary-to-secondary ratios. Results revealed a lack of correlation between threshold of overload and word discrimination in noise at unfavorable primary:secondary ratios for both tests(-6 and 0 dB for the NU-6' s and -12 and -6 dB for the Modified Rhythm Test). Significant correlations (p less than 0.05) were achieved, however, with both word discrimination tests when administered at the more advantageous primary:secondary ratios.


Masking of spondees by interrupted noise in hearing-impaired listeners. The effects of changing the duty cycle of an interrupted-broad band masker on the spondee thresholds of hearing-impaired subjects were explored. Two diagnostic groups of sensorineural hearing loss, presbycusics and cochlear otosclerotics, were investigated. The interruption rate of the masker was 10/sec, and its duty cycle was varied at 25% intervals. Thresholds also were obtained in continuous noise and in quiet. Results when compared with earlier data from normal listeners revealed that subjects with sensorineural impairment exhibited poorer performance in all instances, including the continuous noise condition, when mean masker levels were adjusted to comparable sound pressure levels. The overall pattern of masking was more similar for the normal and hearing-impaired groups when performance was equated in terms of the mean threshold shift each group experienced in continuous noise. Even under these circumstances, however, hearing-impaired subjects demostrated notably greater residual masking under the 25 and 50% noise-on conditions. A major determinant of speech reception in fluctuating noise backgrounds is the dependence of the pattern of masking upon the difference between an individual's masked threshold in continuous noise and his threshold in quiet, rather than upon the sound pressure level of the masker.


Tracheobronchial secretions collected from intact dogs. II. Effects of cholinomimetic stimulation. The effect of cholinomimetic stimulation on the quantity and composition of tracheobronchial secretions (TBS) collected from intact dogs was studied. After the administration of methacholine chloride, the collection rate of TBS increased over threefold within 10 min and returned to the control level by 50 min. Even though there was an increase in the total amount of protein collected, the protein concentration of the secretions decreased initially by 40%, returning to normal by 30 min. This was the result of an apparent dilution of the secretions by nonproteinaceous components. In addition, at 10 min, there was an increase in the protein associated with mucins. These responses were blocked by the preadministration of atropine. When TBS was separated into supernatant and pellet phases by centrifugation, we found that, following methacholine, there was a 35% increase in the ratio of albumin to IgG in the supernatant phase. These results suggest that the increased secretion due to methacholine may result, in part, by a selective filtration of fluid from the interstitial and/or vascular compartment(s) with proteins appearing in the secretory mixture based partially on their size.


Reflex effects of lung inflation and inhalation of halothane, ether, and ammonia. Anesthetized dogs were perfused at constant pressure from an external oxygenator after removing their hearts. Systemic vascular resistance (SVR) and spontaneous breathing effors were monitored while the lungs were ventilated briefly at a low frequency with several tidal volumes and at a low volume with several frequencies. Inflation depressed respiratory activity and transiently lowered SVR to a minimum from which it spontaneously partially recovered. The transient and sustained SVR changes were directly related to peak inflating pressure, independent of the mode of ventilation. Pressurization of the sublaryngeal extrapulmonary airways and addition of ammonia to lung inspirate were without effect. Ventilation with 7% ether had no effect, but at 13% it occasionally increased respiratory activity. Ventilation with 7% halothane depressed SVR and stimulated respiration, whereas 13% caused vasodilation and apnea. The magnitudes of SVR responses to halothane and to inflation were uncorrelated. All effects of inflation and inhalants were eliminated by vagotomy. We condluded that the systemic vessels appear to respond to changes in respiratory minute volume; that this is brought about by intrapulmonary receptors, perhaps of more than one type; that two classical "irritants" have little effect in dogs; and that halothane has effects on breathing and SVR consistent with stimulation of J-receptors.


Comparison of speech detection and spondee thresholds and half- versus full-list intelligibility scores with MLV and taped presentations of NU-6. Several speech audiometric measurements were made on 212 ears with mild sensorineural hearing loss. An 8-dB difference between speech detection and spondee thresholds was observed, which is the same relationship that has been found in normal ears. No significant differences in speech discrimination scores (SDS's) were observed when NU-6 was administered via monitored live voice (MLV) and the Auditec recordings. Although our data support the use of MLV testing, verification with a standardized recording should be considered when unusually poor SDS's are obtained. Half-list and full-list SDS's were analyzed for both taped and MLV presentation modes. This analysis showed that both the MLV and taped stimuli exhibited very similar variability and that about 96% of the half-list scores were within 6% of the full-list scores. The clinician should be cautious, however, because 4% of the ears had half-list/full-list discrepancies ranging from 8 to 14% and differences as large as 28% have been reported by Raffin and Thornton (1977). Furthermore, variability between half-list and full-list SDS's varies as a function of intelligibility impairment, being least for scores approaching the extremes of 0 and 100% and greatest for scores in the 30 to 70% range. Finally, our data suggest that half-list testing can be an effective screening procedure to determine it full-list testing is advisable.


Effects of time and vasoconstrictor tone on O2 extraction during hypoxic hypoxia. To test the role of peripheral vasoconstrictor tone in the efficient use of a limited O2 supply, three groups of anesthetized dogs were ventilated with 9.1% O2 until circulation failed. Two groups were alpha-blocked with phenoxybenzamine and one of those was volume expanded with dextran to restore blood pressure. After O2 utake was lowered in hypoxia, O2 uptake was linearly related to O2 delivery (cardiac output X arterial O2 content) with r = 0.94. The slope of that line was mathematically identical to the extraction ratio and it increased from 0.54 at 10 min to 0.87 at the end of hypoxia (r = 0.99). In both alpha-block groups O2 extraction remained constant with time, and O2 extraction in each alpha-block group was significantly less (P less than 0.01) than in the unblocked group. As further evidence of better O2 extraction, mixed venous partial O2 pressure was significantly less in the unblocked group, 6.2 +/- 3.4 Torr vs. 10.6 +/- 3.2 and 9.9 +/- 2.4 Torr with alpha block (P less than 0.01). Results after alpha-block indicated that a vigorous vasoconstrictor tone during hypoxia conserved O2 by promoting greater extraction by the tissues.


Relation between oxygen uptake and developed tension in dog skeletal muscle. Isometric developed tension of in situ gastro-cnemius-plantaris and semitendinosus muscles of the dog was increased 30% with twitch contraction frequency over the range of 0.5--2.0 tw/sc. Developed tension was decreased 75% after 30--40 min of 11--14 tw/s contractions, fatigue. Recovery from fatigue was slow. Intravenously injected K+ after fatigue increased developed tension 50%. Infusions of epinephrine, norepinephrine, and ouabain did not change developed tension. When the oxygen uptake of gastrocnemius-plantaris muscle was measured during twitch contractions at 0.5, 1.0, and 2.0 tw/s before and after fatigue and after injection of K+, the oxygen uptake was found to be directly related to the developed tension.


Effects of atropine on potentiation of exercise-induced bronchospasm by cold air. The role of vagal efferent activity in the cold air potentiation of exercise-induced asthma was assessed by exercising nine subjects who breathed air at ambient and subfreezing temperatures before and after cholinergic blockade. Lung volumes and maximal expiratory flow volume curves with air and with 80% helium-20% oxygen were obtained before and 5--10 min after each challenge. Isovolume comparisons of maximal expiratory flow rates with the two gases were used to assess relative contributions of large and small airways to flow limitation. Exercise under ambient conditions resulted in the expected airway obstruction and cold air exaggerated the response. Atropine pretreatment had no effect on the cold air potentiation. After atropine with ambient air exercise, there was an increase in the relative contribution of large airways to flow limitation, whereas exercise with cold air resulted in an increase in the contribution of small airways. We concluded that the potentiating effects of cold air are local and suggest that the immediate stimulus is related to cooling of intrathoracic airways.


Behavior of the human pulmonary circulation during head-up tilt. Pulmonary blood volume (PBV), flow, and pressures were measured in 15 cardiopulmonary normal subjects, while supine and at 60 degrees head-up passive tilt. PBV, cardiac index (CI), right ventricular diastolic volume (VDVR), and mean pulmonary arterial (Ppa), venous (Pv), and mean right atrial (Pra) pressures all decreased during the first 4 min of tilting, with little subsequent change thereafter, to 30 min. Relative changes of CI, PBV, and VDVR were of similar magnitude (25--30%; mean Ppa and Pv fell 3.2 and 2.3 Torr, respectively. From measurements of thoracic dimensions and changes in intravascular pressures, we calculate that 11% of the pulmonary vascular bed falls into zone I condition, 34% goes from zone III to zone II, and 55% remains in zone III condition during tilt. A highly significant correlation exists between PBV and CI (P less than 0.001), with no significant change in pulmonary mean transit time during tilt. Significant correlations also exist between PBV and Pra (P less than 0.001), Pv (P less than 0.01), and Ppa (P less than 0.05). We conclude that outflow pressure plays a significant role in determining PBV during head-up tilt, but the major determinant is venous return to the right heart.


Respiratory mechanics in the anesthetized rat. Functional residual capacity (FRC) and pressure-volume (PV) curves of the lung, chest wall, and total respiratory system were studied in 15 anesthetized rats, weighing 307 +/- 10 (SE) g. Pleural pressure was estimated from the esophageal pressure measured with a water-filled catheter. The FRC determined by body plethysmograph was slightly and significantly larger than FRC determined from saline displacement of excised lungs. The difference may be accounted for by O2 uptake by lung tissue, escape of CO2 through the pleura, and abdominal gas. Paralysis in the prone position did not affect FRC, and abdominal gas content contributed only slightly to the FRC measured by body plethysmograph. Values of various pulmonary parameters (mean +/- SE) were as follows: residual volume, 1.26 +/- 0.13 ml; FRC, 2.51 +/- 0.20 ml; total lung capacity, 12.23 +/- 0.55 ml; compliance of the lung, 0.90 +/- 0.06 ml/cmH2O; chest wall compliance, 1.50 +/- 0.11 ml/cmH2O; and respiratory system compliance, 0.57 +/- 0.03 ml/cmH2O. The lung PV curve did not show a consistent change after the chest was opened.


Elasticity of excised dog lung parenchyma. An experimental procedure was developed to measure the stress-strain relationship on rectangular slabs (5.0 X 5.0 X 0.5 cm) of excised dog's lung. The slabs were subjected to biaxial loading and the resulting triaxial deformations were measured. Deformations were measured in the central portion of the specimen by video dimension analysers in order to minimize boundary effects. Specimen thickness was measured with a magnetic reluctance proximeter system. The data were sampled and stored on-line by a PDP-8E computer. An electromechanical servo system was used to control the lateral force. Tests were performed at several pH values and at 20 and 37 degrees C. The tissue exhibited a highly nonlinear stress-strain relationship, compliant at low stress levels and stiff when the stress was high. Hysteresis was observed to be about 28% and was unaffected by a 250-fold change in strain rate. Biaxial loading revealed a new characteristic: there is a change in elastic behavior when the tissue undergoes a compressive strain. When the tissue was in tension increasing the lateral load decreased the compliance, but the opposite was true when compressive strain was present.


Influence of posture on isometric fatigue. The isometric strength of four trained subjects was unaltered by changes in posture. But the endurance of an isometric contraction held to fatigue at 25 and 40% of the maximum voluntary contraction (MVC) was 20% greater in the sitting than in the recumbent posture. This difference was abolished when the exercise was performed with the arm's circulation arrested. At rest, the blood flow through the forearm was greater when the subjects were in the recumbent than in the sitting position but the reverse was true during isometric contractions. In these two postures, there was no difference in the right atrial pressure during the contraction, suggesting that the low-pressure baroreceptors are not responsible for the differences in blood flow during exercise. To date no mechanism is available to explain these observations.


Pulmonary capillary and permeability during hemorrhagic shock. In previous experiments from this laboratory horseradish peroxidase was used to study the structural and functional characteristics of the normal canine pulmonary capillary membrane. The present study used the same technique to try to determine if any change occurred in the pulmonary capillary as a result of hemorrhagic shock. We found that hemorrhagic shock caused a fall (5 expt) or no change (2 expt) in estimated pulmonary transcapillary pressure based on Starling's equation. However, lymphatic flow from the lungs increased. Estimated of filtration coefficients showed a highly significant increase (P less than 0.01) during the hypotensive period. Pulmonary lymphatic protein concentrations were not altered, indicating that water and protein continued to traverse the membrane in the same proportions as under normotensive conditions. These data are consistent with recent observations of minimal changes in the intercellular junctions of the capillary endothelium following hemorrhagic shock made independently on lung tissue from these experiments.


An animal model for the study of regional lung function. An animal model for the study of regional lung function is described. In sheep, the bronchus to the right apical lobe (RAL) of the lung arises directly from the trachea. A tracheal divider, inserted under local anesthesia via a permanent tracheostomy, was used to separate the ventilation of the RAL from that of the rest of the lung. Lobar blood flow was estimated from the RAL contribution to the pulmonary clearance of an intravenous bolus of 85Kr. Gas exchange was measured by conventional methods. Expressed as a percentage of the value obtained for the whole lung, lobar expired volume was 14.7 +/- 4.3%, capillary perfusion was 12.3 +/- 4.2%, oxygen uptake was 14.7 +/- 4.9%, and carbon dioxide production was 13.4 +/- 5.5% (mean +/- SD of 25 studies in 11 animals breathing air). The model permits the study of experimental conditions confined to a single lobe of the lung and offers the advantages of an intact chest wall, spontaneous ventilation and an unanesthetized animal.


Electron microscopy of lung rapidly frozen under controlled physiological conditions. Light microscopy of lung rapidly frozen under controlled physiological conditions has been very successful in correlating pulmonary structure and function. However, to study some aspects of pulmonary capillary morphology, the higher resolution of electron microscopy (EM) is necessary. To date, most EM of lung has involed the instillation of a fixative through the airways or vascular system, techniques that probably alter the normal pressure relationships of the capillaries and therefore their morphology. We describe here a technique for rapidly freezing lung to a depth of 1--2 mm below the pleural surface and preparing sections for EM. Lungs from open-chest rats were frozen at various transpulmonary pressures with cold (--80 degrees C) 70% ethylene glycol. Small pieces were then fixed with a solution containing glutaraldehyde and paraformaldehyde for 24 h at --50 degrees C. Staining was with osmium tetroxide and uranyl acetate. Lung frozen at high volumes showed marked stretching of the alveolar septa with severe deformation of the capillaries. Lung frozen at low inflation pressures revealed open capillaries containing numerous red blood cells; in addition, infolding of the alveolar wall was frequently seen. We conclude that this technique gives a level of preservation of rapidly frozen lung suitable for electron microscopy.


Microscopic tissue alterations following cryosurgery of canine skin. Cryosurgery of cutaneous tissue of 6 dogs was followed by serial biopsies throughout the necrotizing and healing periods. The duration of cold application was controlled by thermocouple monitoring in 3 dogs and by a standard time in 3 dogs. Tissue changes following cryosurgery were consistent in all 6 dogs and were composed of edema, erythema, necrosis, sloughing, granulation, and epithelialization.


Focal cystic hypertrophic gastropathy in a dog. A 6-year-old female dog had a 3-day history of postprandial vomiting of undigested food. Plain and contrast radiography revealed a large mass on the greater curvature of the stomach. On exploratory gastrotomy, the mass was found to consist of hypertrophic rugae resembling cerebral gyri and sulci. The histopathologic findings were similar to those for hypertrophic gastropathy in man and the dog.


Canine coccidiosis attributed to an Isospora ohioensis-like organism: a case report. Coccidiosis due to an Isospora ohioensis-like organism was diagnosed in a 10-week-old pup. The pup had diarrhea, dehydration, and weight loss before it died 7 days after onset of clinical signs. Lesions were limited to the intestinal tract. Beginning in the distal end of the small intestine and extending through cecum and colon were mild histiocytic proliferation in the lamina propria and multifocal cryptitis. Numerous meronts and gamonts of an unidentified coccidium were in or near the lesions. Parasites were in the villous epithelium, lamina propria, and intestinal glands of the distal one-half of the small intestine, cecum, and colon. It was concluded that infection was due to an unidentified coccidium with oocysts structurally similar to those of I ohioensis.


Cauda equina compression from lumbosacral malarticulation and malformation in the dog. Compression of the cauda equina from subluxation, stenosis, or spondylosis of the lumbosacral articulation was found in 20 dogs, 13 of which were German Shepherd Dogs or crossbred German Shepherd Dogs. Pain, posterior paresis, and urinary or fecal incontinence were the most frequent problems associated with the compression. Dorsal laminectomy was performed on 13 cases, with good to excellent results on 10 of these.


Streptoalloteichus, a new genus of the family Actinoplanaceae. A new genus Streptoalloteichus is proposed in the family Actinoplanaceae to distinguish species of actinomycetes which form short or long spore-chains on aerial mycelium, bears oval sporangia with motile spores and has a characteristic cell-wall composition of strain C677-91 type. Strain C677-91 (ATCC 31217, FERM-P No. 4070) was named Streptoalloteichus hindustanus gen. nov. and sp. nov. The actinomycete strain C677-91 produces spore-chain clusters and sclerotia in the aerial mycelium which are morphologically similar to those found in some species of Streptomyces. The cultural characteristics of the strain on agar media also resemble those of Streptomyces species and the colonies have no distinct color. Strain C677-91 produces sporangia or sporangia-like vesicles which contain one to several spores in the vegetative mycelium. The sporangiospores possess a single long polar flagellum and are motile. The cell wall of strain C677-91 contains meso-alpha,epsilon-diaminopimelic acid, alanine, glutamic acid, galactose, mannose, rhamnose and glucosamine. Strain C677-91 has several important characteristics in common with Streptomyces tenebrarius including the production of nebramycin factors but the latter strain does not produce sporangia.


Taxonomic study and fermentation of producing organism and antimicrobial activity of mildiomycin. A taxonomic study of strain B-98891, which produced an antibiotic effective against powdery mildew of barley, identified it as Streptoverticillium rimofaciens. On agar media the antibiotic, which was named mildiomycin, was only weakly active against most fungi and bacteria tested. However, it inhibited some Mycobacterium and Rhodotorula, and it showed excellent control of powdery mildew of barley plants in greenhouse tests at concentrations between 31.2 and 62.5 ppm. Rhodotorula rubra IFO 0907 was selected as the test organism for in vitro assay of mildiomycin.


Isolation and characterization of mildiomycin, a new nucleoside antibiotic. A new antibiotic mildiomycin, strongly active against powdery mildew, was isolated from the culture filtrate of Streptoverticillium rimofaciens B-98891. It is a water-soluble basic antibiotic and was purified by ion-exchange and adsorption chromatography. The molecular formula of the purified compound was determined to be C19H30N8O9(H2O) from physical and chemical data. The UV and NMR spectra suggested that this antibiotic is a nucleoside. On acidic hydrolysis it gave 5-hydroxymethyl cytosine which has not previously been found in nucleoside antibiotics.


Aurantinin, a new antibiotic of bacterial origin. A new polyene class antibiotic, aurantinin (KM-214), was isolated from the fermentation broth of Bacillus aurantinus MASUMA and OMURA sp. nov. The substance is a conjugated triene with a molecular weight of 618 and molecular formula C35H54O9, and melts at 139 approximately 140 degrees C. The antibiotic is active in vitro against Gram-positive bacteria, but not against yeast and fungi.


Funiculosin, a new antibiotic. II. Structure elucidation and antifungal activity. Structure elucidation and some biological properties of an antiviral and antifungal antibiotic, funiculosin, are summarized. Funiculosin is a derivative of N-methyl-4-hydroxy-3, 5-disubstituted-2-pyridone containing a novel substituent, cyclopentanetetrol moiety, that is reported here for the first time in a natural product. The antibiotic protects guinea pigs efficiently against experimental trichophytosis. Toxicity of funiculosin is unique and highly selective for animal species.


Diastereomeric 7-ureidoacetyl cephalosporins. I. Superiority of 7alpha-H-L-isomers over D-isomers. The synthesis and in vitro structure-activity relationship of 7-ureidoacetyl cephalosporins carrying various substituents in the 3-position, compounds that showed an enhanced broad spectrum of antibacterial activity, has been outlined. Contrary to most of the previous observations with diastereomeric isomers of cephalosporins, it has been found that the L-side chain isomers also are very potent antibiotics and are even more active inhibitors of certain beta-lactamase-producing Gram-negative bacteria than the corresponding D-side chain isomers. SQ 69,613, 7beta-[[L-[(aminocarbonyl)amino]-2-furanylacetyl]amino]-3-[[(1-methyl-1H-tetrazol-5-yl) thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, sodium salt, the most active compound tested, except for activity against staphylococci, was as active in vitro as cefamandole.


Diastereomeric 7-ureidoacetyl cephalosporins. II. 7beta [[[(Aminocarbonyl) amino]-2-thienylacetyl] amino]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5-yl) thio] methyl]-8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid. The synthesis and antibacterial activity in vitro of 7-methoxylated cephalosporins having a thienylureidoacetyl or a thienylglycyl C-7 side-chain are described. Acylation of 7 beta-amino-7-methoxycephems with a novel 2-aminooxazolone hydrochloride under neutral conditions gave the thienylureidoacetyl derivatives in good yield with retention of configuration. 7 beta-[[D-[(Aminocarbonyl)amino]-2-thienylacetyl]amino]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5-yl)thio] methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (SQ 14,359) was found to have a broad-spectrum of antibacterial activity in vitro, particularly against beta-lactamase-producing organisms.


Diastereomeric 7-ureidoacetyl cephalosporins. III. Contribution of D- and L-isomers to the growth inhibiting activities of 7alpha-H and 7alpha-OCH3 derivatives for gram-positive and gram-negative bacteria. A series of 7beta-ureidoacetyl, 7alpha-H and 7alpha-OCH3 cephalosporin antibiotics have shown broad-spectrum antibacterial activity in vitro. In the 7alpha-H but not in the 7alpha-OCH3 series, contrary to experience in the antibiotic field, the L-isomers were substantially more active than the D-isomers both in vitro and in vivo particularly, but not exclusively, against Enterobacteriaceae that produce potent chromosomal cephalosporinases. Enhanced resistance to and inhibition of beta-lactamase (s) appeared to be responsible for this effect. Studies in vitro specifically with 7beta-thienylureidoacetyl derivatives showed that D-isomers interacted with L-isomers in the 7alpha-OCH3 series in a synergistic manner against "cephalosporinase-type" enzyme producers while isomers in the 7alpha-H series did not. Examples were presented in which this favorable event resulted in improved efficacy of the racemic mixture over the pure D- or L-isomer alone in appropriate experimental infections.


Semisynthetic cephalosporins with antifungal activity: laboratory studies on 2-pyridinethiol 1-oxide cephalosporins. Cephalosporins are chemotherapeutic agents whose spectrum and use are limited to antibacterial activity. Therefore, it was of interest to find several new semisynthetic cephalosporins which possess in vitro antifungal activity against Trichophyton mentagrophytes, Candida albicans and certain other yeasts. Five new cephalosporins containing the 2-pyridinethiol 1-oxide grouping were examined. Four with this heterocyclic moiety in the 3-position were found to have activity. One with the grouping in the 7-acyl-side chain was inactive. The degree of the antifungal activity was influenced by the substituent at the 7-position. The cephalosporin with a p-hydroxyphenylglycyl side chain was the most potent against the fungal strains studied. Despite this in vitro antifungal activity, none of the compounds protected mice against a systemic Candida albicans infection. All of these cephalosporins had broad spectrum in vitro and in vivo antibacterial activity.


Studies on the biosynthesis of clavulanic acid. I. Incorporation of 13C-labelled precursors. The biosynthesis of clavulanic acid was investigated by feeding 13C-labelled precursors to Streptomyces clavuligerus fermentations. The resulting samples of clavulanic acid were isolated as the benzyl ester and were examined by 13C NMR spectroscopy for 13C-enrichment. The results showed that the carbon skeleton of 1,3-13C2-glycerol was incorporated intact into the three beta-lactam carbons of clavulanic acid. Studies with 1-13C-acetate, 2-13C-acetate and 1,2-13C2-acetate indicated that the remaining five carbons of clavulanic acid were probably derived from alpha-ketoglutarate. 1-13C-Propionate and 3-13C-propionate were not metabolised via the same route as glycerol, but were probably converted to succinate, via methylmalonyl CoA, and hence via the tricarboxylic acid cycle to the clavulanic acid precursors.


Absorption, distribution and excretion of cefsulodin, an antipseudomonal cephalosporin, in mice, rats and dogs. A single dose of 20 mg/kg of cefsulodin [3-(4-carbamoyl-1-pyridiniomethyl)-7 beta-(D-alpha-sulfophenylacetamido)-ceph-3-em-4-carboxylate monosodium salt] was administered subcutaneously to mice, and intramuscularly to rats and dogs. The plasma and tissue levels reached the peak 15 approximately 30 minutes after administration. In mice and rats, no plasma levels were measurable 2 and 4 hours after administration. In dogs, the plasma levels were measurable 6 hours after administration. The level in the kidney of mice was slightly lower than the plasma level, while in rats and dogs, the level in the kidney was higher than the plasma level. The cefsulodin levels in the lung of rats and dogs were relatively high, and the level in mice was relatively low. The hepatic levels were very low in all test animal species. Cefsulodin was mainly excreted into the urine, and the excretion of cefsulodin into the bile was very slight.


Inhibition of mitochondrial functions by the antibiotics, bikaverin and duclauxine. Four cytotoxic antibiotics, bikaverin, duclauxine, PSX-1 and vermiculine, were examined with respect to their interference with glycolysis and respiration and their possible ionophoric or cytolytic activity. Duclauxine was found to be an inhibitor of respiration and bikaverin an uncoupler of oxidative phosphorylation of tumor cells and of isolated rat liver mitochondria. Bikaverin was also found to be an efficient haemolytic agent. Vermiculine and PSX-1 showed no effect in the systems employed.


High pressure liquid chromatography of zearalenone in cereals. A high pressure liquid chromatographic method has been developed for determining zearalenone in cereals. The sample is extracted with phosphoric acid and chloroform. The extract is cleaned up by washing on a silica gel column with benzene-hexane or cyclohexane-ethylene dicholride-ethyl ether and eluting with chloroform, and further purified by extracting into alkaline solution, washing with water, adjusting pH, and back-extracting into chloroform. Zearalenone is determined by using a liquid chromatograph with 2 columns in series packed with Spherisorb 10 micron ODS and with an ultraviolet detector set at 236 nm. The detection limit of the method is 2 microgram/kg. Recovery is about 90% when benzene-hexane is used in the washing step and about 80% when cyclo-hexane-ethylene dichloride-ethyl ether is used.


High pressure liquid chromatographic determination of aflatoxins in peanut products. A precise and sensitive high pressure liquid chromatographic method is proposed for determining aflatoxins B1, B2, G1, and G2 in all types of peanut products. The method is based on acidified aqueous methanol extraction, partition of aflatoxins into dichloromethane, and purification of the extract on a 2 g silica gel column. Aflatoxins in the purified extract are completely resolved on a microparticulate (10 micron) porous silica gel column in approximately 10 min with a water-saturated chloroform-cyclohexane-acetonitrile solvent. The preferred detection system, B1, and B2 by ultraviolet absorbance at 360-365 nm and G1 and G2 by fluorescence, allows accurate and sensitive detection of all 4 aflatoxins at levels as low as 0.3-1.0 microgram/kg. Repetitive assay of 3 samples of naturally contaminated peanut butters containing total aflatoxins (B1 + B2 + G1 + G2) at levels of 5, 10, and 15  microgram/kg gave within-laboratory coefficients of variation of 11, 5, and 5% respectively.


Gas-liquid chromatographic determination of T-2 toxin and diacetoxyscirpenol in corn and mixed feeds. A gas-liquid chromatographic method has been developed for the simultaneous determination of the T-2 toxin and diacetoxyscirpenol (DAS) in corn and mixed feeds. The analytes are extracted with aqueous methanol. Ammonium sulfate is used to denature and precipitate proteinaceous compounds and to salt out low polar compounds. The analytes are selectively concentrated into chloroform, which is washed with aqueous potassium hydroxide to remove acidic compounds. Residual interferences are removed by silica gel column chromatography. Heptafluorobutrylimidazole (HFBI) is added to form esters of the analytes. The HFB-esters are separated on an SE-30 glass column and measured witha 63Ni electron capture detector, using methoxychlor as an internal standard. The method has been applied to corn, livestock feeds, and pet food. The limit of detection is 100 ng T-2 toxin and 25 ng DAS/G. Average recoveries of 105 and 97% and coefficients of variation of 17 and 10% were obtained for T-2 toxin and DAS, respectively.


Confirmatory test for aflatoxin M1 on a thin layer plate. The identity of aflatoxin M1 can easily be confirmed directly on a thin layer plate by reacting aflatoxin M1 with trifluoroacetic acid (TFA). This confirmation reaction is carried out on the thin layer plate which has been developed in 2 dimensions and used for the quantitation of aflatoxin M1 in the sample. TFA is superimposed on the separated M1 spot. The plate is kept in the dark 3 min, heated to 75 degrees C for 5 min, and developed with chloroform-methanol-acetic acid-water (92 + 8 + 2 + 0.8). The Rf value of the blue-fluorescent derivative is compared with that for the M1 standard. The method was used successfully on extracts of milk, cheese, and liver. M1 quantities on the plate as low as 0.5  ng can be confirmed by this method. The method is also suitable for simultaneous confirmation of aflatoxin B1.


Headspace sampling and gas-solid chromatographic determination and confirmation of greater than or equal to 1 ppb vinyl chloride residues in polyvinyl chloride food packaging. The determination and confirmation of residual vinyl chloride (VC) in polyvinyl chloride (PVC) food packaging materials are described. PVC packaging materials are dissolved in dimethylacetamide (DMAC). VC is sparged from solution with helium gas and collected in sealed vials of ethanol. VC is detected and quantitated by using a headspace sampling technique and standard gas-solid chromatography (GSC) with flame ionization detection. GSC peak heights of about 5% full scale deflection are obtained for samples of PVC containing 1 ppb VC. Polymer samples taken from tubing, blood bags, food packaging films, bottles, and unprocessed resin were analyzed. Residual VC levels ranged from 0.3 to 913 ppb. VC was confirmed by GSC-mass spectrometry, using selected ion recording of m/z 62 and 64 in conjunction with full mass scans to identify components eluting near VC.


Gas-liquid chromatography and flame photometric detection of phosphine in wheat. A screening method using gas-liquid chromatography with  flame photometric detection has been developed for determining phosphine in wheat. Phosphine is measured as the sum of physically bound intact phosphine and that derived from residual aluminum phosphide. Wheat is extracted in a closed, partially evacuated glass system by refluxing with 10% sulfuric acid. Liberated gases are swept into a gas-collection flask fitted with rubber septa to permit gas sampling. Aliquots of collected gas are injected into a gas chromatograph. Phosphine is quantitated by peak area as determined by an electronic integrator. Recoveries varied with concentration: 67% was recovered at 0.10 ppm and 98% was recovered at 19 ppm. For concentrations less than 1.5 ppm, the coefficient of variation was 9.95%. Using flame photometric detection, phosphine concentrations as low as 0.04 ppm were easily determined in wheat.


Gas-liqiid chromatographic determination of pydrin, a synthetic pyrethroid, in cabbage and lettuce. Pydrin, alpha-cyano-3-phenoxybenzyl 2-(4-chlorophenyl)-3-methylbutyrate, a synthetic pyrethroid ester, is extracted from vegetables with acetonitrile, partitioned into petroleum ether before cleanup on a Florisil column, and analyzed by electron capture gas-liquid chromatography. Recoveries of 88-102% at 0.01 ppm, 99-102% at 0.1 ppm, and 98-104% at 1.0 ppm were obtained from fortified cabbage and lettuce samples. The method appears applicable to detection of the residues of Pydrin in the foliage of many types of crops.


Oil flotation extraction of light filth from ground capsicums excluding paprika: collaborative study. A collaborative study has been completed on an improved method for the isolation of light filth from ground capsicums other than paprika. The proposed method involves isopropanol pretreatment, wet-sieving, and extraction from cooled 60% ethanol with a mineral oil-heptane mixture. The collaborative tests by the proposed method showed an approximate 2-fold increase in recoveries of insect fragments and rodent hairs with acceptable coefficients of variation and clean filter papers. The proposed method has been adopted as official first action to replace 44.123.


Thin layer chromatographic method for the detection of uric acid: collaborative study. A collaborative study has been completed on an improved method for the detection and confirmation of uric acid from bird and insect excreta. The proposed method involves the lithium carbonate solubilization of the suspect excreta material, followed by butanol-methanol-water-acetic acid thin layer chromatography, and trisodium phosphate-phosphotungstic acid color development. The collaborative tests resulted in 100% detection of uric acid standard at the 50 ng level and 75% detection at the 20-25 ng level. No false positives were reported during tests of compounds similar to uric acid. The proposed method has been adopted official first action; the present official final action method, 44.161, will be retained for screening purposes.


Mineral oil extraction of light filth from rubbed sage: collaborative study. An improved method has been developed for extracting light filth in rubbed sage. The method is similar to 44.A04 and 44.A05, except the light filth is isolated by using 20% isopropanol with mineral oil. Collaborative results show that the proposed method yielded better recoveries than the present official first action method. The method has been adopted as official first action.


Fruits and fruit products. Non-parametric methods for detection of adulteration of concentrated orange juice for manufacturing. The composition of organic constituents (total sugars, reactive phenols, total amino acids, arginine, and gamma-aminobutyric acid) has been measured in a large (360 samples) selection of concentrated orange juice for manufacturing and orange pulp wash in the U.S. trade. The detection of adulteration with sugar, reducing sugars, and citric acid addition has been investigated by using non-parametric nearest neighbor classification techniques in the 4-space of log ratios of the compositions. The results show that such detection is possible with a type 1=type 2 error rate of 10% for 20% adulteration if at least 7 samples are taken. The assumptions of such samplings are discussed.


Improved microbiological assay for penicillin residues in milk and dairy products. The microbiological assay for penicillin residues in milk was improved. Acidification of milk with phosphoric acid to pH 4.5, centrifugation to remove precipitated proteinaceous matter, adjustment of pH to 6.0, a second centrifugation, and the use of a 2% agar base layer resulted in the consistent measurement of 0.01 unit penicillin activity/ml milk. Sarcina lutea was used as the assay organism. Recoveries in milk averaged 94%, in nonfat dry milk 84%, in cottage cheese 97%, and in cream cheese 94%.


Gas-liquid chromatographic determination of selenium in biological materials, using 4-bromo- and 4-chloro 1,2-diaminobenzene as derivatizing reagents. A method is described for the gas-liquid chromatographic determination of traces of selenium in marine biological materials. The method is based on the reaction of Se(IV) with bromo- and chloro-substituted 1,2-diaminobenzenes. The benzoselenadiazoles so formed are sensitive to electron capture detection. The sample is digested in a nitric-perchloric acid mixture and selenium is reduced to the IV oxidation state. Different aliquots of the digest solution are reacted with either 4-bromo- or 4-chloro-1,2-diaminobenzene to quantitatively form the corresponding 2,1,3-benzoselenadiazole. Recovery of added selenite to a fish meal sample was 95% for the bromo derivative and 101% for the chloro derivative. Different portions of a well mixed fish meal sample were analyzed in independent laboratories by the fluorometric method and by atomic absorption spectrophotometry (hydride generation). The following mean values (microgram/g) were found: present method 1.89, fluorometric method 1.91, atomic absorption method 2.1. The lower limit of detection for the method described was 13 ng, using the bromo derivative, and 27 ng, using the chloro derivative.


Leak in evaporated milk by anodic stripping voltammetry and atomic absorption spectrophotometry: cooperative interlaboratory study. The results of a cooperative study on the determination of lead in evaporated milk, using a double blind referee technique, are reported. This study was designed to determine the normal variability of methods currently used for lead analysis by canned food industry laboratories. Twenty-three laboratories participated in this study. Each laboratory was instructed to use atomic absorption spectrophotometry (AOAC 25.065), anodic stripping voltammetry, or carbon rod atomic absorption spectrophotometry. Overall, the results appear to be in close agreement with the spiking levels. The coefficient of variation for all laboratories was 36.0% at the 0.15 ppm lead level and 16.8% at the 0.40 ppm lead level.


Spectrofluorometric determination of atropine-hyoscyamine in tablets and powders. A method is presented for determining atropine-hyoscyamine alkaloids in tablets and powders by spectrofluorometry. The sample is first shaken with 0.1N H2SO4 solution; then and aliquot containing about 0.1 mg alkaloid is transferred to a separatory funnel, made basic with NaOH solution, and extracted with CHCL3. The extract is added to a CEelite-092N H2SO column and the eluate is discarded. The alkaloids are then eluted from the column with ammoniacal CHCl3 and the eluate is evaporated to dryness. The residue is dissolved in CHCL3, EOSIN Y solution is added, and the solution is mixed. Aliqutos of a reference solution of atropine are similarly treated and a blank is prepared from CHCl3 and eosin Y solution, All solutions are scanned with a recording spectrofluorometer in the emission mode from 440 to 700 nm and the excitation set at 475 nm. The emission peaks near 550 nm are used for quantitative estimation of the alkaloids. The method cannot generally be used in the presence of other organic bases but is applicable in the presence of barbiturates.


Differential pulse and AC polarography of reserpine, using an aprotic organic solvent system. Previously demonstrated advantages of aprotic organic solvent for analysis of organic pharmaceuticals have been applied to the assay of reserpine tablets. A procedure using high sensitivity electroanalytical relaxation methods was developed which is simpler and more sensitive than the USP assay. Most data were obtained with a commercial differential pulse polarograph which is widely available. However, less extensive measurements with other instruments established the general applicability of the differential pulse method, as well as various acpolarographic techniques. The results of differential pulse polarographic (DPP) and USP assays of synthetically formulated reserpine tablets compare favorably. The DPP method yields an average recovery for 0.1 mg reserpine/tablet of 100%, with a range of 97.6-102.3%. Corresponding data for 0.25 mg reserpine/tablet are 99.3%, with a range of 97.6-100.6%. Satisfactory results also were obtained in content uniformity studies, using single 0.1 and 0.25 mg tablets.


Fluorometric determination of metaraminol bitartrate injection. A fluorescence assay for metaraminol bitartrate injection has been developed which uses o-phthalaldehyde as a fluorogenic reagent, and dose not require prior isolation of metaraminol. It is 50 times more sensitive than the USP assay. Six analyses of 2 vials of the same lot of metaraminol bitartrate injection gave 96 +/- 0.4% of label declaration.


Semiautomated method for determining ferrous sulfate in pharmaceuticals: collaborative study. A semiautomated colorimetric method is described for determining ferrous sulfate in tablets, capsules, and elixirs. The method involves the formation of an orange-red complex between 1,10-phenanthroline and ferrous ion. Collaborators were supplied with 3 solid composites and one liquid sample. Two of the solid composites were prepared from commerical tablets of different dosage and one from commercial timed-release capsules; the fourth sample was an elixir. The results of the automated analyses for ferrous sulfate agreed well with those of the applicable USP method. No interference was found from ferric iron or from the red and green coloring used on coated tablets. The method has been adopted as official first action.


Determination of fluoride in deboned meat. A rapid analytical procedure was investigated for measuring the fluoride content of deboned meat, using a specific ion electrode. The method involves a defatting procedure followed by chelation with EDTA before final solubilization and measurement. Fluoride content is determined by comparison to a standard curve on 2-cycle semi-logarithmic graph paper of fluoride ion vs. millivolt readings. The method is applicable to hand and mechanically deboned pork, beef, and poultry within a range of from 0.7 to 25.0 ppm fluoride. Higher concentrations may be measured by analyzing additional standards and extending the curve into a third cycle. The curve is not linear below 0.7 ppm. Samples less than 0.7 ppm may be estimated by extrapolation. Studies with fortified samples indicate average recoveries of 92 +/- 4.2%. The standard deviation of duplicates within the laboratory is 0.5 ppm in the range stated above.


Survey for aflatoxin B1 in chicken eggs. Samples of egg products were obtained during January and July 1977 from 35 establishments located in the southern part of the United States. Of the 112 samples analyzed, aflatoxin B1 was found in 1 sample of liquid egg white at a level of 0.06 ng/g. No aflatoxin was found in 101 samples of shell eggs offered for sale to consumers in Alabama, Georgia, South Carolina, and North Carolina in the late fall of 1977.


Improved method for determination of light filth in chocolate products. The recovery of rodent hairs from chocolate has been significantly improved by the introduction of an additional defatting step, substitution of 40% isopropanol for water, and substitution of mineral oil-heptane (85+15) for heptane in the trapping-off step. These changes have no adverse effect on insect fragment recovery. An average recovery of 95% was obtained for rodent hairs, coefficient of variation 7.9%. Insect fragment recoveries were 100%.


Mitochondrial biogenesis during fungal spore germination: effects of the antilipogenic antibiotic cerulenin upon Botryodiplodia spores. Germination of spores of the fungus Botryodiplodia theobromae was inhibited by the antilipogenic antibiotic cerulenin. The spores remained viable in the presence of the antibiotic, however, and after prolonged incubation they were able to overcome the inhibition. Cerulenin inhibition of germination was reversed by Tween 40 and Tween 60 (derivatives of palmitate and stearate, respectively), but not by representatives of a range of free fatty acids or their soaps. Cerulenin abolished incorporation of [14C]acetate into sterols and triglycerides and reduced its incorporation into fatty acids by 69%. Cyanide-sensitive oxygen consumption by spores incubated in the presence of cerulenin was greatly reduced throughout germination, and the activity of cytochrome c oxidase was no more than 13% of the activity in untreated spores, even after prolonged incubation. However, low-temperature difference spectra of mitochondrial extracts showed that the cerulenin-treated spores accumulated a threefold excess of cytochrome a, whereas the cellular concentrations of cytochroms c and b were identical to those of untreated spores. Cerulenin treatment sharply reduced the rates of whole spore protein and RNA synthesis. Cerulenin had no effects upon mitochondrial morphology which could be discerned with an electron microscope.


Biodegradation of acyclic isoprenoids by Pseudomonas species. The ability of various pseudomonads to utilize acyclic isoprenoids as a sole carbon source was investigated. Tests for utilization of acyclic isoprenols such as citronellol and geraniol were complicated by toxic effects of these alcohols, and most species tested were killed by exposure to citronellol or geraniol (0.1%, vol/vol) in liquid culture. In the case of Pseudomonas citronellolis, sensitivity to isoprenols is reduced by prior induction of the isoprenoid degradative pathway via either growth on succinate in the presence of citronellol or growth on citronellic acid. For this species, citronellic acid proved to be the best isoprenoid growth substrate tested. Geraniol utilization as a taxonomic indicator for different subgroups of pseudomonads is discussed. Only a few of the species tested were able to utilize acyclic isoprenoids. Two species which utilize C10 acyclic isoprenoids, P. aeruginosa and P. mendocina, were shown to contain the inducible enzyme geranyl-coenzyme A carboxylase, one of the unique enzymes in the isoprenol degradative pathway known to occur in P. citronellolis. Of the species which utilized geranitol, none showed definite growth on the homologous C15 and C20 isoprenols.


Effect of cultural conditions on the concentrations of metabolic intermediates during growth and sporulation of Bacillus licheniformis. Intracellular concentrations of adenine nucleotides and intermediates of the Embden-Meyerhof pathway and the tricarboxylic acid cycle have been determined during growth and sporulation of Bacillus licheniformis in a variety of different media. The ATP pool was independent of growth rate and nitrogen source, but the use of glucose as a carbon source resulted in a twofold elevation in the ATP pool during exponential growth. The intracellular phosphoenolpyruvate pool was at least twofold higher during gluconeogenesis than during glycolysis. The finding that the use of glutamate as the sole nitrogen source resulted in at least a fivefold elevation of the alpha-ketoglutarate pool suggests a role for alpha-ketoglutarate in the repression of the enzymes of the tricarboxylic acid cycle responsible for alpha-ketoglutarate synthesis. Not one of the metabolites assayed appears to function as a signal of the nutrient deprivation which accompanies the initiation of sporulation.


Membrane phospholipid asymmetry in Bacillus amyloliquefaciens. The phospholipid distribution in the membrane of Bacillus amyloliquefaciens was studied by using phospholipase C (B. cereus), phospholipase A2 (Crotalus), and the nonpenetrating chemical probe trinitrobenzenesulfonic acid. After treatment of intact protoplasts of B. amyloliquefaciens with either phospholipase, about 70%  of total membrane phospholipid was hydrolyzed; specifically, about 90, 90, and 30% of phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin, respectively. Under these conditions, protoplasts remained intact and sealed. However, when protoplasts that were permeabilized by cold-shock treatment were incubated with either of the phospholipases, up to 80% of cardiolipin was hydrolyzed and phosphatidylglycerol and phosphatidylethanolamine were hydrolyzed virtually to completion. In intact cells, 92% of the phosphatidylethanolamine could be labeled with trinitrobenzenesulfonic acid under conditions in which the reagent did not penetrate the membrane to any significant extent. These results indicate that 70% of total phospholipid of this bacillus exists in the outer half of the bilayer. The distribution of phosphatidylethanolamine in this bilayer is highly asymmetric with it being located predominantly in the outer half. The results with phospholipases suggest that the distributions of cardiolipin and phosphatidylglycerol are also asymmetric but independent confirmation of this is required.


Presence of glycerophospholipid: cholesterol acyltransferase and phospholipase in culture supernatant of Aeromonas hydrophila. Human erythrocyte membrane glycerophospholipids are deacylated by Aeromonas hydrophila 13-h culture supernatants, resulting in the production of cholesterol ester, free fatty acid, and water-soluble phosphates. This activity appears to be due to the actions of an acyltransferase (phosphatide:cholesterol acyltransferase, EC 2.3.1 group) and a phospholipase (phosphatide acyl-hydrolase). The enzyme activities are produced simultaneously in late exponential/early stationary phase, are precipitated together from the culture supernatant with 85% ammonium sulfate, and are eluted together near the void volume during gel filtration on Sepharose 6B. These results suggest that A. hydrophila produces a multienzyme complex with an unusual mode of action on membrane lipids. The complex is distinct from the hemolytic factor aerolysin, which is also produced by A. hydrophila.


Bacterial xanthine oxidase from Arthrobacter S-2. Arthrobacter S-2, originally isolated by enrichment on xanthine, produced high levels of xanthine oxidase activity, requiring as little as a 20-fold purification to approach homogeneity with some preparations. Molecular oxygen, ferricyanide, and 2,6-dichlorophenol-indophenol served as electron acceptors, but nicotinamide adenine dinucleotide did not. The enzyme was relatively specific when compared with previously studied xanthine-oxidizing enzymes, but at least one purine was observed to be oxidized at each of the three positions of the purine ring that have been subject to oxidation by this type of enzyme. The enzyme had a relatively high Km for xanthine (1.3 X 10(-4) M), and substrate inhibition was not observed with this compound, in contrast to the enzyme from cow's milk. In fact, an opposite effect was observed, and double-reciprocal plots with xanthine as the variable substrate showed a concave downward deviation at high concentrations. At 2.5 mM xanthine the enzyme had a specific activity approximately 50 times that of the most active preparations of the milk enzyme. The spectrum of the Arthrobacter enzyme resembled that of milk xanthine oxidase, suggesting a similarity of the prosthetic centers of the two enzymes. The bacterial enzyme was relatively small and may be dimeric, with approximate native and subunit molecular weights of 146,000 and 79,000, respectively.


Differences in incorporation of nucleic acid bases and nucleosides by various Mycoplasma and Acholeplasma species. Eight species representative of the serological diversity of the Mycoplasmatales were tested for their ability to incorporate radiolabeled nucleic acid precursors into acid-insoluble material. Cultures in complex growth medium were centrifuged and resuspended in minimal essential medium (Eagle). For Acholeplasma laidlawii, labeling occurred mainly during the first 4 h of incubation, with substrate saturation at 20 micron. All organisms tested incorporated uracil, adenine, and guanine; none incorporated cytosine. Thymine was incorporated only by bovine group 7, Mycoplasma putrefaciens, and Mycoplasma pneumoniae (strain 3546), but deoxynucleosides enhanced thymine incorporation in A. laidlawii, Mycoplasma gallisepticum, M. pneumoniae (strain AP-164), and Mycoplasma hyorhinis. Nucleoside incorporation (adenosine, guanosine, uridine, cytidine, and thymidine) was not observed for the arginine-utilizing species, Mycoplasma hominis and Mycoplasma arginini, whereas all other organisms tested incorporated nucleosides. The incorporation pattern provides additional metabolic evidence to support the biochemical and antigenic diversity of these organisms. The recognition of differences in incorporation of nucleic acid precursors is important not only to the specific labeling of these organisms, but also to the study of metabolism and transport.


Isolation and characterization of morphogenetic mutants of Arthrobacter crystallopoietes. Mutants of Arthrobacter crystallopoietes that exhibited altered ability to undergo the normal sphere-to-rod-to-sphere morphogenetic cycle were isolated. The procedure used to isolate these mutants involved velocity sedimentation in a sterile sucrose gradient to separate morphogenesis-deficient spherical cells from rod-shaped cells capable of normal morphogenesis. Three classes of mutants were obtained: (i) those that cannot form rods, (ii) those that cannot form long rods, and (iii) those that form long rods but exhibit more extensive rudimentary branching than the wild type. The isolation and characterization of these mutants are described, and the use of these mutants in the study of the morphogenetic cycle of arthrobacters is discussed.


Temperature-sensitive multicellular mutants of Wangiella dermatitidis. Three temperature-sensitive morphological mutants of Wangiella dermatitidis were isolated and characterized. The mutants grew in the yeastlike morphology at the permissive temperature (25 degrees C) but expressed a multicellular (Mc) phenotype at the restrictive temperature (37 degrees C). Cultures of Mc 2 and 3 incubated at the restrictive temperature showed rapid reductions in the percentage of budded cells in the population. In contrast, budding continued for several generations in cultures of Mc 1. Incubation of cultures of Mc 2 and 3 at the restrictive temperature for 48 h resulted in nearly total conversion of yeastlike cells to the multicellular form; about 50% of the cells of Mc 1 had converted to multicellular forms after 48 h at the restrictive temperature. Studies using radiolabeled compounds documented that DNA, RNA, and protein synthesis continued at the restrictive temperature. The results suggest that multicellularity is the result of inhibition of bud emergence and cell separation without inhibition of growth nuclear division, and cytokinesis.


Morphological and cell association characteristics of Rochalimaea quintana: comparison of the Vole and Fuller strains. The vole and Fuller strains of Rochalimaea quintana were grown on monolayers of mouse L cells irradiated 7 days previously and examined by light microscopy and scanning and transmission electron microscopy. Most of the bacteria of both strains were shown to adhere to the L cells but remained in an extracellular location. Cell division was frequently seen among the extracellular bacteria. The few intracellular bacteria seemed to be within vacuoles and did not multiply. Attachment to the eucaryotic cell did not seem to involve pili or other bacterial surface structures. The dimensions of the bacteria were approximately 0.45 micron in width by 1.0 to 1.7 micron in length. The cell envelope consisted of the usual trilaminar cell wall and plasma membranes separated by a layer of low electron density, as found in other gram-negative bacteria. No significant differences between the vole and Fuller strains either in morphology or relationship to eucaryotic cells were encountered.


Ultrastructural and physiological changes occurring upon germination and outgrowth of Azotobacter vinelandii cysts. Dormant cysts of Azotobacter vinelandii germinated at 30 degrees C in Burk nitrogen-free media containing 1% glucose. Samples taken at intervals and examined by electron microscopy revealed that as germination progressed, vesicle-like and fibrillar structures became visible in the intine region. Lamellae associated with the cell membrane appeared in the central body at 6 h post-initiation of germination. Both electron micrographic and chemical analysis showed that the poly-beta-hydroxybutyrate content of cysts decreased significantly after 4 h of germination. Dormant cysts were resistant to sonic oscillation, but this property was lost during their conversion to metabolically active vegetative cells.


Rate of major protein synthesis during the cell cycle of Caulobacter crescentus. The rate of major protein synthesis was examined during the synchronous differentiation of Caulobacter crescentus. Total cell proteins were pulse-labeled with [35S]methionine at different times in the swarmer cell cycle and analyzed by sodium dodecyl sulfate- polyacrylamide gel electrophoresis. The rates of synthesis of total cell proteins and of about one-half of the individual major proteins examined increased through G1 and S periods but remained nearly constant during G2 period. The rates of synthesis of the other half of the individual major proteins either increased continuously throughout the swarmer cell cycle or doubled during S period. One stage-specific protein was also detected in late S period. For most of the major proteins examined, the rate of synthesis in the swarmer cell was less than that in the stalked cell. It seemed that, before the onset of G2 period, the Caulobacter cell was already able to synthesize each major protein at the additive rate of the two progeny cells. Compared to the stability of cellular proteins, the functional degradation rate of mRNA coding for individual major proteins was rapid, with half-lives of 0.4 to 5.8 min. It thus seems that the rate of major protein synthesis mainly reflects the transcriptional control of gene expression.


Location of ribosome-binding sites on the tetracycline resistance transposon Tn10. Eight ribosome-binding sites were located on the single-stranded Tn10 DNA loop which was formed after denaturation of lambda phage DNA containing the Tn10 transposon sequence. Ribosomes were bound only to the Tn10 loop contained on the R strand of lambda DNA but not to that on the L strand, suggesting that one of the two strands of Tn10 DNA is selectively transcribed. Six of the eight ribosome binding sites were located in one-half of the DNA loop. The maximum sizes of potential polypeptides were calculated for these genes to range between 9,500 and 84,000 daltons.


Effect of guanosine tetraphosphate on in vitro protein synthesis directed by E1 and E3 colicinogenic factors. The in vitro synthesis of colicin E3 was found to be stimulated by guanosine 5'-diphosphate 3'-diphosphate (ppGpp), while that of several other ColE3 plasmid-specific proteins was reduced in the presence of this nucleotide. The ColE1 plasmid-directed in vitro synthesis of colicin E1 was also found to be stimulated by ppGpp.


Living with the characterologically altered brain injured patient. Characterological defects secondary to brain injury tend to disrupt normal family interaction patterns and create adjustment problems for the patient's close family members. Social and psychological ties, expectations, and vulnerabilities make caretaking spouses and dependent children particularly susceptible to emotional stresses. Unrealistic expectations often compound these problems. Almost all family members experience some depression. Counseling family members can improve the quality of their adjustment and their care of the patient. Counseling goals include helping the family readjust expectations, providing practical management advice, and alerting family members to their own needs. Specific recommendations for dealing with common family problems are offered.


Diagnostic delay in presenile dementia.

Diagnostic delay in presenile dementia. In a retrospective study, patients with presenile dementia and histories of symptoms of depressive illness and of antecedent diagnosis of depressive disorder were found to have significantly longer intervals from initial physician contact to final diagnosis than patients without such histories. Patients with histories of psychiatric treatment prior to final diagnosis also had longer intervals, with differences approaching statistical significance. In contrast, intervals from symptom onset to first physician contact were noted to be shorter for those patients with these antecedent histories. A discussion of these results, a review of the relevant literature and an illustrative case report are presented.


Indicators of E.C.T. response and non-response in the treatment of depression. Two matched groups of depressed patients, one ECT responsive and one ECT non-responsive, were compared for clinical, historical and treatment variables. Three factors predicted good response: (1) duration of illness less than one year; (2) presence of a precipitating event; (3) presence of delusions. A trend suggested that concurrent physical debilitation was a negative indicator. Possible explanations for these findings were proposed.


Lithium in diabetes mellitus: a paradoxical response. An unusual hyperglycemic effect of lithium was observed in a patient suffering from diabetes mellitus and mania. In the presence of constant insulin dosage, high serum lithium levels were correlated with high fasting bloos sugar, while lowering of the lithium level led to a lowering the fasting blood sugar. These observations are discussed in terms of the effects of lithium on glucose metabolism and with particular emphasis on the ability of lithium to increase the release of glucagon.


Capgras syndrome: case report of an adolescent and review of literature. Case report of an adolescent with Capgras Syndrome in the absence of organic brain syndrome is presented. Four cardinal features associated with Capgras Syndrome--(1) moderate-to-severe anxiety or stress, (2) ambivalent love-hate relationships with important love objects, (3) body image distortion, and (4) rejection-abandonment phenomena--were developed vis-a-vis the developmental tasks of adolescence. The literature consisting of 75 case reports is reviewed from the standpoint of differential diagnosis especially with respect to organic brain syndromes and schizophrenia.


Toxoplasmosis masquerading as a psychotropic side effect. When treating a patient with neuroleptics or tricyclic antidepressants, it is usually assumed that complaints of blurred vision can be ascribed to the anticholinergic side effects of these drugs. The authors present a patient treated with imipramine and trifluoperazine whose complaints of blurred vision led to the diagnosis of toxoplasma chorioretinitis.


A comparison of the onset of action and therapeutic efficacy of amoxapine and amitriptyline. A total of 61 moderately to severely depressed outpatients were treated for four weeks with either amoxapine (a dibenzoxapine tricyclic) or amitriptyline. This double blind study showed that amoxapine was as effective as amitriptyline and had an earlier onset of action. Maximum doses used were 300 mg of amoxapine and 150 mg of amitriptyline. Side effects were similar for the two drugs, except for impotence or loss of libido in eight male amoxapine, as against three amitriptyline patients. Laboratory, EKG, and vital signs findings showed no pathological trends.


Incest in psychiatric practice: a description of patients and incestuous relationships. Patients reporting incest resembled the author's other patients in age, sex marital status, education, residence and social class. Both initiating and receptive partners were represented among these patients. In two-thirds of 32 patients, incest had occurred more than 1 year previously. No one clinical picture predominated. Thirty-two patients had been involved in 42 incestuous relationships. Initiating partners, always older, included 32 men and 10 women. Coitus was the most frequent sexual activity; homosexual relations were present in 4 instances. Adults initiating incest had usually lost access to adult sexual partners by death, divorce, illness, or marital partners' refusal to cohabit.


Common errors in the treatment of suicidal patients. As a suicidology consultant to a county coroner and to several hospitals, the author has noted 5 common errors by the mental health caretakers in the treatment and management of suicidal patients. Prototypical cases of these errors are presented and discussed along with suggestions for prevention.


Role interference: an analysis of suicide victims, homicide offenders, and non-violent individuals. The relationship between role interference and suicidal, homicidal and non-violent behavior is investigated. Interference obtains to the extent that enactment of certain roles tends to impede the enactment of others. High degrees of interference in the performance of roles is generally frustrating to individuals and is a potential generator of outwardly directed violence. Low degrees of role interference is likewise disconcerting to persons but in the absence of an external target for aggression may lead to self deprecation and ultimately suicide. Detailed life histories of 98 suicide victims, 62 homicide offenders, and 76 non-violent individuals having no history of homicidal or suicidal behavior were studied. Homicide offenders suffered a higher incidence of role interference than the suicide or non-violent samples. Suicide victims tended to experience low degrees of such interference.


Portrait of a pusher--"mother".

Portrait of a pusher--"mother". The authors describe a case history of a drug pusher who has represented a special problem for their drug treatment program, since he functions symbiotically with this program, and is intimately involved with its clientele. Such a development, in the course of a drug treatment program, has implications for the practicality of the treatment approach. Methods of coping are discussed.


Imipramine and desipramine plasma levels: relationship to dosage schedule and sampling time. Tricyclic antidepressant plasma levels were determined after administration if imipramine (11 patients) and desipramine (ten patients) as a single daily bedtime dose or in divided doses. Plasma levels of the tricyclics were relatively stable on both dosage schedules due to the long plasma half-lives of these drugs. No difference was found between the two dosage schedules and the therapeutic effectiveness or the severity of side effects. The majority of patients preferred the single dose schedule. These findings support the use of a single daily dose of imipramine and desipramine.


Acute phencyclidine (PCP) intoxication: psychopathology and prognosis. In 15 patients hospitalized with phencyclidine (PCP) psychosis, several measures of psychopathology were examined in relationship to urine PCP levels, duration of hospitalization and mode of intoxication. Duration of hospitalization was found to be significantly shorter for smokers than for ingesters of PCP. Impaired ability to estimate 30 seconds duration was found to correlate significantly with a higher urine PCP level and a longer hospitalization. No other measure of psychopathology was found to correlate with either duration of hospitalization or urine PCP levels, now were other measures of psychopathology found to distinguish these patients from patients with acute functional mental illness.