Evaluation of breath alcohol instruments. I. In vitro experiments with alcolmeter pocket model. An Alcolmeter Pocket Model breath alcohol device, on an electrochemical (fuel cell) oxidation principle for ethanol analysis, has been evaluated under in vitro conditions. The result of a test is displayed on an analogue meter within 20--30 seconds after sampling: replicate tests may be made within 3--5 minutes. The electrochemical detector used was found to respond to acetaldehyde, methanol, isopropanol and n-propanol vapours besides ethanol, but it was insensitive to acetone vapour. The Alcolmeter response with a 0--2.0 mg/ml scale was linearly related to ethanol vapour concentration up to 1.0 mg/ml blood alcohol equivalent concentration; above this level the response was curvilinear, the Alcolmeter reading being too low. The standard deviation of an ethanol vapour determination in vitro was +/-0.0175 mg/ml at a mean concentration of 0.902 mg/ml. The accuracy of the device expressed as percent recovery at 0.50, 1.0 and 1.4 mg/ml blood alcohol concentrations was 96.8%, 98.3% and 88.3%, respectively. When the Alcolmeter was was calibrated at 0.50 mg/ml and used occasionally each day over an 18-day period, the drop in initial calibration was 0.01 mg/ml per week.


Evaluation of breath alcohol instruments. II. In vivo experiments with alcolmeter pocket model. The precision and accuracy of an Alcolmeter Pocket Model breath alcohol instrument have been investigated in experiments with human subjects under controlled conditions. The instrument response was zero in all tests with breath samples from alcohol-free subjects. The standard deviations of ethanol determinations in breath were +/-0.0722 mg/ml during ethanol absorption and +/-0.416 mg/ml during ethanol elimination. The standard deviation during the elimination phase increased with ethanol concentration in the sample, being +/-0.0416 mg/ml on average at a mean concentration of 0.420 mg/ml, corresponding to a coefficient of variation of 9.9%. The blood alcohol estimates using the Alcolmeter were somewhat too high during active absorption of ethanol, and too low during elimination, when a constant blood-breath alcohol ratio of 2100:1 was used to calibrate the instrument. During the elimination phase of ethanol kinetics and at a mean blood alcohol concentration of 0.50 mg/ml, the mean Alcolmeter result was 0.456 +/- 0.169 mg/ml with 95% confidence, i.e. varying between 0.287 and 0.625 mg/ml 95 times out of 100 tests at this critical blood alcohol level


Biochemical reconstruction of three cases of death--results of international cooperation. Biochemical serotonin and histamine determinations were applied to the reconstruction of three suspected homicide cases. To distinguish between ante-mortem and post-mortem wounds and to time the ante-mortem injuries the concentrations of free histamine and serotonin in the wound samples and in the control samples from neighbouring intact skin were examined. The results of these biochemical determinations allowed a reconstruction of the events and one of the three cases was shown to be suicide instead of homicide. The methods can be used at least during the first 4--5 days after death and sometimes even longer. This allows for international cooperation when investigating and reconstructing complicated cases of death.


Post-mortem hypoxanthine levels in the vitreous humour. An introductory report. Post-mortem hypoxanthine levels in vitreous humour were determined in 86 consecutive legal autopsy cases. In cases of sudden death caused by trauma or by myocardial infarction, levels ranging from 0 to 540 mumol/l were found. The mean value was about ten times higher than normal in vivo plasma levels. The hypoxanthine levels seem to be independent of time post-mortem, at least during the first 48 hours. It is known that augmentation of the hypoxanthine plasma, cerebrospinal fluid, and urine levels reflects tissue hypoxia. In the present material no elevation of hypoxanthine levels in the vitreous humour was found in cases of strangulation or suspension, while statistically significant elevation was found in cases of drug intoxication. It is concluded that this may reflect the effect of drug-induced prolonged tissue hypoxia caused by respiratory depression.


The possible effect of hashish on leukocytes and plasma lipids. The possible fluctuation of leukocytes and plasma lipids, 30--60 min after smoking hashish, was studied. Total phospholipid content in both leukocytes and plasma was increased in a similar way to the total lipid content after smoking hashish. Differences in most of the phospholipid classes in leukocytes and plasma before and after smoking hashish were observed while the values in controls and chronic users of the drug before smoking hashish were found to be relatively close. Findings are discussed in relation to the pharmacological action of the drug on liver lipid metabolism.


Isolation of drugs from blood and tissues with XAD-2 bags. Nylon bags containing 2-g portions of Amberlite XAD-2 resin were used for systematic analysis of drugs in biosamples. The procedure requires 10 or less grams of material, two XAD-2 bags, and enables rapid and economical isolation of most common drugs. The method was demonstrated on autopsy blood spiked withe 19 of the most common drugs, and was routinely used in cases of fatal and non-fatal poisoning. The eluates were clean and suitable for direct gas chromatographic and ultraviolet spectrophotometric analysis. The procedure used appeared more convenient than XAD-2 column extraction procedures. Classic solvent extraction methods were usually less efficient.


[Adipose tissue and obesity. Part 1: fat cell size and fat cell number]. The role of the number and size of adipocytes in the pathogenesis of obesity is reviewed. The observation that hyperplasia of adipose tissue is associated with child-onset obesity was over-interpreted. Nevertheless most recent studies suggest that fact cell size and number are regulated and can influence food intake. The discovery of preadipocytes has opened a new field or research offering the possibility of new concepts in the prevention and therapy of obesity.


[Is the Atkins diet safe in respect to health?]. The "Atkins dietary revolution" is a ketogenic diet consisting almost exclusively of food from animal origin. A similar diet was introduced as the "Banting diet" more than a century ago by the English physician Harvey. This carbohydrate-free diet is high in fat, cholesterol and purines. The measurable ketosis is a precondition for the effect of this kind of diet. However, the predictable hyperlipacidemia and ketosis are recognized health risks. Additionally, hypercholesterolemia is to be expected in a greater part of the adherents to such a diet. Even children under ketogenic diet develop hypercholesterolemia within a short time. A similar high-fat diet, known as the Sippy diet for ulcer therapy, was found earlier to cause an increased incidence in coronary heart disease. On the basis of the known facts the Atkins diet seems to be potentially hazardous to health, unless a controlled study is performed. However, considering the risks it seems very problematic to perform such a study.


[Prevention of special complications in gynecologic operations. Part 2. Prevention of adhesions in the peritoneal region, prevention of wound healing disorders]. Own experiences in the prevention of certain complications of gynecological operations are presented. Prophylaxis proved to be possible intraoperatively in capillary bleedings and infections as well as postoperatively in disorders of intestinal motility, peritoneal adhesions and healing disturbances. The successful treatment of our patients in 1976 (1583 major operations including 158 patients at high risk) is summarized in suggestions for therapy.


[Percutaneous kidney biopsy. Evaluation of a diagnostic method]. The evaluation of the results of nearly 800 percutaneous renal biopsies, including biopsies in which insufficient renal tissue was obtained or histologic changes were non-specific, indicated that in 85% of the cases a positive diagnosis could be made. The liberal extension of the indication to percutaneous renal biopsy to include oligosymptomatic renal diseases, the nephrotic syndrome and acute renal failure often resulted in therapeutic and prognostic consequences. Renal biopsy does not facilitate the diagnosis of pyelonephritis. Uremia, severe atherosclerosis, small kidneys, advanced age and lack of cooperation are not contraindications to percutaneous renal biopsy nor do they increase its risk. Severe complications are extremely rare and are always secondary to retroperitoneal hemorrhage. Close observation and prompt treatment can always preclude a fatal outcome. Long-term complications are not to be expected. If the technique of percutaneous renal biopsy and its histologic evaluation are efficiently performed, further extension of the indications to biopsy could be medically sanctioned.


[Behavior of the fasting blood sugar values in diabetics treated with oral and parenteral pentoxifylline]. Fasting blood sugar levels in hospitalized diabetics with peripheral vascular disease receiving pentoxifylline i.v. and p.o. did not-in comparison to a control group-show statistically significant changes during the observation period in well-controlled, stable diabetes mellitus (BS is less than or equal to 130 mg%); a decrease did occur in hyperglycemic patients. As a hypoglycemic drug effect was not demonstrable in the well-controlled patient-group, the decrease might well be attributable to improved dietary and antidiabetic care during the hospital-stay.


[Allergic alveolitis. Diagnosis and therapy in practice]. The diagnostic steps for the disease are listed and are discussed. The diagnosis includes: 1. the anamnesis, 2. the clinical examination, 3. the chest X-ray, 4. serological-immunological methods of examination including detection of antibodies, skin tests, allergen inhalation test, 5. lung function tests. The therapy consists in antigen exclusion and corticosteroids.


[Perbronchial lung biopsy and respiratory function tests]. 27 patients with diffuse pulmonary disease according to X-ray findings were examined by lung function and perbronchial lung biopsy. The lung function tests are compared with the extent of impairment of lung function expected according to lung specimen obtained by perbronchial biopsy. It is shown that lung function is a good indicator if the lung specimen is representative for the disease, and that the results of perbronchial biopsy mostly are representing the disease. The use of perbronchial biopsy is discussed and shown in some cases.


[Juvenile hip pain. 2. Femur head epiphysiolysis, hip dysplasia, tumors]. The early symptom in hip joint diseases in children is pain. Pain is localized in the groin and thight, but mostly in the knee. Other important signs are limping and reduced internal rotation. If a hip disease is suspected it is necessary to take X-rays in two planes. If diagnosis is early and special therapy started immediately, the results are usually excellent without deformation of the hip. Otherwise early osteoarthritis can develop. This is important because osteoarthritis in the hip joint is in 75% of the cases due to hip joint diseases in childhood. The problems of diagnosis and treatment of the most common hip joint diseases in children (transient synovitis, rheumatoid arthritis, osteomyelitis, Legg-Perthes disease, slipped capital femoral epiphysis, dysplasia, tumors) are discussed.


[Death and dying in our time].

[Death and dying in our time]. As modern technical medicine more and more progresses, more old and sick people die in hospitals. Therefore, there arise new tasks of dealing with dying people and their families, which the doctors and nurses are not sufficiently prepared for. The question of telling the truth to the patient and accompanying him during the process of dying is a problem of communication between doctor and patient, connected with many emotional conflicts not only for the dying patient, but also for doctor and nurses and people taking care of him. Since the management of dying people is often inhuman, it is necessary to integrate the subject of "death and dying" into the education of doctors and nurses.


[Effect of fractures on the juvenile epiphysis]. In a clinical and experimental study two factors have been found to be most important for the increase of growth after fractures in childhood: 1. Trauma leads to changes of blood circulation in the epiphysis. Changes in oxygen tension cause morphometric alterations of cellular and intercellular constituents of the plate as well as measurable changes in thickness of the plate and length of the cartilage columns. These changes are relatively uniform and nearly independent of the kind of fracture and its treatment. Growth stimulation by fracture alone is moderate, limited to the time of fracture healing and hardly influenced by implantation of metallic material for osteosynthesis. 2. Angular deformity stimulates the epiphyseal plate to spontaneous correction. This correction is mostly stimulated by the change of pressure caused by the muscular coat and only minimally by static load. Correction of angular deformity can only happen under simultaneous increase in longitudinal growth, as long as pressure in the plate is changed. If angular deformity does not lead to changes in pressure by muscular tension on the concavity of the fracture (e.g. recurvation of tibia, varus in supracondylar fracture of humerus, torsion deformity of long bones) no stimulation of the epiphyseal plate will occur. In these cases the increase of growth is insignificant.


[Experiences with replantation of large limbs]. From November 1975 to August 1978 26 limb replantations were performed. All 5 lower extremities had to be reamputated. Of the 21 replanted arms 15 healed without major complications, 9 have regained sufficient function up to now; 3 others show signs of reinnervation.


[Prevention in general practice: what is dangerous for liver cells?]. The liver is engaged in detoxication and elimination of toxic-nutritional foreign material. In addition it is of high affinity to viruses and parasites. Different influences may cause pathological reactions, which are of great consequence to the normal and even more to the previously damaged liver. In general, interactions and cumulative effects of different toxic agents occur. The increase of chronic hepatic disorders in the last years has to be noticed considering economic and social medical aspects. At the first place of agents with a toxic effect to the liver there is alcohol, followed by hyperalimentation and malnutrition. Numerous drugs with different reactions to liver morphology, liver impairment caused by environmental influences and infections retain further places. Prophylaxis and therapy of toxic-nutritional liver damage is based on recognition and elimination of the noxes. Patients with chronic liver disease need a special management and consultation by their physician.


[Rehabilitation in ambulatory coronary groups]. In Western Germany anti-coronary clubs are a relatively new form of organized rehabilitation and secondary prevention after myocardial infarction. At the present time there are operating about 120 such groups, sponsored by hospitals, sports clubs, groups of family doctors, educational organizations and patient selfhelping groups. The groups divided in "good risk", "bad risk" and "preventive" group play an important role in a comprehensive rehabilitation program. Their aims are: changing the risk factors and life style, easing anxiety, depressions, denial and overmotivation, improving life quality, reintegrating the patients into social life, improving physical fitness. Multicenter studies are necessary to find out, if Anticoronary Clubs are able to lower the mortality rate and the reinfarction morbidity.


[BCG vaccination and leukemia. Epidemiologic studies on the effect of BCG vaccination on leukemia]. The possible influence of BCG vaccination on leukemia mortality of infants was investigated by various authors. The methods used were prospective trials, retrospective studies and epidemiological analyses. In some of these studies a protective effect of BCG vaccination was observed, in other studies no influence was found. Up to now this problem is subject to controversial discussions. An epidemiological analysis of the Austrian data using trend analyses, regression analyses and analyses of rank correlation showed an inverse correlation between BCG vaccination rate of newborns and leukemia mortality in the age group 0--5 years. From this correlation a protection rate of 0.77 could be calculated.


[Cytological smears in gynecological practice]. The technique of gynecological smear is discussed. Since poor results, i.e. false-negative findings, are frequently caused by an inadequate preparation, the possible errors in smear taking and their elimination are described. The validity of additional microbiological and cyto-hormonal findings in routine smear is pointed out.


[Post-pill amenorrhea and menarche].

[Post-pill amenorrhea and menarche]. After stopping hormonal contraception, some women have temporarily impaired fertility. In a study on more than 500 women, using the age of menarche as parameter, it was evaluated which would develop post-pill amenorrhea. It was found that women with early menarche are emotionally more stable, get higher school degrees and have a more positive state of mind regarding sexuality, pregnancy and birth than those with late menarche. All these factors should be considered in patient care.


[Possibilities of treatment in acute failure of the right heart]. Acute right heart failure can be influenced positively by means of right-sided respectively left-sided intraventricular balloon pump and the intraaortic counterpulsation. Yet, due to a narrow safety margin one should not try the right-sided intraventricular balloon pump in humans. Left-sided intraventricular balloon pumping immediately enhances cardiac output by 20--30%; the myocardial blood-flow to the subendocardium increases by 50%; finally the myocardial oxygen supply improves. Balloon inflation only during ejection time is demanded. So, up to 6 hours, properly timed left intraventricular balloon pumping does not damage left ventricular dimensions. The use of the intraaortic balloon pump was without problems. This pump device increases myocardial bloodflow to the subendocardium most effectively. There is a broad safety margin, too. Intraaortic counterpulsation should be started as soon as acute right heart failure is evident.


[Clinical and experimental studies on changes in lysosomal enzyme activity in fatty livers (author's transl)]. In order to examine a role of lysosomes in the pathogenesis of fatty livers, analysis was made on possible etiologic factors, clinical signs and symptoms as well as laboratory data of routine liver function tests in 32 subjects with fatty livers. Of 18 cases, enzyme activities of serum acid phosphatase (Acp), beta-glucuronidase (betaG) and n-acetyl-beta-glucosaminidase (nbetaG) were measured and compared with those obtained in 20 normal subjects. Subjective symptoms were observed in 75% of the cases examined, liver swelling in 56%, positive GOT, GPT and BSP retention were in 59, 75 and 68%, respectively. The activity of serum lysosomal enzymes such as Acp, betaG and nbetaG were significantly increased and their incidence was 28, 89 and 78%, respectively. In animal experiments, activities of these enzymes in both serum and liver homogenate were examined in rats with choline-deficient, ethionine-treated, and alcoholic fatty livers. Results obtained were as follows: 1) Lysosomal enzyme activity in sera and livers of choline-deficient rats showed a significant decrease in lysosome-rich fraction and a significant increase in supernatant fraction and sera. 2) The enzyme activity in ethionine-treated rats decreased significantly in lysosome-rich fraction and tended to increase in supernatant fraction. The activity of betaG in sera increased markedly. 3) In rats given ethanol for 4 weeks, the enzyme activity of sera and liver homogenates significantly increased in lysosome-rich fraction. These results indicate that the analysis of serum lysosomal enzyme activity, in the light of clinical features and laboratory data of routine liver function tests, is useful for the diagnosis of the fatty liver. A discussion is given of a possible mode of variation of lysosomal enzymes in rats with fatty livers.


[Effect of physical exercise on glycolysis in human red blood cells (author's transl)]. Untrained healthy male volunteers were studied to observe the effects of physical exercise (bicycle ergometer, 920 kpm/min for 10 min, 15 min and 30 min) upon glycolytic intermediates in red blood cells. The levels of glucose 6-phosphate, fructose 6-phosphate, pyruvate  and lactate increased after each exercise. The levels of fructose 1,6-diphosphate increased and phosphoenolpyruvate decreased respectively only after 30 min of exercise. At the rest period of 30 min after 30 min of exercise the lactate level still remained unchanged, however all the other intermediates returned to the preexercise values. A negative crossover point seemed to exist between fructose 6-phosphate and fructose 1, 6-diphosphate after 15 min of exercise. A positive crossover point was observed between phosphoenolpyruvate and pyruvate after 30 min of exercise. There were significant increases in hexokinase and pyruvate kinase activities, but not in phosphofructokinase activity after 30 min of exercise. These facts suggested that the increase in pyruvate kinase activity was due to the elevated fructose 1,6-diphosphate level after 30 min of exercise. A significant increase in plasma dopamine-beta-hydroxylase activity was found after each exercise. A close positive correlation was observed between pyruvate-phosphoenolpyruvate ratio and dopamine-beta-hydroxylase activity after 30 min of exercise. It was suggested that pyruvate-phosphoenolpyruvate ratio provided a reliable index of physical exercise.


Effect of catecholamines on insulin secretion and liver glycogenolysis in the rat. The relative activities of adrenaline, noradrenaline and isoprenaline in producing hyperglycaemia, hyperinsulinaemia and glycogenolysis in liver and skeletal muscle have been studied in both fed and fasted rats. Independent of prandial state isoprenaline significantly elevates plasma insulin level. Noradrenaline has no demonstrable glycogenolytic action in the liver even at a dose of 1 mg/kg regardless to the prandial state. These observations suggest that in the intact organism enhanced insulin-secretory effect of isoprenaline counteracts and masks the glycogenolytic effect of this drug in the liver.


Recovery of immune system in diabetic mice after treatment with insulin. In mice with alloxan-induced diabetes, humoral and cellular immunological reactivity were weak. The number of leucocytes, and especially lymphocytes, was reduced, and the weight and cellularity of lymphatic organs were lower than in normal mice. Treatment of diabetic mice with insulin reversed morphological and functional deficiency of the immunological system. Observed depression of immunological functions was attributed to impaired transport of glucose into immunocompetent cells, rather than to toxic effects of alloxan.


Adipose tissue cellularity: effect on insulin and thyroxine. The influence of insulin and thyroxine on the cellularity of adipose tissue in the rat epididymal fat pad has been studied. Incorporation of (3H) thymidine into the DNA of fat cells and stroma was measured together with fat cell size and number in rats pre-treated with either one of these hormones. There was an increase in fat pad weight in insulin treated rats which was due to 'lipid filling' of existing adipocytes and not increased proliferation of new fat cells. Thyroxine treated rats showed a decrease in fat pad weight caused by a decrease in size of individual fat cells. Cell number remained unaffected by either treatment.


Effects of pinealectomy and of sham-pinealectomy on blood glucose levels in the alloxan-diabetic rat. Alloxan-diabetic male rats were used to test effects of pinealectomy (PX) and of sham-pinealectomy (SPX) on blood glucose levels at mid-dark in the daily light-dark (LD 12:12) cycle. Animals received a diabetogenic dose of alloxan 10 days postoperatively. Blood glucose was measured on days -1,1,2,3,4,5,7,10 and 15 after 8-hour fasts. Pinealectomized (PX) and non-operated (C) animals were equivalent in their hyperglycemia following alloxan. SPX animals contrastingly showed significantly less response to alloxan than did animals of the other two groups. This difference of the SPX animals was in terms of lower hyperglycemia, better maintenance of body weight and of survival. It is concluded that intracranial surgery alone (SPX) and without visible brain damage can affect mechanisms of glucose homeostasis, and that at least in some circumstances such a surgical effect is not only different from effects of surgical pinealectomy (PX), but is also probably not pineal-dependent.


Action of insulin on thyroid protein synthesis in dogs. The study was performed on dog thyroid slices after in vivo administration of monocomponent insulin (carotid perfusion of a lobe: 25 mIU/kg/min for 15 min, and 2nd lobe perfused as control: saline) and after in vitro hormone addition (100 mIU/ml of glucose-free incubation medium). The 3H-leucine incorporation in thyroid proteins (dpm/mg) was determined following incubation of slices for 1 h and 2 h. Insulin given in situ significantly increased the mean incorporation into homogenate and cytosol proteins: about 130% after 1 h of incubation, for an identical hypoglycaemia in both lobes. When insulin was added to the medium, a similar response was observed after 1 h of incubation, the effect being less pronounced after 2 h but still significant. The sucrose gradient analysis of the soluble fraction showed that the stimulation affected all thyroid proteins. Polyribosomes isolated from in vivo insulin-perfused thyroids exhibited an increased activity when tested in cell-free system. These data suggest that insulin stimulated the thyroid protein synthesis in dogs through a direct action on the thyroid cell.


Prolactin cell adenomas of the human pituitary. Morphologic features of prolactin cells in the nontumorous portions of the anterior lobe. Twenty pituitary glands, harbouring prolactin cell adenomas, obtained from autopsy of male and female patients of various ages and dying of different diseases have been investigated by the immunoperoxidase technique in order to reveal the morphologic changes of prolactin cells in the nontumorous protions of the anterior lobes. For comparison, 40 nontumorous pituitary glands and 20 pituitaries lodging adenomas not consisting of prolactin cells have also been studied. In all four prolactin cell adenoma cases, the anterior lobes contained numberous prolactin cells outside the tumors, indicating a lack of involution of prolactin cells in the nontumorous portions of the pituitary glands.


Specific weight loss in hyperthyroidism. To assess metabolic turnover in patients with hyperthyroidism weight loss in mg . kg-1 . min-1 (specific weight loss, SWL) was measured by means of a new type of scale working on the buoyance principle with a minimal susceptibility of 10 mg per 100 kg. Weight loss rate was significantly increased in hyperthyroid patients (n=22;SWL, 22.0 +/-1.4 (SE) mg. kg-1 . min-1) compared to health controls (n=11; SWL, 10.0 +/-0.4 (SE) mg . kg-1 . min-1). The fall in SWL paralleled the decrease in thyroid hormone levels in serum in the beginning of antithyroid treatment and remained unchanged thereafter in spite of a further drop in thyroxine (T4) and triiodothyronine (T3) levels. After six weeks of treatment SWL reached the upper normal range despite hormone levels in the low range of normal. The findings indicate that the normalization of the basal metabolic rate as assessed by the estimation of SWL is slower than the decline in serum hormone levels. There was no correlation between thyroid hormone levels and SWL before initiating therapy or during antithyroid treatment (T4 vs. SWL, r = 0.22;n = 56). Thus serum levels of T4 and T3 appear to be rather insufficient indices regarding the respective metabolic state as estimated by SWL.


31P magnetic resonance of intact endocrine tissue: adrenal glands of dogs. Intact dog adrenal glands were examined for their phosphate profiles in the 31P nuclear magnetic resonance spectrometer at 36.43 MHz at 31 degrees, and the resultant spectra were compared to those obtained from perchloric acid extracts of the same endocrine tissue. The adrenal gland presents a unique organ for whole tissue spectroscopy in that its phosphate profile shows a number of features which have not been previously observed in similar phosphate profiles obtained from other intact tissue preparations. A number of prominant resonances are observed in the intact adrenal gland which arise from phosphorus-containing substances that are not extractable with perchloric acid. The 31P shifts of these resonances indicate that they may arise from phosphorylated proteins and membrane phospholipids.


Changes in plasma amino acid levels during the human menstrual cycle and in early pregnancy. A preliminary report. Fasting plasma amino acids have been measured in two normal females. In one subject changes in the levels of the amino acids have been followed weekly during four normal menstrual cycles. In the second subject changes in amino acid levels have been followed through a normal cycle, conception and early pregnancy. During a normal menstrual cycle it has been found that not all amino acids behave in a similar fashion. At least three distinct patterns of behaviour in respect of changes of concentration have been identified. In pregnancy it has been found that the depression in amino acid levels, reported by other workers in later pregnancy, in fact occur immediately following conception. The changes in levels of plasma amino acids which occur during the menstrual cycle and in pregnancy appear to be inversely related to the reported changes in progesterone levels which occur during these physiological events.


Actions of salbutamol in late pregnancy. Serum levels of human chorionic gonadotrophin, human placental lactogen, total oestriol and progesterone in diabetic and non-diabetic women during infusion of salbutamol. The acute effects of salbutamol on the serum levels of hPL, hCG, oestriol and progesterone in diabetic and non-diabetic pregnant women in the last trimester were studied. I.V. administration of salbutamol in an increasing dose from 0 to 22.5 ug/min caused no change in hPL or hCG levels in any group. Progesterone showed a significant fall after 40 min in the diabetic group and a less pronounced, although still significant fall after 80 min in the non-diabetic group. The oestriol levels increased significantly after 40 min in the diabetic group while no significant change was noted in the non-diabetic group. It is speculated that the effects in the diabetic women might be explained by a redistribution of the available placental 5-pregnenolone due to an increased fetal conversion into oestriol. This might be due to an increased sensitivity of the adrenal cortex of these foetuses to a supposed stimulatory effect of salbutamol.


Effects of calcium intake and renal function on plasma immunoreactive parathyroid hormone levels in rats. Parathyroid hormone (PTH) was estimated with a radioimmunoassay system which recognised presumably intact rat PTH as the major circulating species. A low calcium diet was found to increase PTH levels in shamoperated and in 5/6-nephrectomized rats. In rats with normal renal function, a low calcium intake resulted in a significant mean increase of PTH levels, while changes in plasma calcium, magnesium, phosphate and protein concentrations were not detectable; at the same time the ability of the duodenum to transport calcium in vitro was accelerated. In 5/6-nephrectomized rats, on the other hand, PTH levels were dramatically increased in hypocalcemic animals kept on a calcium deficient diet, while intestinal calcium transport was impaired. These data support the concept that hypocalcemia is the most important stimulus for increased PTH release. Since increased PTH levels can be seen in situations where increased 1,25-dihydroxy-cholecalciferol synthesis is known to occur, it appears that this metabolite cannot inhibit a rise in plasma PTH levels.


[Covalent C-terminal fixation of cyanogen bromide peptides in the liquid-phase-sequenator (author's transl)]. This paper describes the covalent fixation and hydrophilisation of homoserin lactone peptides enabling complete C-terminal sequencing in the squenator. Dimethylformamide, dimethylsulfoxide and 6M guanidine hydrochloride in water were used as solvents, ethylendiamine, hexamethylendiamine and histamine base as amino components. The diamine peptide derivative was reacted with the hydrophilic isothiocyanates I and IV, the fixed peptide was sequenced to the C-terminal amino acid, Histamine reacted particularly well and the program with 0.1N quadrol and the hydrophobic buffers was especially suitable for this derivative. The phenylthiohydantoin derivative of homoserine was proven in good yields. The application of this method is suggested.


Kinetic investigations of liver microsomal esterases with oxazepam esters. Hepatic microsomal esterases of the mouse responsible for the bioactivation of inactive (prodrug) esters of the centrally acting oxazepam were studied. The enzymes are situated on the cytoplasmic side of the microsomes. The esterases are partly solubilized and partly inactivated by homogenization in aqueous glycerol and treatment with deoxycholate. There is good correlation between the rates of hydrolysis and steric constants of the acyl moiety. Substrate binding increases to an optimum with the number of carbon atoms in the acyl moiety and is of hydrophobic nature. An attempt has been made to classify the esterases on the basis of the effect of inhibitors and activators.


Pentagastrin analogs containing alpha-aminooxy acids, I. Synthesis of analogs substituted at the N-terminus. Nine new pentagastrin analogs containing a free or protected alpha-aminooxy acid at the N-terminus were synthesized stepwise. Analogs containing leucyl, norleucyl, norvalyl, L- and D-2-aminodecanoyl residues instead of methionyl residue were also prepared. The peptides were synthesized by the active ester method with subsequent removal of the protecting groups. The purification of the end products was performed by crystallization or column chromatography on silica gel.


Pentagastrin analogs containing alpha-aminooxy acids, II. Simultaneous substitutions at the N- and C-terminus. Twenty-seven new pentagastrin analogs containing free or protected aminooxy acids at the N-terminus and D-2-aminooxy-3-phenyl-propionic acid, L-(4-chlorophenyl)glycine, L- and D-phenylglycine, or L- or D-cyclohexyglycine instead of the C-terminal L-phenylalanine and analogs extended by aminooxyacetic acid at the C-terminus, were synthesized stepwise. The end products were purified by chromatography on DEAE-cellulose or silica gel.


Pentagastrin analogs containing alpha-aminooxy acids, III. Biological studies and structure-activity relationships. The stimulation of gastric acid flow by thirty-six new pentagastrin analogs administered intravenously, intrajejunally and intrarectally was determined in rats. Some of the analogs are several times more active than the pentagastrin (Peptavlon, I.C.I.) used as control. Unlike the control, some analogs are absorbed in active form from the jejunum. The N-terminal substitution with alpha-aminooxy acids seems to increase the gastric secretory response, probably by improving the enzymatic resistance of the molecule to various enzymes. None of the analogs tested significantly inhibit the gastric acid output in rats induced by pentagastrin. Structure-activity relationships are discussed.


Diazoniobenzenesulfonate as marker for cell surface proteins: study of the surface coat of Trypanosoma congolense. It is possible to label selectively the surface coat of Trypanosoma congolense with radioactive sulfanilic acid diazonium salt. As demonstrated by both sodium dodecylsulfate polyacrylamide gel electrophoresis and isoelectric focusing, radioactivity is incorporated into only one protein, which has a molecular weight of 57 000 and an isoelectric point of 6.25. This indicates that the surface coat of T. congolense is a homogeneous layer, composed of molecules of one type of protein.


Structure and activity of insulin, XVI. Semisyntheses of desheptapeptide-(B24--30)- up to destripeptide-(B28--30)-insulin with lysine or alanine in place of arginine in position B22: influence on the three-step-increase of activity in positions B24--26 (Phe-Phe-Tyr). The desonapeptide-(B22--30)-insulin pentamethyl ester, protected with Boc- at the two N-terminal amino groups, was prepared as described in the preceding XVth communication[6]. The free carboxyl group of the glutamic acid residue B21 of this compound was coupled to the following synthetic oligopeptide esters (X = Lys or Ala): X-Gly-OMe X-Gly-Phe-OMe X-Gly-Phe-Phe-OME X-Gly-Phe-Phe-Tyr-OMe X-Gly-Phe-Phe-Tyr-Ala-OMe After coupling, the semisynthetic products were deprotected and purified. Their biological activities were determined in the mouse fall test and by measurement of blood glucose levels. There were no statistical differences between the values obtained for the lysine B22 and alanine B22 products. The three-step increase in activity due to the amino acids Phe-Phe-Tyr (B24--26) was still recognizable, but compared with the analogues containing arginine B22, the activities were very stronly diminished. These results are in contrast with the assumption that activity of insulin is dependent on the formation of a strong ionic linkage between the asparagine-A21 carboxyl group and any positive charge in B22. The results, however, demonstrate the high specificity of the arginine guanidino group in position B22.


The conformer nature of the multiple forms of beef liver catalase as obtained by biochemical and small-angle X-ray scattering experiments. A model for the quaternary structure of the beef liver catalase molecule. Two of the five multiple forms of beef liver catalase have been extensively studied using biochemical and biophysical analysis techniques. The two molecules, cat I and cat V, have different isoelectric points (pH 6.55 and 5.6), different surface charges (25.8 and 32.7 elementary charges) and display different numbers of primary amino groups on their surfaces. The numbers of tyrosine residues on the surfaces of the two molecules are also different (16 and 10 at pH 10). Since the two forms of catalase can be interconverted, the described changes may be caused by conformational changes of the four protein subunits within the molecule. This mobility of the polypeptide chains is also demonstrated by the different absorption spectra below 390 nm. Using small-angle X-ray scattering, the radii and the volumes of the two catalase forms were shown to be different (cat 1 is smaller than cat V). All five multiple forms of beef liver catalase are concormers of the molecule. A model for the quaternary sturcture of the beef liver catalase are conformers of the molecule. A model for the quaternary structure of the beef liver catalase molecule is suggested. It consists of a regular configuration of four prolate rotational ellipsoids (semiaxes: a = 52 A, b = c = 21 A) in close contact in which the nearest neighbour subunits are shifted by 37 A parallel to each other. Thus the height of the complete molecule is 141 A and the diameter 94 A.


Crystallization, crystal structure analysis and atomic model of the complex formed by a human Fc fragment and fragment B of protein A from Staphylococcus aureus. Crystals of the complex formed by human Fc fragment and fragment B (FB) of protein A from Staphylococcus aureus were prepared and the crystal structure determined at high resolution by multiple isomorphous replacement. Phase were improved considerably by combining these phases with calculated phases from the Fc component. FB is a small globular protein built of three parallel helices arranged in a triangular array. It binds by the first two helices of Fc and is attached to segments of CH2 and CH3. The CH3 module is unchanged between complex and Fc fragment crystals, but CH2 changes its position slightly relative to CH3. In addition, the upper third of CH2 is disordered in the complex crystals. Possible sources of this disorder are discussed.


Synthesis of protected decapeptide (B21--30) of human insulin. The synthesis of the C-terminal decapeptide derivative B21--30 of the B-chain of human insulin (benzyloxycarbonyl-gamma-t-butoxyglutamyl-arginyl-glycyl-phenylalanyl-phenylalanyl-O-t-butyltyrosyl-O-t-butylthreonyl-prolyl-Nepsilon-t-butyl-oxycarbonyllysyl-O-t-butylthreonine t-butyl ester, (XIII) by the fragment condensation of the tripeptide B21--23 with the heptapeptide B24--30 is described. Two new routes for both the tripeptide and the heptapeptide were established. A simplified method for the purification of O-t-butylthreonine t-butyl ester was developed.


Inactivation of boar acrosin by peptidyl-arginyl-chloromethanes. Comparison of the reactivity of acrosin, trypsin and thrombin. A survey of the reactivity of 16 peptidyl-argininyl-chloromethanes with boar acrosin indicated that these compounds as a general group of reagents were highly effective in the inactivation of acrosin since at least half of the reagents tested rapidly inactivated this protease at a concentration of 0.10 micrometer or lower. For example, Dns-Glu-Gly-ArgCH2Cl inactivates acrosin by 50% in 1.8 min at a concentration of 75 nM, whereas in contrast, a 14000-fold higher concentration of Nalpha-tosyllysyl-chloromethane is required to obtain an equivalent rate of inactivation. A comparison of the reactivity of acrosin and trypsin with the peptides of arginyl-chloromethane containing different substituents in the P2 and P3 positions suggests that the secondary binding sites of these two proteases are very similar. Reagents with homoarginine, lysine and D-arginine in the P1 position have also been prepared and evaluated, but these were considerably less effective than the corresponding arginyl-chloromethanes in the inactivation of both acrosin and trypsin.


Further studies on the membrane glycoprotein defects of S--s--and En(a--)-erythrocytes. Sodium dodecylsulfate polyacrylamide gel electrophoretic methods were applied to S--s--and En(a--) red cells as well as to erythrocytes from individuals being heterozygous for these defects. The results demonstrate more conclusively than previous data, that the glycosylated part of the MN glycoprotein is lakcing in En(a--) red cell membranes. S--s--U--erythrocytes either lack the Ss glycoprotein completely or contain a defective molecule which is devoid of the glycosylated part. Conversely, S--s--U+ cells exhibit small amounts of Ss glycoprotein which could only be detected when large amounts of extracted glycoproteins were separated. It is shown that this molecule possesses the 'N' antigenic determinant.


Comparison of the kininogenase activity of human pancreatic trypsins and porcine Kallikrein on Met-Lys-bradykinin and human plasma kininogen. Human trypsins 1 and 2 both converted Met-Lys-bradykinin to bradykinin and released bradykinin from kininogen in human plasma as measured by bioassay with the isolated guinea pig ileum. Porcine kallikrein did not act on Met-Lys-bradykinin and released kallidin from human kininogen. Since human trypsin 1 is only partially and trypsin 2 completely inhibited by soybean trypsin inhibitor, these data show that the criterion of susceptibility to soybean trypsin inhibitor cannot be used to discriminate between trypsin and kallikrein of different species.


Crystalline [A21-desamido]bovine insulin. [A21-Desamido]insulin is the major product formed during mild acid hydrolysis of bovine insulin at low insulin concentration. The derivative was isolated by standard procedures and its purity established by isoelectric focusing, disc electrophoresis and electrophoresis on cellulose acetate strips. The identity of the acid-transformed derivative was determined as [A21-desamido]insulin by the action of carboxypeptidase A, using conditions under which a C-terminal aspartic acid residue would not be removed. The biological activity of this crystalline derivative was found to be 15.9 units/mg as measured by the mouse convulsion assay.


Mental health services: equity, quality, and constraints. The current restrictions on resources for mental health care reflect political priorities rather than immutable economic forces, the author contends. He believes that despite the public cynicism about the motives of professionals who plead for additional resources for mental health, professionals have a continuing obligation to be advocates for patients and to lobby for universal access to effective services. In his view, the best use of resources would take three major programmatic thrusts: a full range of services for psychotic patients, consultation and collaboration with primary-practice physicians to enhance their skill in the management of psychiatric and psychosomatic problems, and comprehensive maternal and child health programs. He emphasizes the need for services for minority groups, who continue to bear a disproportionate health burden as reflected in longevity and mortality rates, and for increased funding for health research.


Developing a psychiatric inpatient service in a rural area. An inpatient psychiatric service was developed in a community hospital in Appalachia by building on the hospital's existing mental health components, a comprehensive alcoholism program and an inservice training program for general-duty nurses. In its first phase, the program offered a consultation service to help physicians deal with emotional problems of medical-surgical patients and a daytime therapy program for selected patients referred by the consultation service. In the second phase, psychiatric patients were admitted directly from the community and placed in wards throughout the hospital. Eventually a separate 23-bed psychiatric unit was opened. Between 1973 and 1976 more than 2200 patients were treated by the therapy service or as psychiatric inpatients. The author discusses problems that occurred during each phase of development, including a continuing shortage of nursing staff.


Meeting the special needs of Appalachian alcoholics. The Appalachian Regional Comprehensive Alcoholism Program in Beckley, West Virginia, was established in 1972 to meet the needs of a four-county catchment area. In designing the program, planners took into account the special cultural and economic characteristics of the population to be served--individualism, isolation, religiosity, conservatism, distrust of newcomers, and economic deprivation--and set up a program they believe respects local norms. For instance, the program has been staffed mainly by local professionals. Individual therapy, family therapy, and crisis intervention are used much more frequently than group psychotherapy, and clients have a strong voice in treatment decisions. Local ministerial associations are involved whenever possible, and outreach efforts include home visits as well as the use of the media to disseminate information.


The alcohol emergency room in a general hospital: a model for crisis intervention. The alcohol emergency room at Denver General Hospital provides a model for crisis intervention for alcoholics. The unit is staffed 24 hours a day by nurses. Patients spend an average of 45 minutes in waiting time and triage. About half the patients require inpatient treatment; the others are referred to outpatient programs. Unit staff emphasize consultation and liaison with other hospital staff to coordinate the treatment system and provide education on aspects of alcoholism. They also explain the patient's condition and needs to his significant others, who provide crucial external support. Staff currently see 400 patients and answer about 270 telephone requests for information about alcoholism treatment monthly.


Mental health training and consultation: a model for liaison with clergy. A chaplain, a psychiatrist, and a social worker developed a program to train clergy in more effective mental health case management, counseling, and referral skills, and to sensitize mental health center professionals to the work of the clergy in mental health. The program consists of 12 two-hour meetings conducted by a clinician and a chaplain once a week. In the first four meetings, clergy are trained to use a self-report personal-data kit with individuals presenting with emotional problems. In the next eight meetings, clergy make case presentations that are used as a basis for consultation and group discussion. The project has established a firm basis for collaboration between clergy and mental health center staff in providing care.


Schizophrenia through the eyes of families. The authors contend that families of schizophrenics have received too little help from mental health professionals, even though in many cases families are the real primary care agents for long-term patients released from mental hospitals. They discuss some of the problems that occur in living with a long-term patient and describe mutual-support and advocacy groups that relatives have formed. They emphasize that mental health professionals can help families of schizophrenics by providing practical, realistic advice on how to deal with the illness, by offering empathy and support rather than placing blame, and by working to ensure that there are adequate treatment and rehabilitation services available.


No-fault malpractice insurance: Swedish plan shows us the way. No-fault medical accident insurance could help solve some of the medical malpractice insurance problems that plague American physicians and hospitals. In Sweden, the Patient Injury Insurance Plan has been working for two years, and, with the statistics that are now available, the claims experiences under the new Swedish system can be compared with the claims experience in the United States under the traditional liability system. The author believes that an insurance system similar to the Swedish plan can work in the United States, but only if certain conditions are created.


Materials management cuts costs with internal, external controls. A hospital's materials management division is helping to contain nonpayroll expenses, particularly purchasing expenses, through centralized purchasing, inventory control, a monitoring and projection system for costs related to procurement activities, and participation in a group purchasing arrangement.


Computerized systems cut dietary department's costs. The department of dietetics at a large teaching hospital has substantially reduced its food and labor costs through use of computerized systems that ensure efficient inventory management, recipe standardization, ingredient control, quantity and quality control, and identification of productive man-hours and appropriate staffing levels.


Synopsis '78: hospital cost containment stalled; few health bills pass. During 1978, hospitals brought to Capitol Hill the heaviest barrage of information that Congress has ever seen about rising health care costs, the causes of those costs, and what is being done about them.


Education and training need to be brought up-to-date. Training of hospital employees consumes a large segment of total hospital dollars, so it is important to document clear proof of value. If training programs are to be successful, they must be relevant to the measured needs of the institution. In addition to this essential characteristic of the training process, the author discusses six others that affect the outcome of such programs.


Hospital takes systematic approach to educational programs. After analysis of its educational programs showed them to be deficient, Deaconess Hospital, Evansville, IN, undertook the development of a comprehensive, systematic program of education and training. First steps were the establishment of an independent department of education and the preparation of an official position on the use and value of educational programs within the hospital.


Emergency department key factor in hospital admissions. A significant proportion of a community hospital's inpatient population is admitted through the emergency department. Administrators who are aware of the frequency of this practice in their institutions can schedule and staff services more efficiently. A 1975 survey conducted by the American Hospital Association shows that the number of emergency department admissions depends on hospital size, location, ownership, and type of physician coverage in the emergency department.


Alternative birth centers offer family-centered care. With the endorsement of the five major health care associations involved in maternity/newborn care and the support of the American Hospital Association, hospitals throughout the country are beginning to offer alternatives to traditional maternity care. Alternative birth centers (ABCs) have sprung up throughout the country and are providing one-room, homelike environments for labor, delivery and recovery; Leboyer atmosphere; natural childbirth techniques; rooming in; and early discharge. Consumer demand has greatly influenced the changes, and hospital costs have been cut in half.


Baptist and Catholic hospitals share to save money, improve services. Highlands Medical Center, Louisville, is the first instance of a Baptist-Catholic sharing arrangement in the nation. Without attempting to alter the religious dedication or legal operation of either of the two hospitals involved, the Highlands Center arrangement helps the facilities and their patients to save money and the hospitals to provide their community with a greater range of services at the lowest possible cost.


Anointing the healers: a consecration ceremony for all staff. All persons who serve the Lord through their work in a health care facility should be considered healers and need to be made aware of the special charisms in healing. Among the means of accomplishing this are pastoral department input into new staff orientation, periodic workshops recalling the Church's commitment to healing, special days of recollection, Masses, Bible services, scriptural sharings, and prayer groups. Especially meaningful may be a ceremoney in which individuals are anointed with oil to dedicate them to the charisms of their particular tasks.


CHA as an ecclesial community.

CHA as an ecclesial community. In discussing the CHA Study Committee's description of the CHA as an ecclesial community, Fr. Clarke explains why the committee chose this particular description and outlines in theological and pastoral terms what accepting this concept means for the present and future activities of CHA.


Theological and scientific understandings of health. A pilgrimage to discover the meaning of "health" within a Christian orientation begins with religious and scientific definitions of the term. The ideas of thinkers such as Tillich, Hiltner, Lapsley, and Illich are briefly explored, as is the unrealistic American insistence on complete and constant physical well-being. This leads to the consideration of a holistic concept of health and the suggestion of theological guidelines for a Christian approach to defining health.


A board searches for a CEO.

A board searches for a CEO. The board of directors of Mercy Hospital, Urbana, IL, recently chose to conduct its own search for a new CEO. Research on other self-directed searchers, delineation of the hospital's needs, and development of criteria for the CEO led to the screening of candidates. Both the board and the new CEO believe that the selection method--a time- and energy-consuming one--contributed greatly to the current mutual satisfaction.


Unions and health care cost inflation.

Unions and health care cost inflation. Since the 1974 Taft-Hartley amendments gave nonproprietary hospital employees the right to organize and bargain collectively, the rate of growth of work stoppages has decelerated and the number of man days lost has been reduced. Wage rates, earnings differential, and fringe benefits have not been significantly improved by unionization. And little evidence supports the contention that unionization has contributed substantially to the inflation experienced in the health care industry.


Needed: cooperation to create a health care data bank. Only the best possible utilization of existing data collection systems and the cooperative development of more comprehensive systems will enable participants in health care delivery to control the vast bulk of health care data and thereby make more rational efforts to control industry costs and quality. Relief from the threat of antitrust action would facilitate such necessary cooperative efforts.


Drug-induced liver injury.

Drug-induced liver injury. In the usual course of events, the liver detoxifies hepatocytotoxic intermediates of drug metabolism. But when protective systems are overloaded by normally harmless drugs, the intermediates can cause massive, even fatal, hepatic necrosis.


The fine structure of human thyroid cancer. This ultrastructural description of human thyroid cancers is based on the available literature and on our own studies of about 150 cases. Electron microscopy is an invaluable diagnostic adjunct to light microscopy, as it may eliminate inaccurate designations such as "small cell malignant tumors of the thyroid," which include tumors of different histogenetic origin with a different prognosis and treatment that share only a similarity in appearance under the light microscope. Ultrastructure is also of diagnostic importance in cases of medullary carcinoma that imitate papillary or follicular patterns or lack amyloid stroma. Its importance in separating follicular adenomas from carcinomas, however, has not been proven. In conjunction with other methods ultrastructural study might throw new light on the controversial classification of papillary and follicular carcinomas and improve our understanding of their different biologic behavior. Immunoelectron microscopy may help in solving the problem of amyloid pathogenesis in endocrine tumors and in charting the subcellular mechanisms involved in the production of multiple polypeptide hormones in a single tumor.


A re-evaluation of diabetic glomerulosclerosis 50 years after the discovery of insulin. Diabetic glomerulosclerosis is a degenerative-proliferative lesion involving most glomeruli in the kidneys of all individuals with hereditary, pancreatic, or experimental diabetes mellitus. The exact nature of the lesion and its etiology remain unknown. Morphologically there appear to be two courses this disease may follow. There is a benign course, which occurred in 89 per cent of our series of adult diabetic patients, consisting of concurrent thickening of the capillary basement membranes and diffuse glomerulosclerosis. The benign course is slowly progressive over many years, and rarely leads to renal failure. The accelerated course, in our experience is always superimposed on the changes of the benign course, and consists of a more rapid progression with the development of glomerulocapillary microaneurysms. Kimmelstiel-Wilson nodules, exudative-deposit lesions, and glomerulocapsular adhesions leading to glomerular obliteration and renal failure. Data are presented to support the concept that large Kimmelstiel-Wilson nodules are formed by the organization of the glomerulocapillary microaneurysms.


Histologic changes in Paget's disease treated with calcitonin. In recent years the use of calcitonin in the medical treatment of Paget's disease of bone has been gaining in popularity. Seventeen patients on long term calcitonin therapy showed objective morphologic evidence of improvement in the quality of bone matrix deposition, in the trabecular organization, and in the parameters used to evaluate the rates of bone turnover. The usefulness of bone biopsy and fine detail radiography of the biopsy core in evaluating the progress of disease and the effects of therapy is discussed. It is concluded that the longer the period of therapy and follow-up, the higher the proportion of patients showing histologic evidence of improvement.


Collision carcinoma of the gastric cardia. A case of squamous cell carcinoma of the lower end of the esophagus colliding with adenocarcinoma of the cardiac end of the stomach is reported. The rarity of the lesion, its diagnostic criteria, and therapeutic implications are briefly discussed.


Localized congenital hepatic fibrosis presenting as an abdominal mass. A six month old male with a liver mass localized to the right lobe that had the histological features of congenital hepatic fibrosis is presented. The mass was resected and the patient was asymptomatic 10 months after surgery. This case serves to further broaden the range of lesions with the histological features of congenital hepatic fibrosis; only one other example of localized congenital hepatic fibrosis has been previously reported.


Tenosynovial chondroma of the hand: a case report with a brief review of the literature. Tumors of hyaline cartilage occurring in the soft tissues of the hands and feet arise by metaplasia from the synovium of tendon sheaths and have a benign clinical evolution. Although recurrences may develop, they show no tendency to metastasize. The majority of these tumors prove to be diagnostic difficulties because of the alarming histologic atypia they display. We report a case of tenosynovial chondroma of the hand and briefly review the salient features of these lesions.


Cardiac amyloidosis in hereditary neuropathic amyloidosis diagnosed by endomyocardial biopsy. A 57 year old male first developed progressive neuropathy of the lower extremities and then similar involvement in the upper extremities. Two years later he developed dyspnea and then complete atrioventricular block requiring the use of a permanent cardiac pacemaker. An older brother had an almost identical clinical course, including the installation of a permanent pacemaker. Both died of left ventricular failure. An endomyocardial biopsy revealed extensive amyloidosis and fibrosis of the myocardium. This is the first report of a myocardial biopsy showing amyloid on electron microscopy.


Immunohistochemistry of human cutaneous sarcoidosis: a study of nine cases. Biopsy specimens of sarcoid skin lesions from nine patients with sarcoidosis were examined for the presence of immunoglobulins, complement (C3 and C4), and fibrinogen. Granulomas in all skin lesions were positive only for fibrinogen. The literature relating to the immunohistochemistry of sarcoidosis is reviewed and its significance discussed.


Yolk sac tumors with pure and mixed polyvesicular vitelline patterns. Four cases of polyvesicular vitelline tumor are presented; two were of a previously unreported pure type, and the other two were mixed with endodermal sinus tumor. The morphologic features of the vesicles favor an endodermal origin, as originally proposed by Teilum. Marked specialization of the vesicular lining cells, seen ultrastructurally, suggests a differentiation toward gut structures and mature yolk sac. One case of pure polyvesicular vitelline tumor showed massive erythropoiesis. We propose that the pure tumor reflects an intermediate degree of differentiation within the selectively endodermal yolk sac tumor group, that is, a further stage of organization than the endodermal sinus tumor. In  our cases of pure polyvesicular vitelline tumor, the marked degree of differentiaiton was correlated with an improved prognosis, as in the case of the possible homologue of this tumor, the yolk sac tumor of the infant testis. In contrast, the two cases of the tumor admixed with endodermal sinus tumor illustrated the low survival rate expected in the pure endodermal sinus tumor; in these cases the metastases had no polyvesicular component. Because of the significance of such a difference in prognosis we emphasize the importance of an accurate diagnosis, suggesting that a large number of sections be taken in order to demonstrate any endodermal sinus tumor component that may be present, and that the possibility of pure polyvesicular vitelline tumor always be considered in the differential diagnosis of multicystic ovarian tumors.


Genital actinomycosis and intrauterine contraceptive devices. Cytopathologic diagnosis and clinical significance. This study from a community hospital documents the relatively frequent occurrence of Actinomyces in 36 women diagnosed by Papanicolaou stained cervicovaginal smears. Actinomyces was identified exclusively in patients wearing an intrauterine or vaginal foreign body (intrauterine devices and pessaries). By using fluorescein isothiocyanate labeled antiserum, the organisms were shown to be Actinomyces israelii in eight randomly selected smears. In five cases Actinomyces was demonstrated in tissues obtained by endocervical and endometrial curettage. Anaerobic microbiologic studies performed in 11 of the 36 cases showed a relatively high rate of recovery of Actinomyces (36.3 per cent) with a yield of four positive cultures. In two cases the isolated organisms were shown to be Actinomyces israelii. In the remaining two cases the exact species of Actinomyces has remained uncertain. Of the 36 patients with smear evidence of Actinomyces, nine (25 per cent) had pelvic inflammatory disease, a rate that appears to be much higher than the already high rate among all intrauterine device users. This observation underscores the importance and clinical significance of the finding of Actinomyces in the vaginal smears of intrauterine device users. Papanicolaou stained cervicovaginal smears provide a relatively easy, inexpensive, fast, and highly specific method for the morphologic diagnosis of Actinomyces and allows us to recognize at a relatively early stage a group of patients who are potentially at risk for the development of pelvic inflammatory disease and its associated more severe complications. The hope is that prompt identification of Actinomyces in cervicovaginal smears can help to prevent the more serious infectious complications in intrauterine device users.


Histogenesis of cloacogenic carcinoma. Fine structure of anal transitional epithelium and cloacogenic carcinoma. The fine structural morphology of the anal transitional zone is described and is compared to the ultrastructure of urothelium and anal squamous epithelium. Electron microscopic observations in eight cases of cloacogenic carcinoma are also presented. Anal transitional epithelium is not highly specialized and incorporates features of both urothelium and squamous epithelium; slight urothelial differentiation is considered vestigial. Cloacogenic carcinoma is a specific morphologic entity, which can be distinguished from urothelial and squamous carcinoma. The "transitional" variant is composed of cells similar to the intermediate zone cells of anal transitional epithelium, an observation confirming the concept that the transitional form of cloacogenic carcinoma is derived from the anal transitional zone. The "basaloid" and pleomorphic variants appear to represent less differentiated forms of cloacogenic carcinoma.


Infarction of the lymph nodes: a cause of a palisading macrophage reaction mimicking necrotizing granulomas. A case of multiple cholesterol atheromatous emboli with infarction in various parts of the body is described. The histologic appearance of infarction in the lymph nodes, a rare condition, mimicked that seen in necrotizing granulomas usually secondary to micro-organisms.


Long term post-traumatic retrograde corticospinal degeneration in man. The spinal cord and brain of a man who died 18 years after a crush injury of lumbar segments contained some unusual lesions. There was a reduced number of myelinated axons in the corticospinal tracts as high as the fifth cervical segment. Such retrograde degeneration has been described in human pyramidal tracts only a few times. The results of reported studies of experimental retrograde degeneration have been inconsistent. The course of the fasciculus  gracilis, as delineated by gliosis, was atypical, and an unusual glial nodule, possibly neoplastic, was present in the dorsal columns at C8.


Partial deletion of 4p16 band in a ring chromosome and Wolf Syndrome. A new case of ring chromosome 4 in a 2-day-old female child with multiple malformations is described. By means of the GTG-banding technique, a karyotype 46,XX,r(4), (p16 leads to q35) was determined. The characteristics of the child's karyotype and the relationship with the structure of the chromosome, especially the location of the deletion that produces the syndrome, are compared with previous reports.


Chromosome 7 short-arm interstitial deletion (p14). A 13-year-old girl presented with microcephaly, short and broad neck, low posterior hairline, congenital heart disease, limitation of joint movement, and mild mental retardation. Chromosomal analysis showed interstitial deletion of band p14 of the short arm of chromosome 7. The patient's physical and cytogenetic findings are compared with those of five other patients with 7p- deletions.


Small metacentric nonsatellited extra chromosome: report of five mentally retarded individuals and review of literature. Contribution to further delineation of a new syndrome. Five mentally retarded male patients with a supernumerary small metacentric nonsatellited chromosome were found to have many clinical features in common. The face showed characteristic small crowded features, the bodily habitus was asthenic, and the hands and feet had minor abnormalities. Renal anomalies were present in two patients. One patient had a myelomeningocele. Cytogenetic studies employing Q, R, and C banding in four patients showed the small extra chromosome to have staining properties compatible with an isochromosome of the short arm of chromosome 18. A comparison with previous case reports suggests a new syndrome. However, the identity of the extra chromosome has not yet been determined.


Male pseudohermaphroditism: genetics and clinical delineation. The genetics and clinical delineation of male pseudohermaphroditism are reviewed. These disorders are categorized initially by their genetic etiology--cytogenetic, Mendelian, or teratogenic. It is especially important to distinguish cytogenetic forms, usually associated with 45,X/46,XY mosaicism, from Mendelian (genetic) forms because in the former the prevalence of gonadoblastomas or dysgerminomas is about 15--20%. Genetic forms include (1) those associated with a multiple malformation pattern, (2) those due to an error in adrenal or testicular hormonal biosynthesis, (3) complete testicular feminization, (4) incomplete testicular feminization, (5) Reifenstein syndrome, (6) pseudovaginal perineoscrotal hypospadias, and (7) agondia, and possibly other conditions. Incomplete testicular feminization and the Reifenstein syndrome may or may not represent varied expressivity of the same trait. The designation pseudovaginal perineoscrotal hypospadias is appropriate only if constellations of clinical features are present and if no metabolic abnormalities are demonstrable. Etiology and available genetic data are reviewed for each of these disorders.


Partial trisomy for the short arm of chromosome 2 due to familial balance translocation. A partial trisomy for the short arm of chromosome 2 (p21 leads to pter) was observed in a severely retarded infant with facial, skeletal, genital, renal, and CNS anomalies. The phenotypically normal mother and older brother had a balanced translocation between the short arm of chromosome 2 and the long arm of chromosome 14: 46, XX-XY,t(2;14)(p21;q32).


Humoral immune responses in foetal sheep. A total of fifty-two foetal sheep between 49 and 126 days gestation were injected with polymeric and monomeric flagellin, dinitrophenylated monomeric flagellin, chicken red blood cells, ovalbumin, ferritin, chicken gamma-globulin and the somatic antigens of Salmonella typhimurium in a variety of combinations. Immune responses were followed in these animals by taking serial blood samples from them through indwelling vascular cannulae and measuring the circulating titres of antibody. Of the antigens tested, ferritin induced immune responses in the youngest foetuses. A short time later in gestation, the majority of foetuses responded to chicken red blood cells, polymeric flagellin, monomeric flagellin and dinitrophenylated monomeric flagellin. Only older foetuses responded regularly to chicken gamma-globulin and ovalbumin. However, antibodies to all these antigens were first detected over the relatively short period of development between 64 and 82 days gestation and this made it difficult to define any precise order in the development of immune responsiveness. Of the antigens tested only the somatic antigens of S. typhimurium failed to induce a primary antibody response during foetal life. The character and magnitude of the antibody responses in foetuses changed throughout in utero development. Both the total amount of antibody produced and the duration of the response increased with foetal age. Foetuses younger than 87 days gestation did not synthesize 2-mercaptoethanol resistant antibodies or IgG1 immunoglobulin to any of the antigens tested, whereas most foetuses older than this regularly did so.


Correlation between intestinal synthesis of specific immunoglobulin A and protection against experimental cholera in mice. The importance of locally and systemically formed antibodies of various classes for protection against experimental cholera has been studied in mice immunized with cholera toxin. Groups of mice were given various numbers of peroral or intravenous immunizations, or a combination of both. Serum antibodies and antibodies synthesized by spleen and small intestine in vitro during tissue culture were measured by the enzyme-linked immunosorbent assay, and protective immunity against intestinal toxin challenge was determined by means of a small-bowel loop assay. Regression analyses showed a close correlation between the magnitude of intestinal synthesis of specific immunoglobulin A (IgA) antibodies and protection (r = 0.98), whereas neither the local formation of IgG or IgM nor the production of antitoxin antibodies of any immunoglobulin class by spleen showed any significant correlation with protection. The serum titers of IgG and IgM antibodies did not show any such relation, whereas the level of specific IgA in serum, probably mainly derived from the intestine, correlated significantly (r = 0.90).


Increased tissue conductance and ion transport in guinea pig ileum after exposure to Staphylococcus aureus delta-toxin in vitro. Prior studies had shown that Staphylococcus aureus delta-toxin was able to inhibit water absorption in guinea pig ileum and to elevate the cyclic AMP content of this tissue, but was unable to elicit certain cyclic AMP-mediated changes in Y-1 adrenal or Chinese hamster ovary cells. Because water movement passively follows the net movement of electrolytes in the gut, this study investigated the effect of delta-toxin on ion transport in guinea pig ileum maintained in vitro. The transmural potential difference (PD) of guinea pig ileum was measured and nullified with an automatic voltage clamp. The short circuit flowing under these conditions (I(sc)) was measured, and the conductance was calculated (I(sc)/PD). Unidirectional (22)Na(+) and (36)Cl(-) fluxes were measured. In a glucose-free Ringer solution, delta-toxin caused an immediate spike in PD and I(sc), and the extent and duration of the spike generally increased with increasing toxin concentration. The conductance of ileum was increased by delta-toxin, and this effect on conductance could be blocked by lecithin, a known inhibitor of delta-toxin. Tissue in the presence of glucose did not exhibit a spike in PD or I(sc) when exposed to delta-toxin. In a glucose-free medium, delta-toxin caused a 1.5- to 2.5-fold increase in both the unidirectional absorption and secretion of Na(+) and Cl(-), whereas the net secretion of Na(+) increased above basal levels. The observation that delta-toxin causes a prompt increase in intestinal ion flux lends credence to the concept that the elevation in cellular cyclic AMP, which occurs later, is a secondary response to the toxin. The rapid increase in ion flux may reflect the ability of delta-toxin to augment intercellular movement of ions across the mucosa rather than the stimulation of transcellular processes.


Pulmonary infection of mice with Staphylococcus aureus. The survival of Staphylococcus aureus in the lungs of mice was studied under various conditions. Doses of 10(7) to 10(9) washed staphylococci were quantitatively introduced into the lungs after intratracheal inoculation in mice under either ether or sodium pentobarbital anesthesia. Mice were sacrificed at intervals, the lungs were excised and homogenized, and the cocci were enumerated by plate count. The 50% lethal dose was 6 x 10(8) cocci per mouse, and mice died within 24 h but without proliferation of the inoculum. Mice given 10(8) cocci intratracheally under pentobarbital anesthesia regularly survived and eliminated the organisms over a 48-h period. The use of ether anesthesia resulted in persistence of the inoculum for up to 48 h, but the organisms were then eliminated. Inability to proliferate did not appear to result from a lack of iron because pretreatment of the mice with ferric ammonium citrate or Imferon did not alter inoculum survival. Staphylococci inoculated intratracheally in mice infected with influenza virus 3 to 21 days previously showed no enhanced persistence or multiplication. Cocci preclumped with fibrinogen, inocula mixed with 10 times the number of Formalin-killed staphylococci, or inocula of the encapsulated Smith strain did not survive any better than conventional inocula, suggesting that phagocytosis might not be the sole mechanism for elimination. However, a sedimentable fraction from normal or infected lung homogenates proved either inhibitory or cidal for staphylococci in vitro.


Immunoprophylaxis with BCG of experimental Echinococcus multilocularis infections. Previous studies have demonstrated that prophylactic treatment with BCG protects cotton rats (Sigmodon hispidus) against experimental infections with Echinococcus multilocularis; this treatment can, however, induce granulomatous reactions. In an attempt to identify a minimum prophylactic dose of BCG which would not induce granulomas, cotton rats were treated intraperitoneally with various doses of BCG (10(1) to 10(7) colony-forming units [CFU]) and then inoculated intraperitoneally with one brood capsule of the parasite. Consistent and complete protection was obtained by the inoculation of as few as 10(3) CFU of BCG. A dose of 10(1) CFU gave no protection whatsoever, and 10(2) CFU gave only partial protection. Doses larger than 10(3) (10(5), 10(7) CFU) also afforded complete protection but gave rise to granulomatous lesions. At the time of the inoculation of the parasite, protection coincided with a general elevation of leukocytes, especially cells of the monocyte/macrophage series. It is proposed that these results support evidence for the macrophage being the principal potential effector cell in hydatid disease.


Morphological and cytochemical characterization of cells infiltrating mouse lungs after influenza infection. To initiate evaluation of the cell-mediated immunological response to influenza virus in a major site of disease, lung cells were obtained by transpleural lavage from lungs of uninfected mice and from those infected 3 or 6 days previously with 5 50% mouse infectious doses (MID(50)) of avirulent (P3) or virulent (P9) influenza A Hong Kong (H3N2) virus. The number of cells recovered by lavage was dependent on the dose, time after inoculation, and the type of virus used for inoculation. Although lavage pools were shown to contain peripheral blood leukocytes, this contamination was shown to be consistently less than 5% of the total leukocytes harvested. Among the ca. 0.75 x 10(6) lavage cells obtained from each uninfected mouse, about 90% were macrophages or lymphocytes in approximately equal proportion. T, B, and null (lyphocytes lacking theta or surface immunoglobulin markers) lymphocytes averaged 23, 9, and 7% of cells in these suspensions, respectively. After infection with either P3 or P9 virus, increased numbers of activated macrophages and lymphoblasts were observed. The major change during P3 infection was an increase in absolute numbers of null lymphocytes. In contrast, during P9 infection, T and B lymphocytes and macrophages progressively increased in absolute numbers while null cells decreased. These data suggest that cell-mediated immunological responses to influenza virus occur in the lung during infection, but that the responses to virulent and avirulent variants may differ both qualitatively and quantitatively.


Analyses of glucans from cariogenic and mutant Streptococcus mutans. The extracellular, water-soluble and cell-associated, 1 N NaOH-soluble glucans from cariogenic Streptococcus mutans 6715-13 "wild type" (WT) and glucan synthesis-defective mutants with diminished virulence have been quantitatively and qualitatively analyzed by methylation analysis and gel chromatography. The mutants synthesized more of a highly branched alpha-(1 --> 6)-rich extracellular polymer than WT, and some of this glucan was also found to be cell associated in all but one case. WT, in distinction to the mutants, also synthesized a highly branched, alpha-(1 --> 3)-rich, cell-associated polymer. Treatment of these two distinct polymer types with dextranase or an alpha-(1 --> 3)-hydrolyzing enzyme indicated they were composed of both alpha-(1 --> 3) and alpha-(1 --> 6) linkages and of alpha-(1 --> 6) with branches at the 3-position, rather than of separate alpha-(1 --> 3) and alpha-(1 --> 6) homopolymer mixtures. Gel chromatography before enzymatic hydrolysis disclosed a high degree of polydispersity in both glucan classes. After hydrolysis polydispersity was reduced, again without resolution of two glucan populations. These findings suggest that (i) there are two distinct glucan classes, one alpha-(1 --> 3) rich and the other alpha-(1 --> 6) rich in WT, (ii) diminution of virulence in the mutants is probably ascribable to a failure to form the alpha-(1 --> 3)-rich component, (iii) both alpha-(1 --> 6)- and alpha-(1 --> 3)-rich glucans are found in association with the cell, and (iv) both highly branched glucan types are dextranase and alpha-(1 --> 3)-hydrolase sensitive, and methylation analysis and gel chromatography suggest polymers with highly polydisperse molecular weights which contain mixtures of linkage types.


Induction of a mucosal antitoxin response and its role in immunity to experimental canine cholera. The induction of a jejunal antitoxin response was studied in dogs immunized with cholera toxin or toxoid. Single doses of toxoid given subcutaneously (s.c.) or of toxin given intraluminally (i.l.) were each effective in priming the mucosal immune system, whereas toxoid given i.l. was much less effective. In contrast, toxin and toxoid given i.l. were each effective as booster antigens. The local secondary response was rapid and brief, the peak occurring at about 7 days after i.l. boosting and declining by 90% after 2 more weeks. After s.c. priming and i.l. boosting with toxoid, antitoxin-containing plasma cells appeared predominantly in the portion of jejunum exposed to the i.l. booster. The appearance of antitoxin-containing plasma cells in jejunal lamina propria correlated with the amount of antitoxin recovered in jejunal washings which, in turn, correlated with protection against challenge with cholera toxin. Thus, lamina propria antitoxin-containing plasma cells appeared to be the source of protective antitoxin. However, after sequential s.c.-oral immunization with toxoid, protection against challenge with Vibrio cholerae far outlasted the major systemic and local antitoxin responses and was not obviously explained by either. These studies reveal methods for induction of a mucosal antitoxin response, but leave in question the mechanism of prolonged protection induced by s.c.-oral immunization of dogs.


Genetic mapping of mutations in independently isolated nontoxinogenic mutants of Vibrio cholerae. Conjugal mating experiments were performed between donor strains of Vibrio cholerae carrying the vibrio sex plasmid P and recipient strains lacking the P plasmid. Donor and recipient genotypes differed with respect to toxinogenicity (tox), nutritional requirements, and antibiotic susceptibilities. Recombinants carrying selected donor and recipient markers were produced at low frequencies in conjugal matings. Mapping of tox markers was accomplished by scoring for the frequency of coinheritance of the donor tox allele with selected and unselected donor markers. Four independently isolated tox markers were analyzed. Each of these four tox markers was shown to be linked to the his-1 site in linkage group I on the genetic map of V. cholerae. In matings between a recipient strain carrying the tox-1 marker and donor strains carrying either tox-2, tox-3, or tox-4, all selected his+ recombinants remained nontoxinogenic. Matings between multiply marked strains demonstrated that the position of tox-1 with respect to other genetic loci in linkage group I is as follows: met-2--trp-1--asp-1--nal-1--his-1--tox-1. These findings demonstrate that a chromosomal determinant linked to his-1 in linkage group I on the genetic map of V. cholerae is essential for toxinogenesis and suggest that tox-1,tox-2, tox-3, and tox-4 may be alleles of a single tox gene.


Local and systemic immunity to influenza infections in ferrets. To establish whether immunity to influenza infection in the ferret is local or systemic, two sites of challenge were utilized: the nose and the anatomically isolated tracheal pouch. Infection of either site did not spread to the other site, and challenge of either site resulted in seroconversion by 13 days. Simultaneous challenge of both sites 21 days after the primary infection revealed that prior infection of the pouch prevented subsequent reinfection of the pouch, but not infection of the nose. Thus, systemic immunity did not prevent the initiation of nasal influenza infection in the ferret. However, the duration of virus shedding from the nose was reduced to half of that seen when ferrets were infected for the first time, showing that the prior pouch infection did lead to a more rapid recovery from the subsequent nasal infection. Passively administered anti-influenza antibody did not prevent or modify the nasal infection, but it did prevent the pouch infection. This is consistent with the observation that an initial infection of the nose prevented pouch infection upon challenge 21 days later. The prior nasal infection also prevented the subsequent nasal infection. These data suggest that immunity to acquisition of influenza infection in the ferret is a local phenomenon, whereas recovery from active infection is influenced by systemic immune mechanisms.


Demonstration of three different subtypes of adenovirus type 7 by DNA restriction site mapping. Restriction site mapping of the genomes of eight different isolates of adenovirus serotype 7 (Ad7) has been performed with six different restriction endonucleases. In this analysis, 37 different restriction sites were localized. Three distinctly different cleavage patterns of the genomes of the Ad7 strains were observed. These strains could not be distinguished by serological techniques. The following three subtypes were defined on the basis of their restriction site patterns: the Ad7 prototype, represented by strain Gomen originally isolated from a case of pharyngitis; subtype Ad7a, represented by the Ad7 vaccine strain and strains isolated from undifferentiated respiratory disease and from a healthy carrier; and a third subtype of Ad7, represented by three strains which were isolated from Swedish patients, all having pronounced clinical symptoms indicating severe systemic infection. A comparison of the restriction site maps of the protype of Ad3 and the three subtypes of Ad7 indicated greater differences in the position of restriction sites between strains of Ad7 than between strains of the two serotypes. This technique is consequently recommended to obtain a more precise definition of distinct entities of viruses.


Scanning electron microscopy of experimental Trichophyton mentagrophytes infections in guinea pig skin. Trichophyton mentagrophytes invasion of guinea pig skin was examined by scanning electron microscopy. Biopsies were obtained daily for 12 days from experimental infection sites. Dermatophyte invasion, examined in detail by scanning electron microscopy of cross-sectioned, prefixed skin was evidenced by: the appearance of hyphae within the stratum corneum; follicular invasion by hyphae, which remained initially within the follicle wall; emergence of the hyphae from the wall into the follicular canal; proliferation of the fungus down the follicle, with furrowing of the follicle wall and hair shaft cuticle; penetration of hyphae into the hair shaft by subcuticular and transcuticular routes; and massive peripilar hyphal proliferation with arthrosporogenesis. A three-dimensional perception of the invasion sequence of a dermatophyte in guinea pig skin was obtained by scanning electron microscopy.


Characterization of glucosyltransferase-deficient, plasmid-containing mutants of Streptococcus mutans LM-7. The possibility that glucosyltransferase (GT)-mediated insoluble-glucan synthesis from sucrose is controlled by the 3-megadalton plasmid pAM7 in Streptococcus mutans LM-7 has been examined. A low-sucrose agar medium was developed to readily detect and quantitate presumptive GT-negative mutants. Such mutants were isolated from Todd-Hewitt broth cultures grown either with or without sodium dodecyl sulfate (10 microgram/ml) or acriflavine (0.5 microgram/ml) at frequencies ranging from about 0.01 to 1%. Independently isolated mutants had the following characteristics: (i) cells were virtually devoid of cell-associated GT and did not aggregate upon addition of sucrose; (ii) cell-free culture fluids synthesized 10X less insoluble glucan than those of the parent; and (iii) cultures grown with sucrose did not form adherent deposits on the wall of the culture tube, as is typical of S. mutans. Both parent and mutants formed relatively little soluble glucan in 1-h assays. Three independently isolated mutants and the parent were found to contain similar amounts of plasmid DNA. Analysis by sucrose density gradient centrifugation and agarose gel electrophoresis did not reveal a size difference between the plasmids from parent and mutants. These results show that (i) S. mutans LM-7 generates GT-deficient mutants at relatively high frequency that still contain a 3-megadalton plasmid; (ii) both cell-associated and extracellular GT levels are depressed in the mutants, which suggests that these activities are directly or indirectly controlled by the same gene or by genes that segregate as a unit.


Ciliated respiratory epithelial monolayers: new model for Mycoplasma pneumoniae infection. Hamster respiratory epithelial cells were cultured in a monolayer format, and 20% of the cells were ciliated. Mycoplasma pneumoniae attached to the epithelial cells in a neuraminidase-specific fashion and induced ciliostasis and cytonecrosis.


Long-term fecal excretion and resistance induced in mice infected with Yersinia enterocolitica. European isolates of Yersinia enterocolitica serotypes O3 and O9 were shown to infect but not kill mice, which became fecal excreters for up to 135 days. The mice challenged with 500 50% lethal doses of the virulent WA strain of Y. enterocolitica serotype O8 survived, and some excreted the virulent strain. This rodent model may be of value in assessing the ecological significance of rodents in the maintenance of Y. enterocolitica.


Enhanced glucosyltransferase activity in penicillin-treated cultures of Streptococcus mutans. Penicillin-treated cultures of Streptococcus mutants GS-5 produced elevated levels of extracellular glucosyltransferase activity.


Protective effect of vaccination against Mycoplasma pulmonis respiratory disease in rats. Intravenous vaccination of rats with either viable or Formalin-inactivated Mycoplasma pulmonis reduced the incidence and severity of lower respiratory tract lesions after intranasal challenge with viable organisms. Intranasal vaccination with killed organisms reduced the severity of rhinitis, but did not affect lesions in any other region of the respiratory tract. The maximum protection against upper tract lesions (rhinitis, otitis, and laryngotracheitis) was provided by intravenous immunization with viable organisms. Dual vaccination (intraperitoneal plus intranasal) with killed organisms provided no significant protection in any segment of the tract. However, these ineffective vaccine regimens did not potentiate the lesions. These results conclusively demonstrate that vaccination of rats against mycoplasma respiratory disease is feasible and also suggest that systemic vaccination may provide greater protection for the lungs than intranasal vaccination, at least when equivalent antigen doses are used.


Lung clearance of Staphylococcus aureus strains with differing protein A content: protein A effect on in vivo clearance. We have studied the clearance from murine lungs of two strains of Staphylococcus aureus, one possessing high and the other possessing low levels of protein  A.S. aureus FDA 209 and S. aureus Wood 46 were assayed for their ability to bind mouse immunoglobulin G, using an indirect radioimmunoassay. S. aureus FDA 209 binding of mouse immunoglobulin was significantly greater than that of S. aureus Wood 46 (118,909 versus 37,845 cpm). Clearance of these two strains from the lung after a 30-min aerosol inoculation period was not significantly different. The percentage of bacteria remaining in the lung was 49.2 and 55.0% at 2h, 31.8 and 33.2% at 3 h, and 25.4 and 17.2% at 4 h for protein A-rich and protein A-poor strains, respectively (P greater than 0.20 at each time). These data suggest that the previously demonstrated in vitro antiphagocytic effect of protein A may not be relevant to pulmonary clearance mechanisms.


Cross-protection in mice after immunization with H2N2, H3N2, and Heq2Neq2 influenza virus strains. Mice were vaccinated with the influenza viruses A/Japan/57 (H2N2), A/Hong Kong/68 (H3N2), and A/Equi/Miami/63 (Heq2Neq2) and the hemagglutinin and neuraminidase recombinants derived from these viruses. After infection with the parent viruses, protection was compared with serological findings. It was found that influenza vaccine protects not only against infection with a strain identical or closely related to the vaccine strain, but against heterologous strains as well. Vaccination with Hong Kong/68 and its neuraminidase recombinant resulted in a heterologous neuraminidase inhibition titer against Japan/57 and in a protection against infection with Japan/57. By contrast, after vaccination with Japan/57 and its neuraminidase recombinant, no relevant heterologous neuraminidase inhibition titer against Hong Kong/68 was observed, whereas a protection against infection with Hong Kong/68 did exist. A cross-protection between Hong Kong/68 and Miami/63, but no relationship in the hemagglutination or neuraminidase inhibition tests, was established in the preinfection sera. A one-way antigenic relationship between these viruses was confirmed by the rise of hemagglutinin or neuraminidase antibodies against Hong Kong/68 in the postinfection sera. No cross-protection or serological relationship existed between Miami/63 and Japan/57. Besides the hemagglutinin and neuraminidase, a third factor, the "mouse-protecting antigen," was considered to contribute to the protection obtained. According to the protection observed, the mouse-protecting antigen of Hong Kong/68 virus is related to that of Japan/57 as well as Miami/63 virus. The mouse-protecting antigens of both Japan/57 and Miami/63 are related to that of Hong Kong/68.


In vitro colonization of Streptococcus mutans on enamel. An in vitro model consisting of enamel from extracted human molars, suspended from wires in inoculated culture tubes, was used to study the adhesion of bacteria to enamel. Under conditions in which there was no macroscopically visible plaque formation, electron micrographs showed no bacterial deposits on the enamel surface. In samples where Streptococcus mutans attached to enamel, an extracellular, pellicle-like material was associated with the bacteria adjacent to the enamel. This material appeared to bind to the enamel surface and to mediate bacterial attachment. Membrane-filtered (Millipore Corp.) saliva deposited a thin surface layer on the enamel, but there were no observable alterations of S. mutans attachment to enamel pretreated with saliva. It was noted that Bratthall serotype c and e strains of S. mutans, when grown in glucose-containing medium, attached, although less tenaciously, to enamel and nichrome wires. Chemical and gas chromatographic analyses of cell-associated materials formed by serotype c and e strains cultured in glucose-containing medium revealed low amounts of glucose-positive material and no polymer linkages characteristic of glucan; yet the same strains cultured in sucrose-containing medium had relatively high amounts of glucose-positive material, with polymer linkages-characteristic of glucan. Serotype a, b, and d strains could attach only in sucrose-containing media.


Specific binding of staphylococcal alpha-toxin to isolated rabbit vagus nerves in vitro. The binding of staphylococcal [125I]alpha-toxin to rabbit vagus nerves in vitro was a saturable process. The radiolabeled alpha-toxin binding was reduced by the coaddition of added navive alpha-toxin, indicating that the binding is specific. Sucrose gradient analysis of detergent-extracted complexes of [125I]alpha-toxin-rabbit vagus nerves showed both high and low S-value peaks analogous to those observed with similarly treated alpha-toxin-rabbit erythrocyte preparations (P. Cassidy and S. Harshman, Biochemistry, in press).


Requirement for a radiosensitive lymphoid cell in the generation of lipopolysaccharide-induced rejection of a murine tumor allograft. Lipopolysaccharide-initiated rejection of a tumor allograft requires the cooperation of a radiosensitive nonadherent lymphoid cell population and an adherent cell, presumably a macrophage.