New insights into the classification and mechanisms of hereditary, chronic, non-haemolytic hyperbilirubinaemias. Gilbert's syndrome is typically associated with a deficiency in hepatic bilirubin UDP-glucuronosyltransferase activity (B-GTA). The overproduction of bilirubin that is often found in this condition could be a fortuitous coincidence that leads to the unmasking of the disease, which otherwise often remains latent. Some cases of chronic unconjugated hyperbilirubinaemia could, however, be related to a defect in hepatic uptake, as reflected by alterations in BSP kinetics. Severe deficiencies of hepatic B-GTA exist in all types of Crigler-Najjar disease. An increased proportion of bilirubin monoglucuronide is always found in bile when a B-GTA deficiency is present. This observation strongly suggests a common biochemical defect in Gilbert's syndrome and in Crigler-Najjar disease, and thus renders the suggestion that the latter condition may be separated into two groups somewhat inappropriate. There is, however, no doubt that further knowledge of the conjugating enzyme, or enzymes, is required: such information may lead to the characterisation of several types of enzymic defects. Whereas little is new as far as the Dubin-Johnson syndrome is concerned, Rotor's syndrome can no longer be considered to be a variant of the former. The transport defect which is involved in most cases of Rotor's syndrome, if not in all, is an impairment of hepatic storage, thus distinguishing it from the impairment of excretion which is involved in the Dubin-Johnson syndrome. The distinct patterns of urinary coproporphyrin excretion, which were recently reported in Dubin-Johnson and Rotor's syndromes, offer additional evidence for a clear differentiation between these two entities.


Alpha heavy chain disease (report of 18 cases from Iraq). The clinical and pathological features of 18 new patients with alpha heavy chain disease seen at two referral centres in Baghdad, Iraq, are described. The series included 14 males and four females ranging in age from 14 to 47 years. Almost all patients presented because of long-standing abdominal pain and diarrhoea. The tissue diagnosis and extent of the disease were established at laparotomy in most patients. Peroral jejunal biospy was used in a number of patients, mainly for follow-up. The serological abnormality was confirmed by immunoselection technique. Most of the patients had extensive thickening of the bowel wall and/or tumour masses of the small intestine and mesenteric nodes. Histopathological sections showed muscularis. Preliminary results of the treatment, including two long remissions, are reported. In general, our observations agree with those made by other authors, mostly from the Middle East and Africa. We believe that a high index of clinical suspicion, routine use of the immunoselection, and recognition of the early pathological changes may hopefully lead to the detection of more cases before the frank neoplastic phase of the disease.


Twenty-five years of progress in bilirubin metabolism (1952-77). This review deals with the development of our understanding of the chemistry of bilirubin and its glucuronide derivatives during the years 1952-1977. It examines the relation between haem metabolism and bilirubin formation and our present knowledge of hepatic transport of bilirubin. The heterogeneity of familial hyperbilirubinaemia is discussed.


Effect of alrestatin on arginine-induced secretion of glucagon and insulin in the rat. Alrestatin, a lens aldose reductase inhibitor, decreased i.v. arginine-induced glucagon levels and augmented arginine-stimulated insulin release in the ether anesthetized rat. Alrestatin may then be useful in the treatment of diabetes mellitus, due to its actions on insulin and glucagon, and its capacity to delay the onset of sugar-induced cataracts in the rat.


Preliminary studies of plasma growth hormone releasing activity during medical therapy of acromegaly. The in vitro growth hormone releasing activity of plasma obtained from six acromegalic subjects was measured before and during therapy. In five subjects, plasmas were obtained before and during successful medical therapy with medroxyprogesterone acetate (MPA). The sixth subject was sampled before and after transphenoidal Sr90-induced hypopituitarism. All subjects had a decrement in fasting growth hormone levels with respective therapies (29-88%). The in vitro growth hormone released from Rhesus monkey anterior pituitaries was assessed after incubating one lateral half in control plasma (pre-therapy) and the contralateral pituitary half in plasma obtained during or after therapy. Studies with plasmas obtained from the five patients successfully treated with MPA showed a decreased in growth hormone releasing activity during therapy in all (18-57%). Plasma obtained after Sr90 pituitary ablation in the sixth subject had 35% more growth hormone releasing activity than obtained before therapy. These results suggest that active acromegalics who respond to MPA with significantly lowered growth hormone levels may actually achieve this response because of a decrease in growth hormone releasing factor measured peripherally. The opposite response in one acromegalic subject, following Sr90 pituitary ablation and hypopituitarism, suggests that growth hormone releasing factor secretion may increase when growth hormone levels are lowered by ablative therapy.


Thyroxine binding globulin (TBG) assay by radial immunodiffusion. Thyroxine-binding globulin was measure by radial immuno-diffusion using ten microliters of serum. The mean value in a normal group was 1.95 +/- 0.38 mg/dl. The values obtained by this method were in good agreement with the thyroxine-binding capacity determined by paper electrophoresis (r = 0.956).


Preparing tube feedings in-house.

Preparing tube feedings in-house. Tube feeding is a therapeutic technique that is widely used in various medical specialties. This article describes the economic and microbiological considerations used by Goldwater Memorial Hospital, New York City, in its decision to prepare tube feedings in-house.


[Clinical, parasitological and serological investigations in sarcosporidiosis (sarcocystis suihominis) of man (author's transl)]. Eight medical students ate for a test raw meat from a pig experimentally infected with sporocysts of Sarcocystis suihominis. The meat had been officially controlled before and was found free of Trichinella and qualified for consumption. 6 to 24 hours after the meal all persons suffered from acute clinical symptoms, above all diarrhoea and vomiting, coldness and sweating which decreased, however, within 12 to 24 hours. Clinical-chemical investigations made during this time suggested an acute infection combined with an exsiccosis; the specificity of the infection, however, could only be established by serological methods and by microscopical demonstration of sporocysts of the agent. The pathogenetic relations are discussed.


In vitro immune response of spleen cells from mice genetically selected for high or low antibody production. The aim of this study was the identification of the cell type in which genes selected for high or low response to SRBC express their functions. Spleen cells from high (H) and low (L) responder mice were immunized with SRBC in the Mishell and Dutton system. An antibody response of different magnitude was found in cultures of H and L spleen cells, the difference being at least as great as that observed in vivo. This finding under experimental conditions allowing the exclusion of any influence of the animal milieu during the immune response, suggest macrophages, B, and T lymphocytes as possible target cells of gene action. In vitro cell separation and recombination experiments in which spleen cells were immunized with SRBC, TNP-LPS, or TNP-HRBC indicate that the genetic differences between H and L responders brought about by selective breeding are expressed in lymphocytes to greater extent than in macrophages. The role of histoincompatibility in the recombination experiments in unlikely but cannot be excluded. Among lymphocytes, B cells but not helper T cells were found more responsive in cultures of spleen cells from H than from L mice.


[Insulin concentration in polytraumatized patients during infusion of glucose, fructose and sorbitol]. Serum insulin concentration was measured during infusion of glucose, fructose or sorbitol for several days in polytraumatized patients. The patients are divided in two groups, one group with normal glucose tolerance and a second group, where an extreme disturbance of the glucose utilization was found. In patients with normal glucose tolerance the glucose substitutes had the same metabolic effects as in metabolically healthy volunteers. In patients with disturbed glucose tolerance the glucose substitutes (fructose as well as sorbitol) effected an increase in blood glucose concentration and in serum insulin concentration. It is concluded that the increase in blood glucose concentration causes the increase in serum insulin concentration. Obviously, in a certain group of polytraumatized patients a "metabolic insulin resistence" exists. Therefore, glucose utilization is decreased despite an increase in serum insulin. In most cases the metabolic disturbance in these patients is mastered, if glucose substitutes are used instead of glucose as energy source. However, in many cases glucose can be administered only if insulin is given additionally.


[Metabolic behavior in pre, intra, and postoperative total parenteral feeding with a carbohydrate combination and amino acid solution]. Ten metabolically healthy patients who had undergone gastric resection were fed intravenously from the preoperative to the 5th postoperative day. No metabolic alternations were observed by using a low dosis of 0.12 mg/kg.h of glucose, fructose and xylitol in combination with amino acids. We were able to limit the so-called postaggressive syndrome by eliminating the fasting state. These results were proven by evaluating the most important metabolic parameters like triglycerides, free fatty acids, lactate and by measuring the carbohydrate-nitrogen balances.


[Postoperative feeding of patients after maxillofacial surgery with Sokoham--a new parenteral nutritive solution]. 20 patients--15 to 71 years old--, who underwent maxillofacial operations received Sokoham for 10 days via a nasogastric tube. In 0.8% of all applications vomiting occurred. Wound healing and time of hospitalisation were the same as in a control group. Mean frequency of defecation was 0.5/24 hours. Body weight, serum electrolytes, fat content of the blood, blood gases, pH, base deficit, serum transaminases, urea-nitrogen and creatinine in the serum did not change significantly.


[The behavior of free serum amino acids and their urinary excretion after major abdominal surgery]. 32 patients with major abdominal surgery received parenteral nutrition with 360 g carbohydrate mixture (levulose: glucose: xylitol = 2:1:1)/24 hours and 1000 ml of a 10% L-amino acid mixture (potato-egg-pattern)/24 hours. Daily nitrogen balances were balanced or slightly negative. Daily we determined serum amino acid concentration; essential amino acid levels were normal. Of non-essential amino acid levels glutamine acid and aspartic acid were elevated. Amino acid loss in urine was 0.2 to 3%, only tryptophan and histidine were excreted in 6.6 and 7.0%.


In vitro susceptibility of gentamicin and/or tobramycin resistant gram-negative bacilli to seven aminoglycosides. The in vitro activity of gentamicin, tobramycin, sisomicin, netilmicin, amikacin, kanamycin and streptomycin was tested simultaneously by the agar dilution method against 584 clinical isolates of gram-negative bacilli that were resistant to gentamicin and/or tobramycin. About half of the gentamicin-resistant Pseudomonas were susceptible to tobramycin but cross-resistance was virtually complete between gentamicin and tobramycin for Enterobacteriaceae. Sisomicin was much more active than gentamicin against Klebsiella, Escherichia and Citrobacter species. Only 18.9%, 27.4% and 27.9% of Klebsiella, Enterobacter and Serratia respectively were resistant to netilmicin. Amikacin was the most effective aminoglycoside with an overall resistance of 15.6%. Kanamycin was effective against 40% of Proteus and Providencia species. Surprisingly, more than half of Klebsiella and Enterobacter species and 85.3% of Serratia species were susceptible to streptomycin.


Stimulatory effect of cigarette smoke on the metabolism and covalent binding of benzo(a)pyrene in the trachea of the rat. The activity of aryl hydrocarbon hydroxylase (substrate: benzo(a)pyrene) was increased in the tracheas of rats exposed to cigarette smoke for 1 h daily for either 1 or 10 days. However the degree of increase in activity was lower in the trachea than in the lung. After a single exposure, activity in the trachea was at its highest level 12 h following exposure (3.2-fold compared to the control), but had returned to the control level within 24 h. Also, the amounts of covalently bound metabolites of benzo(a)pyrene were increased (2-fold) in nucleic acid and protein fractions of the trachea, when the rats were killed 12 h after a single cirgarette smoke exposure. No significant changes in activity of epoxide hydratase (substrate: styrene oxide) could be detected in the trachea. After repeated exposures the activity of UDP-glucuronosyltransferase (substrate: methylumbelliferone) was increased (1.7-fold) in the trachea.


Experimental fractures of monkey mandibles. Forty vervet monkey half mandibles, 20 with unerupted third molars and 20 with erupted third molars, were partially embedded in acrylic and loaded in an Instron tensiometer. Mandibles with unerupted third molars fractured at 15.8 +/- 2.5 kg while those having erupted third molars fractured at 26.4 +/- 4.2 kg. Mandibles having unerupted third molars were shown to be significantly weaker than the mandibles with erupted third molar (t = 8.45, P less than 0.001).


Periodontal and pulpal healing of monkey incisors preserved in tissue culture before replantation. The effect of delayed replantation and tissue culture as a prevention against root resorption was examined in green Vervet monkeys (Cercopithecus aethiops). Extracted incisors were kept in tissue culture medium (Eagle's medium) for 5--14 days before replantation. The extra-alveolar dry period before tissue culture ranged from to 0 to 60 min. Incisors not subjected to tissue culture served as controls. The animals were sacrificed 8 weeks after replantation. The following histologic parameters were registered for each tooth: surface resorption, inflammatory resorption, replacement resorption (ankylosis), downgrowth of pocket epithelium, and periapical inflammatory changes. The evaluation of the pulp included the extent, recorded in mm, of pulp survival. Histometric evaluation showed that teeth immediately placed in tissue culture medium for 5--14 days showed improved periodontal healing, exhibiting significantly less inflammatory resorption than control teeth immediately replanted. Also the extent of pulp survival was significantly increased in teeth replanted after tissue culture. Teeth bench-dried for 60 min and thereafter placed in tissue culture medium also showed a significant reduction in the amount of inflammatory resorption and increased amount of pulp survival compared with control teeth replanted after the 60-min dry extra-alveolar period. Replacement resorption (ankylosis) was found in all teeth in both groups.


Angiolithiasis in the parotid gland.

Angiolithiasis in the parotid gland. Angiolithiasis in salivary glands is a very rare finding, and the knowledge of the morphology and histogenesis of these stone formations is therefore limited. Six small angioliths from the left parotid gland of a 19-year-old girl were examined histologically, microradiographically and diffractometrically. The angiolithiasis developed from a regressed hemangioma which had been observed since the patient was 3--4 years old. The angioliths, which were encapsulated, exhibited in some areas diffuse laminations with shells of alternatingly high and low mineral content. In other areas, however, the sturcture of the stones was rather homogeneous. The central part of all angioliths was highly mineralized. All the examined specimens exhibited a mineral pattern of pure apatite. Due to presence of erythrocytes as well as of fibroblasts in the matrix of the examined noduli, the diagnosis of angiolithiasis was established.


Acute leukemia. An atypical case presenting with gingival manifestations. A case of acute myeloblastic leukemia is reported in which the mode of presentation was an atypical gingival lesion. A definite diagnosis was not made until the patient was in a final stage due to non-diagnostic histologic material. One must be suspicious of any gingival lesion, particulary if there is a sudden onset of bleeding or hyperplasia. Leukemia must be considered even if initial investigations are negative, as in the case presented.


Syndrome of multiple mucosal neurofibromas, pheochromocytoma and medullary thryoid carcinoma. Report of a case. The present article reports a case of multiple endocrine neoplasia (MEN) syndrome, type IIb, including the following components: pheochromocytoma, medullary thyroid carcinoma, hyperplastic corneal nerves and multiple mucosal oral and ocular neurofibromas. The patients, a 35-year-old male also exhibited a series of other pertinent findings, i.e. thickened eyelids, marfanoid habitus and widened mandibular canal and mental foramina. In the discussion the clinical and histopathologic characteristics of the present case are compared with similar findings in other previously published cases. The multiple oral mucosal neurofibromas are described clinically and histopathologically and are stated as a pathognomonic early sign of the syndrome.


Vestibulo-lingual sulcoplasty with skin graft in the mandible. A surgical technique for vestibulo-lingual sulcoplasty in the mandible is described. The lingual bone surface is entirely covered by a mucoperiosteal flap, when the muscles have been cut and the lingual sulcus deepened. A split-thickness skin graft taken from the inner surface of the upper arm is placed vestibularly in the region 38--48. An essential increase of the denture-wearing area is obtained and the dislocating effect of the muscles on the denture is eliminated.


Nasopalatine duct cyst. An aggressive condition in adolescent Negroes from South Africa? A retrospective study of the clinical and radiographic features of 51 nasopalatine duct cysts has been undertaken. Of these cysts 33 were in males (64.7%) and 18 in females (35.3%). The age range was 9--67 years (mean 32 years) with most cases being detected between the second and fourth decades. All of the subjects aged less than 20 years were South African Negroes and the lesions found in this group were invariably over 3.5 cm in diameter and rapidly growing. The average size for all of the cysts (after taking in 19% radiographic enlargment factor into account) was 3.5 cm in greatest diameter. The most common signs were: palatal swelling (88.2%), tooth displacement (78.4%) and sub-labial swelling (17.6%). Root resorption was found in only two of the 40 cysts which encroached upon lamina dura. No recurrences occurred following marsupialization or enucleation.


Lip lesions in Nigerian Igbos.

Lip lesions in Nigerian Igbos. Lesions of the lip seen over a 7-year period at a central laboratory serving the Igbos of Nigeria are reviewed. In approximately 8,500 surgical specimens, there were 63 jaw tumors but only 16 lip lesions. Two of lip lesions alone were carcinomas. Both were (a) squamous celled, (b) associated with sunlight-induced degeneration of the dermal collagen, and (c) found in young adult albinos. Accordingly, albinism should be recognized as one of the etiologic factors in lip cancer. The appropriate preventive action is the wearing of wide-brim hats by albinos.


Histiocytosis X. An analysis of 50 cases. Evidence is presented to support the author's view that Letter-Siwe disease, Hand-Schüller-Christian disease and eosinophilic granuloma are but variants of a single disease entity--histiocytosis X. The clinical behaviour of the disease and its prognosis are related to the age ofonset of the disease. Details of 50 cases of histiocytosis X have been analyzed. The mean age of presentation was 15 years 4 months. The male:female ratio was 3:2, 54% of patients in this series had disease confined to the jaws and oral cavity, males being more likely than females to have disseminated disease.


A clinical study of 32 patients with oral discoid lupus erythematosus. Thirty-two patients (26 females, six males) with discoid lupus erythematosus of the oral mucosa have been studied. Patients with signs of dissemination were classified according to accepted criteria. Four patients had systemic lupus erythematosus as well as oral discoid lesions. Twenty-one patients had discoid skin lesions. Eleven patients had oral discoid lesions without involvement of the skin. The time of onset of the oral discoid lesions rnaged from 6 to 75 years with a mean of 41 years. The mean druation of the oral lesions was 4.2 years. Symptoms from the oral discoid lesions were present in 75% of the patients. The clinical appearance varied and some lesions were difficult to differentiate from oral leukoplakia or from the lesions seen in oral lichen planus. The oral discoid lesions were most often seen on the buccal mucosa, the gingiva, labial mucosa and vermilion border. The oral lesions were infected by yeasts in more than half of thepatients.


Case for ligation of the external carotid artery in composite operations for oral carcinoma. The ligation of the external carotid artery has been performed routinely in 186 executive composite operations in patients suffering from carcinoma of the tongue, floor of the mouth, mandibular gingiva and buccal mucosa. The ligation is indicated because it controls the hemorrhage, ensures a clean operating field and prevents the facial edema and hematoma formation without having any unwanted effect on the healing of the wound.


Biochemical effects on radioprotective agents on the liver microsomal hydroxylating system: in vitro studies. The action of two radioprotectors--cysteamine and cystamine--on the liver microsomal multi-enzyme hydroxylating system, a key stem in drug and biological compounds metabolism, has been studied. Their effects have been systematically analysed at the level of individual enzyme activities and global functions. The two compounds are quite inactive on NADPH and NADH cytochrome c reductase activities, but slightly denature the cytochrome P450 into cytochrome P420. Furthermore, they do inhibit to some extent (30 per cent at 10(-2) M) the rate of codeine hydroxylation and totally suppress ((at 10(-2) M) the NADPH-induced lipid peroxidation which occurs during enzymatic functioning. These results are discussed in the light of the toxicity of radioprotectors.


Morphological and physiological changes in the monkey visual system after short-term lid suture. The effects of short-term unilateral form vision deprivation by suturing the lids in macaques were evaluated in terms of cortical neuronal electrophysiology and cell sizes in the lateral geniculate nucleus (LGN). The results show that cell shrinkage in the LGN is proportional to the duration of lid closure, with the parvocellular laminae being somewhat more sensitive than the magnocellular layers. Conversely, the electrophysiologically demonstrable shift of cortical dominance in favor of the nondeprived eye occurs after only 1 week of deprivation, with equal severity in short-term and long-term deprived monkeys, and precedes the appearance of histological abnormalities in the LGN. This increased cortical vs. geniculate sensitivity to visual deprivation is further demonstrated by the findings that (1) lid closure near the end of the second month of life does not affect the LGN cell size but causes the cortical electrophysiological deprivation effect and (2) the cortical eye dominance but not the size of the geniculate cells can be reversed in favor of the previously deprived eye by surgical closure of the formerly dominant eye.


An identification scheme for gram-negative nonfermentative bacilli. A series of six flow charts have been developed to identify the Gram-negative nonfermentative bacilli most commonly isolated from clinical specimens. Colonial morphology and oxidase reactivity determine the pathway to be followed on the flow chart which then indicates the specific test to be performed. Most isolates can be identified within 24 hours using 3 to 5 tests. Each pathway was selected on the basis of 94-100 per cent confidence limits otherwise an alternate pathway is shown. Results of 2,788 nonfermenters identified by this scheme are discussed.


Tracheobronchial secretions collected from intact dogs. I. Protein and mucous glycoprotein composition. Tracheobronchial secretions (TBS) were collected in situ from intact, anesthetized dogs, and the protein composition and distribution were studied. TBS were separated into supernatant and pellet phases by centrifugation. The supernatant phase comprised 85%, by weight, of the secretory mixture; yet it contained only 58% of the total protein. Peptides of apparent molecular weights ranging from 16,000 to 154,000 daltons were present in the supernatant phase. Electrophoretic and immunochemical comparisons with canine serum suggested that certain of the peptides were of plasma origin including albumin, IgG, IgA, and IgM. Albumin and IgG comprised approximately 70% of the supernatant phase protein. Three percent of the total TBS protein was contributed by the peptide portion of mucous glycoproteins (mucins), and 97% of the mucin protein separated into the pellet phase. When compared to mucus collected from a canine tracheal pouch, TBS was less concentrated in mucins, and differences were observed in electrophoretic profiles between the two samples.


High pressure liquid chromatography with ultraviolet absorbance or fluorescence detection of carbaryl in potato and corn. Carbaryl (1-naphthyl-N-methylcarbamate) was extracted from corn and potato with acetone. The acetone extract was partitioned into methylene chloride-hexane and concentrated for cleanup on a 5% water-deactivated Florisil column. The fraction containing the carbamate was subjected to high pressure liquid chromatographic (HPLC) analysis on a 25 cm LiChrosorb Si60 (5 micron) column (2.2 mm id) by various means. First, the pesticide was analyzed directly with ultraviolet (UV) absorption detection at 254 nm and a mobile phase of trimethyl-pentane-isopropanol (96 + 4). After this, a fluorescent derivative was prepared, using dansyl chloride (5-dimethylaminonaphthalene-1-sulfonyl chloride), and analyzed by HPLC, using trimethylpentane-dioxane (95 + 5) as the mobile phase, with both UV absorption (254 nm) and fluorescence (excitation 365 nm, emission greater than 400 nm) detection. The response/ng for the dansyl derivative with fluorescence detection was 1.3 times greater than that for UV detection and 8 times more sensitive than direct UV detection of the carbamate. About 10 ppb carbaryl could be detected after derivatization in the foods studied; for direct analysis, 30-50 ppb could be detected at a 2:1 signal:noise ratio. Recoveries at 0.1 ppm averaged 90% by direct analysis compared with 78% after derivatization.


Evaluation of potential for aflatoxin occurrence on celery, cauliflower, lettuce, and taro root inoculated with Aspergillus flavus and A. parasiticus. Culture samples of lettuce, cauliflower, celery, and taro root (Colocasia esculenta) were assayed for the presence of aflatoxin after inoculation with Aspergillus flavus and A. parasiticus. Cultures of A. flavus produced both aflatoxins B1 and G1 on taro root, but produced by B1 on lettuce, cauliflower, and celery. For taro root, the percentage of aflatoxin G1 produced was considerably greater than that of B1. While A. parasiticus did produce mycelia and spores on the lettuce and taro root samples, there were not detectable levels of any aflatoxin produced. All the samples studied were successfully extracted and analyzed qualitatively and quantatively for the presence of aflatoxin by using official AOAC thin layer chromatographic procedures. There is sufficient evidence that Aspergilli  can grow on some leafy produce and one strain produced aflatoxins.


Aerobic growth thermograms of Streptococcus lactis obtained with a complex medium containing glucose. With different culturing methods both simple and complex thermograms were obtained with Streptococcus lactis grown aerobically in a complex medium containing growth-limiting concentrations of glucose. The thermogram profiles have been interpreted in relation to growth rate, glucose degradation, and molar growth yields calculated for different time intervals during growth.


Positive regulation of amidase synthesis in Pseudomonas aeruginosa. Mutants of Pseudomonas aeruginosa were isolated that were acetamide-negative in growth phenotype at 41 degrees C and constitutive for amidase synthesis at 28 degrees C. Two mutants were derived from the magno-constitutive amidase mutant PAC111 (C11), and a third from a mutant that had enhanced inducibility by formamide, PAC153 (F6). The three temperature-sensitive mutants produced amidases with the same thermal stabilities as the wild-type enzyme. Cultures growing exponentially at 28 degrees C, synthesizing amidase constitutively, ceased amidase synthesis almost immediately on transfer to 41 degrees C. Cultures growing at 41 degrees C were transferred to 28 degrees C and had a lag of about 0.5 of a generation before amidase synthesis became detectable. Pulse-heating for 10 min at 45 degrees C of a culture growing exponentially at 28 degrees C resulted in a lag of about 0.5 of a generation before amidase synthesis recommenced after returning to 28 degrees C. Acetamide-negative mutants that were unable to synthesize amidase at any growth temperature were isolated from an inducible strain producing the mutant B amidase PAC398 (IB10). Two mutants were examined that gave revertants producing B amidase but with novel regulatory phenotypes. It is suggested that amidase synthesis is regulated by positive control exerted by gene amiR.


Pleiotropic effect of a rifampin-resistant mutation in Bacillus subtilis. Rifampin-resistant (Rifr) mutants were isolated spontaneously from Bacillus subtilis strain 168. A fraction of the mutants did not grow on a minimal medium. A high concentration of one of the L-amino acids (glutamic acid, glutamine, arginine, proline, aspartic acid, or asparagine) was required to restore their growth on the medium. Further analysis of one of the mutants (strain RF 161) suggested that the mutant is unable to use ammonia as a nitrogen source and requires amino acids instead. Activity of glutamate synthase was not detected in the crude extract of the mutant. The Rifr mutation was closely located to cysA and the drug resistance was cotransformed with the property of amino acid requirement at 100% frequency. All revertants to prototrophy tested showed the rifampin-sensitive (Rifs) property. The activity of the DNA-dependent RNA polymerase of the mutant was resistant to rifampin. It is concluded that some alteration of RNA polymerase may cause absence of the activity of an enzyme involved in the nitrogen metabolism.


Formylation of initiator tRNA methionine in procaryotic protein synthesis: in vivo polarity in lactose operon expression. Eucaryotic and procaryotic organisms differ in two aspects of their translation machinery: polycistronic messengers are expressed as a sequence of individual proteins only in procaryotes, and the initiation of protein synthesis proceeds with an initiator tRNA which is found to be modified (formylated) in procaryotes and not in eucaryotes. In the present study, we show that formylation is required in vivo for the coordinate expression of the Escherichia coli lactose operon. Our experiments are consistent with a translation mechanism using dissociated ribosomes at the 5' end of the mRNA in a reaction that is only weakly dependent on formylation at this initiation step; the ribosomes then travel along the messenger and can reinitiate after the intracistronic barrier without dissociation. This latter initiation step is strongly dependent on the level of formylation: a low level of the formyl group, obtained by the antifolic agent trimethoprim, induces a strong polarity in the expression of the lactose operon. There exist mutant strains in which this polarity is much less apparent than in the wild type. We show here that such is the case of rpsL mutants. Ribosomes mutated in the S12 protein (rpsL) are found to be much more easily dissociated than the wild type. This might explain why the expression of the lactose operon on rpsL strains remains coordinated when the intracellular level of formylation is decreased.


Evidence that the regulation of diphtheria toxin production is directed at the level of transcription. It has been known for several decades that iron inhibits the production of diphtheria toxin by Corynebacterium diphtheriae by preventing expression at maximal levels. We examined the inhibition kinetics of toxin production after the addition of either iron or rifampin to iron-limited cultures of C7 (betatox+). Iron-mediated inhibition of toxin production was found to be linear within the range of 16 nM to 16 micron. The inhibition kinetics following the addition of iron or rifampin was almost identical. [3H]RNA extracted from iron-limited toxigenic C. diphtheriae was found to hybridize to a greater extent to corynephage beta DNA than either [3H]RNA extracted from toxigenic C. diphtheriae before the onset of toxin production or [3H]RNA extracted from nonlysogenic, nontoxigenic C. diphtheriae.


Altered mRNA metabolism in ribonuclease III-deficient strains of Escherichia coli. The metabolism of mRNA from the lactose (lac) operon of Escherichia coli has been studied in ribonuclease (RNase) III-deficient strains (rnc-105). The induction lag for beta-galactosidase from the first gene was twice as long, and enzyme synthesis was reduced 10-fold in one such mutant compared with its isogenic rnc+ sister; in the original mutant strain AB301-105, synthesis of beta-galactosidase was not even detectable, although transduction analysis revealed the presence of a normal lac operon. This defect does not reflect a loss of all lac operon activity galactoside acetyltransferase from the last gene was synthesized even in strain AB301-105 but at a rate several times lower than normal. Hybridization analyses suggested that both the frequency of transcription initiation and the time to transcribe the entire operon are normal in rnc-105 strains. The long induction lag was caused by a longer translation time. This defect led to translational polarity with reduced amounts of distal mRNA to give a population of smaller-sized lac mRNA molecules. All these pleiotropic effects seem to result from RNase III deficiency, since it was possible to select revertants to rnc+ that grew and expressed the lac operon at normal rates. However, the rnc-105 isogenic strains (but not AB301-105) also changed very easily to give a more normal rate of beta-galactosidase synthesis without regaining RNase III activity or a faster growth rate. The basis for this reversion is not known; it may represent a "phenotypic suppression" rather than result from a stable genetic change. Such suppressor effects could account for earlier reports of a noninvolvement of RNase III in mRNA metabolism in deliberately selected lac+ rnc-105 strains. The ribosomes from rnc-105 strains were as competent as ribosomes from rnc+ strains to form translation initiation complexes in vitro. However, per mass, beta-galactosidase mRNA from AB301-105 was at least three times less competent to form initiation complexes than was A19 beta-galactosidase mRNA. RNase III may be important in the normal cell to prepare lac mRNA for translation initiation. A defect at this step could account for all the observed changes in lac expression. A potential target within a secondary structure at the start of the lac mRNA is considered. Expression of many operons may be affected by RNase III activity; gal and trp operon expressions were also abnormal in RNase III- strains.


Occurrence of squalene in methanol-grown bacteria. The nonpolar lipids of methanol-grown bacteria which utilize one-carbon (C1) compounds via the RMP pathway (Pseudomonas C, Pseudomonas methylotropha, and Methylomonas methanolica) were found to contain squalene in concentrations between 0.1 to 1.16 mg/g of cell (dry weight). Squalene could not be detected in lipid extracts of methanol-grown bacteria which utilize C1 compounds via the serine pathway.


Enzymatic excision of free hypoxanthine from polydeoxynucleotides and DNA containing deoxyinosine monophosphate residues. A DNA glycosylase that releases free hypoxanthine by selective cleavage of dIMP residues in polydeoxynucleotides and DNA containing this nonconventional nucleotide is present in Escherichia coli cell extracts. The partly purified enzyme, termed hypoxanthine-DNA glycosylase, does not require divalent cations or phosphate for activity, and it acts more efficiently on double-stranded than on single-stranded substrates. The enzyme has properties different from either uracil-DNA glycosylase or 3-methyladenine-DNA glycosylase and is present at normal levels in E. coli mutants deficient in either of the latter two DNA glycosylases.


Evolution of immunoglobulin subclasses. Primary structure of a murine myeloma gamma1 chain. The amino acid sequences of the first six CNBr fragments (303 residues) of MOPC 21 heavy chain have been determined, using conventional techniques. This completes the determination of the covalent structure of MOPC 21 immunoglobulin (IgG1, K). MOPC 21 gamma1 chain is a unique sequence of 440 amino acid residues. The variable region is homologous with human VHIII sequences. A comparison of the constant region with homologous sequences shows that mouse gamma1 and gamma2 proteins differ much more than the gamma subclasses in man and in some other species and supports earlier suggestions that mouse gamma1 and the four human gamma subclasses have a common ancestor different from the mouse gamma2 ancestor. Unlike some other subclasses, but like those in the guinea pig, mouse gamma1 and gamma2 genes have diverged before speciation. An expansion-contraction mechanism is proposed for the evolution of gamma genes.


Spontaneous somatic mutations. Structural studies on mutant immunoglobulins. The precise alterations in the protein amino acid sequences of the immunoglobulin heavy chains of spontaneously arisen MOPC 21 mutant clones IF2 and IF1 have been determined. All the cyanogen bromide fra-ments of both heavy chains have been isolated and compared to the wild type CNBr fragments. For IF2, there is an internal deletion, from the wild type sequence, of 96 amino acids, from residues 121 to 215 inclusive. Moreover, in IF2, there are no disulfide bonds formed between heavy and light chains, presumably because of the deletion of the CH1 pseudosubunit. There are no other alterations in its covalent structure. For IF1, there is a deletion of the COOH-terminal 83 amino acids (residues 358 to 440, inclusive). Although IF1 heavy chain behaves on dodecyl sulfate-polyacryl-amide gels as if it were only 10 to 15 residues shorter than wild type, no other amino acid sequence differences from wild type are found. IF1 arose most likely by a nonsense mutation of a serine codon. For IF2, whose deletion is like that seen in some human heavy chain disease proteins, the most likely explanation is an error of recombination. The structure of IF2 suggests that the heavy chain variable region ends at a position homologous to residue 120 of the MOPC 21 heavy chain.


Effect of cold exposure on rat erythrocyte membrane-bound acetylcholinesterase. Role of thyrotropin in the thyroid hormones interplay. The influence of cold exposure at 4 degrees C for different periods of time (from 12 h to 42 days) on the allosteric inhibition by F- of the rat erythrocyte membrane-bound acetylcholinesterase from rat fed a corn oil diet was studied. The cold exposure decreased the values of the Hill coefficient n from 1.6 to 1.0. When the cold-exposed rat was transferred from the cold environment to 23 degrees C, the values of n reached the control values. The factors that play in the allosteric desensitization phenomenon were characterized as L-triiodothyronine, L-thyroxine, and thyrotropin. The relationship betwen changes in the values of n and physiological concentrations of thyroid hormones and thyrotropin in cold-exposed rat is shown. Thyrotropin showed a facilitatory action on the blocking action of thyroxine on the triiodothyronine effect. The intravenous injection of thyrotropin-releasing hormone (TRH) yields confirmatory results for this regulatory mechanism since the values of n for acetylcholinesterase shifted as predicted.


Dihydrofolate reductase from amethopterin-resistant Lactobacillus casei. Sequences of the cyanogen bromide peptides and complete sequences of the enzyme. The complete amino acid sequence of dihydrofolate reductase from an amethopterin-resistant strain of Lactobacillus casei has been determined by sequence analysis of peptides produced by cleavage with cyanogen bromide, trypsin, staphylococcal protease, and myxobacter protease. Comparison of this sequence with those of reductases from other bacterial sources shows that the enzymes are homologous. The Lactobacillus casei reductase sequences shows a 29% sequence identity with that of the Escherichia coli enzyme and a 34% identity with the sequence of the enzyme from Streptococcus faecium. The NH2-terminal 68 residues of the L. casei reductase show a 54% sequence identity with that of the enzyme from S. faecium.


Formation and function of N-acetyloglucosamine-linked phosphoryl- and pyrophosphorylundecaprenols in membranes from Bacillus cereus. Membranes from Bacillus cereus AHU 1356 incorporated radioactivity from UDP-N-acetyl[14C]glucosamine into three alkaline-stable and acid-labile lipids which were extracted into chloroform:methanol (2:1) and separated from each other by thin layer chromatography on silica gel plates. The major labeled lipid (Lipid 1) and a minor one (Lipid 2) were identified as N-actetylglucosaminyl phosphorylundecaprenol from several analytical criteria involving mass spectral data and from reversal of their formation by UDP. These two lipids appear to differ in geometry of their polyprenol moieties. The third labeled lipid (Lipid 3) was identified as N-acetylglucosaminyl pyrophosphorylundecaprenol. Antibiotic 24010, a tunicamycin-like antibiotic, at 1 microgram/ml was found to inhibit almost completely the formation of Lipid 3, whereas it inhibited the formation of Lipid 1 much more weakly and rather enhanced the formation of Lipid 2. Radioactivity was also incorporated into a polymer from UDP-GlcNAc and from Lipid 3. UDP-N-acetylmannosamine, UDP-N-acetylgalactosamine, and UDP-glucose supported the incorporation. Antibiotic 24010 strongly inhibited the incorporation of radioactivity from UDP-GlcNAc into polymer, whereas it did not affect the incorporation from Lipid 3. Thus, it is concluded that N-acetylglucosaminyl pyrophosphorylundecaprenol serves as a precursor in the synthesis of a polymer presumed as the cell wall polysaccharide of this bacterial strain.


L-asparaginase bound to collagen membranes: effect of glutaraldehyde  crosslinking on stabilization of catalytic activity. Reconstituted bovine collagen has been used extensively in our laboratory as a carrier for immobilized E. coli L-asparaginase. The activity and catalytic stability of these collagen-asparaginase membranes can be altered substantially by conditions used in membrane crosslinking with glutaraldehyde. As the concentration of glutaraldehyde used in tanning is increased, the initial specific activity of collagen-asparaginase membranes decreased asymptotically to a limiting value. Similar results occurred when membranes were subjected to increasing time periods of tanning at a constant glutaraldehyde concentration. These observations point to a time-concentration relationship for glutaraldehyde tanning and its effect on the specific activity of collagen-asparaginase membranes. Specific activities of membranes tanned a glutaraldehyde concentrations of 5% or higher appear to be very stable over long periods of alternate storage and assay. This result, however, is not observed with membranes tanned at glutaraldehyde concentrations lower than 5% for short periods of time (approximately 30 sec to 1 min). It is not clear whether the instability of membranes tanned at lower concentrations of glutaraldehyde or shorter intervals of tanning is due to enzyme elution from the membrane or denaturation of the bound enzyme.


Fat macroglobulemia caused by fractures or total hip replacement. The numbers of fat macroglobules in peripheral venous blood were recorded for 136 prospectively studied patients with fractures of the pelvis, femur, or tibia. Groups of healthy volunteers and patients undergoing total hip replacement or abdominal surgery served as controls. The highest incidence of fat macroglobulemia occurred in patients with fractures of the femur who also had these clinical signs: petechiae, hypoxia, and depression of the central nervous system. The peak incidence occurred within the first eight hours after fracture. The over-all incidence of fat macroglobulemia in the other surgical patients was roughly equivalent to that in the fracture patients.


Calcium-binding protein of the chick chorioallantoic membrane. I. Immunohistochemical localization. The preparation of a specific antiserum (anti-CaBP) against the calcium-binding protein (CaBP) of the chorioallantoic membrane (CAM) is described. The anti-CaBP appeared to be specific for the CaBP by immunodiffusion and immunoelectrophoresis. Application of the anti-CaBP in immunofluorescence histochemistry revealed that the CaBP is present in the CAM only at developmental ages corresponding with the expression of the calcium transport function of the membrane. Furthermore, the CaBP is localized to the ectoderm of the CAM, appears to be exposed to the entire external surface of the ectoderm, and can be shown to be associated with cells enzymatically dissociated from the CAM. These results are consistent with a functional role of the CaBP in the CAM calcium transport process.


Surface organization and composition of Euglena. II. Flagellar mastigonemes. The surface of the Euglena flagellum is coated with about 30,000 fine filaments of two distinct types. The longer of these nontubular mastigonemes (about 3 micron) appear to be attached to the paraflagellar rod whereas the shorter nontubular mastigonemes (about 1.5 micron) are the centrifugally arranged portions of a larger complex, which consists of an attached unit parallel to and outside of the flagellar membrane. Units are arranged laternally in near registration and longitudinally overlap by one-half of a unit length. Rows of mastigoneme units are firmly attached to the axoneme microtubules or to the paraflagellar rod as evidenced by their persistence after removal of the flagellar membrane with neutral detergents. SDS-acrylamide gels of whole flagella revealed about 30 polypeptides, of which two gave strong positive staining with the periodic acid-Schiff (PAS) procedure. At least one of these two bands (glycoproteins) has been equated with the surface mastigonemes by parallel analysis of isolated and purified mastigonemes, particularly after phenol extraction. The faster moving glycoprotein has been selectively removed from whole flagella and from the mastigoneme fraction with low concentrations of neutral detergents at neutral or high pH. The larger glycoprotein was found to be polydisperse when electrophoresed through 1% agarose/SDS gels. Thin-layer chromatography of hydrolysates of whole flagella or of isolated mastigonemes has indicated that the major carbohydrate moiety is the pentose sugar, xylose, with possibly a small amount of glucose and an unknown minor component.


[Surgery of bimalleolar fractures. Results of a series of 70 cases with a follow up of 3 to 8 years (author's transl)]. The authors analyse a series of 70 bimalleolar fractures treated surgically with a follow up of 3 to 8 years. The late functional and radiological result was favourable in 85.7% of cases. They were able to determine from this series a certain number of unfavourable factors on which surgery has a better influence than immobilisation: e.g. fractures with a large displacement, impaction of the roof of the mortar, open fractures. Complications were rare; in 50% they were the result of technical errors avoidable with experience. The indications are noted; the authors consider that certain fracture should be operated straight away, that others should not be operated on, and that finally, an intermediate group should first be treated by immobilisation.


Identification of low chlorinated biphenyls in human milk by gas chromatography--mass spectrometry. Trichlorobiphenyl and tetrachlorobiphenyls in human milk were analysed by gas chromatography--mass spectrometry and mass fragmentography using silicone OV-1 and Apiezon L grease as stationary phases. Low-chlorinated biphenyls had been considered to be excreted rapidly from the body, but in this work quantities of tri- and tetrachlorobiphenyls amounting to approximately 30% of the total of polychlorinated biphenyls (PCBs) accumulated in the milk were identified. This result is important in relation to the quantification of PCBs and in studies of the mechanism of biological degradation.


Phenylboronic acid as a ligand for biospecific chromatography of serine proteinases. Via attachment of p-(omega-aminoethyl)phenylboronic acid to CH-Sepharose in the presence of water-soluble carbodiimide, a new sorbent for the biospecific chromatography of serine proteinases was obtained. The sorbent was shown to be suitable for the purification of subtilisn, alpha-chymotrypsin and trupsin. It is assumed that the serine hydroxyl group at the active site of the enzyme forms, with the boronic acid moiety of the ligand, a structure that imitates transition enzyme--substrage complex. The presence of glycerol selectively improves the binding of serine proteinases, presumably because of stabilization of the tetrahedral state of the boron atom. Direct isolation of subtilisin from a Bacillus subtilis cultural filtrate on phenylboronic acid-containing sorbent gives a virtually homogenous enzyme (42-fold purification) in a nearly-quantitative yield.


Rats defend different body weights depending on palatability and accessibility of their food. Normal adult rats lived on powdered diets adulterated to contain as much as 1.6% quinine sulfate, on a palatable high-fat diet, or in Skinner boxes with 45-mg Noyes pellets available on fixed-ratio (FR) schedules as high as FR 156. They maintained lower body weights over periods of months in proportion to the percentage of quinine adulteration or the fixed ratio. Rats exposed to the high-fat diet overate as much and gained weight as rapidly as rats recovering from food deprivation, and became moderately obese. Rats having become lean or obese contingent on the palatability or accessibility of their diet defended body weight by eating more in the cold, less when force-fed by gavage, and more to restore weight after food deprivation. Yet on chow they restored and defended body weights typical of rats whose diet had been confined to commercially prepared chow. These results are interpreted to be inconsistent with motivational models that rigidly distinguish drive from incentive, that treat body weight changes as evidence for failure to regulate energy balance or body weight, or that rely exclusively on deprivation of food or reduction of body weight for definitions of need for calories. Instead, caloric homeostasis in rats may incorporate ecological constraints.


Immunization with dextransucrases, levansucrases, and glycosidic hydrolases from oral streptococci. II. Immunization with glucosyltransferases, fructosyltransferases, and glycosidic hydrolases from oral streptococci in monkeys. The feasibility of immunizing monkeys with enzymes from oral streptococci in an attempt to reduce dental caries was investigated. Forty rhesus monkeys, Macaca mulatta, were used. Cariogenic streptococci, S mutans, were implanted into all the monkeys' mouths. There was no pathological effect resulting from immunization. Of the 40 animals, 30 retained the implanted flora throughout the experiment; the remaining 10 were reimplanted until the streptococci remained. In six months, gross carious lesions were evident with plaque. Inhibitiors present in the monkey sera after immunization inhibited glucosyltransferase, fructosyltransferase, and neuraminidase activities. It was presumed the inhibitors were antibodies. There was a reduction of 68.6% in the total carious lesions in the animals immunized intraorally with glucosyltransferase, 62.4% reduction in those injected with fructosyltransferase, and 57.4% reduction in total lesions in those immunized with glycosidic hydrolases after 19 months, as compared to the control group. There were no gross lesions apparent in the group immunized with glycosidic hydrolases. It appears that immunization with enzymes significantly reduces carries and is feasible in a primate model.


The bilateral response of the temporal muscle in the rhesus monkey (Macaca mulatta) to short-term detachment of the muscle and increased loading of the mandible. The electromyographic background activity of the temporal muscle was observed after short-term detachment of the muscle and with increased loads on the mandible. Detachment of the muscle increased its mean spike discharge within the first 24 days but without significant changes in the spectral components. Loading the mandible decreased both the spike discharge and spectral components.


Selective habitation of bacteria on "intermediate soft" keratin surfaces of the rat tongue. Scanning and light microscopy reveals a selective and restrictive pattern of indigenous bacterial colonization on the tongue of rats which is accounted for by the preferred attachment of bacteria to an "intermediate soft" keratin substrate. Total intravenous feeding of rats for 10 days did not alter this colonization pattern indicating that oral intake of food is not responsible for maintenance of this pattern.


8-Hydroxyquinolines with the potential for long-term anticaries activity: design, synthesis, and in vitro evaluation. 8-Hydroxyquinolyl benzoate and 8-hydroxyquinolyl para-(fluorosulfonyl)benzoate have been synthesized and evaluated in vitro as antimicrobial and antiplaque agents. Both compounds inhibited the growth of cultures of the following genera: Streptococcus, Staphylococcus, Actinomyces, Lactobacillus, Veillonella, and Candida. Minimum inhibitory concentrations ranged from 0.98 to 62 microgram/ml. Extracted human teeth pretreated with 1% solutions of either compound and rinsed with distilled water exhibited reduced in vitro plaque formation for 48 hours. These results indicate that the in vitro antiplaque activity of 8-hydroxyquinolines can be enhanced by attaching the appropriate side chain in the 8-position.


Dental instrument sterilization by glass bead conduction of heat. Long handle dental instruments were inoculated with oral organisms or with B subtilis and immersed in a glass bead sterilizer at 400 F (204 C). It required from 12 to 14 minutes to sterilize the instruments at that temperature.


Remodeling of cancellous bone in young monkeys. Remodeling of the trabeculae in cancellous bone was studied in young monkeys that received multiple weekly injections of lead acetate as a marker. Resorption of old bone and deposition of new bone were demonstrated. The rate of deposition of new bone varied at different sites of the same trabeculae and also at different time intervals of the same site. The total formation time of new bone to fill the trabecular space was estimated.


The cranial base in fetal Macaca nemestrina: a quantitative analysis of size and shape. The applicability of Fourier analysis to quantitate the midsagittal cranial base has been demonstrated utilizing fetal Macaca nemestrina. This methodology accurately measures the irregular form of the endocranial profile, minimizes the effects of size, and maximizes shape differences. This method provides a quantitative dimension to the descriptive analysis of shape change previously observed with a combined histologic and cephalometric analysis.


Dentinoenamel junction area in primary teeth affected by Morquio's syndrome. Separation of dentin from enamel in the dentinoenamel junction area was achieved in primary teeth affected by Morquio's syndrome. Enamel and dentin junctional surfaces were examined by means of scanning electron microscope. These two tissues were found to be separated by a layer of amorphous material. Recurrent treatment of the amorphous layer with NaOCl caused its gradual removal, exposing the rod tips on the enamel side and tubules on the dentin side. It is reasonable to suppose from this finding that the amorphous layer is rich in organic matrix. It is suggested that this amorphorus layer is analogous to the enamel-dentin membrane, that in this syndrome is only partially calcified.


Randomised controlled trial of early discharge for inguinal hernia and varicose veins. A randomised controlled trial has been conducted into the effects of discharging patients from hospital either 48 hours or six to seven days after operations for inguinal hernia and varicose veins. There was no statistically significant difference in major postoperative complications between the two lengths of stay for either of the two conditions. Similarly there was no difference between the two groups of hernia patients in relation to eventual recurrences. There was no significant difference in length of convalescence between long-stay and short-stay patients in full-time occupations. The savings to the statutory services of discharging patients early were estimated at 25.72 pounds per patient. Patients appeared to approve of the type of care they experienced, regardless of length of stay. However, the families of short-stay patients were significantly less enthusiastic in their attitudes towards the policy of early discharge than the families of long-stay patients.


Visual-field displacements in human beings evoked by accoustical transients. Sixty-two of 133 subjects reported visual-field displacements when they were exposed to intense (125 dB SPL) repetitive audiofrequency transients. This phenomenon was investigated in three experiments. Frequency (100-5000 Hz) was varied in experiment I; repetition rate (0.5/s--6.0/s) was varied in experiment II; acoustical transient onset/offset time (0.2--25 ms) was examined in experiment III. The results of these three experiments indicated that the largest proportion of displacement reports and the largest perceived motion magnitudes followed stimulation in the 500- to 1000-Hz frequency range at repetition rates of about 1/s. Response differences as a function of onset/offset time were erratic. The pattern of results obtained in this study, in conjunction with the results of previous investigations of acoustical vestibular stimulation, suggests that the visual-field displacments resulted from stimulation of the receptors of the vestibular system. These experiments may account for discrepancies in reports of infrasound-evoked eye movements. Finally, it is suggested that intense sound exposure may damage the vestibular receptors with or without concomitant damage to the auditory portion of the membranous labyrinth.


Antigen-induced neutrophil chemotactic activity in man. Correlation with bronchospasm and inhibition by disodium cromoglycate. We have previously reported increased neutrophil chemotactic activity in sera obtained after positive antigen inhalation responses in atopic subjects. This report describes the kinetics of appearance of this serum activity and the effects of antigen dose and disodium cromoglycate pretreatment on the response in 10 ragweed-sensitive subjects. Significantly increased chemotactic activity was present as early as 1 min, peaked at 10 min, and persisted through 24 hr after inhalation of antigen. The increased chemotactic activity correlated with the degree of bronchospasm induced by antigen inhalation and the amount of antigen administered. The increased chemotactic activity and bronchospasm were blocked by administration of disodium cromoglycate prior to antigen challenge. These findings are consistent with a postulated antigen-induced anaphylactic release of chemotactic activity. The correlation of this activity with the degree of bronchospasm and its appearance after administration of even small doses of antigen suggest that this activity may be important in antigen-mediated bronchospasm.


Nutritional management of the adult patient undergoing peritoneal dialysis. Until the last few years, maintenance peritoneal dialysis (PD) often was associated with progressive wasting due to frequent episodes of peritonitis, loss of considerable amounts of protein into the dialysate, and poor nutritional intake. Recently, available techniques have made PD a feasible alternative for the long-term care of the patient with end-stage renal failure. The incidence of peritonitis has been markedly reduced, and protein loss is only 4 to 20 gm. per dialysis treatment. Preliminary studies have shown no differences in the nutritional status of patients undergoing PD or hemodialysis, although both groups have evidenced malnutrition. In the patient undergoing PD, daily intakes of 1.2 to 1.5 gm. protein and 35 kcal per kilogram body weight are recommended. During times of stress, parenteral administration of nutrients may be necessary. Dietary supplements may often be required chronically. Careful studies are needed to difine the nutritional needs of the patient undergoing PD.


Peripheral nerve hamartoma with macrodactyly in the hand: Report of three cases and review of the literature. Two patients with hamartomas of the median nerve and one with hamartoma of the ulnar nerve associated with macrodactyly are described. Nineteen similar cases of this association have been found in the literature. There is evidence that these two conditions are part of a localized regional growth disturbance. This type of macrodactyly should be differentiated from neurofibromatosis.


An experimental study of the central gustatory pathways in the monkey, Macaca mulatta and Cercopithecus aethiops. 1. The cortex at the base of the central fissure, that is the fronto-parietal operculum, represents primary sensory receptive cortex for gustatory modalities. 2. This primary receptive cortex for taste is linked to the anterior Island of Reil by short inter- and intracortical association fibers. Thus the anterior island is a gustatory association area involved in the subjective recognition of gustatory modalities. 3. The nucleus ventralis posteromedialis pars parvocellularis of the dorsal thalamus is the thalamic receptive nucleus for gustatory impulses in the macaque. 4. Fibers which originate at rostral glossopharyngeal levels from the dorsal visceral gray terminate in the contralateral mucleus ventralis posteromedialis pars parvocellularis.


Bacteria induce lymphokine synthesis polyclonally in human B lymphocytes. We have studied the ability of various bacteria to stimulate human lymphocytes to produce leukocyte migration inhibitory factor (LIF). Mononuclear cells from adult and cord blood as well as purified T and B lymphocytes were stimulated with killed bacteria. The culture supernatants were tested for the presence of LIF by the agarose migration method. All nine bacterial strains tested activated unseparated mononuclear cells and B lymphocytes but not T cells to produce LIF. LIF was also present in cord blood cell cultures suggesting that the stimulation of lymphocytes was polyclonal rather than antigenic. Therefore, we propose that one of the physiologic functions of B lymphocyte lymphokines might be to form part of the nonspecific defense mechanisms against microbial invasion.


Allogeneic restriction of acquired antimicrobial resistance in the rat. Allogeneic restriction of acquired antimicrobial resistance was demonstrated by infusing Listeria-immune thoracic duct lymphocytes (TDL) into syngeneic, semisyngeneic, or allogeneic recipients that had been infected i.v. with Listeria monocytogenes (LM). Syngeneic and semisyngeneic recipients enjoyed a high level of protection (3 to 5 log10 reduction in viable organisms within 64 hr, whereas allogeneic recipients were poorly protected (0.5 to 2.5 log10 reduction). Sharing one haplotype between TDL-donors and challenged recipients resulted in a 4- to 8-fold increase in the level of resistance transferred. Histoincompatibility reactions could be excluded as a cause of the observed allogeneic restriction. Therefore, it is likely that LM-immune TDL are unable to respond efficiently to the antigens of this organism in an allogeneic environment. But allogeneic immune TDL do confer a low level of protection that cannot be ascribed to B cell activity nor to GvH and/or HvG reactions. Evidence is provided that the modest protection afforded by allogeneic lymphocytes is conveyed by the same T cell subset that operates with much higher efficiency in genetically related rats.


Antibody-dependent lymphocytotoxicity: an analysis of effector cell-target cell interactions. Antibody-dependent lymphocytotoxicity (ADL) was studied with human peripheral blood lymphocytes as effector cells and P815 mouse mastocytoma cells, sensitized with rabbit IgG antibodies, as target cells. Enzyme-substrate-like kinetics were used to describe ADL inhibition induced by two types of inhibitors. Human IgG, both native and heat-aggregated, proved to be a competitive inhibitor of ADL at the target cell level. Human peripheral blood monocytes inhibited ADL in an apparently irreversible fashion, without appreciable evidence for competition. The data obtained provide strong support for the validity of an enzyme-substrate-like mechanism of ADL. Moreover, our results indicate that in order to measure the lytic capacity properly, cell populations, well defined as to their composition, should be used. In the second part of the study, it was investigated whether the two types of cells capable of lysing sensitized target cells, i.e., null cells and T cells, differed with respect to their affinity for the target cells. Application of enzyme-like kinetics revealed considerable differences in maximal killing rate between the two subsets of K cells. However, the parameter related to the affinity toward the target cells was found to be of the same magnitude for the two types of effector cells.


Evidence that bovine conglutinin reacts with an early product of C3b degradation, and an improved conglutination assay. When EAC43b were treated with heated serum in EDTA, reactivity with bovine conglutinin appeared rapidly, even at 0 degrees C, and almost simultaneously with the loss of C3b rosetting capacity. At the time conglutinability first appeared, there was no detectable decrease in I-A or hemolytic C3 activity, and no detectable C3 antigen release from the cells. With prolonged exposure to heated serum in EDTA, I-A (immune adherence) and hemolytic C3 activity were lost. If this exposure was at 37 degrees C, C3 antigen became strongly detectable in the supernatant fluid, and eventually conglutinability was markedly reduced or lost, whereas C3d rosettes were unaffected. We suggest that bovine conglutinin reacts with some early product of C3b degradation, rather than with C3d, and propose that this intermediate be designated C3k. We have developed a semi-quantitative assay for bovine conglutinin, utilizing a Coulter Counter to register the decrease in total particles due to red cell aggregation. By using this method, we have detected conglutination with mouse complement (C) as well as with that from man and the guinea pig.


Characterization and genetic control of the immune response to synthetic polypeptide antigens of defined geometry. Both humoral and cell-mediated immune responses to the synthetic helical hapten-carrier conjugate poly-Glu-Tyr-Lys(TNP)-(Glu-Tyr-Ala)5 were found to be linked to the major histocompatibility locus in mice and guinea pigs. The responder mouse strains (H-2d haplotype) showed a primary IgM response with an IgG component appearing after the secondary immunization. The antibody response was accompanied by a positive DTH reaction in responder strains. Nonresponder mice (H-2b or H-2k haplotypes) showed neither IgM nor IgG antibodies and the DTH reaction was negative. Administration of the antigen as a complex with an immunogenic carrier was not effective in inducing a response in nonresponder mice. In guinea pig studies, it was found that strain 2 animals were able to mount an antibody response against the TNP-hapten and a DTH response against the polypeptide backbone. Strain 13 animals gave no anti-TNP antibodies at the lower dose levels and DTH activity was entirely negative for all doses of immunizing antigen. Replacement of the TNP hapten by the arsanilazo dipeptide derivative, BOC-gly-ARA-tyrosine, converted the nonresponder strain 13 guinea pigs into complete responders showing antibody and DTH reactions to both the hapten and the polypeptide backbone.


Osmometric studies of human immunoglobulin G. Osmometric studies of solutions of IgG and its H and L chains were performed in various solvents. It has been found that in aqueous solutions and in guanidinium chloride solutions of low and moderate concentrations IgG molecules are associated which prevents molecular weight determination. On the other hand, in 6 M guanidinium chloride the molecular weight of IgG was determined to be 145,000. Addition of sodium dodecyl sulfate to aqueous solutions of IgG does not prevent association.


The use of iodinated antibody in immunodiffusion analysis to detect yeast nonsense termination fragments. A sensitive procedure based  on the use of iodinated antibody allows the detection of antigenic material by double-diffusion analysis. An autoradiographed immunodiffusion pattern reveals distinct radio-labeled precipitation lines for an extract of yeast with wild type HIS4 protein and for extracts of mutants containing HIS4 nonsense fragments.


Listeria monocytogenes infection in neonates: Investigation of an epidemic. From March 10 through October 29, 1975, Listeria monocytogenes infection occurred in seven neonates born at a 401-bed general hospital in Greenville, S.C. In the preceding 19 months, there had been only one case. Six of the seven isolates from the infected infants were of serotype JB. Risk of listeriosis in neonates was associated with being born to mothers of low socioeconomic status and being born to mothers who had had vaginitis during pregnancy. Nosocomial transmission of L. monocytogenes appeared unlikely, but the source of the infection is unknown. Of the media used in this study for isolation of L. monocytogenes from mixed cultures, McBride's agar and McBride's agar with nalidixic acid were most useful.


Alterations of body fluid compartments and distribution of tissue water and electrolytes in rhesus monkeys with rocky mountain spotted fever. Chair-restrained rhesus monkeys (Macaca mulatta) were inoculated subcutaneously with 10(2)--10(3) plaque-forming units of virulent Rickettsia rickettsii. The latent period for fever and rickettsemia was three to four days; death occurred six to eight days after infection. Total circulatory electrolyte levels and fluid volumes, including plasma, red blood cell, true circulatory blood, and extracellular fluid, increased. The expansion of the extracellular and plasma volumes resembled findings reported during severe Rocky Mountain spotted fever in humans, guinea pigs, and rabbits. Total water content of the liver also increased. Intracellular concentrations of K+, as well as total Na+ and K+, decreased in the diaphragm. Both the lung and medulla oblongata showed increased levels of intracellular Na+ and water with simultaneously decreased levels of extracellular Na+ and water. Such an intracellular overhydration of the medulla oblongata could contribute to death as a result of depression of the cardiovascular and respiratory centers. On the basis of the findings in monkeys, the intravenous infusion of fluids and electrolytes during clinical therapy of severe rickettsial infections should be considered extremely dangerous.


Antibodies to proteases and exotoxin A of Pseudomonas aeruginosa in patients with cystic fibrosis: Demonstration by radioimmunoassay. Sera from 33 patients with cystic fibrosis and two pediatric patients being treated for chronic pulmonary infections not related to cystic fibrosis and six sera or serum pools from uninfected individuals were tested with a microtiter radioimmunoassay for reactivity against exotoxin A and two proteases from Pseudomonas aeruginosa. Exotoxin A was purified from a low-protease strain of P. aeruginosa and shown to have adenosine diphosphate-ribose transferase activity and mouse lethality. Proteases were purified from an isolate of P. aeruginosa from a patient with cystic fibrosis and had proteolytic activity against elastin and collagen in an assay employing dimethylated protein substrates. The antibody responses of the patients detected using 125I-labeled antibody to human immunoglobulin were correlated with clinical evaluations expressed as a composite score based on pulmonary findings, case histories, growth and nutrition, and chest X rays. Values in the radioimmunoassay for patients' sera were compared with those of a control serum pool and expressed as the ratio of counts per minute (cpm) in patient serum to the cpm in the control pool. Inverse correlations were found between these ratios for each of the pseudomonas exoproducts and clinical scores; highest ratios occurred in patients showing the lowest clinical scores. These results confirm that proteases and exotoxin A of P. aeruginosa are produced in cystic fibrosis pulmonary infections due to P. aeruginosa and suggest that they may serve as significant virulence factors in these chronic infectious states.


Quantitative determination of the serum antibody response to the capsular polysaccharide of Bacteroides fragilis subspecies fragilis in women with pelvic inflammatory disease. The antibody response to the capsular polysaccharide of Bacteroides fragilis subspecies fragilis in women with acute pelvic inflammatory disease was quantitated using a sensitive radioactive antigen-binding assay. Antibody detected by this technique correlated highly with antibody measured by quantitative precipitin analysis (r = 0.99). Paired acute- and convalescent-phase sera obtained from women from whom B. fragilis subspecies fragilis was isolated by culdocentesis showed significantly greater changes in antibody concentration than corresponding sera obtained from women with gonococcal or nongonococcal pelvic inflammatory disease when B. fragilis subspecies fragilis was not isolated from the cul-de-sac. These data indicate that the steady state of immunity to this capsular antigen is perturbed in patients with acute nongonococcal pelvic inflammatory disease from whom this organism can be cultured. This finding may indicate that B. fragilis can play a significant role in acute pelvic inflammatory disease and can be involved early in the infectious process.


Antibacterial functions of macrophages in experimental protein-calorie malnutrition. I. Description of the model, morphologic observations, and macrophage surface IgG receptors. An experimental protein-calorie malnutrition was produced in weanling Sprague-Dawley rats. The model resembles human malnutrition with respect to weight loss, inanition, angular stomatitis, anemia, lymphopenia, hypoproteinemia with hypoalbuminemia, and marked thymic involution. In addition, systemic invasion by gram-negative rods was documented. However, no edema was produced, and animals did not survive for longer than six weeks on the protein-deficient diet. One percent glycogen was found to be a satisfactory nonprotein stimulus for induction of a peritoneal exudate consisting primarily of young macrophages. Electron microscopy showed that morphologic events of phagocytosis and degranulation proceeded normally in macrophages from protein-deficient animals. In addition, cell surface receptors for IgG were preserved under these experimental conditions. These data indicate that weanling rats may be employed as a small animal model for servere, fulminant protein-calorie malnutrition in humans.


Antibacterial functions of macrophages in experimental protein-calorie malnutrition. II. Cellular and humoral factors for chemotaxis, phagocytosis, and intracellular bactericidal activity. Cellular and humoral aspects of the antibacterial activity of macrophages during experimental protein-calorie malnutrition were studied. There were no defects in chemotaxis or bactericidal activity of cells from protein-deficient animals, although phagocytosis-associated oxygen consumption and hexose monophosphate shunt activity were depressed. However, marked impairment of humoral chemotactic factors generated in the peritoneal cavity by glycogen injection and of heatlabile serum opsonins for Staphylococcus aureus, Escherichia coli, Salmonella typhimurium, and Salmonella enteritidis was found. The studies suggested that some macrophage antibacterial functions measured in vitro are not altered in experimental acute protein-calorie malnutrition, but that serum factors, presumably complement-derived, would limit their in vivo function. Thymic involution and lymphocyte depletion would further impair in vivo cellular immune reactions affected by macrophages. This model may therefore prove useful for the study of specific aspects of cellular immunity in malnourished hosts and of specific rehabilitation strategies.


Phenotypic and epidemiologic correlates of auxotype in Neisseria gonorrhoeae. Previous studies from Seattle, Wash., suggested that strains of Neisseria gonorrhoeae which require arginine, hypoxanthine, and uracil (Arg-Hyx-Ura- auxotype) are unifomly highly susceptible to penicillin G, are relatively resistant to complement-dependent killing by heated, pooled human serum, and are associated with disseminated gonococcal infection. For further study of the epidemiology of these strains and for analysis of the susceptibility to penicillin, serum sensitivity, and the nutritional requirements of gonococcal isolates from other cities, a survey was made of urethral and cervical strains isolated in 1972--1974 from 50 randomly selected pateit-s with uncomplicated gonorrhea from each of nine cities. Arg-Hyx-Ura-strains represented greater than 50% of isolates from Seattle and Des Moines, Iowa, 22% of isolates from Denver, Colo., and Dayton, Ohio, and less than or equal to 12% of the isolates from Boston, Mass., Newark, N.J., Norfork, Va., Miami, Fla., and Oakland, Calif. Arg-Hyx-Ura- strains were recovered from 42% of white and 9% of black patients (P less than 0.001), and clincis with the highest incidences of these strains had the highest proportion of white patients among those with gonorrhea. Arg-Hyx-Ura- strains were all susceptible to less than or equal to 0.125 microgram of penicillin G/ml and were more resistant than strains with other auxotypes to killing by heat-inactivated human serum plus complement (P less than 0.001).


Host-parasite relationships in experimental airborne tuberculosis. VII. Fate of Mycobacterium tuberculosis in primary lung lesions and in primary lesion-free lung tissue infected as a result of bacillemia. Evidence suggests that in humans tuberculous disease usually arises at apical or subapical sites in the lungs seeded as a consequence of an early bacillemic phase of the infection. This study examined the fate of bacilli transported via the bloodstream to metastatic sites in the lungs of guinea pigs after aerosol infection with approximately two viable virulent Mycobacterium tuberculosis. The results revealed that, even after logarithmic-phase multiplication of bacilli in primary lesions had been terminated, bacilli seeded via the bloodstream to metastatic sites in the lung were able to multiply. These observations, made in an animal model that mimics the conditions under which tuberculosis develops in human subjects, challenge the relevance of systemic macrophage activation in experimental airborne tuberculosis in guinea pigs.


Serum bactericidal action and activation of the classic and alternate complement pathways by Neisseria gonorrhoeae. In order to more fully understand the host defense mechanisms against gonococcal infections, we decided to define the role of the classic and alternate complement pathways in gonococcal BA. Sera and infecting isolates were collected from several patients with genital and disseminated gonococcal infections. Sera from two never-infected subjects and a hypogammaglobulinemic patient were also collected. Sera from patients with genital gonorrhea and never-infected controls demonstrated marked BA for gonococci after 30 min incubation. Chelation of these sera with MgEGTA delayed the expression of BA. Consumption of C3, but not C4, was observed in chelated samples. BA could not be demonstrated in any of the sera from DGI patients or the patient with hypogammaglobulinemia. Aliquots of fresh and chelated hypogammaglobulinemic serum to which IgM or IgG antigonococcal antibodies were added showed marked BA by 30 and 60 min, respectively. Absorption of chelated serum with a serum-sensitive isolate eliminated the previously observed delayed BA. The findings suggest that gonococcal serum BA is primarily associated with activation of the classic complement pathway. Activation of the alternate pathway also occurs; however, its expression is delayed and appears to be antibody-dependent.


Model for disseminated intravascular coagulation: bacterial sepsis in rhesus monkeys. DIC is a hemorrhagic syndrome frequently encountered as a complication in severe gram-negative bacterial sepsis. An animal model for sepsis-associated DIC was developed in order to permit study of the appearance and development of this syndrome in relation to the entire disease process. Rhesus monkeys (4 to 6 kg) were infected by intravenous injection of 10(9) Salmonella typhimurium organisms and studied for a period of 7 to 10 days following infection. Ten of 23 infected monkeys developed petechial rash characteristic of DIC, which appeared on days 1 to 2 infection and lasted 4 to 5 days. In the group of monkeys developing rash, activation of coagulation was suggested by an 80% decrease in platelet count and 20% to 30% increases in PT and APTT. Fibrinolytic system activation was indicated by the appearance of FDP. Kinin system activation was evidenced by decreases in both prekallikrein nad kininogen. Changes in laboratory tests suggestive of subclinical DIC were also noted in infected monkeys which did not develop a rash. Pathologic evidence of DIC was obtained through observation of numerous fibrin thrombi in the kidneys of the only monkey which died in the course of infection. Occurrence of DIC in association with this experimental infection in rhesus monkeys was established on the basis of clinical, laboratory, and pathologic criteria. Expression of the syndrome on days 1 to 2 following infection correlated with the period of increasing bacteremia.


Aerosolized antigen dose-response studies in asthmatic monkeys. This study used rhesus monkeys which had consistent respiratory responses to a standard (single dose) aerosolized concentration of A. Beginning at lower concentrations of A which did not produce an airway response, fourfold increases in concentrations of A were aerosolized until an airway response occurred. Compared with the single-dose SAC after the TAC, some consistently reactive animals did not respond to the SAC, the maximal concentration of A used for testing. Thus the TAC may result in an acute form of airway desensitization. Animals vary in the threshold concentrations of A required to produce an airway response. A repeat challenge with the threshold antigen concentration during the same experiment resulted in a second response generally greater than the initial response at the same concentration. When the SAC system was similarly evaluated by repeat challenges with the same A concentration, the second response was generally less than the first. Different methods for evaluation of pharmacologic inhibitors of IgE-mediated airway responses may be developed from this rhesus model of asthma.


Purification and some properties of ovine placental lactogen. A method has been described for the purification of ovine placental lactogen (oPL) involving the use of freshly obtained sheep foetal cotyledons. Tissue was extracted with 0.1 M-ammonium bicarbonate and the supernatant fraction, adjusted to pH 7, was brought to 60% saturation with ammonium sulphate. The resulting precipitate was then subjected to a sequence of chromatographic steps using columns of Sephadex G-100 and carboxymethylecellulose. During each stage of the purification, the lactogenic activity was monitored with a pregnant rabbit mammary gland radioreceptor assay. The yield of oPL corresponded to 8 mg/kg wet foetal tissue and the oPL possessed lactogenic activity equivalent to 1 mg ovine prolactin/mg protein and GH-like activity equivalent to 0.8 mg human GH/mg protein. The biological activity of oPL was confirmed using a rabbit intraductal mammary gland assay in vivo. After polyacrylamide gel electrophoresis at pH 8.9, oPL was resolved into one major band (isoelectric point 8.2--8.4) and four minor components, which were thought to be deamidation products of oPL. Microimmunoelectrophoresis and immunodiffusion studies confirmed that the preparation of oPL was free from serum protein contaminants.


The control of responding by auditory stimuli: interactions between different dimensions of the stimuli. Sounds have position in addition to other dimensions, such as intensity and frequency. Position rapidly gains control of spatially contiguous responses and this may interfere with control of responding by other acoustic dimensions. These experiments investigated interference of a tone-noise discrimination by the discrimination of acoustic position. Squirrel monkeys were studied when responding was differentially reinforced in the presence of both spectral content (tone-noise) and positional differences between the stimuli, and when responding was differentially reinforced only in the presence of spectral differences. Under the first condition, responding rapidly came under the control of the position of the noise in the two monkeys tested. The position of the tone controlled responding in one monkey; in the second monkey, responding came under the control of the spectral content of the tone. Under the second condition, responding was initially under the control of the noise in all three monkeys tested. This persisted for the duration of the condition for two of the monkeys; in one monkey, responding came under the control of the spectral content of the noise. Under the second condition, responding was also initially under the control of the position of the tone for all monkeys, but control by spectral content of the tone relatively rapidly developed in two of three monkeys.


The role of H-2-linked genes in helper T-cell function. III. Expression of immune response genes for trinitrophenyl conjugates of poly-L(Tyr, Glu)-poly-D,L-Ala--poly-L-Lys in B cells and macrophages. Using lymph node T cells from poly-L(Tyr,Glu)-poly-D,L-Ala--poly-L-Lys[(TG)-A--L]-primed animals and B cells from animals primed with trinitrophenylated (TNP) protein or lipopolysaccharide, we have obtained anti-TNP-(TG)-A--L direct plaque-forming responses in vitro. Response to this antigen was shown to be controlled by the H-2 haplotype of the animal studied. The strain distribution of in vitro response was very similar to that previously reported by others for in vivo secondary IgG responses to (TG)-A--L. We investigated the cell types expressing the Ir gene(s) for (TG)-A--L in our cultures. F1, high responder x low responder mice were primed with (TG)-A--L. Their T cells were active in stimulating anti-TNP-(TG)-A--L responses of high responder but not low responder B cells and macrophages (MPHI), even though both preparations of B cells and Mphi were obtained from mice congenic at H-2 with one of the parents of the F1. For three low responder strains tested, of the H-2h2, H-2k, and H-2f haplotypes, the anti-TNP-(TG)-A--L response of low responder B cells and Mphis in the presence of high responder, F1 T cells could not be improved by the addition of high responder, antigen-bearing Mphis to the cultures. In one strain of the H-2a haplotype, it was shown that neither the B cells nor Mphis could be functional in anti-TNP-(TG)-A--L responses. Our results therefore suggested the Ir genes for anti-TNP-(TG)-A--L responses were expressed at least in B cells in all the low responder strains we studied, and, in mice of the H-2a haplotype, in Mphis too.


Cellular and genetic control of antibody responses in vitro. III. Immune response gene regulation of accessory cell function. The possibility was investigated that Ir genes regulate the function of cells other than T or B cells in the primary IgM responses to the synthetic antigens trinitrophenylated poly-L-(Tyr,Glu)-poly-D,L-Ala--poly-L-Lys [TNP-(T,G)-A--L]and trinitrophenylated poly-,-(His,Glu)-poly-D, L-Ala--poly-L-Lys [TNP-(H,G)-A--L]. The primary responses of (B10 x B10.A)F(1) spleen cells to both antigens were abrogated by Sephadex G-10 passage, and restored by the addition of spleen adherent cells. The cell type in the spleen adherent cell population active in reconstituting the responses to TNP-(T,G)-A--L and TNP-(H,G)-A--L was a non-T, non-B, radiation-resistant, glass-adherent spleen cell. The responses of Sephadex G-10-passed (responder x nonresponder)F(1) spleen cells to TNP-(T,G)-A--L or TNP-(H,G)-A--L were reconstituted by spleen adherent cells from only responder strains. Spleen adherent cells from F(1) mice reconstituted the responses to both antigens. Spleen adherent cells from each of the strains tested reconstituted the non- Ir gene-controlled response to a third antigen, TNP-keyhole limpet hemocyanin. The inability of spleen adherent cells from nonresponder strains to reconstitute the responses to either TNP-(T,G)-A--L or TNP-(H,G)-A--L was not a result of active suppression induced by the presence of nonresponder adherent cells, since a mixture of responder and nonresponder spleen adherent cells reconstituted the responses to both antigens. The use of spleen adherent cells from recombinant strains demonstrated that the autosomal dominant genes controlling the ability of spleen adherent cells to function as accessory cells in the responses to TNP-(T,G)-A--L and TNP-(H,G)-A--L are located in the K or I-A regions of the responder H-2 complex, the same region(s) of H-2 as the Ir genes controlling overall in vitro and in vivo responsiveness to these antigens.


Pattern of shedding of two noncultivable viruses in stools of newborn babies. Noncultivable viruses have been associated with diarrhea affecting newborn babies in obstetric hospital nurseries. Persisting infection in a special care nursery in Melbourne, Australia, permitted a study of the pattern of excretion of these viruses. Ten babies admitted to the nursery within 2 hr of birth were randomly selected for prospective study. Feces were collected daily for 14 days and were examined by electron microscopy. All ten babies excreted detectable amounts of duovirus (rotavirus, HRVL agent, IGV) for at least 1 day. Age at onset of excretion varied from 2 to 13 days. Eight of the ten babies developed diarrhea. Excretion of duovirus preceded the onset of diarrhea by 12--72 hr and persisted for at least 3 days. Seven of the ten babies also excreted detectable amounts of 28-nm virus-like particle for 3--8 days. The identity of this particle is unknown. Morphologically it resembles Norwalk agent and "astrovirus." Excretion of this 28-nm particle coincided with symptoms of diarrhea in four babies, all of whom were also excreting duovirus. It is concluded that most newborn babies admitted to a nursery where duovirus infection is endemic will excrete this virus at least once during the first 2 weeks of life. Excretion of virus particles will either precede development of diarrhea or be asymptomatic. Selective isolation of babies with diarrhea is thus unlikely to control spread of duovirus infection within a hospital nursery.


A method for surgical management of focal epilepsy, especially as it relates to children. A method of surgical management for intractable epilepsy is described. The essential features are: 1) all surgical manipulation is carried out under general, rather than local, anesthesia; 2) the sensorimotor region is readily identified in the anesthetized patient by recording cortical sensory evoked responses; and 3) the epileptogenic focus is localized by extraoperative electrocorticography via indwelling epidural electrode arrays, localization deriving from recordings made during spontaneously occuring clinical seizures. Cases are presented to demonstrate that: 1) in some instances, recording of sensory evoked responses is the only means of sensorimotor localization in both the awake and anesthetized patient, and 2) spontaneous and electrically induced electroencephalographic seizure activity may provide false localization of the focus, the correct localization requiring recordings made during spontaneous clinical seizures. The outcome of surgery and the various epileptogenic lesions encountered are described. A good result has been achieved in 61% of patients followed 1 to 10 years. When the results obtained in children are analyzed alone, 70% have benefited from surgery.


Measurement of folates in human plasma and erythrocytes by a radiometric microbiologic method. A radiometric microbiologic assay for the determination of folic acid in human plasma and red blood cells is described. The assay is based upon the measurement of 14CO2 produced from the metabolism of [1-14C] gluconate by Lactobacillus casei. The 14CO2 evolved was shown to be proportional to the amount of added DL-N-5-methyltetrahydrofolage (DL-N-5-methyl FH4). A total of 26 normal plasma and 57 blood hemolysates were assayed in parallel by this radiometric and the standard (turbidimetric) microbiologic assay. The correlation coefficients for the two assays were r = 0.96 for plasma and r = 0.98 for red-cell folate. Lyophilization of L. casei was found to simplify this radiometric assay by eliminating routine maintenance and culture of this microorganism.


A new germanium-63/gallium-68 generator. A germanium-gallium generator producing EDTA-free Ga-68 would permit the synthesis of a broad range of Ga-68 radiopharmaceuticals and thus facilitate the widespread application of positron tomography. We have investigated a solvent-extraction generator system producing the weak complex of Ga-68 with 8-hydroxyquinoline (Ga-68 oxine), free of EDTA. The conditions for optimum Ga-68 yield and minimum Ge-68 breakthrough involve extraction with chloroform from pH 5 buffer containing Ge4+ carrier, followed by evaporation to dryness. This produces Ga-68 yields of 70--80%, with Ge-68 breakthrough of less than 0.003%. The Ga-68 oxine is readily converted to other radiopharmaceuticals such as Ga-68 EDTA, Ga-68 EDTMP, or Ga-68 colloid, and is conveniently delivered dry. The extraction system is simple and amenable to automation, and the low loss rate and 280-day half-life of Ge-68 provide a generator with a long useful lifetime.


In vivo distribution of carbon-11 phenytoin and its major metabolite, and their use in scintigraphic imaging. Curie quantities (0.3--1.5 Ci) of H11CN were used in the synthesis of C-11-tagged phenytoin (C-11.DPH) and 5-(p-hydroxyphenyl)-5-phenylhydantoin (C-11.HPPH), using a modified Bücherer-Bergs reaction. The H11CN was produced from a mixture of 95% nitrogen and 5% hydrogen by a 45-min bombardment with 10-MeV protons at 10 muA. Following i.v. infusions of C-11 DPH (13.7 mg/kg at a rate of 29 mg/min) into the left femoral vein of Rhesus monkeys, DPH shows persistent concentration in the brain and liver fields. Extravascular administration shows significant retention at the site of administration. Intravenous bolus injection of [11C]-HPPH into a Rhesus monkey, at a dose of 6.4 mg/kg, resulted in localization of this compound in the liver, gallbladder, urinary bladder, and intestinal fields. Loss of activity from the liver region, with appearance of this activity in the intestinal field, suggests that [11C]-HPPH is secreted into the intestine via the bile. Further investigation is needed to study the potential of [11C]-DPH as a brain-scanning agent and [11C]-HPPH as a possible cholescintigraphic agent.


Facial growth after condylectomy and alloplastic condylar replacement. Condylectomy or condylectomy with immediate alloplastic condylar replacement was performed in six young and six adult rhesus monkeys to investigate the role of the mandibular condyl in facial growth. The experimental findings support the hypothesis that the condyle does not have a primary influence on mandibular growth and that alterations in biomechanics are mainly responsible for the observed alterations in mandibular morphology after condylectomy.


Experimental transmission of Sarcocystis from icterid birds to sparrows and canaries by sporocysts from the opossum. Cowbirds (Molothrus ater) and grackles (Cassidix mexicanus) infected with muscle cysts of Sarcocystis were fed to opposums (Didelphis virginiana) and fecal sporocysts from the latter were given to sparrows (Passer domesticus, Family Ploceidae), canaries (Serinus canarius, Family Fringillidae) and ducks (Anas platyrhynchos, Family Anatidae). Asexual parasites were found in the endothelium of sparrows and canaries but not in ducks. When birds were kept 10 weeks or more after infection, muscle cysts were found grossly and microscopically in the majority of sparrows, and in 1 canary, but not in ducks. Muscle zoites were found in digests of all sparrows and canaries but not in that of ducks. Metrocytes and forms dividing by endodyogeny also were found in the digest. Thus, avian Sarcocystis was transmitted experimentally from 2 genera of 1 family (Icteridae) to 2 different families of passerine intermediate hosts by sporocysts from the definitive host. This is the broadest intermediate host spectrum known for a species of Sarcocystis.