Acetylated methylmannose polysaccharide of Streptomyces griseus. Locations of the acetyl groups. The positions of esterification of the 4 to 5 acetyl residues in the acetylated methylmannose-containing polysaccharide from Streptomyces griseus have been established by the methyl replacement technique, wherein ester substituents are specifically replaced with methyl ether substituents. The newly incorporated methyl groups were distinguished from 3-O-methyl groups by the use of polysaccharide containing radioactively labeled endogenous methyl groups. The positions of methyl group localization were established by a proton magnetic resonance study of the intact methyl-replaced polysaccharide combined with an analysis of the constituent monosaccharides by gas-liquid chromatography-electron impact mass spectrometry of their alditol acetate derivatives. These studies demonstrate that the acetyl groups are located at position 6 of approximately half of the 10 contiguous alpha(1 leads to 4)-linked 3-O-methyl-D-mannose residues. Purification of the polysaccharide was accomplished by an added step involving affinity chromatography on a column containing immobilized palmitoyl residues. The affinity of the polysaccharide for this long chain lipid suggests that its plays a role similar to the methylmannose-containing polysaccharide of Mycobacterium smegmatis in its regulation of the bacterium's fatty acid synthetase.


Evidence for the location of high mobility group protein T in the internucleosomal linker regions of trout testis chromatin. Antibodies against the trout testis non"histone chromosomal protein, high mobility group protein T (HMG-T), have been elicited in goats. The antiserum was shown to be specific for HMG-T and did not cross-react with histone 1 or with the other two trout testis HMG proteins, H6 and ubiquitin. Purified anti-HMG-T IgG was used to determine the location of HMG-T within chromatin subunits separated on sucrose gradients. Binding of fluorescent labeled anti-HMG-T to these subunits clearly supports the notion that this protein is associated not with the nucleosome core but rather with the internucleosomal linker regions, and previously suggested (Levy W., B., Wong, N.C.W., and Dixon, G. H. (1977) Proc. Natl. Acad. Sci. U.S.A. 74, 2810-2814).


Thyrotropin suppression by 3,5-dimethyl-3'-isopropyl-L-thyronine in man. The thyromimetic activity of 3,5-dimethyl-3'-isopropyl-L-thyronine (DIMIT), a nonhalogenated thyroid analog, was studied in adult men using suppression of TRH-induced TSH release to assess this activity. In nine men, aged 30-58 yr, the TSH increment after 500 microgram TRH iv was compared to the TSH response to TRH 24 h after oral administration of 1 mg DIMIT. Eight euthyroid subjects had normal baseline TSH levels of 1.5 +/- 0.2 (SE) microunit/ml that fell significantly to 0.7 +/- 0.2 microunit/ml 24 h after DIMIT (P less than 0.005). Their TSH increments after TRH fell from 15.3 +/- 2.8 to 6.7 +/- 1.6 microunit/ml 24 h after DIMIT (P less than 0.001). One subject with probable Hashimoto's thyroditis had an elevated TSH of 18 microunit/ml, with an exaggerated TSH response to TRH of 72 microunit/ml. His basal TSH fell to 7.6 and his TSH increment fell to 14.3 microunit/ml 24 h after DIMIT. The suppression of TSH was relatively prolonged. In four subjects, the TSH response to TRH was still blunted from 5-12 days after DIMIT. In one subject, the TSH increment returned to normal 15 days after DIMIT. DIMIT had no significant effect on PRL secretion. There was no evidence of toxicity in patients receiving DIMIT. DIMIT has effective thyromimetic activity in man, as shown by its significant and prolonged suppression of TSH secretion.


Menstrual cycle characteristics in chronically hemiovariectomized cynomolgus monkeys (Macaca fascicularis). Circulating levels of LH, FSH, estradiol, and progesterone were measured by RIA in daily serum samples throughout the menstrual cycle in five regularly cycling, chronically hemiovariectomized, adult cynomolgus monkeys. The hormonal patterns, the lengths of the follicular and luteal phases, and the overall cycle length were nearly indistinguishable from those observed in intact cycling monkeys. The findings accord with the notions 1) that in intact monkeys, the contralateral ovary contributes little, if at all, to the regulation of the function or lifespan of the corpus luteum, and 2) that the corpus luteum after spontaneous luteolysis has no local residual effect inimical to new follicle growth.


Plasma clearance and plasma half-disappearance time of exogenous thyrotropin-releasing hormone and pyroglutamyl-N3im-methyl-histidyl prolineamide. The plasma clearance rate (PCR) and plasma half-disappearance time (t1/2) of TRH was compared to the PCR and t1/2 of pyroglutamyl-N3im-methyl-histidyl prolineamide (methyl-TRH), a more potent analog of TRH in normal subjects. The PCR for TRH was 1500 +/- 329 (SD) ml/min, which was significantly greater than the PCR of methyl-TRH (783 +/- 96 ml/min). The t1/2 of TRH was 6.2 min compared to a t1/2 of 11.5 min for methyl-TRH. The slower clearance of methyl-TRH is probably due to the increased resistance to degradation by serum enzymes of methyl-TRH compared to TRH.


Thyrotropin-releasing hormone activity in the human placenta. TRH immunological and biological activities were found in extracts of human placentas. Eight term placentas delivered by cesarean section were extracted with 2 N acetic acid, followed by glacial acetic acid. Lyophilized samples were resuspended in phosphate-buffered saline. TRH activity, identified in each placental extract by specific RIA, averaged 19.8 +/- 3.3 pg/mg protein. TRH in the placental extracts was similar to synthetic TRH by four criteria. 1) Serial dilutions of placental extracts defined an immunoassay inhibition curve with a slope (-1.93) identical to synthetic TRH (-1.88). 2) When placental extract or synthetic TRH was chromatographed on Sephadex G-10, TRH immunoreactivity was found in similar fractions of the eluate. 3) Both placental extract and synthetic TRH stimulated TSH release from rat pituitaries in vitro. 4) Human serum degraded placental extract and synthetic TRH in a qualitatively similar manner. Placental minces did not accumulate exogenous [3H]TRH, suggesting that the placental TRH may originate endogenously.


Detection of a thyroxine-binding protein physicochemically similar to serum thyroxine-binding globulin in normal human urine. Urine samples from 10 normal adult males were analyzed for T4-binding globulin (TBG) employing a sensitive and specific RIA for serum TBG. Urine TBG was detected at a concentration of 1.74 +/- 0.87 microgram/100 ml (mean +/- SD). The excretion rate was 13.2 +/- 6.5 microgram/g creatinine. There was no correlation between the daily excretion rate of TBG and the total urinary protein (r = 0.52). Analyses of this T4-binding protein by gel filtration and on both reverse flow paper and gel electrophoresis demonstrated similar characteristics to those of serum TBG and purified TBG. The maximum T4-binding capacity of urine TBG, determined by reverse flow paper electrophoresis and equilibrium dialysis, was similar to that of serum TBG. The association constant for T4 of urinary TBG (0.46 +/- 0.20 X 10(10) M-1), determined by equilibrium dialysis, was significantly less (P less than 0.01) than that of serum TBG at 1.43 +/- 0.23 X 10(10) M-1). The Scatchard analysis of the urinary data demonstrated two T4-binding sites, one of high affinity consistent with TBG and one of low affinity consistent with albumin. It is concluded that the TBG present in urine has the major physical and biochemical features of the intact molecule, but that minor conformational and/or charge alterations occurred during renal excretion. Interpretation of data on thyroid hormones in normal urine should take the presence of TBG into consideration.


Plasma cortisol response to thyrotropin-releasing hormone and luteinizing hormone-releasing hormone in Cushing's disease. In 3 of 6 patients with pituitary-dependent Cushing's disease, a paradoxical increase of plasma cortisol was observed both after LRH (delta max, 13.9 +/- 3.7 microgram/100 ml; 113 +/- 52%) and TRH (delta max, 8.0 +/- 2.9 microgram/100 ml; 53 +/- 18%) administration, the maximum values being achieved 30--60 min after the iv bolus injection. In the remaining 3 patients and in 15 control subjects, plasma cortisol levels did not rise in response to either LRH or TRH administration but rather showed a slight to distinct decrease during the study period. The paradoxical cortisol response in half of the patients with Cushing's disease may be the consequence of loss of specificity of the pituitary receptor or alteration in the hypothalamo-pituitary pathways.


Sensitive microplate enzyme-linked immunosorbent assay for detection of antibodies against the scrub typhus rickettsia, Rickettsia tsutsugamushi. A microtiter enzyme-linked immunosorbent assay (ELISA) has been developed for the titration of antibodies against scrub typhus in human and animal sera. Scrub typhus rickettsiae were grown in monolayers of irradiated mouse LM3 cells and separated from host cell materials by differential centrifugation, filtration through a glass filter (AP-20, Millipore Corp.), and isopycnic banding in Renografin density gradients. The scrub typhus ELISA antigens were obtained from the purified viable rickettsiae by French pressure cell disruption and addition of 0.2% Formalin to the soluble extract. Antisera prepared in rabbits against the prototype Karp, the Kato, and the Gilliam strains of scrub typhus were used to standardize the ELISA and to compare its sensitivity and specificity to that of the indirect fluorescent antibody test (IFA). ELISA titers were measured as the greatest serum dilution showing an optical density 0.25 above controls or by the optical density achieved at a fixed serum dilution. The IFA and ELISA end point titers were quite similar, and all three measures of titer had comparable specificity for the strains of scrub typhus. No cross-reactions between the typhus and scrub typhus wera were observed by ELISA. Both the immunoglobulin M (IgM) and IgG antibody titers of 12 sequential sera from four patients with scrub typhus were obtained by IFA and ELISA. The IFA and ELISA end point titers for IgM and IgG had correlation coefficients of 0.91 and 0.97, respectively, whereas the ELISA optical density values at a serum dilution of 1:100 had slightly lower correlations with IFA titers (0.80 and 0.94). Early rising IgM titers followed by rising IgG titers were demonstrated by ELISA in three patients with primary scrub typhus infections, whereas the IgG response predominated in a patient with a reinfection. It is concluded that the ELISA for scrub typhus is a very satisfactory alternative to the IFA test.


Serotypes and antibiotic susceptibilities of Pseudomonas aeruginosa isolates from single sputa of cystic fibrosis patients. A phenotypic characterization of Pseudomonas aeruginosa from single sputum samples of 21 typical cystic fibrosis patients indicated a high frequency of heterogeneity among isolates on the basis of differences in antibiotic resistance, colony morphology, pigmentation, and serotype. Two or more isolates with different but stable susceptibilities to carbenicillin, gentamycin, streptomycin, tetracycline, chloramphenicol, and sulfamethoxazole plus trimethoprim were detected in 38% of the sputa. Differences generally were independent of the mucoid state of the strain. O-antigen group determination with the Difco typing set showed that two or more serologically distinct strains were present in 10/21 sputum specimens. Nonmucoid derivatives of mucoid isolates almost always retained both the antibiotic susceptibilities and serotype of their parent strain. These data suggest that cystic fibrosis patients may be cocolonized/coinfected by different strains of P. aeruginosa more frequently than generally believed. Alternatively, phenotypically distinct strains from a single patient might arise as phenotypic dissociants from a single infecting strain. Because of the frequency and multiplicity of phenotypically distinct P. aeruginosa isolates which we obtained from our cystic fibrosis patients, it is important to select multiple isolates from sputum cultures for antimicrobial susceptibility testing so as to assess adequately the susceptibility of this organism to antibiotic therapy in cystic fibrosis. We recommend that several colonies of each distinguishable colony type of P. aeruginosa be pooled for the antibiogram.


Electron capture gas-liquid chromatographic-mass spectral identification of acids produced by Neisseria meningitidis in a defined medium. The acid metabolites and the cellular fatty acids of three strains of Neisseria meningitidis grown in a chemically defined liquid medium were determined with computerized frequency-pulsed electron capture gas-liquid chromatography. Five acids not previously reported were subsequently identified: isobutyric, octanoic, decenoic (C10:1), dodecenoic (C12:1), and tetradecenoic (C14:1). These acids were produced during active metabolism and were not detected as cellular constituents. The frequency-pulsed electron capture gas-liquid chromatography methods which we used provide a rapid, reliable, sensitive means of detecting both these and other metabolic and cellular acids in spent culture medium.


Serotypes of Neisseria meningitidis isolated from patients in Norway during the first six months of 1978. During the first 6 months of 1978, 114 strains of Neisseria meningitidis isolated from patients in Norway were serotyped. Among 27 group C strains, type 2 was most common, whereas 82% of the 82 group B isolates did not react with antisera to the standard serotypes 1 to 12. These strains were shown to belong to a new serotype, type 15. Also some group A and C strains had the type 15 antigen. Investigations on a possible immunoprophylaxis against group B meningococcal disease in Norway should accordingly proceed with type 15 rather than with type 2 meningococci.


Halophilic, lactose-positive Vibrio in a case of fatal septicemia. A halophilic Vibrio species was isolated from blood cultures from a 59-year-old male with enteritis. The strain differed from Vibrio parahaemolyticus and Vibrio alginolyticus by its ability to ferment lactose, its production of beta-galactosidase, and its lower NaCl tolerance. A report of this infection and a description of the isolate is presented.


Rapid identification of nonfermentative gram-negative rods by the Corning N/F system. A total of 1,298 nonfermentative gram-negative rods were used to evaluate the performance of the individual biochemical tests in the N/F System and to determine whether the observed results included sufficient key reactions for rapid identification. The system included an oxidase test, two screening tubes providing 5 test reactions designed primarily to identify pigment-producing strains of Pseudomonas aeruginosa, and a plate providing 12 test reactions designed to identify other nonfermenters. With both the tubes and plates, most results were consistent with expected conventional test reactions. Use of the tubes permitted the identification of 90% of the strains of P. aeruginosa in 24 h and 97% in 48 h. Use of the plates permitted the identification of 95% of the other oxidative nonfermenters within 24 h and 96% within 48 h. Only 26% of weak and nonoxidative nonfermenters were identified because of the nonreactivity of these organisms in this system. There were no misidentifications based on misleading test results.


Rapid diagnosis of Pseudomonas aeruginosa urinary tract infections by radioimmunoassay. A solid-phase radioimmunoassay designed to detect serotype 6 Pseudomonas aeruginosa antigens was evaluated for its ability to rapidly diagnose urinary tract infections. Twelve P. aeruginosa serotypes were easily differentiated in the assay from eight other gram-negative bacterial species. During log-phase growth, the assay detected antigens in culture when approximately 10(6) or more serotype 6 P. aeruginosa organisms were present. Both cell-associated and solubilized antigens were detected. The assay detected antigens in 13 of 17 urine specimens which grew greater than 10(5) P. aeruginosa, 3 of 38 which grew other gram-negative rods, and none of 83 with no growth. Two of the three positive specimens from the other gram-negative rod group probably also contained P. aeruginosa. No preincubation of the urine specimens was required, and results were available within 2.5 h. The assay represents an improvement over other procedures for rapidly diagnosing urinary tract infections in that it allows diagnosis by species and should be adaptable to semiautomation.


Sensitive neutralization test for rubella antibody. A modified rubella virus plaque neutralization test for measuring rubella antibody was developed based on the potentiation of the virus-antibody complex by heterologous anti-immunoglobulin. The test is highly sensitive, yielding titers on the average 50 to 100 times higher than the haemagglutination inhibition test or the conventional plaque neutralization test. The sensitivity of this enhanced neutralization test is somewhat limited by the existence of a prozone phenomenon which precludes testing of low-titered sera below a dilution of 1:16. No prozone effect was observed with cerebrospinal fluids. The specificity of the enhanced neutralization test was determined by seroconversion of individuals receiving rubella vaccine. Although the rubella hemagglutination inhibition test remains the test of choice in routine diagnostic and surveillance work, the enhanced rubella neutralization test is particularly useful in monitoring low-level antibody in the cerebrospinal fluid in patients with neurological disorders and in certain instances of vaccine failure.


Serological study of trichloroacetic acid extracts of Bacteroides fragilis. Immunodiffusion techniques were used on trichloroacetic acid extracts from 10 strains of Bacteroides fragilis in detecting precipitating antibodies against this species in immune rabbit sera. Species and even strain specificities were observed in these precipitin reactions. Multiple antigens were detected in the extracts from some strains, whereas only one precipitin band per extract developed during agar-gel diffusion tests of others. The antigen extracts were found to be both heat stable and resistant to hydrolysis by alpha-chymotrypsin. Four serological patterns were demonstrated in homologous and heterologous reactions with the B. fragilis. antigen-antibody systems used. The results showed that some strains were serologically distinct from others, indicating that the strains tested are of more than one serotype.


Discrepancies in Weil-Felix and microimmunofluorescence test results for Rocky Mountain spotted fever. Only 4.2% of 284 single specimens and 17.6% of 51 pairs of sera reactive in Weil-Felix agglutination tests for Rocky Mountain spotted fever were confirmed by a specific Rickettsia rickettsii microimmunofluorescence test.


Acquired cell-mediated immunodepression in acute Chagas' disease. In this study two groups of patients with acute Chagas' disease were identified. Group one consisted of five patients with apparent acute Chagas' disease. These patients showed symptoms and signals of an acute illness, such as high fever and enlarged spleen. One of these patients developed severe myocarditis and heart failure. Group two consisted of seven patients with inapparent acute Chagas' disease. This was a nonclinical entity, not perceived by the patient who did not seek medical care. The diagnosis was made by the shift of a serologic test which indicates the presence of immunoglobulin M antibodies to Trypanosoma cruzi. The patients with apparent acute Chagas' disease showed positive delayed-type skin response to T. cruzi antigen. Also, their leukocytes showed significant inhibition of migration in the presence of this antigen. By contrast, the patients with the inapparent acute Chagas' disease did not show positive delayed-type skin response to T. cruzi antigen and no significant inhibition was observed when their cells migrated in the presence of this antigen. Of interest, none of these patients was capable of developing contact sensitivity to 2,4-dinitrochlorobenzene. However, three out of five patients with the apparent acute disease and all the normal control subjects showed positive contact reaction after sensitization to this drug. The results of these experiments would suggest that the thymus-derived (T)-lymphocyte function is depressed in patients with the clinically inapparent acute Chagas' disease. This immunodepression seems to be acquired in the course of the T. cruzi infection because all patients showed positive delayed-type skin response to at least one ubiquitous microbial extract, thus indicating previously normal T-cell function. We hypothesize that T. cruzi antigens may directly stimulate T cells with the concomitant release of factors that might become supressive for T-cell responses. Furthermore, the suppressive effect might interfere with the T-cell response to other antigens, such as to 2,4-dinitrochlorobenzene.


Gonococcal interactions with polymorphonuclear neutrophils: importance of the phagosome for bactericidal activity. Gonococci are capable of attaching to the surface of polymorphonuclear leukocytes (PMN). In this location they resist phagocytosis and are not killed by PMN. To delineate the factors involved in the survival of these gonococci, we investigated the interaction of virulent gonococci, which adhere to cells and resist phagocytosis, and avirulent gonococci, which are phagocytized and killed by PMN. In the presence of serum, both virulent and avirulent gonococci associate equally well with PMN and stimulate increases in oxidative metabolism. In the absence of serum virulent gonococci attached to PMN and stimulated PMN oxidative metabolism to a greater extent than avirulent gonococci which did not attach to PMN (P = 0.0009). Therefore, the survival of virulent gonococci attached to the PMN surface is not a result of failure to activate oxidative and bactericidal mechanisms. Both virulent and avirulent gonococci stimulated equivalent PMN specific granule release as measured by the appearance of lactoferrin in the media. Phagocytosis of avirulent gonococci stimulated significantly greater beta-glucuronidase release (P = 0.01) and myeloperoxidase-mediated iodination of protein (P = 0.001) by PMN than attachment of virulent gonococci. In the absence of serum neither type of gonococci stimulated beta-glocuronidase release or protein iodination by PMN. Thus, virulent gonococci fail to stimulate primary granule release by PMN. To further assess the role of attachment versus ingestion on the survival of gonococci, PMN were treated with cytochalasin B to block ingestion. Cytochalasin B-treated PMN were unable to kill either virulent or avirulent gonococci despite normal degranulation stimulated by the latter. The failure of PMN to kill surface-attached gonococci appears to be a consequence of the failure of PMN to enclose the virulent gonococci within a phagosome. The phagocytic vacuole thus plays a critical role in normal PMN bactericidal activity by providing a closed space in which the proper concentration of substances may be achieved to generate microbicidal activity.


Paraprotein estimation: a comparison of immunochemical and densitometric techniques. Paraproteins have been estimated by both immunochemical methods and densitometric analysis of electrophoretic strips. Correlation between the results obtained by these two methods, although generally good for assessing trends, varied between patients.


Determination of antibody levels to Candida albicans in healthy and hospitalised adults using a radioimmunoassay. A radioimmunoassay for antibody to Candida albicans is described. The test used whole, killed organisms as the antigen and radiolabelled sheep anti-human globulins to quantitate different classes of antibody to C. albicans. The assay has been compared with an Ouchterlony precipitin method and found to be simpler, more rapid, and more sensitive than the latter. Results obtained from two groups of symptomless adults indicated that the range of antibody level was wider for a hospitalised group than for a group of blood transfusion donors, particularly in respect of IgG and IgA antibody. The reason for the increase of antibody in hospital patients was not clear but may have been related to antibiotic therapy. The difficulties in interpretation of Candida serology have therefore been re-assessed in the light of more detailed knowledge of the range and type of antibody to be expected in normal individuals.


Carriage of group D streptococci in the human bowel. Sixty faecal samples, 39 from adults and 21 from neonates, were investigated by means of a simple scheme to isolate and identify any group D streptococci present. A number of differences were found between the two groups. Group D streptococci were absent from 43% of the neonates compared with only 13% of the adults; Streptococcus bovis was commoner in the neonates (23.8%) than in the adults (5%), and Streptococcus faecium was not isolated from any of the infant samples although it was common in the adult samples (25%). The viable counts of Strep. faecium were found to be, on average, 100-fold lower than those of Streptococcus faecalis. The methods and results are discussed with reference to the clinical significance of group D streptococci, especially in bacterial endocarditis and carcinoma of the colon.


Biosynthesis and characterisation of IgMlambda in a case of chronic lymphocytic leukaemia with intracellular immunoglobulin inclusions. Lymphocytes from a patient with chronic lymphocytic leukaemia were found to contain crystalline inclusions in the cytoplasm. These crystals were characterised as IgMlambda immunoglobulin. The cells did not have detectable surface immunoglobulin. No paraprotein was detected in the serum and no excess of immunoglobulin was detected in the urine. The cells incorporated radioactive leucine into mu and lambda chains during an 18-hour culture. Studies of labelled lysate immunoglobulin revealed a molar excess of heavy chain over light chain while in contrast free light chain was the only detectable secreted immunoglobulin product during the culture period. These findings are discussed in relation to immunoglobulin synthesis by normal and other neoplastic lymphocyte preparations.


Severe clinical conditions associated with Bacillus cereus and the apparent involvement of exotoxins. Twenty-one cases of infection with Bacillus cereus are summarised. The histories supplied showed that at least 15 of these were associated with severe or potentially severe symptoms including two deaths. Analysis of the production of exotoxins, including haemolysin and phospholipase, by these strains is given, and the relevance of these metabolites to the severity of the condition is discussed. Three incidents of bovine mastitis resulting from B. cereus and involving three deaths are also included. The observations presented here together with those of previous reports which are reviewed indicate that B. cereus may be of clinical importance, not just an opportunist but also as an agent of potentially severe infections in its own right.


Projections from cortex to tectum in the tree shrew, Tupaia glis. Sensory neocortex of the tree shrew was divided into three main areas: the visual field, the auditory field, and the somatic field which includes motor cortex. Cortical cells which project to the tectum were identified by injecting HRP into superficial or deep layers of the superior colliculus and into various parts of the inferior colliculus. The main result is that these descending projections are well organized according to their origin in the three main sensory fields of the cortex. (1) Auditory field: labeled cells are found only in the core or auditory koniocortex, after injections of HRP in the pericentral area of the inferior colliculus; labeled cells are found in auditory belt areas after injections in posterior parts of the intermediate and deep layers of the superior colliculus, adjacent to the inferior colliculus. (2) Somatic field: labeled cells are also found in the somatic field after injections in the intermediate and deep layers of the superior colliculus, so that auditory and somatic fields probably overlap to some extent. The results do not exclude the possibility that somatic koniocortex has an exclusive target in the intermediate or deep layers of the superior colliculus. (3) Visual field: labeled cells are found only in the core or striate cortex after injections in the superficial layers of the superior colliculus. Labeled cells are found in the visual belt after injections in the rostral parts of the intermediate layers of the superior colliculus. When these results are related to ascending sensory pathways a picture emerges of a series of circuits or loops which interconnect parallel sensory pathways. These loops eventually reach the deep layers of the superior colliculus which of course have indirect access to motor neurons.


Connections of areas 3b and 1 of the parietal somatosensory strip with the ventroposterior nucleus in the owl monkey (Aotus trivirgatus). Anatomical tracers were injected into electrophysiologically defined sites in somatosensory cortical Area 3b (SI proper) and Area I (posterior cutaneous field) of owl monkeys after these cortical subdivisions had been extensively explored in microelectrode mapping experiments. These mapping experiments revealed that both Areas 3b and 1 contain complete and separate representations of the body surface (Merzenich et al., '78). Restricted injections of the retrograde tracer, horseradish peroxidase (HRP), into either Area 3b or Area 1 labeled neurons within a band of cells in the ventroposterior nucleus (VP). The location of the labeled band in VP varied with the location of the injection site in both representations, and the labeled region of VP was overlapping for injections in corresponding body parts in the two representations. Neurons projecting to the hand and foot cortical representations were in architectonically identified subnuclei. Because injections into either Area 3b or Area 1 labeled over half of the neurons in the appropriate regions of VP, it appears that some neurons in VP project to both cortical representations. Finally, injections of HRP combined with the anterograde tracer, 3H-proline, indicate that VP neurons are reciprocally interconnected with both Areas 3b and 1.


Immunologic and biochemical evaluation of the potency of whole insect body extracts. Recent studies have indicated that currently available whole body extracts have little potency and are ineffective for diagnosis and treatment of stinging insect allergy. Pure venom is a potent effective allergen but is difficult to obtain in sufficient quantities from all Hymenoptera species. In these studies, an attempt was made to prepare a potent whole body extract. Whole bee body extracts were prepared with different extraction periods and at cold and room temperatures. Potency was examined biochemically by measurements of phospholipase A (PLA) activity and immunologically by PLA and bee venom radioallergosorbent test (RAST) inhibition experiments and gel diffusion studies with the use of rabbit antisera. All extracts prepared in the laboratory had some potency, indicating that it is possible to make a whole body extract containing small quantities of PLA or bee venom. However, the potency of these extracts was minimal as compared with bee venom. Three commercial extracts were almost devoid of detectable immunologic activity. While further attempts may be made to prepare a potent whole body insect extract, these results suggest that it is necessary to obtain venom in relatively pure form for the diagnosis and treatment of stinging insect allergy.


Auditing the process of care in a new geriatric unit. To assess whether the objectives of a new Extended Care Unit were reflected in the care of the patients and in the outcome of that care, an audit of the patients' records was performed. The audit sample involved 101 geriatric patients who had been admitted to the Unit for rehabilitation therapy, and then discharged. The study included assessment of the records for compliance with individual audit criteria, examination of the records in terms of a composite audit score, and analysis of the relationship between these scores and three outcome indices. Although the audit did not show a significant correlation between recorded care processes and treatment outcomes, it did reveal the extent to which the Unit's goals were reflected in the care process.


Hyperthyroidism in the elderly.

Hyperthyroidism in the elderly. In 65 elderly hyperthyroid patients (age range 50-78 years), sex differences, signs and symptoms and thyroid function were studied and the data were compared with those on 48 young hyperthyroid patients (age range 20-29 years). The incidence of hyperthyroidism was 3.5 times higher in females than in males among the young patients, whereas it was approximately equal in males and females among the elderly patients. Signs and symptoms conformed with the textbook description in the young subjects but not in the elderly ones. Measurements of serum triiodothyronine (T3) and thyroxine (T4) proved useful in preventing false diagnoses in elderly patients with atypical symptoms. In some elderly subjects with marginal increases of serum T3 and T4 concentrations, measurement of serum thyroid-stimulating hormone (TSH) after administration of thyrotropin-releasing hormone (TRH) was required to achieve an accurate diagnosis. Serum T3 and T4 levels and the thyroidal uptake of radioiodine were slightly but not significantly lower in the elderly than in the young patients. A high titer of circulating thyroid auto-antibody in the elderly may be related to this slight decrease in thyroid function. Serum T3 levels were significantly lower in the elderly than in the young subjects; this suggests impairment of peripheral monodeiodination of T4. Any abnormal serum levels of T4, T3, and TSH before and after administration of TRH could easily be restored to normal by treatment with antithyroid drugs in the elderly patients.


d-Amphetamine in elderly patients refractory to rehabilitation procedures. This report concerns the use of d-amphetamine in 88 elderly inpatients who initially failed to respond to rehabilitation procedures. These patients exhibited characteristics of the Poor Motivation Syndrome (PMS), not associated with depression or dementia. The syndrome was seen five times more frequently in women. d-Amphetamine was given for three weeks in increasing dosage (2.5--10.0 mg twice daily). The responses were scored according to mobility, self-care and motivation. Of the 88 patients, 48 improved and another 28 were discharged who would otherwise have remained dependent nursing cases. An unexpected finding was that 17 patients showed an age-related resistance to the effects of the drug (P less than 0.05). The likelihood of a beneficial response also diminished with increasing age (P less than 0.01). Side effects were in the psychiatric sphere; they occurred in 23 patients early in treatment and were not age-related; the drug was withdrawn. Thus treatment with d-amphetamine should be restricted to selected patients who satisfy the diagnostic criteria of PMS, and administration should be carefully supervised. With these safeguards, a substantial proportion of patients previously refractory to rehabilitation will show improvement, become more independent and may be discharged from the hospital in a much more active, less dependent state of mine and body.


Effectiveness of hypertension screening in an urban optometric clinic. 461 randomly selected eyecare patients were screened for systemic hypertension at the clinic of the University Optometric Center, State College of Optometry, State University of New York. The results of this survey showed 52 screening failures, 30 of whom had previously been identified as having hypertension. The study showed that the screening and follow-up added $2.00 per patient to the chair cost. The authors concluded that the information provided to our patients, students and practitioners warrants this cost.


Occult thyroid disease in an elderly hospitalized population. Measurements of thyroxine (T4), triiodothyronine (T3), thyrotropin (TSH), and T3 talc uptake (T3TU) were performed on 425 hospitalized patients over 60 years of age. Unsuspected thyroid disease was found in 10 patients (2.4%); 9 were hypothyroid and 1 hyperthyroid. Another 11% of the population had abnormal T4 or TSH levels but were not proved to have thyroid disease. Repeat measurements in 10 of 40 patients with low serum T4 concentrations showed a return of the tests to normal. The low T4 levels occurred in patients who were seriously ill and were usually associated with low T3TU values. Low serum T3 concentrations were observed in 32 of the patients. Although we found TSH to be the most useful test in the diagnosis of hypothyroidism, it is a relatively expensive test and is not helpful in screening for hyperthyroidism. The present results suggest that the best single test for screening for occult thyroid disease is a serum T4 measurement. We recommend a T4 test on all hospitalized patients over age 60 years. Serum should be held in the laboratory for measurement of TSH concentration if the T4 result is below 6.5 micrograms/dl. A T3 resin or talc uptake and a serum T3 measurement should be performed to rule out hyperthyroidism if the serum T4 concentration is elevated. This regimen will identify virtually all cases of occult thyroid disease.


When a solution is not a solution: Medicaid and Health Maintenance Organizations. Health Maintenance Organization (HMOs) are repeatedly described as a general solution to the health care crisis and a specific solution to the problems confronting Medicare and Medicaid. The potential incorporation of HMOs into Medicaid has promised to improve the states' ability to accurately budget program expenditures, simplify management, eliminate abusive practices directly linked to fee-for-service (e.g., billing for undelivered services for providing unnecessary care) and, most important, contain costs. At the same time it has promised to increase access to mainstream medicine. Experience has shown, however, that the promise has often been severely compromised by unethical HMO marketing practices, inadequacy of services and excessive administrative costs. This article examines some of the reasons: the political process which created HMOs, the rhetorical claims which exaggerated the strengths of prepaid group practice and distorted the formulation of policy, and the peculiar characteristics of the Medicaid program which have caused Medicaid HMOs to deviate considerably from theoretical expectations.


Rationales for public health insurance coverage of geriatric day care: issues, options, and impacts. America's long-term care system has been widely criticized for many shortcomings. It relies too heavily upon institutional services, it is too costly, it forces inappropriate levels of care upon patients by offering too few options, in too many instances it offers inferior quality care, and it places too much emphasis on caring for physical ills without concern for enhancing patients' quality of life. Alternative modes and settings now under consideration could solve one or more of these problems, but the choices would be constrained by financial and technical barriers. Different choices have profoundly different consequences for costs and numbers and types of patients served. This paper considers different roles and their consequences for one alternative: geriatric day care. The issues raised apply to other alternatives.


Sero-epidemiological survey of scrub typhus. A total of 1017 human sera from ecologically different areas in the states of Rajasthan and Jammu & Kashmir were subjected to the CF test using the Gilliam strain antigen of R. tsutsugamushi. Results revealed that 2.4% and 3.5% of the populations examined in the two states respectively, were found to possess antibodies to the test antigen. The titres of positive sera varied from 1:8 to 1:32. Findings of this survey could be correlated with the local topography, presence or absence of vector and the exposure of susceptible population to the rickettsial agent. However, the presence of hidden foci in the neighbouring areas not covered by the current survey cannot be ruled out.


Ultrastructural localisation of lipopolysaccharide-binding sites with peroxidase-conjugated lipopolysaccharides. The localisation of lipopolysaccharide-binding sites on erythrocytes with peroxidase-coupled LPS is described. LPS was isolated from Fusobacterium nucleatum (Fus MC-8) by phenol-water extraction. The LPS was coupled to horseradish peroxidase by the two-step method of Avrameas and Ternynck (1971). The biological and serological activities of the conjugated LPS were compared with those of the native material. Peroxidase could be coupled to LPS without significant loss of endotoxic or serological activity. The LPS-peroxidase conjugate could be demonstrated on erythrocytes by light and electron microscopy.


Molecular properties of T-lymphoma immunoglobulin. II. Peptide composition of the heavy chain. In order to obtain structural information and to facilitate studies of the covalent structure of T-cell immunoglobulin, we have performed investigations of the peptide fragments of the heavy chain of this molecule, on a comparative basis, with heavy chains of serum immunoglobulins. T-cell immunoglobulin was isolated from 125I-labelled culture medium of monoclonal continuously cultured T-lymphoma cells by means of immunoadsorbents. Peptides were prepared from the purified 125I-labelled heavy chain by means of cleavage with cyanogen bromide or digestion with trypsin. We then resolved these peptides and compared them with those peptides derived from 131I-labelled murine mu, gamma and alpha chains separately and in mixed label experiments by means of polyacrylamide gel electrophoresis in sodium dodecyl sulfate containing buffers, 2 dimensional peptide mapping and ion exchange chromatography. The profiles of the radiolabelled peptides obtained from the T-lymphoma heavy chains were quite distinct from those of murine gamma chains but indicated a structural similarity to both mu and alpha chains, which share some common peptides. These results are consistent with the antigenic and physicochemical data available and suggest that T-cell immunoglobulin is a new isotype that shows similarities to IgA and IgM, but of which the precise nature of the constant region has yet to be delineated.


Molecular properties of T-lymphoma immunoglobulin. III. Peptide composition of the light chain. Structural studies of T-cell immunoglobulin light chains have been carried out in order to ascertain whether they possess a unique structure or if they resemble standard kappa or lambda isotypes. T-cell immunoglobulin was isolated from 125I-labelled culture medium of monoclonal, continuously cultured T-lymphoma cells, and the purified 125I-labelled light chains were subjected to either cleavage by cyanogen bromide or digestion with trypsin. These peptides were then resolved and compared with those derived from 125I- and 131I-labelled murine kappa and lambda chains in mixed label experiments, by means of polyacrylamide gel electrophoresis in sodium dodecyl sulfate-containing buffers and ion exchange chromatography. The profiles obtained suggested that T-lymphoma immunoglobulin light chains are immunoglobulin polypeptides and most closely resemble kappa chains. These results support available antigenic and mRNA hybridization data, which also suggest that T cells bear kappa-like light chains.


Genetic control of the immune response to mammalian chymotrypsins in mice. I. The immune response to high doses of bovine alpha-chymotrypsin. The primary and secondary immune response to the antigen bovine pancreatic alpha-chymotrypsin was investigated in inbred mice. It was found that strain differences in the immune response only became apparent after secondary immunization. The genetic control of the immune response was investigated in twelve different strains of mice, F1, F2 and F1 backcross hybrids, following secondary immunization. A continuous distribution for the mean antibody responsiveness was obtained. High responsiveness was associated with both the H-2 haplotype and three non-H-2 loci. Furthermore the F1 hybrids produced a greater quantitative antibody response to chymotrypsin than either of the corresponding parental strains.


Genetic control of the immune response to mammalian chymotrypsins in mice. II. The immune response to low doses of bovine alpha-chymotrypsin. The immune response to the antigen bovine pancreatic alpha-chymotrypsin was investigated in ten recombinant strains of mice. Using a fixed antigen-percentage bound isotope technique, it was found that the quantity of antibody produced was related to the H-2 haplotype of the responding animal. A continuous distribution for the mean antibody responses was obtained for the ten strains of mice. High responsiveness was associated with the H-2 haplotype gamma2. The genetic control of the immune response to this immunogen was found to be both quantitative and qualitative.


Inability to demonstrate hydroxylation of tyrosine by murine melanoma "tyrosinase" (L-DOPA oxidase), using the tritiated water assay technique. Validity of the tritiated water assay technique for tyrosine hydroxylase activity as a qualitative method was demonstrated with mushroom tyrosinase. Using this method, isolated murine melanoma "tyrosinase" (L-dopa oxidase) showed no tyrosine hydroxylase activity. This finding supports previous studies in our laboratory which used a variety of histochemical and biochemical methods. The nonenzymatic production of tritiated water caused by tritium exchange with hydrogen peroxide complicates the use of the tritiated water assay technique with crude systems, since hydrogen peroxide is generated by a variety of oxidase reactions. For this reason, previous studies using the tritiated water assay technique with crude systems are ambiguous.


Activation of the alternative pathway of complement by antiserum to factor B. Monospecific rabbit and goat antisera to human complement proteins and human immunoglobulins were tested for their ability to activate the alternative complement pathway. This activation was detected by two methods where classical pathway activation was blocked with EGTA and alternative pathway activation was promoted with added magnesium ions. These two methods consisted of lysis of GSHE and conversion of factor B into split products. C1q-depleted serum was used in a third assay system. Only antiserum to human factor B was able to activate the alternative pathway in the various systems used. None of the other anticomplement sera showed such activity. When antiserum to factor B was fractionated by ammonium sulfate and column chromatography, activation of the alternative pathway was found in the IgG fraction, and this activity was completely removed by absorption with purified factor B but not with other purified complement components.


Pseudomonas aeruginosa bacteremia: relationship of bacterial enzyme production and pyocine types with clinical prognosis in 100 patients. Strains of Pseudomonas aeruginosa were studied from 100 patients with bacteremia. In vitro quantitation of extracellular enzymes (lecithinase, protease, and elastase) and pyocine typing of these isolates were performed. No significant difference was found in the quantity of the enzymes produced or in the pyocine types by isolates obtained from patients dying from bacteremia or surviving this serious infection. Quantitation of the extracellular enzymes and pyocine types of blood isolates were contrasted with similar data obtained from sputum, urine, and skin isolates. One third of all strains produced minimal or no extracellular enzymes regardless of their source. However, the highest enzyme-producing strains were observed in the blood isolates. Although a greater variability of pyocine types was found in blood culture isolates, there was no significant difference between the pyocine types found in blood, urine, sputum, or skin isolates. The lack of correlation between the in vitro quantity of lecithinase, protease, and elastase produced by P. aeruginosa strains isolated from bacteremic patients and the prognosis of these patients supports the possible local rather than systemic significance of these extracellular enzymes in the pathogenesis of P. aeruginosa bacteremia.


Differences in attachment antigens of gonococci in reinfection. On the basis of bacterial typing, pyocin typing, and antibiotic sensitivity tests, two consorts appeared to have been reinfected 34 and 41 days later, respectively, with the same gonococcal strain, suggesting a failure in these patients to develop effective immunity to reinfection. However, these tests do not measure antigens which mediate attachment, a function which may correlate with virulence. When the above infecting strains were retested in an inhibition-of-attachment assay using rabbit gonococcal antisera, the antigens mediating attachment were found to be different. Homologous antisera inhibited attachment of the homologous strain at a high titer. Absorbing the antisera with the initial infecting strain did not remove any of the blocking activity of the antisera raised to the reinfecting strai, but the latter strain did share some attachment antigens in common with the initial infecting strain. Homologous antisera also bound preferentially to pili purified from the homologous strains.


Divalent cations and angiotensin-stimulated sodium transport. The presence of calcium in the fluid in which intestinal or kidney tissue is incubated is required for the tissue to respond to angiotensin. Everted sacs of rat colonic mucosa exhibited an increased rate of fluid transport in the presence of angiotensin; this response was lost when the serosal fluid, but not the mucosal fluid, was calcium-free. Angiotensin-stimulated transport was maintained when calcium was replaced with strontium or barium, but was lost when calcium was exchanged for magnesium. Similarly, calcium ions were required in the incubation fluid of rat kidney cortex slices to demonstrate angiotensin-enhanced sodium transport. These observations are discussed in relation to the possible roles of divalent cations in the mechanism of action of angiotensin.


Shared idiotypic determinants on antibodies and T-cell-derived suppressor factor specific for the random terpolymer L-glutamic acid60-L-alanine30-L-tyrosine10. T-cell derived suppressor factors (TsF) specific for the random copolymers L-glutamic acid60-L-alanine30-Ltyrosine10 and L-glutamic acid60-L-alanine40, referred to as GAT and GA, respectively, were prepared and partially purified on the approprate antigen immunoadsorbents. GAT-TsF obtained from nonresponder DBA/1 (H-2q) and SJL  (H-2s) mice were passed over immunoadsorbents prepared from normal guinea pig serum (NGPS) or guinea pig anti-idiotype antiserum (anti-CGAT) specific for a common cross-reactive idiotype found on most anti-GAT antibodies in all mouse strains tested. Both the directly suppressive activity of the GAT-TsF and the ability of GAT-TsF to induce new suppressor T cells (Ts2) in vitro were adsorbed to and fully recoverable from the guinea pig anti-CGAT-Sepharose immunoadsorbent, while the TsF passed through the control NGPS-Sepharose without appreciable binding. The SJL GAT-TsF specifically eluted from anti-CGAT-immunoadsrobents was shown to still posses I-J determinants. These data provide evidence suggesting a sharing of V region structures between B-cell antibody and T-cell suppressor factor specific for an antigen (GAT) under Ir gene control, in agreement with earlier studies on T and B-cell alloreceptors, T-cell helper factors, and T and B-cell receptors for conventional antigens.


Quantitative differences in the expression of parentally-derived H-2 antigens in F1 hybrid mice affect T-cell responses. Quantitative absorption with specific anti-H-2 sera has shown that the H-2Kb and H-2Dd antigens coded by the B10.A(5R) haplotype are expressed in about fourfold lower amount on the spleen cells of [B10.A(5R) X B10.A(2R)]F1 hybrids than on parental B10.A(5R) cells. In contrast, the H-2Kk and H-2Db antigens of B10.A(2R) are expressed equally on parental and F1 cells. These quantitative differences are reflected in cytotoxic T-cell (Tc-cell) function. Macrophage target cells from F1 mice are killed less efficiently than B10.A(5R) targets by alloreactive or H-2 restricted Tc cells specific for H-2Kb or H-2Dd, and spleen cells of F1 mice are less efficient stimulators of alloreactive Tc cells specific for B10.A(5R) H-2 antigens, whereas F1 and B10.A(2R) cells are equal as targets and stimulators for Tc cells recognizing B10.A(2R) H-2 antigens.


The role of H-2-linked genes in helper T-cell function. VI. Expression of Ir genes by helper T cells. We examined the expression of (TG)-A--L specific Ir genes in helper T cells using  T cells from low responder leads to (B10, high responder x low responder) F1 chimeric mice. In this paper, the low responder strain studied was B10.M, H-2f. B10.M T cells from these chimeric animals do not help anti-TNP-(TG)-A--L responses, even though they have matured in a high responder thymus and been primed and challenged with antigen on high responder Mphi and B cells. These findings indicate that in the H-2f haplotype an Ir-gene controlling anti-(TG)-A--L activity is expressed in helper T cells. The findings are in contrast to those we have obtained and previously reported with T cells of another low responder haplotype, H-2a. Taken together with our previous findings that (TG)-A--L specific Ir genes are expressed by B cells and Mphi of both the H-2a and H-2f haplotypes, the results indicate two sites of action for Ir genes, and suggest two different gene products acting at different stages of the response, both of which are defective in H-2f cells, and only one of which is defective in H-2a cells.


H-2-linked control of cytotoxic T-cell responsiveness to alphavirus infection. Presence of H-2Dk during differentiation and stimulation converts stem cells of low responder genotype to T cells of responder phenotype. Secondary Tc cells generated against Sindbis virus (SIN) are restricted to Dk. All other H-2K or D regions tested show low specific responsiveness. F1 hybrids between low and high responders show dominance of responsiveness but lack complementation. When BALB/c (KdIdDd) low responder fetal liver stem cells were allowed to mature in irradiated high responder recipients C3H.OH (KdIdDk) a response to Dk plus SIN could be generated with Tc cells of BALB/c origin. This result, together with the failure of complementation in the F1 hybrids, implies that the lesion of low responsiveness is in the inability of viral antigen to stimulate a Tc-cell response in association with any self H-2K or H-2D molecule (of those tested) other than H-2Dk. Hypotheses compatible with these data are discussed.


Isoprenoid quinones in the classification of coryneform and related bacteria. Menaquinones were the only isoprenoid quinones found in 85 of the 95 coryneform bacteria examined. Dihydromenaquinones having nine isoprene units were the main components isolated from Corynebacterium bovis, from other glutamic acid-producing strains, and from Arthrobacter globiformis and related species. Dihydromenaquinones with eight isoprene units were found in Brevibacterium linens, the remaining Corynebacterium species and strains probably belonging to the genus Rhodococcus. Tetrahydromenaquinones with eight isoprene units were found in Arthrobacter simplex and Arthrobacter tumescens, and with nine isoprene units in Cellulomonas and Oerskovia. Kurthia and Curtobacterium were characterized by menaquinones with seven and nine isoprene units, respectively, and Microbacterium lacticum and Corynebacterium aquaticum had comparable amounts of menaquinones with 10 and 11 isoprene units. Strains received as Brevibacterium leucinophagum, Corynebacterium autotrophicum, Corynebacterium nephridii, Mycobacterium flavum, Mycoplana rubra and Protaminobacter ruber contained uniquinones as their sole isoprenoid quinones. The isoprenoid quinone data correlate well with major trends in coryneform taxonomy and are of value in the classification of coryneform and related bacteria.


Late transcription and simultaneous replication of simian adenovirus 7 DNA as revealed by spreading lytically infected cell cultures. Miller's technique of spreading DNA was applied to monkey cells productively infected with simian adenovirus 7. This permitted the visualization of cellular DNA transcription, both nucleolar and non-nucleolar, and of late transcription and replication of virus. Virus double-stranded DNA, thin fibres with very few nucleosome-like particles, were observed carrying either transcription or replication complexes. In addition, both RNP transcripts and replication forks were found on some virus duplex DNA. Virus single-stranded DNA replicative intermediates were identified on the basis of their increased thickness and contrast which results from the presence of a DNA binding protein.


Radioimmunoassay of hepatitis B e-antigen (HBeAg): identification of HBeAg not associated with immunoglobulins. A radioimmunoassay for hepatitis B e-antigen (HBeAg) is described. Polystyrene beads coated with IgG prepared from a human serum containing antibodies to HBeAg (anti-HBe) and anti-HBe IgG labelled with 125I-p-hydroxyphenylpropionic acid N-hydroxysuccinimide ester were used in the test. The radioimmunoassay was approximately 1,000-fold more sensitive than immunodiffusion. At least a transient presence of HBeAg in serum appears to be a common feature of infections by hepatitis B virus. The radioimmunoassay was instrumental in establishing conditions for identification of apparently free monomeric HBeAg. The HBeAg has an approximate mol. wt.of 35,000 and was recovered after isoelectric focusing in fractions with a pH between 4.25 and 4.8. Polyacrylamide gel electrophoresis revealed the presence in HBeAg of a polypeptide with an apparent mol. wt. of 17,000.


The measurement of haemagglutinin and matrix protein present on the surface of influenza virus infected P815 mastocytoma cells. A thermodynamic approach has been used to measure the amount of haemagglutinin and matrix protein expressed at the surface of P815 cells infected for periods between 4.5 and 11 h with either WSN (H0N1) or JAP (H2N2) strains of type A influenza virus. This involved measuring the interaction of different concentrations of labelled (Fab)2 preparations of specific antibody with normal and infected cells. Assuming that one molecule of (Fab)2 bound to one molecule of antigen, values for the number of molecules of antigen/infected cell ranged from 7.6 X 10(5) to 1.7 X 10(7) for haemagglutinin and 1.3 X 10(5) to 1.1 X 10(6) for matrix protein. The ratio of haemagglutinin/matrix protein was lower for WSN-infected cells (1.7) than for JAP-infected cells (10). The same reagents were reacted with three purified A type virions; WSN, JAP and Port Chalmers (H3N2). Each preparation bound anti-matrix protein (Fab)2 though the value for haemagglutinin/matrix protein was much higher (66) than for infected cells and suggested that a virion may have a small number (about 12) of matrix protein molecules exposed though it was not excluded that the matrix protein detected was exposed only on damaged virions. Pre-treatment of infected cells with unlabelled reagent (anti-haemagglutinin) reduced the subsequent binding of the same labelled reagent but not the binding of the labelled matrix protein reagent and vice versa, suggesting that the haemagglutinin and matrix protein were not very close to each other on the cell surface.


Two cases of mucopolysaccharidosis type III (Sanfilippo). A biochemical study. The mucopolysaccharide and lipid composition of human nervous tissue and viscera from one case of Sanfilippo disease type A and one case of Sanfilippo disease type C, were investigated. In the brain a moderate increase of acid glycosaminoglycans occurred. This phenomenon was much more pronounced in the viscera, especially in the liver. In all tissues this increase was mainly due to an accumulation of heparan sulphate. Changes in lipid composition were noted, but can be regarded as secondary effects. The biochemical results reported also suggest some general conclusions. (a) AGAG and lipid analyses do not permit differentiation between the subtypes of Sanfilippo disease. (b) The differences in lipid composition can probably be considered as consequences of variation in secondary effects. (c) The severe demyelination in brain correlates well with the biochemical lipid analysis. However, in other instances it remains difficult to bridge the gap still existing between some morphological and biochemical data.


Characteristics of motor programs underlying arm movements in monkeys. 1. The experiments described here are addressed at identifying some of the processes underlying arm movements in monkeys. 2. We used three adult monkeys that were trained to point to a target light with the forearm and hold at that position for about 1 s in order to obtain a reward. During the experimental sessions the monkey was seated in a primate chair and its forearm was fastened to an apparatus that permitted flexion and extension of the forearm about the elbow in the horizontal plane. 3. We tested their performance prior to and after bilateral dorsal rhizotomy (C2--T3). Forearm movements were performed without the sight of the arm both before and after the surgical intervention. In intact animals we unexpectedly displaced the arm prior to movement initiation (150--200 ms) and observed the outcome of this displacement on movement termination. Our results indicated that the arm moved accurately to the target. The same procedure was used in the deafferented monkeys, yielding qualitatively the same results; i.e., a displacement of the initial position did not affect the attainment of the intended final position. 4. These results are relevant to the question of what is being controlled by motor commands. It appears that the controlled variable is an equilibrium point resulting from the interaction of agonist and antagonist muscles. Consequently, a change in the equilibrium leads to movement and the attainment of a new posture. The fact that both intact and deafferent monkeys display essentially similar motor behavior in our highly practiced task should not obliterate the dramatic difference in motor performance that exists between intact and rhizotomized animals. In fact, the successful execution of the learned motor performance in the deafferented animal is contingent on the animal's body being in a fixed relation to the arm apparatus. Whenever we changed the usual spatial relationship between the monkey's body and the arm apparatus, the animal's pointing response to the target was inaccurate. All of our intact monkeys, in contrast, were able to compensate quickly for any variations in their accustomed position with respect to the arm apparatus. The dramatic inability of the deafferented monkey to execute accurate pointing responses in an unusual postural setting underscores the great importance of the afferent monkey to execute accurate pointing responses in an unusual postural settiing underscores the great importance of the afferent feedback. These findings suggest that, in the performance of visually evoked learned movements, one of the major functions of the afferent feedback is in the adaptive modifications of learned motor programs.


Photoreceptor coupling in retina of the toad, Bufo marinus. I. Anatomy. 1. Red rod photoreceptors in the toad retina, which are known to be physiologically coupled, were examined for interreceptor contacts. 2. A dense network of large gap junctions was found between the inner segments of red rods, this being the only specialized site of contact that was observed between rods. Each red rod contacts an average of about four neighboring red rods with a junctional area of approximately 0.75 micrometer2. From freeze-fracture micrographs, the density of junctional particles was found to be 5 X 10(3)/micrometer2. 3. The large gap junctions were found only to connect red rods to each other in agreement with physiological data. Only small focal gap junctions were seen between red rods and cones and no junctions were observed between red and green rods. 4. It is concluded that these gap junctions are the site of coupling between the red rods and that the coupling is electrical.


Photoreceptor coupling in retina of the toad, Bufo marinus. II. Physiology. 1. Intracellular recordings were made from red rods in the isolated toad retina in order to measure the extent of the rod coupling. 2. The network space constant, lambda, was determined from edge-response profile measurements on single cells (lambda = 19 +/- 7 micrometer). 3. The network input resistance was estimated to be between 45 and 100 M omega. 4. Both discrete and continuous resistance networks, which were based on the anatomical observations, were analyzed. For lambda/D greater than 1, a continuous network provides an accurate description of the discrete network (D, cell spacing for a square grid). 5. The network analysis was used to estimate the specific resistance of junction membrane (0.75--1.5 omega.cm2) and the conductance provided by a single junctional channel (2.0--3.9 X 10(-12) mho). 6. The anatomical and physiological results provide strong evidence that the rods are electrically coupled. It is concluded that photoreceptor coupling provides averaging of the voltages in adjacent cells. The consequences of this averaging are discussed.


Supplementary motor area in the monkey: activity of neurons during performance of a learned motor task. 1. Recordings were made of the natural discharges of neurons in the supplementary motor area (SMA) of conscious monkeys trained to perform stereotyped motor task, pulling a horizontal lever, with either hand. 2. Of the total population of cells, 80% showed modulation of their activity during particular movements of either limb. Many cells had a similar pattern of modulation regardless of whether the contralateral or ipsilateral hand was used. Of the remaining 20%, some cells were related to leg or body movements or to visual experience. 3. Cells whose activity was related to movements of distal joints were found in approximately equal numbers to those whose discharges occurred with proximal movements. 4. Only 5% of cells tested sent their axons into the pyramidal tract, and only 14% of units investigated showed responses to passive manipulation of the limbs. The effective afferent input usually was of a rather complex kind. 5. The findings suggest that the discharges of a large number of neurons in SMA are changing during particular movements of either arm, and that only a small number of cells receive peripheral afferent sensory input. These results contrast with those obtained in the primary motor area and suggest a different role for SMA in the control of movement.


The effects of ischemia on long-tract neural conduction in the spinal cord. In this experiment, the effects of ischemia on neural conduction in the monkey spinal cord were studied. In six monkeys generalized ischemia of the spinal cord was created by bleeding the animals to a hypotensive level below the lower limits of autoregulation in the spinal cord. The progressive development of spinal cord ischemia was documented by blood-flow measurement using the hydrogen clearance method. Physiological integrity of the spinal cord was monitored and recorded by the spinal evoked response. The spinal evoked response did not disappear until at least 10 minutes of profound ischemia. At levels of ischemia 20% to 25% of normal blood flow, the spinal evoked response was unchanged. It is concluded that long-tract neural conduction in the spinal cord is relatively resistant to the effects of ischemia.


Detection of a large arteriovenous fistula between the internal lliac vessels by radionuclide angiography. A patient evaluated for heart failure was found by routine radionuclide angiography to have a large internal iliac arteriovenous fistula of presumed postoperative origin. The value of radionuclide angiography is described with a review of the literature on such unusual cases.


Tables to estimate total binding capacity of thyroxine-binding globulin from the in vitro thyroid function tests. Equations for an estimate of the total binding capacity of serum thyroxine-binding globulin (TBG-TC) were developed relating this parameter to serum thyroxine concentration (T4) and in vitro uptake (T3U). This extimate demonstrated a highly significant, positive correlation with TBG-TC as measured by electrophoresis in ten normal subjectdividuals with altered TBG-TC but without thyroid dysfunction (ten normal pregnant women, ten healthy women receiving anovulatories, ten nephrotics, and ten patients with severe malnutrition). True-positive (sensitivity) and true-negative (specificity) ratios were calculated for total and free T4 in serum, T3U, Free T4 Index, and both measured and calculated TBG-TC. False-positive results for free T4 index (12%) were due to altered TBG-TC. In such cases, 93% were recognized by the calculated TBG-TC from the calues of the invitro tests. It is concluded that this estimates should be added to the in vitro thyroid tests for their proper interpretation in cases where altered TBG-TC could be misleading. This estimate applies only to the particular in vitro testing system used herein.


Radionuclide computed tomography of the body using routine radiopharmaceuticals. I. System characterization. A whole-body computed tomography system for single-photon emitters was evaluated from the standpoint of spatial resolution, sensitivity, and thresholds for count densities with reference to standard doses of currently used radiopharmaceuticals in patients. In air and tissue equivalents, spatial resolution was relatively constant throughout the field of view and attentuation correction algorithms returned uniformity of response to within 10%. In a phantom of the human abdomen 1.5-cm spherical "cold" lesions and 1-cm "hot" lesions could be resolved. Aspects of the partial-volume effect were observed and investigated. To detect 1.5 cm-cold lesions in an abdominal phantom, five million events were required.


Radionuclide computed tomography of the body using routine radiopharmaceuticals. II. Clinical applications. A whole-body computed tomography system for single-photon emitters was used to investigate the clinical utility of this imaging modality. We have explored its application in brain, lung, liver, kidney, cardiac, bone, and gallium imaging in over 200 patients. Brain images demonstrated better lesion contrast than that in standard scintiphotos. Images of the lung and liver showed radionuclide distribution that was not readily apparent in standard scintiphotos. Moderate or strongly positive pyrophosphate myocardial images demonstrate the potential for infarct quantitation. ECG-gated cardiac blood-pool images in cross section, displayed in cine formate, permit evaluation of segmental motion of the free and septal walls of both ventricles. These results suggest significant clinical potential for this imaging modality using standard radiopharmaceuticals, but some increase in system sensitivity will likely be necessary to realize the full benefit of the technique.


Effect of portal versus jugular methionine infusion on circulating amino acids and nitrogen metabolism in sheep. A double cross-over experiment with 2 rumen-, bilateral jugular vein-, and portal vein-cannulated sheep was performed to determine the influence of route of infusion of methionine on nitrogen balance and blood amino acids. Nutrients were provided solely by intravenous (glucose, amino acids, water soluble vitamins) and intraruminal (VFA, minerals) infusion. Treatments consisted of the infusion of methionine plus serine via either the jugular vein (J) or portal vein (P). The respective order of treatments for the sheep were J, P, J, P and P, J, P, J. Urinary nitrogen was lower and nitrogen balance was greater when methionine was infused intrajugularly. No differences in plasma or blood urea nitrogen were noted. Plasma amino acid (PAA) concentrations showed no significant differences due to treatment. However, whole blood amino acid (BAA) concentrations revealed significantly greater levels of methionine when methionine was administered via the jugular vein. Expression of PAA as percentages of the total resulted in no treatment effects, while percentage expression of BAA showed that methionine was greater and phenylalanine was lower during intrajugular methionine infusion. The central and peripheral methionine infusion routes appeared to influence slightly the subsequent utilization of methionine as determined by effects on BAA levels of methionine. It was demonstrated that results from analysis of circulating amino acids may differ according to whether plasma or whole blood samples are used, and whether the data are expressed as concentrations or proportions.


The influence of dietary protein concentration and energy intake on mitogen response and tumor growth in melanoma-bearing mice. The effects of energy deprivation and low or high dietary protein levels upon lymphocyte transformation of spleen cells from syngeneic tumor-bearing and control mice were studied in a murine model of malignant melanoma. Both T- and B-lymphocyte transformation were significantly stimulated by the presence of a growing melanoma. T-cell responses however, were dependent only upon dietary protein concentrations, not the level of energy intake; whereas, the converse was true for B cells. Moreover, mice fed stock diet had the lowest response to mitogens of all diets tested. Except for mice receiving a 15% casein diet, tumor weights were generally not affected by level of intake or the amount of dietary protein. Others have demonstrated that melanoma cells have a greater need for tyrosine or phenylalanine than other tumor cells; thus we hypothesized that lymphocyte transformation may be depressed by relatively low phenylalanine or tyrosine levels in the diet when protein intakes are limited by either a low dietary concentration a restricted intake of a diet containing adequate protein, or both.


Tumor-host responses to various nutritional feeding procedures in rats. Parenteral and enteral nutrition are being used as adjuncts to cancer therapy. A liquid diet formulation containing a 27% solution of glucose and 3.9% crystalline amino acids with electrolytes and vitamins was given continuously for a week via parenteral (iv), and via intragastric (ig) routes and also was given ad libitum via the oral or per os (po) route to groups of Buffalo rats with and without a Morris No. 7777 transplantable hepatoma to find out how these feeding procedures affect tumor-host interactions. Other groups of rats with and without the hepatoma were given solid food ad libitum. The following parameters were examined: mortality, carcass and organ weights, body and tumor growth, nitrogen balance, energy intake, fluid balance, urinalysis, hematology values, and serum protein levels. The results are considered with respect to the influence of the tumor on the host and the influence of the feeding procedure on the animal with and without a tumor. The presence of the hepatoma was associated with: higher mortality, a decrease in carcass mass, leucocytosis, anemia, a decrease in serum IgG, transferrin and albumin, and an increase in serum alpha fetoprotein. The iv and ig feeding procedures alone resulted in some mortality which was exacerbated by the presence of the tumor. Mortality was especially high in the tumorous rats on the ig feeding procedure. The degree of positive nitrogen balance and carcass mass was similar in non-tumorous rats fed the same liquid diet formula when given iv, ig, or po. Tumorous rats fed the liquid diet ad libitum showed anorexia and a significantly lower nitrogen balance. The iv and ig feeding of tumorous rats at a level which was well above those of the tumorous rats given solid or liquid diet ad libitum maintained the same degree of positive nitrogen balance as non-tumorous rats. Even though the iv feeding of tumorous rats maintained about the same degree of positive nitrogen balance as non-tumorous rats, these tumorous rats still suffered loss of carcass mass. It appears that the large rapidly growing hepatoma has priority for available nutrition over the host. It is further suggested that the rapidly growing hepatoma places an ever increasing demand on the available nutrients. Thus, a point is eventually reached where even supplemental nutritional support can no longer meet the needs of the growing hepatoma and the host.


Aggressive ossifying fibroma of the maxilla: review of the literature and report of case. A case of a large aggressive ossifying fibroma of the maxilla has been presented. Review of the literature shows the literature shows the difficulty in establishing a definitive diagnosis through any single diagnostic modality. Much of this is due to the confusion surrounding classification of the tumor. Adjunctive radiographic and nuclear medicine diagnostic aids were utilized, as were clinical, laboratory, and histopathologic studies, in resolving the diagnostic questions posed by this large and aggressive tumor. Surgical intervention through conservative enucleation produced few operative or postoperative difficulties and minimal residual deformity. Differentiation of ossifying fibroma from other benigh and malignant neoplasms as well as fibrous dysplasia is important in the correct management of this lesion.


Antidiuretic hormone excess.

Antidiuretic hormone excess. Known physiologic mechanisms explain the elevated blood ADH levels observed in most patients with the syndrome of inappropriate ADH. Therefore the word "inappropriate" is a misnomer. It implies that the mechanisms that regulate ADH release are not functioning normally--which is not true. The term misleads the physician who, ideally, should determine why a patient has an excessive blood ADH level and initiate appropriate treatment. Patients with ectopic production of ADH and hyponatremia should be so labeled: "Hyponatremia due to ectopic ADH production." The term SIADH, if used at all, should be reserved for the rare patient with CNS injury or disease that causes increased ADH release and in which the hypothalamic center does not respond normally to afferent peripheral stimuli.


A clinical study of young infants after small intestinal resection. We prospectively studied 15 infants who, before 3 months of age, underwent resection of the small intestine-proximal in 3 infants, mid in 6, and distal in 6. Two died before one year of age. Many required prolonged parenteral nutrition, but by one year, 12 of the 13 survivors were on oral feedings only, and seven were above the third percentile for height and weight. Developmental delay occurred in the early postoperative period but diminished with time. There was compensatory adaptation of the remaining gut, shown by improving fat and B12 absorption and duodenal bile-salt concentrations. Bacterial contamination complicating end-to-side anastomoses occurred in two cases (P), gastric hyperacidity in four of 12 (1P, 3M), and hyperoxaluria in eight of 14 (1P, 5M, 2D). Studies of immune competence revealed normal cellular immune function (11/11), transient hypogammaglobulinemia (3/14), hypocomplementemia (1/12), and serum autoantibodies (3/10). Thus, massive resection of the small intestine did not preclude spontaneous improvement in absorptive function, growth, and development.


Thyroid function in preterm infants with respiratory distress syndrome. Thyroid function was evaluated in premature infants 30 to 35 weeks EGA with severe RDS and was compared to that of healthy control subjects of similar EGA. The infants with RDS had significantly lower serum levels of T3, T4, and FT4I during the first 60, 90, and 30 days, respectively, and had elevated T3UR during the first 10 days. The mean (+/- SD) postdelivery TSH peak was lower in the RDS group (32.8 +/- 9.6 muU/ml) than in the control group (60.9 +/- 21.8 muU/ml; P less than 0.005). After TRH injection the increment in serum TSH was inversely related to the basal FT4I in the control infants but not in the infants with RDS.


Dual effects of ether on end-plate currents. 1. The effects of diethyl ether (ether) on miniature end-plate currents (m.e.p.c.s) and on acetylcholine-activated end-plate channels were measured in toad sartorius muscle fibres with voltage-clamp and extracellular recording techniques. 2. At low concentrations (less than 20 mM) either made m.e.p.c.s decay faster than normal. At high concentrations (greaster than 40 mM), the decay of m.e.p.c.s was slower than normal. With all concentrations, the cecay remained exponential with single time constant, tau D. 3. At low concentrations ether did not affect the growth phase of m.e.p.c.s and only slightly reduced the amplitude of m.e.p.c.s. At the higher concentrations, the growth phase was slowed and m.e.p.c.s were significantly reduced in amplitude. 4. Ether at all concentrations (5--70 mM) reduced end-plate channel lifetime, the effect increasing with ether concentration. Ether did not significantly alter the elementary channel conductance or the actylcholine null (reversal) potential. 5. Curare reduced tau D which had been prolonged by high concentrations of ether. Ether itself at high concentrations caused a reduction in tau D increased by neostigmine. It is proposed that high concentrations of either inhibit acetylcholine hydrolysis by acetylcholinesterase. 6. The effect of ether in reducing end-plate channel lifetime and reducing m.e.p.c. amplitude, without significantly altering the normal voltage and temperature sensitivity of channel lifetime, is consistent with the proposal that either reduces the stability of open end-plate channels.


Central effect of taurine and its analogues on fever caused by intravenous leukocytic pyrogen in the rabbit. 1. Taurine infused I.C.V. after I.V. injection of leukocytic pyrogen (LP) inhibited the initial rise in body temperature and prolonged fever when infusion was stopped. 2. Similar infusion of taurine also inhibited the hypertermic effect of I.C.V. PGE2 (0.5 microgram) but did not cause prolonged hyperthermia. 3. I.C.V. administration of the taurine analogues hypotaurine and beta-alanine, compounds which have been shown previously to compete with taurine for facilitated transport in C.N.S. tissue, also inhibited the initial increase in body temperature and prolonged LP fever. 4. These results suggest that taurine prolongs LP fever by preferentially occupying a carrier system normally required for termination of the effects of endogenous pyrogens or related central mediators of fever. There was no evidence that taurine prolongs fever by blocking inactivation of central PGE2, a substance proposed previously to be a central mediator of fever.


Aminoglycoside antibiotics. 2. N,N-Dialkylkanamycins. N6',N3''-Dialkyl derivatives of kanamycins A and B were prepared regiospecifically from the parent antibiotics. Although the dimethyl and diethyl derivatives of kanamycin A were inactive in standard antibacterial assays, the dimethyl derivative of kanamycin B showed moderate activity, especially against various strains of Pseudomonas aeruginosa. A method for the selective dimethylation of the 3''-amino group of kanamycin A also was developed.


High-performance liquid chromatographic analysis of penicillin V benzathine oral suspensions. A rapid high-performance liquid chromatographic assay for penicillin V content of penicillin V benzathine bulk drug and oral suspensions is described. Dilution of the oral suspension with methanol containing 1,3,5-trimethoxybenzene as the internal standard allowed for direct analysis on a reversed-phase column with a mobile phase of 53% methanol in 0.05 M aqueous pH 3.5 phosphate buffer. A relative standard deviation of less than 1% was obtained on commercial formulations, and the system was linear over a range 10-40 microgram injected.


Percutaneous nitroglycerin absorption in rats. Percutaneous nitroglycerin absorption was studied in shaved rats by monitoring unchanged plasma drug concentrations for up to 4 hr. Drug absorption from the neat liquid state or from an alcoholic solution was considerably poorer than that from a commercial ointment. This observation was unanticipated since the driving force for percutaneous drug absorption was assumed to be drug thermodynamics. Potential artifacts such as drug volatilization from the skin, reduction of surface area through droplet formation, and vehicle occlusion were investigated, but they did not appear to be responsible for the observed results. Two experimental aqueous nitroglycerin gels were prepared with polyethylene glycol 400. One gel contained just sufficient polyethylene glycol to solubilize the nitroglycerin; the other had excess polyethylene glycol to solubilize nitroglycerin far below saturation. Both gels gave extremely low plasma nitroglycerin levels. The composite data suggested that percutaneous nitroglycerin absorption is highly vehicle dependent and that this dependency cannot be explained by simple consideration of drug thermodynamic activity.


Development of antibodies to meningococcal protein and lipopolysaccharide serotype antigens in healthy-carriers. The nasopharyngeal acquisition of meningococci was followed in healthy military recruits during their primary training. The production of antibodies to meningococcal serotype protein antigens and serotype lipopolysaccharide antigens accompanied the carrier state. Bactericidal antibody to protein serotype 2 was generated in response to the carriage of meningococci of low virulence that carried this antigen.


Spore antigens of Clostridium sporogenes. By means of spore-agglutination of fluorescent-antibody techniques, three serological types were identified among 84 strains of Clostridium sporogenes. Four spore antigens were identified, designated A, B, C and D.  A, B and C were specific for the respective types whilst D was a group antigen shared by strains of the three types. The spore antigens had corresponding somatic antigens; the type-specific somatic antigens were designated I, II, III, and the shared somatic antigen IV. The flagellar antigens were found to be type specific and were designated 1, 2 and 3; no common flagellar antigen was detected. The results of precipitation tests with spore extracts depended on the method of testing. By a capillary-tube ring method there were cross reactions among the three types of C. sporogenes, whilst by immunodiffusion in agar layers the reactions were generally type specific. In a disintegrated spore extract, two non-protein antigenic components, possibly polysaccharide, were detected by means of immunoelectrophoresis. This extract showed cross reaction with antisera to strains of all types of all types of C. sporogenes by the ring test and by immunodiffusion.


Scanning electron microscopy of uncoated human metaphase chromosomes. Human metaphase chromosomes were processed with a 3% glutaraldehydetannic acid technique and examined in a scanning electron microscope at 20 kV either without added metal coating or with 2 nm of sputtered gold coating. Several substrates--aluminum mnium foil, silver mirror deposit and sputtered gold-provided good conductive backgrounds for chromosomal spreads. Silver mirror deposit was the best conductive substrate tested. This method should prove to be a useful tool for monitoring the three-dimensional morphology of mitotic chromosomes with the possibility of studying various banding techniques, chromosomal uncoiling and secondary constrictions currently being examined in chromosomal studies.


Deoxythymidine nucleotide metabolism in Bacillus subtilis W23 infected with bacteriophage SP1Oc: preliminary evidence that dTMP in SP10c DNA is synthesized by a novel, bacteriophage-specific mechanism. Despite the fact that mature SP10c DNA contains dTMP, the acid-soluble fraction of infected cells contained no dTTP during the interval of phage replication. However, infected cells contained normal cellular levels of dATP, dGTP, and dCTP. Upon infection of deoxythymidine-starved Bacillus subtilis M160 (a deoxythymidine-requiring mutant of B. subtilis W23), mature phage DNA with a normal dTMP content was made. SP10c codes for an enzyme that seems to catalyze the tetrahydrofolate-dependent transfer of 1-carbon fragments to the 5 position of dUMP. The transfer of 1-carbon fragments is not accompanied by oxidation of tetrahydrofolage to dihydrofolate, implying that the enzyme in question is not a dTMP synthetase. It is proposed that dTMP in mature SP10c DNA is derived by the postreplicational modification of some other nucleotide and not by the direct incorporation of dTTP into DNA.


Order of assembly of the lower collar and the tail proteins of Bacillus subtilis bacteriophage phi 29. Extracts obtained after restrictive infection of Bacillus subtilis with mutants in cistron 11 of bacteriophage phi 29 are complemented in vitro by extract donors of the lower collar protein (p11). Purified 11- heads, containing the major capsid protein (p8), the fiber protein (p8.5), the upper collar protein (p10), and the virus DNA, can be also complemented in vitro to produce infective virus. This result suggests that 11- heads are intermediates in phage phi 29 morphogenesis. The order of assembly of the lower collar protein p11 and the tail protein p9 was determined in vitro in two complementation steps. The results obtained indicate that the lower collar protein is assembled before the tail protein.


Electron microscopic study of measles virus infection: unusual antibody-triggered redistribution of antigens on giant cells. Vero cells infected with measles virus fuse to form multinucleated cells which incorporated virus-specific antigens in their membrane. The distribution of these antigens was analyzed after a brief treatment with human anti-measles immunoglobulin G, using autoradiography and immunoperoxidase labeling combined with transmission and scanning electron microscopy. Virs-specific antigens were distributed over the entire surface of giant cells treated at 4 degrees C with human anti-measles immunoglobulin G and labeled Protein A. When cells were shifted to 37 degrees C, labeled antigen-antibody complexes were redistributed in two stages. Patch formation occurred in 5 to 15 min. Later, antigen-antibody complexes became concentrated in a paracentral "ring" rather than typical caps. Patch formation occurred in the presence of metabolic inhibitors, whereas ring formation was inhibited by metabolic inhibitors. These rings contained membrane folds, villi, and viral buds, whereas the rest of the membrane was smooth. In addition, shedding, endocytosis of antigen-antibody complexes, and reexpression of antigens were observed. Antibodies to nonviral membrane antigens induced the same pattern of redistribution. Infected cells treated with anti-measles Fab' fragments maintained a homogenous distribution of label throughout the experiments. In conclusion, intact immunoglobulins, but not Fab' fragments, were able to induce a dramatic redistribution of viral antigen on the membrane of giant cells infected with measles virus.


Vaccinia virus replication. I. Requirement for the host-cell nucleus. Using cytochalasin B-induced enucleation techniques, we examined the ability of vaccinia virus to replicate in the absence of the host-cell nucleus in several mammalian cell lines. It was found that virus-infected enucleated cells (cytoplasts) prepared from BSC-40, CVC, and L cells were incapable of producing infectious progeny virus. The nature of this apparent nuclear involvement was studied in detail in BSC-40 cells. Modulations designed to maximize cytoplast integrity and longevity, such as reduction of the growth temperature and initial multiplicity of infection, did not improve virus growth in cytoplasts. Sodium dodecyl sulfate-polyacrylamide gel analysis of the [(35)S]methionine pulse-labeled proteins synthesized in vaccinia virus-infected cytoplasts demonstrated that both early and late viral gene products were being expressed at high levels and with the proper temporal sequence. Vaccinia virus cytoplasmic DNA synthesis, as measured by [(3)H]thymidine incorporation, peaked at 3 h postinfection and was 70 to 90% of control levels in cytoplasts. However, in the cytoplasts this DNA was not converted to a DNase-resistant form late in infection, which was consistent with the failure to isolate physical particles from infected cytoplasts. Treatment of vaccinia virus-infected cells with 100 mug of rifampin/ml from 0 to 8 h to increase the pools of viral precursors, followed by subsequent removal of the drug, resulted in a threefold increase virus yield. This treatment had no effect on virus-infected cytoplasts. Finally, vaccinia virus morphogenesis was studied under an electron microscope in thin sections of virus-infected cells and cytoplasts which had been prepared at various times during a single-step growth cycle. It was apparent that, although early virus morphogenetic forms appeared, there was no subsequent DNA condensation or particle maturation in the cytoplasts. These results suggest that vaccinia virus requires some factor or function from the host-cell nucleus in order to mature properly and produce infectious progeny virus.


Carcinoma of the prostate: local control with external beam radiation therapy. Local clinical control of the primary disease was evaluated in 209 patients with stage C adenocarcinoma of the prostate treated with definitive external beam radiation therapy and followed for a minimum of 2 years. Of these patients 92 per cent required no further prostatic operations for obstruction. Prostatectomy before therapy did not necessarily prevent later prostatic obstruction from occurring. Of 129 patients who had only a needle biopsy before irradiation 90 per cent had improvement of the obstructive and/or irritative symptoms as tumor regression occurred with therapy and these patients did not require a later prostatic operation for obstruction. Stricture formation occurred in 8 per cent of the patients and was not influenced by the type of preirradiation prostatic operation done. If transurethral resection was reuqired after irradiation it was technically more difficult but the morbidity was acceptable. The incidence of hematuria and incontinence was far less than that reported in non-irradiated patients with this disease. Most tumors exhibited a down-grading effect after irradiation. There were no deaths attributable to the treatment. Over-all, 83 per cent of the 209 patients had no urinary complaints after completion of therapy. From a urological viewpoint, good clinical local control is achieved in the patient with stage C adenocarcinoma of the prostate treated with external beam radiation therapy.


Are excretory urograms necessary in evaluating women with urinary tract infection? Of 164 excretory urograms obtained from women with a history of recurrent urinary tract infections 88 per cent were perfectly normal. Eleven (6.7 per cent) had minor, normal, anatomic variations. Nine (5.5 per cent) were considered to have positive findings but in no case a significant finding present that required surgical intervention or altered the therapeutic approach. The total cost of the studies to the patients involved was $17,930. Thus, an extremely negative cost-benefit ratio emerged. The routine use of excretory urograms as part of the evaluation of a woman with a urinary tract infection is expensive, unrewarding and has little justification.


The physician's role in the cost-containment problem. The seemingly uncontrolled increase in the cost of medical care is one of the major problems facing the American physician. Although the reasons for escalated costs are multifactorial, the physician does play an important role in the problem. Several studies have suggested that physicians, the cost generators, and unaware of the economic impact of the care they provide. In the preparation of this report, 70 medical students, 316 interns and residents, and 41 faculty members at the University of Miami School of Medicine were surveyed on their knowledge of frequently ordered hospital billing items. A common lack of knowledge was noted in all three groups of respondents, with the faculty scoring as poorly as the medical students. Major efforts must be directed toward the cost education of physicians.


Single-antibiotic therapy for streptococcal endocarditis. Ninety-nine patients treated with penicillin G potassium, cephalothin sodium, or vancomycin hydrochloride were studied to evaluate single-drug therapy for nonenterococcal streptococcal endocarditis. Eighty-six patients survived; of these, 66 received penicillin alone. The maximum serum bactericidal titer obtained at the expected nadir of serum antibiotic concentration was 1:8 or greater in 70 (95%) of the 74 patients studied. No relapse occurred among the 66 patients treated wih penicillin. Only one patient experienced a relapse that occurred following therapy with suboptimal doses of vancomycin. Distinct second episodes of endocarditis occurred in five patients. While 13 patients died, none died of intractable infection. The treatment of endocarditis due to penicillin-susceptible streptococci with high-dose parenteral penicillin or a bactericidal penicillin substitute for four weeks or longer results in bacteriologic cure rates comparable with those achieved with combined penicillin-streptomycin regimens.