Distribution of antibodies to streptococcal esterases in patients with scarlet fever. Repetitive counterelectrophoresis (RCE), which has been described (Hayano and Tanaka, 1977), was used to assay the contents of antibodies to streptococcal esterases (STE) in sera from patients with scarlet fever. The levels of antibodies to STE were expressed semiquantitatively by reading the intensity of the colored spot developed by RCE with a densitometer. The present study deals with the determination of anti-STE in sera drawn at intervals from 54 patients diagnosed as suffering from scarlet fever. The STE used in this study were prepared from the streptococcal strains as follows. STE-AI was prepared from SS379 (group A, type 40), STE-AII from strain 69882 (group A, type 49), STE-B from strain H36B (group B, lb), and STE-C from strain Austin (group C). Of the 54 cases studied, 32 (59.3%) showed anti-STE-AI, 24 (44.4%) showed anti-STE-AII, 5 (9.3%) showed anti-STE-B, and 23 (42.6%) showed anti-STE-C. Comparison of the titer of anti-streptolysin O (ASLO) with the type of specific reaction of anti-STE-AI and -AII, determined in the same specimen, showed a marked correlation. Of 20 cases giving ASLO titers of 12 or less, 18 showed no sign of anti-STE-AI and -AII and two showed signs of anti-STE-AI. Of 34 cases giving ASLO titers of more than 12, 33 showed signs of anti-STE-AI and/or -AII, 22 showed signs of anti-STE-AI and -AII, 7 showed signs of anti-STE-AI, and 4 showed signs of anti-STE-AII. Of all the cases, 26 showed coincident rises in titers of ASLO and levels of anti-STE-AI and/or -AII during the course of the disease. One case showed the presence of only anti-STE-B. All of the 23 cases that showed the presence of anti-STE-C showed signs of anti-STE-AI. Some of these cases showed a marked increase in levels of anti-STE-C during the course of the disease.


Autoimmunity induced by injection of virus-modified cell membrane antigens in syngeneic mice. C3H/Bi mice developed autoantibodies after repeated inoculations of isolated membranes from primary tissue cultures of a syngeneic ascites lymphoma in which Newcastle disease virus had grown. This was in addition to the tumor transplantation resistance and cytotoxic antibodies previously demonstrated. The complement-fixing antibodies were completely removed from sera by adsorption with ascites tumor cells but only partially by normal mouse liver powder or C3H/Bi erythrocytes. With continued immunization, antibodies to deoxynucleoprotein and heterophile reagins also appeared. After several months, mice showing these serological reactions died with a wasting disease characterized by loss of lymphoid tissue and scarred, atrophied kidneys. No significant antibody response or autoimmune disease occurred in mice receiving membranes from uninfected syngeneic ascites lymphoma.


A modified leukocyte adherence inhibition test in the laboratory investigation of gastrointestinal cancer. A modification of the leukocyte adherence inhibition (LAI) technique has been used to test 60 hospital patients with gastrointestinal symptoms, for specific immunoreactivity against extracts of tumours of colon, pancreas and stomach. The modified technique employed mononuclear cells and soluble tumour extracts in glass test tubes, the non-adherent cells being enumerated in an electronic counter. Laboratory tests were completed before the eventual diagnosis was known. Groups of patients with adenocarcinoma of colon or rectum (11), carcinoma of pancreas (3) and adenocarcinoma of stomach (6) were clearly distinguished from each other and from patients with non-malignant diseases or with neoplasms of different histological type.


Somatic cell hybrids between human lymphoma lines. III. Surface markers. Hybrids between two human lymphoma lines, Raji and Daudi (8A) and Raji BJAB (83) were examined for genetically determined and/or differentiation-related surface markers. HL-A B ce-l alloantigens, Fc and complement receptors, EBV receptors and beta2 microglobulin showed an autonomous ("co-dominant") expression in the hybrid. This is in contrast to most previous studies on other differentiation markers, involving as a rule crosses between cells of different lineages, where the differentiated pattern usually became "eclipsed" in the hybrid. Staining of activated complement and complement consumption tests showed intermediate or partially suppressed expression in the hybrids. This may be viewed in relation to the fact that these reactions do not merely depend on complement binding to the receptor, but also on subsequent activation and binding of the activated complement. A more complex interaction is also suggested for immunoglobulin production. Surface immunoglobulin showed a suppressive or intermediate pattern in both hybrids, whereas intracellular kappa chain production showed an amplification in the 83 hybrid. The beta2 microglobulin deficiency of the Daudi parent was corrected in the Raji/Daudi hybrid. Two new HL-A specificities,A10 and BW17, appeared on this hybrid which were not present on the parental lines. This suggests that the HL-A deficiency of the Daudi cell is due to its lack of beta2 microglobulin.


Increased prescribing of Valium, Librium, and other drugs--an example of the influence of economic and social factors on the practice of medicine. Drug prescriptions per capita in the United States have more than doubled since 1950 without a commensurate improvement in health. Drugs are often prescribed for clinical conditions in which therapeutic benefits do not outweigh the risk of adverse drug reactions. Deaths due to adverse drug reactions are roughly as frequent as deaths due to automobile accidents. Valium and Librium are the first and fourth most commonly prescribed drugs in the U.S., used by one ten adults each year. The rapid rise in use of these drugs has occurred during a period of rising social stress, as indicated by increases in alcohol consumption, suicide, and homicide, Valium and Librium are frequently prescribe for patients who go to doctors with social or other nonmedical problems, often in lieu of attempts to resolve these underlying problems. Overprescribing occurs because the decision to prescribe is influenced not only by consideration of therapeutic benefit, but also by nonmedical factors, for example the widespread expectation by both patient and doctor that the doctor will provide a drug or some other technological treatment. Prescribing decisions are also influenced by the profit-motivated activities of drug companies, including the expenditure of almost one-quarter of every sales dollar on drug promotion. The most widely used source of drug information for doctors is the industry-sponsored Physicians' Desk Reference, which overrates the therapeutic value of Valium and Librium as compared to disinterested medical sources. Drug companies also contribute to overprescribing by introducing numerous minor variants of existing drugs. The therapeutic benefits of such new drugs are often overestimated in the early years of use when adverse side effects are not well known and apparent efficacy is enhanced by placebo effects in uncontrolled observations.


The impact of colonialism on health and health services in Tanzania. This article is about the problems of malnutrition and disease in a rural area of an underdeveloped country, Tanzania. The particular way in which health problems were conceptualized during the colonial era, the structure of the medical services established, and the effects of health care on the health status and size of the rural population of Songea District in Tanzania are shown in the article to have been determined by the economic, social, and political requirements of German and British colonial rulers rather than by the health needs of the African population. Colonial economic policy emphasized the production of cash crops for export, whether by African peasant farmers or by European plantation owners. To provide workers for the plantations, a system of labor migration was instituted. Songea District became an area that supplied male workers to other parts of the country, with grave consequences to the health and nutrition of the women and children left behind. Domestic food production was neglected by Africans forced to migrate in search of cash to pay taxes and by those enganged in the cultivation of cash crops. Extensive malnutrition and persistent ill health related to poor diet are thus traced directly to capitalist underdevelopment of th Tanzanian economy and the structural distortions of a dependent relationship between Tanzania and the metropolitan power.


Mechanism of steroid action in ocular inflammation: Inhibition of prostaglandin production. Prostaglandin E (PGE) concentration the aqueous humor of an intact rabbit eye was less than 0.1 ng. per milliliter and increased to 19 +/- 3 ng. per milliliter 60 minutes following paracentesis. The rise in PGE level was associated with clinical signs of ocular inflammation. Pretreatment with triamcinolone reduced both the accumulation of PGE  in the aqueous humor and the inflammatory response following paracentesis. intravitreal injection of E. coli endotoxin into rabbit eyes increases PGE level in the anterior chamber to 72 +/- 17 ng. per milliliter and induced acute uveitis. slices of iris and ciliary body (ICB) derived from either rabbit eyes with endotoxin-induced uveitis or normal eyes were incubated for 60 to 240 minutes and the rate of PGE release into the medium was measured by radioimmunoassay. after a 4 hour incubation, the PGE release from inflamed ICB was threefold higher than that of normal ICB. incubation of inflamed ICB with hydrocortisone, or Millicorten (100 mug per milliliter) for 4 hours reduced PGE accumulation in the medium by 50 and 81 per cent, respectively. Aldosterone had no effect on the rate of PGE release from inflamed ICB throughout the incubation period. Hydrocortisone or Millicorten also reduced PGE tissue content of inflamed ICB by about 74 per cent during the period of incubation. Indomethacin (100 mug per milliliter) abolished PGE accumulation. The suppressive action of hydrocortisone on PGE release into the incubation medium was prevented by the addition of arachidonic acid (2 mug per milliliter), a substrate for prostaglandin synthesis. By contrast , the inhibitory action of indomethacin was not affected by provision of arachidonic acid. We suggest that glucocorticosteroids reduce PGE accumulation by limiting the availability of the substrate for prostaglandin biosynthesis and thus suppress the inflammatory response.


Actin filaments in normal and migrating corneal epithelial cells. The distribution of actin filaments in the cytoplasm of normal and migrating rat corneal epithelium was studied by in situ labeling with myosin subfragment-one. In normal epithelium, actin filaments are present as an apical network under microplicae of superficial cell layers. This apical actin network is possibly a cytoskeleton responsible for maintenance of the regular array of microplicae. A dramatic difference in distribution of actin filaments was noted between the normal cells and cells migrating to cover an abrasion. In migrating epithelial cells, actin filaments are concentrated is basal regions of the cells. They are present as bundles of parallel filaments near the basal plasmalemma and in networks of filaments in leading edges and cell processes. These basally located actin filaments may play a role in migration of the cells during wound healing.


Pineal reactivity of anti-retina sera.

Pineal reactivity of anti-retina sera. Specific immunofluorescence has been demonstrated in the guinea pig pineal gland by homologous sera from guinea pigs injected with an extract of retina homogenate. This fluorescence appears to be in the cytoplasm and is evenly distributed among the cells of the pineal gland. Specific immunofluorescence in the retina has been previously demonstrated by these sera.


Significant bacteriuria among the mentally retarded. Identification and antibiotic sensitivity of isolates. A study was carried out to determine the prevalence of significant bacteriuria in a mentally retarded population. With 100,000 or more bacteria per ml. of urine used as a criterion for significant bacteriuria, 24 of 665 persons studied were considered to have significant bacteriuria for an overall prevalence rate of 3.6%. Seventeen of the 24 positive urine cultures were either from clean catch or catheter specimens. All patients undergoing catheterization were found to have significant bacteriuria. The individual prevalence rates of significant bacteriuria for males and females were 1.7 and 5.6 respectively. In females, an increasing prevalence of bacteriuria was found primarily in the 55-64 age group. The predominant bacteria isolated from bacteriuric patients were gram-negative bacilli, Escherichia coli, being isolated in 50% of all positive cultures. Antiobiotic sensitivity patterns of the isolates indicated a high degree of in vitro resistance to both ampicillin and cephalothin.


Recovery of Chlamydia and Genital Mycoplasma transported in sucrose phosphate buffer and urease color test medium. Urethral swabs from 75 males with urethritis were extracted into tryptose phosphate broth and then equal aliquots were dispensed into vials containing sucrose phosphate buffer (2SP) and urease color test medium (U-9). No antibiotics were present in the media. After transport to the laboratory, the recovery of Chlamydia trachomatis and Ureaplasma urealyticum was evaluated after inoculation into McCoy's cell cultures and agar medium, respectively. C. trachomatis was recovered from significantly more patients (17 versus 12, P = 0.03) with higher inclusion counts (P less than 0.01) in specimens transported in 2SP as compared with those in U-9 medium. No significant differences between the isolation rate of U. urealyticum and that of Mycoplasma hominis were found with the two media. The rate of inactivation of C. trachomatis and U. realyticum at 4 C was examined by means of reference strains. The inactivation of C. trachomatis was similar in both 2SP and U-9 media, but the number of inclusions was consistently greater in the 2SP medium. In contrast, the number of colony-forming units of U. urealyticum actually increased over a 24-hour period in both media. We conclude that 2SP is the best medium for the combined recovery of C. trachomatis and genital Mycoplasma. The use of one transport medium and hence a single swab culture has the obvious advantages of saving time and expense for both physician and laboratory, and for the patient it will eliminate the possible discomfort of having multiple cultures taken.


Comparison of enteric identification systems. An evaluation of methods for identification of Enterobacteriaceae was made employing the new commercial Micro-Media Enteric System (MMES) with that of the Analytab Products Incorporated (API) and the Conventional tube media schema as suggested by the Center for Disease Control (CDS). The MMES system employed 20 biochemical tests, the API 21, and the CDC procedure 25. Sixteen of these were identical biochemical tests. Two hundred clinical isolates of Enterobacteriaceae were tested employing procedures recommended by the manufacturers of MMES and API, and methods suggested by CDC. Among the sixteen identical biochemical tests the agreement was 98.0% (Conventional), 98.2% (API), and 97.98% (MMES). Bacteria misidentified by the API system totaled 5 (2.5%), 12 (6%) for the Conventional, and 13 (6.5%) for the MMES. Five of the bacteria misidentified with the MMES procedure were due to false positive citrate tests. This problem was subsequently eliminated. The results of this study indicated that the new MMES method for identification of Enterobacteriaceae compared favorably with both the API and Conventional procedures. However, significant advantages of the MMES method were evident in initial purchase price, utilization of technology time, and less tedium performing the test.


Vasoactive intestinal peptide (VIP) cells in the pancrease and gastro-intestinal mucosa. An immunohistochemical and ultrastructural study. Using antibodies against pure porcine VIP in immunoperoxidase and immunofluorescence tests, VIP-immunoreactive cells have been detected in the pancreas-especially in the islets-and gastrointestinal mucosa of the dog, guinea-pig and man. VIP immunoreactive cells were widely distributed in these tissues, never being numerous at any site. Some parallelism has been noted between such cells and ultrastructurally identified D1 cells fo the pancreas and gastrointestinal mucosa. The presence of VIP cells in normal pancreas may help explain the occurrence of pancreatic endocrine tumors producing VIP.


Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. II. Species-specific and species-non-specific antibodies to smooth and striated chicken muscle actin. The injection of rabbits with insoluble or soluble G-actin from chicken smooth or striated muscle will produce antibodies that are equally reactive, and species and tissue non-specific in immunoprecipitation, immunofluorescence and actin-activated Mg2+-ATPase inhibition tests. These antibodies have been used for the identification of actin-containing fibrils in a variety of tissues. When G-actins from chicken smooth or striated muscle are immobilized by chemical linkage to Affi-Gel 702 microbeads, their immunogenicity is increased, but the antibodies obtained against them are species-specific and will only react with actin and actin-containing structures from chicken and are therefore limited in use. It is concluded from this work that insoluble G-actin is the preferable immunogen to obtain precipitating antibodies for wide use.


The identification of the F-cell in the dog pancreas as the pancreatic polypeptide producing cell. The endocrine cells of the processus uncinatus in the dog pancreas were investigated with special reference to the formerly known F-cell. The F-cell was detected frequently in the periphery of pancreatic islets as well as among exocrine tissue. In both localizations the F-cell shows similar ultrastructural features. Membrane-bound irregularly shaped secretory granules of variable electron density were seen. The cell possesses all features of an endocrine polypeptide secreting cell. Using the immunofluorescence and immunoperoxidase technique in the uncinate processus of the dog, we could reveal that the anti-sera against bovine pancreatic polypeptide (BPP) reacts with the cell which is localized at the same sites as the F-cell. We therefore conclude that the pancreatic F-cell is identical to the pancreatic polypeptide-producing cell. The other endocrine cell types of the dog pancreas are glucagon-producing A-cells, insulin-producing B-cells, and somatostatin-producing D-cells, as well as serotonin-producing EC-cells which are regularly present in the dog pancreatic islets and also scattered among exocrine tissue and the duct epithelial cells.


Immunoelectron cytochemical localization of motilin and substance P in rabbit bile duct enterochromaffin (EC) cells. Using an immunoreactive technique the two peptides, motilin and Substance P, have been localized at the ultrastructural level in enterochromaffin (EC) cells. Motilin occurs in cells containing a mixed population of biconcave and round secretory granules whereas Substance P is found in cells with exclusively round granules. These observations confirm the existence of at least two functionally and morphologically different types of EC cell in rabbit bile duct, both of which contain 5-hydroxytryptamine. Classification of the endocrine cells of the gut on a purely morphological basis is clearly impossible, however.


Effect of oral inoculation of Escherichia coli on colostral antibody production in cattle. Oral inoculation of approximately 1.2 x 10(9) viable Escherichia coli to pregnant cows resulted in increased blood serum and colostral whey titers to the "O" antigen. The antibody titers were more pronounced in colostral whey and were correlated with the inoculum strain of Escherichia coli. There was no correlation between antibody titers of the colostrum ingested and the resulting serum antibody titers of the calves. The incidence of diarrhea in calves did not correlate with the antibody titer in the colostrum. The occurrence of diarrhea was significantly greater in calves that did not ingest colostrum until they were 12 hours old, compared with calves that had free access to their dams and suckled within an hour of birth.


Stability of patulin to sulfur dioxide and to yeast fermentation. The affinity of patulin for sulfur dioxide (SO2) is much less than was previously reported and is of little significance at the SO2 concentrations (below 200 ppm) used in the processing of apple juice and cider. However, at concentrations of 2000 ppm SO2 and 15 ppm patulin, combination was 90% complete in 2 days. Removal of SO2 liberated only part of the patulin, which suggests that 2 mechanisms are involved: one reversible (opening the hemiacetal ring) and one irreversible (SO2 addition at the double bond). Test with 2 yeasts used in English commercial cider making confirmed that patulin is effectively removed during yeast fermentation.


Regulation of phospholipid metabolism in differentiating cells from rat brain cerebral hemispheres in culture. Serine incorporation into serine phosphoglycerides: base exchange and decarboxylation patterns. The patterns of serine metabolism into phospholipids of cultured brain cells was examined. Labeled serine was incorporated predominantly into serine- ad ethanolamine-containing phospholipids and sphingolipids. The highest rates of labeling were observed in the (1)acyl-(2)acyl- and (1)alkyl-(2)acyl-serine phosphoglyceride fractions. Serine incorporation into both compounds appears to proceed via a base exchange mechanism. A decrease in the rate of serine phosphoglycerides labeling and a depletion of the ATP levels were observed when oligomycin or the calcium ionophore A23187 was added to the incubation medium. The inhibition of serine incorporation by A23187 could be partially reversed following addition of 10 mM CaCl2. Based on these findings it is suggested that in addition to demonstrating the energy-independent calcium-stimulated pathway, there may also be an energy related pathway. Formation of ethanolamine phosphoglycerides, as a result of serine phosphoglycerides decarboxylation, has been analyzed by using a simplified compartmental model. Of the 0.67 nmol/mg of protein turned over per h in the diacylserine phosphoglyceride compartment, 0.14 nmol/mg of protein are converted into the ethanolamine phosphoglycerides. In a similar manner, of the 0.09 nmol/mg of protein turned over per h in the (1)alkyl-(2)acyl-serine phosphoglyceride compartment, 0.014 nmol/mg of protein is converted into the (1)alkyl-(2)acyl-ethanolamine phosphoglyceride. These figures provide a first indication that a considerable portion of the ethanolamine phosphoglycerides in cultured brain cells is formed via a direct decarboxylation of the serine phosphoglycerides. In estimating the rates of (1)alkenyl-(2)acyl-ethanolamine phosphoglyceride formation from (1)alkyl-(2)acyl-ethanolamine phosphoglyceride the precursor-product specific activity crossover point could not be established. Mathematical analysis, however, enabled us to estimate the flux from the former into the latter as 0.04 nmol/mg of protein per h. A scheme for the possible metabolic interconversions of the ether bond containing serine and ethanolamine phosphoglycerides is proposed.


Codon-acticodon recognition in the valine codon family. An in vitro protein-synthesizing system completely dependent on added valine tRNA (valyl-tRNAval) and programmed with RNA from the phage MS2 has been used to investigate the incorporation into MS2 coat protein of valine from isoaccepting valyl-tRNAsval with the anticodons U AC (U represents 5-oxyacetic acid uridine monophosphate), GAC, and IAC in response to the four valine codons GUU, GUC, GUA, and GUG. By examining the incorporation of valine into NH2-terminal and internal positions of three tryptic peptides from the MS2 coat protein it has been established that these anticodons each recognize all four valine codons. We therefore conclude that under our conditions of in vitro protein synthesis the genetic code, as far as the valine codons are concerned, is operationally a two letter code, i.e. the third codon nucleotide has no absolute discriminating function.


On the role of ATP in phosphodiester bond hydrolysis catalyzed by the recBC deoxyribonuclease of Escherichia coli. The deoxyribonuclease specified by the recB and recC genes of Escherichia coli (recBC DNase; exonuclease V) has been purified to near homogeneity by a new procedure. Although hydrolysis of even a single nucleotide from a duplex DNA molecule by the pure enzyme is absolutely dependent upon ATP, the extent of phosphodiester hydrolysis is strongly inhibited by ATP concentrations of 0.2 mm or greater, and the initial rate is unaffected. Under these conditions, the extent of DNA hydrolysis is proportional to enzyme concentration. In contrast, neither the rate nor the extent of hydrolysis of single-stranded DNA nor ATP is affected by high concentrations of ATP. The amount of large single-stranded polynucleotide generated by the action of the recBC DNase increases as the ATP concentration increases and, at 0.5 mM ATP, becomes equivalent to the amount of acid-soluble nucleotide formed. These findings suggest that high intracellular concentrations of ATP affect the mechanism of the recBC DNase so as to limit the extent of hydrolysis of duplex DNA, while at the same time favoring the formation of single-stranded regions within the duplex. Such regions may be essential intermediates in the recombination process.


Antagonistic action between spermidine and putrescine on association and dissociation of purified, run-off ribosomes from Escherichia coli. The effects of polyamines on the equilibrium between prokaryotic ribosomal subunits and 70 S ribosomes have been studied as a function of concentration of Mg2+ from 2.5 to 7.5 mM. Run-off ribosomes were obtained from Escherichia coli and were washed with buffered 1 M NH4C1. Spermidine at 1 mm favors association of subunits at all concentrations of Mg2+. Putrescine, at concentrations above 8 mM, favors net dissociation at concentrations of Mg2+ below 4.5 mM. Streptomycin behaves like spermidine, while putrescine behaves like initiation factor 1 and initiation factor 3. The effect of putrescine on dissociation is time-dependent and appears to have a half-life of about 3.5 min at 30 degrees. When added after the effects of spermidine or streptomycin on association have occurred, putrescine still causes dissociation. The data suggests that putrescine may reduce net formation of vacant 70 S ribosomes. Another possibility is that putrescine and spermidine may act antagonistically to maintain a labile equilibrium between ribosomal subunits and vacant 70 S ribosomes. It may be significant that the putrescine effect is observed at the concentration of Mg2+ found to be optimum for initiation.


Reaction of bone to the acute chemical trauma of bone cement. The acute chemical trauma of bone cement was investigated in the rabbit tibia. With the experimental model used, the surgical trauma and the thermal effects of the cement could be minimized. Each animal received one implant of polymerized cement as a control and one implant of polymerizing dough as the test material. The animals were observed for up to seventy days and the specimens were then studied by various techniques of histology, microangiography, fluorochrome labeling, and microradiography. With this experimental method the monomer trauma did not add to the minimized surgical trauma. Moreover, no detectable influence on bone regeneration was noted. In view of this minor effect, we suggest that the surgical preparation of the implant bed and the very procedure of inserting bone cement are more important for the acute tissue trauma than is monomer leakage alone.


Molecular events during membrane fusion. A study of exocytosis in rat peritoneal mast cells. We have used thin section and freeze-fracture electron microscopy to study membrane changes occurring during exocytosis in rat peritoneal mast cells. By labeling degranulating mast cells with ferritin-conjugated lectins and anti-immunoglobulin antibodies, we demonstrate that these ligands do not bind to areas of plasma membrane or granule membrane which have fused with, or are interacting with, granule membrane. Moreover, intramembrane particles are also largely absent from both protoplasmic and external fracture faces of plasma and granule membranes in regions where these membranes appear to be interacting. Both the externally applied ligands and intramembrane particles are sometimes concentrated at the edges of fusion sites. The results indicate that membrane proteins are displaced laterally into adjacent membrane regions before the fusion process and that fusion occurs between protein-depleted lipid bilayers. The finding of protein-depleted blebs in regions of plasma and granule membrane interaction raises the interesting possibility that blebbing may be a process for exposing the granule contents to the extracellular space and for the elimination of excess lipid while conserving membrane proteins.


Ultrastructure of a periodic protein layer in the outer membrane of Escherichia coli. Matrix protein (36,500 daltons), one of the major polypeptides of the Escherichia coli cell envelope, is arranged in a periodic monolayer which covers the outer surface of the peptidoglycan. Although its association with the peptidoglycan layer is probably tight, the periodic structure of the peptidoglycan. Although its association with the peptidoglycan later is probably tight, the periodic structure is maintained in the absence of peptidoglycan, and is therefore based on strong protein-protein interactions. A detailed analysis of the ultrastructure of the matrix protein array by electron microscopy and image processing of specimens prepared by negative staining or by freeze-drying and shadowing shows that the molecules are arranged according to three fold symmetry on a hexagonal lattice whose repeat is 7.7 nm. The most pronounced feature of the unit cell, which probably contains three molecules of matrix protein, is a triplet of indentations, each approx. 2 nm in diameter, with a center-to-center spacing of 3nm. They are readily penetrated by stain and may represent channels which span the protein monolayer.


Immunocytochemical localization of secretory proteins in bovine pancreatic exocrine cells. The bovine exocrine pancreatic cell produces a variety of enzymes and proenzymes for export. Biochemical studies by Greene L.J., C.H. Hirs, and G.E. Palade (J. Biol. Chem. 1963. 238:2054) have shown that the mass proportions of several of these proteins in resting pancreatic juice and zymogen granule fractions are identical. In this study we have used immunocytochemical techniques at the electron microscope level to determine whether regional differences exist in the bovine gland with regard to production of individual secretory proteins and whether specialization of product handling occurs at the subcellular level. The technique used is a modification of one previously reported (McLean, J.D., and S.J. Singer. 1970. Proc. Natl. Acad. Sci U.S.A. 69:1771) in which immunocytochemical reagents are applied to thin sections of bovine serum albumin-imbedded tissue and zymogen granule fractions. A double antibody technique was used in which the first step consisted of rabbit F(ab')2 antibovine secretory protein and the detection step consisted of sheep (F(ab')2 antirabbit F(ab')2 conjugated to ferritin. The results showed that all exocrine cells in the gland, and all zymogen granules and Golgi cisternae in each cell, were qualitatively alike with regard to their content of secretory proteins examined (trypsinogen, chymotrypsinogen A, carboxypeptidase A, RNase, and DNase). The data suggest that these secretory proteins are transported through the cisternae of the Golgi complex where they are intermixed before copackaging in zymogen granules; passage through the Golgi complex is apparently obligatory for these (and likely all) secretory proteins, and is independent of extent of glycosylation, e.g., trypsinogen, a nonglycoprotein vs. DNase, a glycoprotein.


Ca++ regulation in caffeine-derived microplasmodia of Physarum polycephalum. Caffeine-derived microplasmodia possess a Ca++-sequestering system which can initiate motility. The experiments presented here suggest that this system is membranous and nonmitochondrial in  nature. Therefore, it is proposed that the shuttle streaming in the plasmodium is controlled by the localized release and uptake of free Ca++ from an intracellular storage system analogous to the sarcoplasmic reticulum.


LH and FSH levels in urine and serum of prepubertal and pubertal children receiving a 3 hour infusion of LH-RH. LH-RH was administered to 17 normal prepubertal and 14 pubertal children as well as to 6 GH deficient prepubertal patients. Urinary immunoreactive LH and FSH were measured in acetone extracts of 3 h collections prior to, during, and immediately after a 3 h infusion of 100 mug LH-RH. Peak LH excretion in response to LH-RH was higher in pubertal than prepubertal children. Girls excreted larger quantities of FSH after LH-RH than did boys. Serum LH and FSH increments evoked by LH-RH correlated significantly (P less than 0.01) with peak urinary gonadotropin excretion. These data suggest that measurement of urinary immunoreactive LH and FSH prior to and after LH-RH administration is clinically useful in evaluation of the reproductive endocrine system of young children and of individuals with low basal levels of gonadotropin.


Enzyme-labelled immunoassay for plasma cortisol. An enzyme-labelled immunoassay for plasma cortisol was developed. For this alkaline-phosphatase was conjugated through 21-hemisuccinate of cortisol using water-soluble carbodiimide. An antibody was raised in rabbits against corisol-21-hemisuccinate bovine serum albumin. Before assay 10 mul samples of plasma were mixed with glutamate buffer, pH 3.3, and boiled to denature endogenous cortisol-binding globulin and alkaline-phosphatase. Separation of "bound and free" cortisol was done by the double antibody method. A linear relationship was obtained between the volume of plasma and the amount of cortisol. The minimal detectable level of plasma cortisol was 1 mug/dl and the coefficients of variation were 2.7%-4.4% (within assays) and 4.7%-6.0% (between assays). Cortisol values determined by the present method correlated well with those determined by radioimmunoassay. The present method of enzyme-labelled immunoassay is suitable for routine clinical analysis of plasma cortisol.


Inhibition of immune hemolysis: serological assay for the heat-labile enterotoxin of Excherichia coli. Sheep erythrocytes sensitized by incubation with the heat-labile enterotoxin (LT) of Escherichia coli are hemolyzed in the presence of anti-LT antiserum and complement. The Microtiter (Cooke Laboratory Products) technique was used to titrate anti-LT antibody in serum by this immune hemolysis reaction. Immune hemolysis was inhibited by preexposure of the anti-LT antiserum to soluble LT before addition of the LT-sensitized sheep cells. E. coli mini-extract preparations were obtained by the polymyxin release technique and assayed for LT by the lysis inhibition test (LIT) and by the adrenal cell assay. All 75 adrenal cell-positive E. coli isolates were positive in the LIT assay. Eight of 318 adrenal cell-negative isolates tested were positive in the LIT assay, possibly indicating the presence of biologically inactive toxin.


Reduction of variation in results of rheumatoid factor tests by use of a serum reference preparation. Standaridizing test results for rheumatoid factor by comparing results obtained for an unknown with results obtained for a serum reference preparation decreased variance between laboratories, as measured in the Center for Disease Control proficiency testing program, by 77%. The amount of improvement was also estimated by the type of test and by the manufacturer's product. Standardization resulted in an increase in the number of reported results that were within a twofold dilution of the median value. The percentage increased from 50.3 to 93.7% for the slide tests and from 78.1 to 91.2% fro the tube tests. Decrease in variance by manufacturer's product ranged from 94 to 27%. The study demonstrated that adopting a reference serum standard could substantially improve the comparability of rheumatoid factor test results and that proficiency testing programs can be used to estimate improvement which could be expected as a result of standardization.


Rapid carbohydrate fermentation test for confirmation of the pathogenic Neisseria using a Ba(OH)2 indicator. The Ba(OH)2 indicator system was demonstrated to be a practical procedure in assisting clinical bacteriologists in the accurate and rapid identification of the pathogenic Neisseria from clinical specimens. This system measured the release of CO2, resulting from the metabolism of fermentable carbohydrate, as the precipitated BaCO3, by means of a spectrophotometer, The method was uncomplicated and can be performed in most clinical bacteriology laboratories.


Development of a lysis-filtration blood culture technique. A lysed-blood culture system that quickly lyses patients' blood near neutrality and is relatively noninjurious to more delicate pathogens such as Haemophilus influenzae and Bacteroides fragilis is reported. The lysing solution includes culture medium, 0.004 M sodium carbonate and bicarbonate, 0.04% Triton X-100,and 0.6% Rhozyme (a mixture of proteases). Most of the pathogens tested multiplied in the lysing solution. The lysed blood normally is immediately filtered. The membrane is transferred to culture broth. The greatest advantage realized from this blood culture technique is separation of pathogens from antibiotics, bactericidal antibodies, complement, opsonins, and phagocytic systems. Another advantage is the concentration of organisms into a small volume of clear medium for faster growth and visualization of growth. It was observed that both gram-negative and -positive organisms were attracted during filtration to the filter material and were not removed from it by backwashing with buffer. Thus, filter membranes with porosities much larger than would nominally be expected to retain bacteria retained all or part of light and heavy Escherichia coli and Staphylococcus aureus suspensions. Advantage may be taken of this phenomenon to use filters with larger pore sizes and avoid filter clogging by poorly lysed specimens. Porr lysis may result from addition of too much blood to the lysing solution, blood with elevated numbers of erythrocytes or leukocytes, or blood from some people whose blood is naturally more resistant to lysis.


Occurrence and significance of Cryptococcus neoformans in the respiratory tract of patients with bronchopulmonary disorders. Cryptococcus neoformans was cultured from 13 (3%) of 469 clinical specimens examined from the respiratory tract of patients with bronchopulmonary diseases. These isolations came from 5 (2%) of 207 patients; 11 isolates were from sputum and 1 each were from bronchoscopic aspirate and empyema pus. The fungus was not cultured from the oropharyngeal washings of 101 apparently healthy volunteers. Of the 5 patients, 3 had pulmonary tuberculosis, including one with pyopneumothorax and 2 with allergic bronchopulmonary aspergillosis as the underlying disease. In the tuberculosis patient with pyopneumothorax and C. neoformans in empyema pus, the fungus was presumably a tissue invader, whereas its role could not be unequivocally ascertained in the remaining 4 patients from whom it was isolated from sputum or bronchial aspirate on at least two consecutive occasions. The question of C. neoformans being a transient resident, commensal, or incitant of benign minimal lesions in the tracheobronchial tree is discussed. A comprehensive laboratory and clinical follow-up is warranted in patients from whose sputum or bronchial aspirate C. neoformans may be cultured even though definitive signs of cryptococcosis may be lacking.


Rapid automated disgnosis of bacteremia by impedance detection. Lysis and filtration of blood culture specimens were combined with impedance detection of bacterial growth to facilitate the diagnosis of bacteremia. A blood lysis-filtration technique (Zierdt et al., 1976) was coupled to a simple, inexpensive automated detection system. The practical and technical aspects of the impedance detection system are discussed. This new blood culturing system was compared to a conventional system for 264 aerobic blood cultures. A 30-ml sample of the blood-broth mixture was withdrawn from the conventional aerobic blood culture bottle and processed in parallel. Excluding the isolation of the commonly recognized contaminants, the detection efficiency was 36% greater in the new system. A total of 53 blood cultures from 107 patients were positive by one or both methods. The new system detected 92% of the total number of positive cultures, compared with 56% detected by the routine method. The explanation of the differences is discussed.


Multiple biotypes of Klebsiella pneumoniae in single clinical specimens. The occurence of multiple biotypes of Klebsiella pneumoniae within single specimens was determined in 59 clinical specimens. Biotyping was performed on five colonies of K. pneumoniae from each specimen, using the API 20E system (Analytab, Inc., New York) for identification of Enterobacteriaceae with strict adherence to the manufacturer's instructions. Multiple biotypes of K. pneumoniae were present in 31% (18) of the clinical specimens. Twenty-eight colonies representative of specimens with single and multiple biotypes were tested further for biotype reproducibility. Whereas genus and species identification was 100% reproducible, variation of one or more biochemical tests on serial transfers resulted in biotype reproducibility of only 64%. The greatest variation in biochemical tests occurred with urease (14%), indole production (10%) and citrate utilization (9%). Multiple biotypes in single specimens appear to be due to both inherent differences among the colonies in the specimen and variability in the system used to determine biochemical reactions. The presence of multiple biotypes limits the usefulness of biochemical typing for epidemiological surveilance of K. pneumoniae.


Bacteriophage typing of Shigella sonnei. A bacteriophage-typing schema was developed for differentiating strains of Shigella sonnei. Sixty-seven bacteriophages were obtained from other collections, and 36 bacteriophages were isolated from sewage. From these 103 bacteriophages, a provisional set of 12 was chosen by computer analysis as being the most sensitive in differentiating strains of S. sonnei isolated in the United States. The provisional schema was used to type 265 strains from different geographical areas. It divided them into 87 different lysis patterns, and all 265 strains were typable. Smooth and rough colonial variants of the same strain had different lysis patterns, so the technique was standardized to type rough colonies only. Reproducibility was difficult to obtain until all conditions were carefully standardized. Changes in results were noted even on different lot numbers of Trypticase soy agar, which was defined as the standard medium. So that the medium would not be a variable, 100 pounds (ca 453.5 kg) of the same lot number was purchased. Bacteriophage typing was very useful in differentiating strains, and work should continue on establishing a standarized schema.


Detection of Haemophilus influenzae type b antigens in body fluids, using specific antibody-coated staphylococci. Protein A-rich staphylococci coated with Haemophilus influenzae type b antiserum agglutinate specifically with homologous bacterial cells or with cell-free supernatant fluids of cultures of the organism. Antibody-coated staphylococci were used to detect soluble antigens in body fluids of patients infected with H. influenzae type b. Cerebrospinal fluid from 36 cases of meningitis caused by this orgainsm showed positive coagglutination tests in 86% of patients prior to initiation of therapy. Antigens could be detected in 46% of sterile cerebrospinal fluid specimens obtained from the same cases 1 to 10 days after therapy. Soluble antigens were also detectable in sera (58%) and urine specimens (67%) of patients with H. influenzae type b septicemia, when such specimens were tested within 10 days of onset of illness. No antigen could be detected in body fluids beyond 10 days. The coagglutination test was positive in 57% of all body fluids examined; contercurrent immunoelectrophoresis (CCIE) was positive in only 27%. All specimens positive by CCIE were also positive by coagglutination. No false-positive reactions were noted by either test in body fluids from controls. The coagglutination test is simple, specific, and more sensitive than the CCIE method and could be a valuable tool for detecting antigens in body fluids of patients with various infections.


Human figure drawings of sex offenders.

Human figure drawings of sex offenders. The D-A-P was administered to 60 male sex offenders from a correctional institution. The drawings of the inmates, taken as a whole, seemed to be indicative of narcissism, assertion, sex disturbance, aggression, and anxiety. Significant differences in the drawings of rapists, child molesters, and incest offenders were found. The following D-A-P aspects appeared to be prominent in the drawings of the Ss, in descending order: reinforcement, long nose, large head, legs wide apart, too much hair, waistline emphasis, slash mouth, long feet, pointy fingers, distorted heads, and opposite sex drawn first.


The relationship of A-B, field dependency, and emotional openness in paranoid and nonparanoid schizophrenics. The relationship between A-B score, Field dependency (EFT), and paranoid-nonparanoid status in 24 male acute psychiatric inpatients was explored. It was hypothesized that paranoids would score as Bs and extreme field independents (conceptual orientation), while nonparanoids would score as As and field dependents (perceptual orientation). As predicted, nonparanoids were significantly more field dependent than paranoids and controls, but no difference was found between paranoids and controls. Contrary to our hypotheses, nonparanoids were significantly more B than paranoids and controls. The only difference between paranoids and controls was that paranoids were less feeling reactive. The results were interpreted in terms of the "normality= of the conceptually oriented paranoid S.


Indoprofen in knee joint osteoarthritis: a double-blind, crossover clinical trial. A double-blind, crossover clinical trial has been carried out in subjects with knee joint osteoarthritis to assess the activity of two nonsteroidal antiinflammatory drugs. Eighteen outpatients with a severe systemic form were given orally indoprofen (800 mg/day), indomethacin (80 mg/day), and placebo for one week without interval between treatment periods. Significant improvement was seen in subjective and objective signs and symptoms after both drugs, which gave similar results. No significant differences between drugs were noted as to patients' opinions and preferences, which were in agreement with clinical indexes. No improvement in most cases and deterioration in a few subjects followed placebo administration, probably because the majority of the sample was made up of placebo non-reactors. Consequently, the activity data of the trial might be interpreted as expression of the pure pharmacologic activity of the tested drugs. Safety was very satisfactory: patients complained only rarely of trivial and clinically unimportant side effects; no variations in laboratory tests were noted.


Treatment of rheumatoid arthritis with flurbiprofen or ibuprofen. Flurbiprofen and ibuprofen were compared in a six-week double-blind randomized study in 208 patients with rheumatoid arthritis. Daily dosages were 120 mg flurbiprofen and 2400 mg ibuprofen for six weeks. Both drugs were effective in providing partial control of RA symptoms. Either or both drugs produced statistically significant improvement in mean values of time of onset of fatigue, grip strength and tender and swollen joint counts. All other standard endpoints of efficacy (except ESR) were improved but not at a statistically significant level. Slightly more than half of the patients improved during the trial. There was no statistically significant difference in the efficacy of the drugs. The incidence of side effects was low with both drugs. Most side effects were related to gastrointestinal tract irritation.


Rheoencephalographic and other studies of betahistine in humans. III. Improved methods of diagnosis and selection in arteriosclerotic dementia. Thirty-six institutionalized geriatric patients with particularly severe and longstanding dementia were selected by chart review and were studied neurologically. Criteria for clinical diagnosis are derived, and it is shown that arteriosclerosis had caused or contributed to their dementias in 33 patients (91.7%) and probably did so in two others (97.2%). Arteriosclerotic dementia may not be rare and may be much more prevalent than may now be considered the case. As 23 patients (63.9%) showed evidence of carotid and/or vertebrobasilar involvement, large-vessel disease also may be more prevalent in geriatric dementia than is now thought and should be considered in all such patients. The results further confirm that careful review of ward records without reference to psychiatric diagnoses of etiology is a simple, rapid, and reliable method of screening a large institutionalized population for possible arteriosclerotic dementia. The selectees should then be surveyed appropriately to confirm the presence of the disease. Since a careful review of the complete hospital records and a thorough neurologic examination proved sufficient, more definitive but more dangerous invasive diagnostic procedures may not be necessary for clinical pharmacologic and similar investigations.


Aerosolized terbutaline sulfate--an evalution of efficacy and side effects in patients with reversible airway disease. Aerosolized terbutaline sulfate at a dose of 0.50 mg has been shown to produce significant bronchodilation in patients with reversible airway disease. The purpose of this study was to evaluate the safety and efficacy of 0.50 mg terbutaline aerosol used on a regular dosage schedule over a six-week period. Sixteen ambulant patients with chronic obstructive pulmonary disease with a component of reversible airway disease were evaluated. The patients were tested at two-week intervals during a six-week period. The patients abstained from all bronchodilatory medications for at least 10 hours prior to the time of evaluation. The evaluation consisted of baseline pulmonary function tests, ECG, CBC, urinalysis, and renal and liver function tests. After the terbutaline was administered, a rhythm strip and pulmonary function tests were repeated at 5, 15, 30, 60, 120, and 180 minutes. Throughout the six-week study, there was a statistically significant increase in FEV1.0 and MMEFR (P less than 0.001): deltaFEV1.0 (ml) 740 (63%) 550 (45%) 340 (25%) 320 (25%) deltaMMEFR (liters/min) 42 (74%) 29 (46%) 28 (43%) 36 (42%). No abnormal laboratory results or paradoxical bronchospasm were noted during the study period; however, sympathomimetic side effects were observed. Aerosolized terbutaline sulfate (0.50 mg) when used on a regular schedule over a six-week period is effective in the treatment of reversible airway disease.


Controlled trial of nomifensin (HOE 984) and viloxazine in the treatment of depression in the elderly. The new antidepressant nomifensin (Hoechst 36984), a drug which showed less cardiotoxicity than tricyclics, was compared in a single-blind comparative trial with viloxazine. Forty elderly depressed patients were treated during four weeks. Nomifensin, N=21, 75 mg/day; viloxazine, N=19, 150 mg/day). Antidepressant effects were assessed by the Hamilton Rating Scale (HRS) for depression, and physical disability, by the Northwestern University Disability Scales. The HRS disclosed statistically significant improvement with both treatments. However, the scores in the nomifensin group fell distinctly form the second week on (P less than 0.01). There were no side effects reported spontaneously by the patients. Sinus tachycardia was recorded in two instances (one viloxazine and one nomifensin patient).


A Golgi study on the habenular nucleus of the cat. The habenular nucleus of kittens was studied using the rapid Golgi and the Golgi-Kopsch silver impregnation methods. The neurons of the medial habenular nucleus (MH) were classified into two types. The type I neurons, the main constituents of the MH, had piriform cell bodies with a mean diameter of 12 mum by 18 mum and two to five primary dendrites; dendrites had many spines. The type II neurons (14 X 23 mum) were fusiform in shape and one to three primary dendrites arose from each pole of the cell bodies; dendrites had few spines. The axons of both types of neurons were traced into the fasciculus retroflexus Meynerti (FR), and intranuclear axon-collaterals arose from axons of the type I neurons. The neurons of the lateral habenular nucleus (LH) were divided into four groups. Type I, II and III neurons were projection neurons of large, medium and small size, respectively. The type I neurons (27 X 43 mum) with four to seven primary dendrites were located mainly in the rostral and ventral areas of the LH. The type II neurons (15 X 33 mum) with two to four primary dendrites, the main constituents of the LH, were distributed throughout the LH. The type III neurons (15 X 25 mum) with two primary dendrites emerging from each pole of the soma were localized to the mediocaudal areas of the LH. The vast majority of axons of these projection neurons passed ventrally or ventrocaudally to enter the FR; only a few axons of these neurons were traced into the stria medullaris thalami (SM). The type IV neurons (12 X 25 mum) were small cells with short axons, suggesting the existence of a neural circuitry intrinsic to the LH. Bundle formation and glomerular arrangement of dendrites were observed in the medium-sized LH neurons. The afferent fibers terminating within the MH coursed in the most part of the SM. These afferents were classified into medium-caliber type I and fine type II fibers; both of these fibers emitted many intranuclear collaterals. There were also observed many fibers of passage which ran between the SM and FR, or between the habenular commissure (HC) and FR. The afferent fibers to the LH were divided into three groups; afferents via the HC, ascending and descending afferents. Most of the descending afferents entered the LH through the SM; some of them traversed the LH to join the FR or HC, or to extend to the pretectal region. The vast majority of the ascending afferents entered the LH via the FR; some of them extended rostrally to enter the SM. Some fibers in the HC also terminated within the LH. In addition to these afferents, many fibers of passage were seen to run through the LH and to bridge over between the SM and FR, or between the SM and HC.


Gastrointestinal blood loss study with a new analgesic compound: nefopam hydrochloride. A new analgesic compound, neopam HCl, was studied for possible gastrointestinal blood loss liabilities by Cr-51 red cell tagging. It was compared with aspirin in usual therapeutic doses of 1.8 Gm per day. Dosage of nefopam HCl was 180 mg per day (two tablets t.i.d). Twenty healthy male volunteers had fecal blood loss determined after one-week crossover periods of drug investigation, each preceded by one-week no-drug control periods. Results show a significant (P is less than 0.01) increase in occult bleeding in the aspirin-treated subjects from a mean of 0.5 ml to 1.63 ml per 24 hours, the mean increase for the group being 1.13 ml per 24 hours. Nefopam HCl treated subjects had an insignificant change from a control mean of 0.55 ml per 24 hours to 0.60 ml per 24 hours; a third of this group actually had a decrease in measurable blood loss. Reported side effects were minimal in both drug groups.


A double-blind comparison of meptazinol with placebo in postoperative pain. In a double-blind clinical trial, meptazinol (400-mg capsules) was shown to be significantly better than placebo in relieving postoperative pain in patients who had under gone total abdominal hysterectomy operations. Analgesic activity was assessed by patients rating their pain before and 1 hour after the administration of each treatment, by sequential analysis of patient and investigator preference for treatment, and by calculating the time interval between doses of the two treatments.


Rheoencephalographic and other studies of betahistine in humans. IV. Prolonged administration with improvement in arteriosclerotic dementia. The effects of prolonged betahistine administration were studied in institutionalized geriatric patients with particularly severe and long-standing arteriosclerotic dementia. Thirty received betahistine hydrochloride (SERC) or placebo orally in fixed dosage for six months on a double-blind basis and were followed by ward behavioral and psychometric ratings. Six others received active medication and were additionally followed by intracranial rheoencephalography (IREG). Thirty patients successfully completed the two studies. No adverse effects ascribable to the drug were encountered. The results show that betahistine caused definite, strong, and highly significant cerebral and scalp arterial vasodilation and circulatory improvement and that these caused equally definite, strong, and highly significant global improvement in the patients' dementias. Bethahistine, thus, acts in humans as a  potent and efficacious cerebral and peripheral microcirculatory and arterial vasodilator which can significantly improve cerebrovascular insufficiency and any associated dementia, no matter how severe either may be and despite the possible presence of large-vessel disease. These improvements sometimes were detected within two weeks or less, developed rapidly to maxima by about 90 days, and were sustained quite well there-after by continued therapy, They may, however, be drug dependent as they regressed in two patients following withdrawal. Various mechanisms whereby a microcirculatory vasodilator can cause arterial vasodilation are considered. The evidence apparently favors that prolonged microcirculatory vasodilatation can cause secondary passive arterial responses.  The possibility that the IREG contains external carotid contamination is again examined. Significant discrepancies between the scalp and IREG effects of bethahistine indicate, however, that the response of any such component was negligible within experimental error and that the component, itself, must have comprised but a small percentage of the IREG. Since the circulatory responses detected by the IREG thus arose essentially completely from the effects of betahistine on the cerebral circulation alone, this appears to be the first direct demonstration that cerebral circulatory improvement can cause improvement in mental function in patients with even severe, longstanding, and apparently clinically irreversible arteriosclerotic dementia and that such improvements can be effected pharmacologically.


Acid etch and enamel bond composite restoration of permanent anterior teeth affected by enamel hypoplasia. We have found that this procedure, using acid etching, can be very successful for providing an intermediate restoration of teeth disfigured by enamel hypoplasia. The technique is easy, relatively inexpensive for the patient, and quite effective for improving the appearance of the tooth.


Harmful effects of near-ultraviolet radiation used for polymerization of a sealant and a composite resin. An electronic device used to polymerize a sealant and a composite resin has been found to emit 365-nanometer radiation at levels sufficient to kill the bacterium Escherichia coli rapidly (greater than 14,000 ergs per square millimeter per second). Because of the absence of shielding on the probe, significant amounts of energy (up to 45% of that at the probe tip) were measured at the sides of the probe. These findings--supported by the well-documented findings of biological damage caused by near-ultraviolet radiation, including skin cancer, damage to the lens of the eye, and mutagenic effects--suggest that clinicians take appropriate precautions to avoid potential hazards to themselves and their patients.


Occlusal migration of a filled root: report of case. A mandibular premolar was treated by root canal therapy. The root migrated 11 mm in an occlusal direction beneath the anterior retainer of a fixed partial denture during eight years after treatment. Caries occurred at the junction of the root and the anterior retainer; and, as caries continued, the root apparently migrated to compensate for the lost tooth structure.


Antibody-dependent cellular cytotoxicity in asthmatics. Using a 51Cr-labeled, antibody-coated chicken red blood cell assay system, mononuclear cell and granulocyte preparations from infectious asthmatics, noinfectious asthmatics, and normal controls were tested for antibody-dependent cellular cytotoxicity (ADCC) capacity. The corrected cytotoxic indices of mononuclear cell preparations from infectious asthmatics were reduced (34 +/- 10 SD) as compared to noninfectious asthmatiics (47 +/- 7 SD) or normal controls (47 +/- 6 SD). Granulocyte preparations from infectious asthmatics and normal controls had a severity of the disease or in vivo drug treatment.


Bronchocentric granulomatosis: a complication of allergic bronchopulmonary aspergillosis. Hypersensitivity to the fungal antigens of Aspergillus fumigatus may result in a spectrum of immune injury collectively known as allergic bronchopulmonary aspergillosis (ABPA). This report describes a 14-yr-old boy who presented clinical findings consistent with ABPA,including a history of asthma, blood eosinophilia, serum precipitins, and IgE antibodies to Aspergillus fumigatus. Sputum Aspergillus, pulmonary infiltrates, and dual types I and III skin reactions to Aspergillus fumigatus were observed also. Pathology of the resected right upper lobe revealed severe bronchial destruction with the findings of bronchocentric granulomatosis. Noninvasive septate fungal hyphae compatible with Aspergillus were identified. Cultures from sputum and surgical specimens grew Aspergillus and Mycobacterium intracellulare avium. The PPD-B (purified protein derivative-Batty) intradermal skin test produced a 6 mm induration (PPD-S was negative). The patient's condition has been well controlled with prednisone and several antituberculous drugs. In addition, inflammatory and immunologic parameters have begun to return to normal. The relationship between ABa and the atypical mycobacterial infection is not clear. The association of ABPA with the severe bronchial destruction seen in bronchocentric granulomatosis is emphasized to alert physicans to this serious sequelae of ABa seen in the asthmatic.


Assessing bronchodilator responsiveness. This report concerns the difficulty in distinguishing variation due to drug effects from the spontaneous daily variation in baseline levels of pulmonary function in the evaluation of bronchodilator responsiveness in asthmatic children. Both prospective and retrospective studies were employed to assess the importance of the influence of the baseline level of function from which change is measured on the degree of responsiveness observed. It was found that for baseline levels up to approximately 90% of the predicted normal value, all patients had the capacity to respond significantly to a bronchodilator. However, there was no limit of baseline function below which a significant response did not occur.


Comparison of subcutaneous terbutaline with epinephrine in the treatment of asthma in children. Comparison of the bronchodilator and cardiovascular effects of subcutaneous terbutaline and epinephrine in the treatment of acute asthmatic attacks in children disclosed that bronchodilation occurred with 5 min after subcutaneous injection of either drug (0.01 mg/kg, maximum 0.25 mg). Bronchodilation was maintained for 4 hr after administration of either drug. Small increases in pulse rate followed administration of terbutaline but not epinephrine. No clinically significant side effects were noted with either drug.


Histologic responses in human skin test reactions to ragweed. IV. Effects of a single intravenous injection of steroids. A controlled study has been carried out dealing with the early effects of a single intravenous dose of either methylprednisolone or placebo or newly developed and ongoing cellular inflammatory responses in immediate hypersensitivity skin test reactions. There was no significant difference in the tissue eosinophil responses to ragweed injected 2 hr before and 2 hr after placebo; there was a significant rise (101% +/- 39) from the second to fourth hour after antigen injection. By contrast, there was a marked decrease in the tissue eosinophil response to antigen injected 2 hr after steroids as compared to the pattern seen in the presteroid reaction. In addition, the eosinophil numbers not only did not increase from the second to fourth hour when steroids were injected at the second hour but decreased markedly. These findings suggest early suppressive effects on tissue eosinophil responses within 2 hr after steroid were administered intravenously. Also, there may be trafficking of eosinophils both into and out of these inflammatory sites during the first hours after intradermal antigen injection.


Severe persistent biphasic local (immediate and late) skin reactions to insulin. A patient with adult-onset insulin-requirging diabetes mellitus had persistent severe local reactions to all available insulins of animal origin. Skin reactions were biphasic in nature with both immediate and late characteristics. An extensive immunologic investigation of this problem was undertaken, revealing evidence of reaginic antibody involvement in the reactions. Routine histologic studies suggested the possibility that Arthus-type mechanisms played a part, although this impression was not confirmed by immunofluorescent microscopy. A program of medical management provided some relief of symptoms.


Clinical investigation of fenoterol, a new bronchodilator, in asthma. In a randomized, double-blind, noncrossover, study, 28 asthmatic patients received either ephedrine or fenoterol (Th1165a) a new beta 2-sympathomimetic bronchodilator, for 90 days. Forced expiratory flows and volumes and plethysmography were performed after a single test dose on days 1, 45, and 90. The subjects' clinical course was followed for patient acceptance and side effects. Compared to ephedrine, Th1165a was demonstrated to be a potent bronchodilator with a prolonged duration of action. Adrenergic side effects were minimal, and no tolerance or toxicity was demonstrated.


Precautions in the application of immunohistochemical techniques to tissues of the pig hypothalamo-neurohypophysial system. Immunohistochemical procedures were used to localize neurophysin in the hypothalamo-neurohypophysial axis of the domestic pig. The topographical distribution of neurophysin as revealed by the immunofluorescence "sandwich" technique was similar to that found when either the immunoglobulin-peroxidase bridge method or the peroxidase-labeled gamma-globulin technique was employed. However, application of the peroxidase-anti-peroxidase (PAP) complex procedure resulted in nonspecific staining of the magnocellular structures. This phenomenon was attributed to the action of PAP on the tissue and after screening a number of other vertebrate species was found to be unique to the pig. Minimal nonspecific binding of the PAP could be achieved either by reducing the reaction time of PAP to 5 min or, by the addition of 1% (v/v) normal serum to all reagents and wash solution. That the PAP-binding protein is a component of the hypothalamo-neurohypophysial axons is discussed.


The low prevalence of Clostridium botulinum in the lakes, marshes and waterways of the Camargue. Mud samples collected in June 1975 from the lakes, marshes and waterways of the Camargue were examined for Clostridium botulinum. The Grand Rhône and Petit Rhône were shown to contain types B and E, but of 44 samples taken from well distributed sites on the Ile de la Carmargue, only two (4-5%) were positive and these contained type E alone. The survey indicated a much lower prevalence of Cl. botulinum than any encountered in recent surveys of inland aquatic environments elsewhere.


A comparison of the distribution of Clostridium botulinum in soil and in lake mud. In 1975, 25 soil samples were collected from the London area. Of these, 20 were obtained 200-300 yards from 20 lakes that had been shown in 1974 to contain mud contaminated with one or more of types of B, C, D, and E of clostridium botulinum. By means of a technique comparable with that use for the examination of mud, the 20 soil samples were found negative. The remaining 5 soil samples, obtained from sites that were not in close proximity to lakes, were also negative except for one that contained type B.


Incidence of salmonella infection in animals in England and Wales 1968--1974. Of a total of 23,609 incidents of salmonella infection reported during the period 1968--74, 20,326 occurred in cattle, 1744 in poultry and other birds, 675 in sheep, 558 in pigs and 306 in other species of animals. Despite the large number of serotypes isolated (153), 88% of incidents were due to only two stereotypes: S. dublin (15,929 incidents of which 15,446 occurred in cattle ) and S. typhimurium (4842 incidents of which 3785 occurred in cattle and 732 in birds). S. choleraesuis was the third (314 incidents which with only 5 exceptions occurred in pigs) and S. abortusovis (243 incidents all in sheep) was the fourth most commonly isolated serotype. The following six serotypes were each recorded in more than 100 incidents: S. newport (177), S. agona (170), S. virchow (169), S. anatum (152), S. enteritidis (150) and S. montevideo (111). The other 143 serotypes accounted for only 5-8% of total incidents. The trends of annual incidence of salmonella infection in cattle, sheep, pigs, poultry and other birds during the 17-year period 1958-74 and the distribution of the main serotypes in the individual species of animals from 1968 to 1974 are discussed.


Bacteriostasis of Escherichia coli by milk. I. Colonization of breast-fed infants by milk resistant organisms. Human milk has a bacteriostatic effect on Escherichia coli in vitro. The milks of 40 mothers were tested for this effect against E. coli isolated from their stools, from those of their own babies, and from those of babies not breast-fed. The milks had a direct bacteriostatic effect, not dependent on complement, on some but not all the strains of E. coli. Breast-fed babies receiving supplementary bottle feeds were colonized with milk-resistant strains, whereas bottle-fed babies and, surprisingly, babies completely breast-fed were colonized equally with milk-sensitive and milk-resistant strains, as were the mothers. These results suggest that the bacteriostatic effect of human milk, demonstrable in vitro does sometimes operate in vivo. The antibacterial activity of human milk is not infleunced by the O, H, or K antigens of E. coli and is effective against other Gram-negative organisms, e.g. Salmonella, Klebsiella, Proteus.


Escherichia coli serotypes in the faeces of healthy adults over a period of several months. The faeces of nine subjects eating mainly at home were collected at regular intervals over periods ranging from 2--5 months. Although a large number of serotypes of E. coli were isolated, the variety per subject was lower than is usually found. In most subjects only a limited number of serotypes persisted over most of the periods of study while many serotypes were only isolated on single occasions.


Phagocytosis measured as inhibition of uridine uptake by Candida albicans. Inhibition of 3H-uridine incorporation into Candida albicans can be used as a sensitive index of phagocytic function because: 1) there is a linear correlation between uridine incorporation and yeast number; 2) phagocytic cells do not incorporate significant amounts of uridine in short term cultures; and 3) C. albicans replicating inside phagocytic cells does not take up uridine from culture medium. Appropriate conditions for measuring phagocytic capacity of human polymorphonuclear cells (PMN's) were 5 x 10(5) C. albicans and 5 x 10(4) PMNs in 0.5 ml of medium containing 2.5% AB serum. This mixture was incubated for 30 min at 37 degrees C. Aliquots were then transferred into microtiter wells  and incubated for a further 60 min in the presence of 3H-uridine. Under these conditions PMN leucocytes from 25 healthy individuals caused suppression of uridine incorporation ranging from 33 to 75% (50 +/- 12).


Effects of preincubating human peripheral blood lymphocytes on their ability to bind sheep red blood cells. The effect of preincubating human peripheral lymphocytes at 37 degrees C or 4 degrees C various lengths of time on their subsequent ability to form rosettes with sheep red blood cells (SRBC) was investigated. Lymphocytes suspended in either medium or medium containing fetal calf serum (FCS) and preincubated at 37 degrees C for 30 min exhibited marked decrease in rosette formation with a return to normal values by 120 min. However, neither lymphocytes suspended in medium and preincubated at 4 degrees C nor lymphocyte suspended in medium containing normal human serum (HS) and preincubated at 37 degrees C exhibited this phenomenon.


Separation of epidermal layers of the newborn rat. A method is presented for the separation of epidermal strata by the successive elimination of either the basal or basal and spinous cells with 0.24 M NH4Cl at pH 9.5. Histologic evidence suggests that the residual epidermal strata obtained after incubation of the skin with NH4Cl are reproducible; hence, this technique circumvents loss of granular layer histidine-rich protein inherent with trypsin separation and provides an effective procedure for biochemical analysis of arginine-rich and lysine-rich proteins in the various differentiating epidermal cells.


A new technique for cross sectioning free hairs. A new method of sectioning hair specimens has been devised that employs Araldite as an embedding medium. This has greater rigidity than paraffin and eliminates the possibility of compression on sectioning with resultant distortion of the hair shaft. Semithin sections could be sectioned at the perpendicular and examined with phase contrast optics. By correlating the cross section with the longitudinal position of the hair shaft within this translucent medium, exact cross sections of various hair types could be obtained.


Ex vivo hemodialysis culture of microbial and mammalian cells. A small hemodialysis culture unit was developed which can be attached to an arterial-venous shunt and worn by an animal for days. The unit consists of a blood channel separated by a membrane from a dialysate chamber in which microbial or mammalian cells can be cultured. Bacterial multiplication proceeded first exponentially at the maximal rate and then arithmetically at a lesser, dialysis-limited rate. In a survey of 16 pathogenic microorganisms and five types of mammalian cell, results indicated that most of the aerobes grew well, but none of the obligate anaerobes grew at all. The separation of the culture from cellular and macromolecular host defense mechanisms allowed the cultivation of parasitic cells on an animal that was not naturally susceptible.


Patterns of exogenous and endogenous hematopoietic repopulation following radiation injury. Patterns of hematopoietic recovery in mice given an LD50/30 radiation exposure were different from those in animals given an LD100/30 exposure and a marrow transplant sufficient to effect 50 per cent 30-day survival. Death occurred later, and recovery of some peripheral blood elements was delayed in the LD50/30 mice. Over the period of study, splenic hematopoiesis and stem cell renewal were insignificant in the LD50/30 mice yet were active in the marrow-transplanted animals. The slow recovery of erythropoiesis in the LD30/30 mice is explained in part by the lack of early splenic hematopoiesis. This effect appeared to be attributable to the splenic microenvironment which, although being suitable for growth of unirradiated hematopoietic cells, was not conducive to repair of radiated splenic hematopoietic cells.


Detection of antilymphocyte antibodies in patients with systemic lupus erythematosus by indirect immunofluorescence on acetone-fixed lymphocytes. Antilymphocyte antibodies in normal and SLE sera were found to react with lymphocytes fixed in acetone at -30 degrees C. for 10 minutes. At a dilution of 1:40, 3.7 per cent of normal and 90.9 per cent of SLE sera were positive with the use of indirect immunofluorescence with a pepsin-digested, FITC-labeled anti-Fab serum. The reaction was independent of temperature between 4 degrees and 37 degrees C. Three patterns of staining were seen in the lymphocyte surface: reticular, ring, and globular. The ring pattern appeared to correlate with IgM, and the reticular and globular patterns with IgG antilymphocyte antibodies. Absorption with lymphocytes, insolubilized extracts of rat liver and kidney, and human brain revealed that antilymphocyte and ANA activity could be independently removed. Variation in reactivity with cells from different normal donors was similar to that seen with the microcytotoxicity test. Acetone-fixed lymphocytes appear to be a much more sensitive target than viable cells in suspencion in IIF tests for antilymphocyte antibodies. The IIF test with fixed cells also appears slightly more sensitive than cytotoxicity testing and has the advantage of allowing storage of target cells.


Antitumor antibodies in human breast cancer sera as detected by fixed cell immunofluorescence and living cell membrane immunofluorescence assays. The reactions of patients' sera with cultured human breast cancer cells were studied by fixed cell immunofluorescence  (FCF) and living cell membrane immunofluorescence (MF) tests. The results suggested that the FCF reaction detected antibodies that were cell or organ specific, whereas the MF test was more indicative of an antineoplastic immune response.


Biological activities of tritiated endotoxins: correlation of the Limulus lysate assay with rabbit pyrogen and complement-activation assays for endotoxin. Tritiated endotoxins were prepared by three different methods. The biological activities of the tritiated endotoxins were determined by the Limulus amebocyte lysate assay, a rabbit pyrogen assay, and a complement-activation assay and were compared to native, unlabeled endotoxin. All three tritiated endotoxin preparations manifested adequate biological activity in each of the three assay systems, and all three assays ranked the biological activity of the different endotoxin preparations in the same order. Endotoxin tritiated by the Wilzbach procedure retained most of its biological activity and also had the highest specific radioactivity. The good correlation between the Limulus lysate, rabbit pyrogen, and complement-activation assays suggests that the same active site of the endotoxin molecule is identified by the three different assays.


The platelet factor 3 immunoinjury technique re-evaluated. Development of a rapid test for antiplatelet antibody. Detection in various clinical disorders, including immunologic drug-induced and neonatal thrombocytopenias. A modified version of the platelet factor 3 immunoinjury technique for the detection of antiplatelet antibody is described in detail; heat-inactivated sera and frozen platelets are employed. The modification allows the results of the test to be available within 4 to 5 hours after drawing the blood sample, in contrast to the 3 days required in the original method. The technical work involved is significantly reduced. The reproducibility of the technique is improved. Results obtained with the two tests correlate well. Data accumulated in 458 cases of clinical disorders associated with antiplatelet antibody are reported. Drug-dependent antiplatelet antibody was demonstrated in 26 cases of thrombocytopenia associated with 19 different drugs. Data are also presented for 23 cases of neonatal thrombocytopenia.


The treatment of acromegaly by Yttrium implantation. Yttrium implantation is one of the many ways of treating acromegaly. The advantages are the minor nature of the procedure and the fact that pituitary replacement is not as commonly required as after hypophysectomy. Thus in young female patients menstruation may be resumed following treatment and pregnancy has occurred. The procedure is not as free from complications as external irradiation but the response is more satisfactory.


Hypothalamic, pituitary and testicular function during sexual maturation of the male rat. Hypothalamic content of gonadotrophin-releasing hormone (GnRH), serum LH and FSH, capacity of the testis to synthesize testosterone in vitro, and testicular 5-ene-3 beta-hydroxysteroid dehydrogenase-isomerase and 17 beta-hydroxysteroid dehydrogenase were measured in groups of rats at approximately 5 day intervals from birth to day 64 and at days 74 and 89. The capacity of the testes to synthesize testosterone in vitro was measured in the presence of a saturating dose of rat LH. Gonadotrophin-releasing hormone increased steadily from 0-17 ng per hypothalamus at birth to a maximum of 7 ng at day 52 and then remained constant. LH concentrations were highly variable and often exceeded adult values between days 10 and 32. After day 32 a steady rise was observed which reached adult values between days 37 and 42. FSH concentrations markedly increased from 255 ng/ml observed at birth and day 10 to a peak value of 1000 ng/ml at day 32. Subsequently there was a steady decline in FSH values until day 74 when the concentration returned to values found at birth. 5-ene-3 beta-Hydroxysteroid dehydrogenase-isomerase activity exhibited a rapid increase between days 12 and 19 followed by an even greater rate of increase between days 19 and 32 when adult levels were attained. 17 beta-Hydroxysteroid dehydrogenase activity was very low between birth and day 22. Enzyme activity began to increase at day 22 with a rapid increase in activity observed between days 37 and 58. The increase in capacity to synthesize testosterone closely followed the increase in 17 beta-hydroxysteroid dehydrogenase activity. The study demonstrates that during sexual maturation in the male rat, changes in serum LH and FSH do not reflect changes in hypothalamic GnRH. The appearance of Leydig cells as monitored by 5-ene-3 beta-hydroxysteroid dehydrogenase-isomerase activity precedes by approximately 20 days the increase in testicular capacity to synthesize testosterone in vitro. The latter coincides with the increase in 17 beta-hydroxysteroid dehydrogenase activity. These results suggest that 17 beta-hydroxysteroid dehydrogenase is a limiting factor in the ability of the testis to respond to LH stimulation.


Luteinizing hormone release after two injections of synthetic luteinizing hormone releasing hormone in the ewe. Anoestrous ewes were given two injections of 30 mug synthetic luteinizing hormone releasing hormone (LH-RH) separated by one of the following intervals: 1-5, 3, 6, 12 or 24 h. The first injection caused an increase in the plasma LH concentration in each animal. The response to the second injection was dependent on the interval between the injections. When the second injection was administered 1-5 h after the first it caused a further increase in the LH concentration to maximal levels which were significantly greater than those induced in the other anoestrous groups. When the second injection was administered 3 h after the first, there was no significant difference between the responses to the two injections although the time to reach the maximal LH concentration was shorter and the height of the LH peak was greater in each animal following the second injection. When the second injection was administered 6, 12 or 24 h after the first, the LH response was significantly less, in terms of height and area of the induced peak, than following the first injection. The LH response to the second injection was particularly low in the 12 and 24 h groups. Two injections of 30 mug synthetic LH-RH were also administered at 1-5 h intervals to ewes on either day 10 of the oestrous cycle or at onset of oestrus. The pattern of LH responses in all these animals was similar to that observed in anoestrous ewes injected at 1-5 h intervals. The total LH release, as assessed in terms of the induced peaks, was significantly greater in the onset of oestrus group than in the day 10 group or any of the anoestrous groups. Presumably the sensitization-desensitization sequence of the pituitary gland to LH-RH which has been demonstrated, together with the effects of sex steroid hormones, must play an important part in the development and decay of the natural preovulatory LH peak.


The nature of the proteins in 'chloramphenicol particles' from Escherichia coli A19 (Hfr rel met rns). The unusual particles which accumulate in cell-free extracts from Escherichia coli A19 during chloramphenicol inhibition ('chloramphenicol particles') have been isolated by large-scale rate-zonal density gradient ultracentrifugation. The proteins and RNA species composing these particles have been examined. The rRNA species present are precursor and mature forms of 16S and 23S rRNA which accumulate during inhibition. The proteins prepared directly from the particles give strong multiple immunoprecipitates with antisera specific to 30S and 50S ribosomal proteins. The soluble proteins of the cell prepared in the same manner do not give this immunological reaction. Two-dimensional electrophoresis patterns of the proteins from the 'chloramphenicol particles' strongly resemble those for 30S and 50S ribosomal proteins, i.e. they are predominantly basic low molecular weight proteins, and are dissimilar to the patterns for the soluble proteins of the cell. It is concluded that the 'chloramphenicol particles' are a heterogeneous group of ribonucleoproteins comprising the bulk of the rRNA accumulating during inhibition in association with variable amounts of some of their corresponding ribosomal proteins. The particles are therefore not artefacts of preparation, as previously thought, but arrested ribosome precursors.


Excystment of axenically prepared cysts of Hartmanella culbertsoni. Axenically prepared cysts of Hartmannella culbertsoni readily excysted in the presence of heat stable factors prepared from Escherichia coli, Klebsiella aerogenes, Staphylococcus aureus, Sarcina lutea, Bacillus subtilis, Bacillus megaterium and several fungi. Peptone, proteose peptone, tryptone or amino acids also promoted excystment. Crowding of the cysts and dilution of bacterial extracts adversely affected the excystment. Continual presence of the factors in the medium was essential for excystment.


Microbial metabolism of amino alcohols. Control of formation and stability of partially purified ethanolamine ammonia-lyase in Escherichia coli. Induction of ethanolamine ammonia-lyase formation in Escherichia coli required both the ethanolamine and vitamin B12, and was gratuitous during growth on glycerol. Ethanolamine analogues inhibited enzyme activity and inhibited growth with ethanolamine as the the nitrogen source, but did not act as inducers. Enzyme formation was more rapid when ethanolamine was added to cultures containing vitamin B12 rather than the reverse. Enzyme formation was subject to catabolic repression, glucose and acetate being particularly effective. Chloramphenicol, I-aminopropan 2-01 and 1,3-diaminopropan-2-01 prevented enzyme induction. Ethanolamine ammonia-lyase, resolved from its cobamide coenzyme, was purified 35-fold. The apoenzyme was stable for several days in the presence of ethanolamine, dithiothreitol, glycerol and K+ ions. Enzyme formation therefore requires both substrate and cobamide coenzyme to be present simultaneously as inducers.


The nature of the proteins present in the 'relaxed particles' from methionine-starved Escherichia coli A19 (Hfr rel met rns). The 'relaxed particles' formed during methionine starvation of Escherichia coli A19 (Hfr rel met rns) have been isolated by large-scale rate-zonal density gradient ultracentrifugation. The proteins and rRNA species associated with these particles have been examined. The rRNA species present are precursor and mature forms of 16S and 23S rRNA. The bulk of the rRNA which accumulates during starvation is found within the particles. The proteins prepared directly from the particles give strong multiple immunoprecipitates with antisera specific to 30S and 50S ribosomal proteins. The soluble proteins, prepared and examined in the same manner, do not give this immunological reaction. Two-dimensional electrophoresis patterns of the proteins from the particles show that the proteins co-migrate with proteins from 30S and 50S ribosomes and are entirely dissimilar to the proteins prepared by the same methods from the soluble fraction of the cells. On the basis of these and other observations, it is concluded that the 'relaxed particles' are not artefacts but are arrested ribosome precursors containing both rRNA and certain ribosomal proteins. The free pool of ribosomal proteins is low in exponential-phase cells and is not significantly increased by a 2 h period of starvation for glucose. The implications of these observations concerning the proteins associated with 'relaxed' and 'chloramphenicol particles' are discussed in raltion to ribosome biogenesis and the stabilization of rRNA.


Fluctuations in buoyant density during the cell cycle of Escherichia coli K12: significance for the preparation of synchronous cultures by age selection. The buoyant densities of Escherichia coli K12 were investigated by isopycnic centrifugation in gradients of colloidal silica (Ludox) and polyvinylpyrrolidone. Bacteria from an exponential culture in a defined medium supplemented with hydrolysed casein banded at densities between 1-060 and 1-115 g ml-1; the mean density was 1-081 g ml-1. At the higher densities, two populations of cells were present: smaller cells were approximately twice as numerous as, and half the modal volume of, the population of larger cells. A homogeneous population of cells of intermediate volume equilibrated in the least dense region of the density band. Synchronous cultures were established by inoculating cells selected from the most or least dense regions of the band into spent growth medium. The results are consistent with a fluctuation between maximal density at cell birth and division, and minimal density near the middle of the cell cycle. In synchronous cultures prepared by continuous-flow age selection, the first division occurred after a period that was significantly shorter than the length of subsequent cell cycles. Cells selected by this procedure were of similar mean density to those in the exponential culture but were more homogeneous with respect to size. The possibility that the smallest (and densest) cells in an exponential culture are retained in the rotor, and are thus excluded from the synchronous culture, is discussed.


Synthesis of ribosomal and transfer ribonucleic acids in yeast during a nutritional shift-up. The growth rate of Saccharomyces cerevisiae was increased by adding a mixture of amino acids to cultures containing proline as the sole nitrogen source. The transition from balanced growth in the basal medium (doubling time 4 h) to balanced growth in the enriched medium (doubling time 2 h) took about 2-5 h. The rate of RNA accumulation increased soon after the enrichment to almost its final value. This increase began after a short lag of 10 to 15 min, therefore synthesis of new RNA polymerase molecules may be required before stable RNA production can increase. The different stable RNA species were not stimulated at different times after the enrichment, but all increased continuosly throughout the transition. The rRNA species accumulated in a co-ordinate fashion at a rate faster than the rate of tRNA accumulation.


Aerobic and anaerobic bacterial respiration monitored by electrodes. A technique is described by which both oxygen and nitrate (or nitrate or chlorate) levels were continuously monitored during bacterial respiration. Paracoccus (Micrococcus) denitrificans and Escherichia coli oxidizing succinate rapidly ceased to reduce nitrate when oxygen was available, and equally rapidly commenced nitrate reduction when all the oxygen had been consumed. By contrast, membrane vesicles isolated from P. denitrificans reduced oxygen and nitrate simultaneously. The respiratory nitrate reductase in intact cells of P. denitrificans appeared to be inacessible to chlorate present in the reaction medium, and it is suggested that the nitrate reductase is orientated on the plasma membrane so that nitrate gains access from the inner (cytosolic) face.


Introduction of bacteriophage Mu into Pseudomonas solanacearum and Rhizobium meliloti using the R factor RP4. Phage Mu-1 and a thermoinducible derivative, Mu-1 cts 62 were inserted into the broad host range R factor RP4. These hybrid plasmids were transferred by conjugation to a phytopathogenic bacterium Pseudomonas solanacearum GMI 1000 and a legume-root nodule bacterium Rhizobium meliloti 2011. The Mu genome is transcribed and tranlated in these new hosts: P. solanacearum (RP4:Mu cts) cultures have a spontaneous production of about 5 X 10(5) plaque-forming units ml-1 which is similar to the frequency of spontaneous Mu production in E. coli; the Mu production of R. meliloti is lower (about 10(2) plaque-forming units ml-1).


Taxonomy of the genus Serratia.

Taxonomy of the genus Serratia. One hundred and fifty-six strains of Serratia and related bacteria including representatives of Enterobacter liquefaciens, Enterobacter cloacae, Enterobacter aerogenes, Erwinia carotovora, Erwinia chrysanthemi, Erwinia herbicola and Erwinia nimipressuralis were studied using 223 morphological, physiological, biochemical and carbon source utilization tests. The results were subjected to computer analysis. At the 80% similarity level all strains, except two, grouped into eight phenons representing: (A) Serratia marcescens with the neotype CCM303 (ATCCI3880); (B) S. marinorubra with the monotype NCTC10912 (ATCC27614); (CI) S. liquefaciens with the type ATCCI4460; (C2) S. plymuthica with the monotype CCM640 (ATCC183); (D) Erwinia herbicola with the neotype of Enterobacter agglomerans NCTC9381; (E) Enterobacter cloacae with the neotype NCTCI0005 and Erwinia nimipressuralis; (F) Erwinia carotovora with the type ATCC495, Erwinia atroseptica and Erwinia chrysanthemi; (G) Klebsiella mobilis with the neotype NCTCI0006. At the 70% similarity level the phenons formed two groups: (A, B, CI, C2) and (D, E, F, G). The following conclusions were drawn. (I) There are three species of enterobacteria producing prodigiosin: S. marcescens, S. plymuthica and S. marinorubra. (2) There are four species of Serratia, one colourless (S. liquefaciens). (3) Subphenons (biovars) are described within the four species of Serratia. (4) Non-pigmented wild-type strains of S. marcescens can generally be differentiated from pigmented strains by characters other than pigmentation, because subphenons are homogeneous with respect to pigmentation. This survey raised some problems of nomenclature because old descriptions could be found that could loosely fit the present phenons. Comparison with an authentic culture was considered to be the most objective way of identifying these phenons with earlier named species.


Naloxone-induced reversal of schizophrenic hallucinations. In a single-blind pilot study 0.4 mg naloxone i.v. was found temporarily to reduce or abolish auditory hallucinations in four cases of chronic schizophrenia whereas saline was without effect. In one of these patients there was a similar reversal also of her visual hallucinations. Two additional cases who denied hearing voices before the injections, reported no subjective effects.


Effect of positive end expiratory pressure ventilation on intracranial pressure in man. THsi study was designed to define the effect of positive end expiratory pressure (PEEP) ventilation on intracranial pressure (ICP). In 25 patients with severe head trauma with and without associated pulmonary injury the following parameters were simultaneously monitored under mechanical ventilation with and without PEEP:ICP, arterial blood pressure, central venous pressure, arterial blood gases, and cardiac rate. In addition, the volume-pressure response (VPR) was evaluted in each patient to assess cerebral elastance. The results indicate a significant increase in ICP with the application of PEEP only in the 12 patients who manifested increased cerebral elastance by VPR. Half of this latter group manifested impairment of cerebral perfusion pressure to levels less than 60 mm Hg. Return to baseline CIP levels was observed with termination of PEEP. No significantly consistent changes in other parameters were noted.


Correction of the short-face syndrome-vertical maxillary deficiency: a preliminary report. Careful clinical studies and cephalometric and model analyses are required in the planning for treatment of vertical maxillary deficiency. The maxilla is repositioned inferiorly to elongate the face and to achieve facial balance. Simultaneous Le Fort I osteotomy with bone grafting is a helpful adjunct in the treatment of vertical maxillary deficiency in either dentulous or edentulous patients.


Oral surgery program university of california, san francisco. The two cases presented here demonstrate a successful therapeutic modality with resection followed by immediate reconstruction with bone grafts. At least one of the cases presented seems to indicate that the tumor is relatively less aggressive than the simple ameloblastoma. The histopathology of the two cases suggests that this odontogenic tumor has a wider histologic variation than originally described by Pindborg.