bluuebunny/crossref_metadata_2025 huggingface dataset:
Oriented Immobilization of Bacteriophages for Biosensor Applications
Authors: Tolba M. , Minikh O. , Brovko L. Y. , Evoy S. , Griffiths M. W.
Published: December 2009

A method was developed for oriented immobilization of bacteriophage T4 through introduction of specific binding ligands into the phage head using a phage display technique. Fusion of the biotin carboxyl carrier protein gene (
bccp
) or the cellulose binding module gene (
cbm
) with the small outer capsid protein gene (
soc
) of T4 resulted in expression of the respective ligand on the phage head. Recombinant bacteriophages were characterized in terms of infectivity. It was shown that both recombinant phages retain their lytic activity and host range. However, phage head modification resulted in a decreased burst size and an increased latent period. The efficiency of bacteriophage immobilization with streptavidin-coated magnetic beads and cellulose-based materials was investigated. It was shown that recombinant bacteriophages form specific and strong bonds with their respective solid support and are able to specifically capture and infect the host bacterium. Thus, the use of immobilized BCCP-T4 bacteriophage for an
Escherichia coli
B assay using a phage multiplication approach and real-time PCR allowed detection of as few as 800 cells within 2 h.
Septins AspA and AspC Are Important for Normal Development and Limit the Emergence of New Growth Foci in the Multicellular Fungus
Aspergillus nidulans
Authors: Lindsey Rebecca , Cowden Susan , Hernández-Rodríguez Yainitza , Momany Michelle
Published: December 2009

Septins are cytoskeletal proteins found in fungi, animals, and microsporidia, where they form multiseptin complexes that act as scaffolds recruiting and organizing other proteins to ensure normal cell division and development. Here we characterize the septins AspA and AspC in the multicellular, filamentous fungus
A
spergillus nidulans
. Mutants with deletions of
aspA
,
aspC
, or both
aspA
and
aspC
show early and increased germ tube and branch emergence, abnormal septation, and disorganized conidiophores. Strains in which the native
aspA
has been replaced with a single copy of
aspA-GFP
driven by the native septin promoter or in which
aspC
has been replaced with a single copy of
aspC-GFP
driven by the native promoter show wild-type phenotypes. AspA-GFP and AspC-GFP show identical localization patterns as discrete spots or bars in dormant and expanding conidia, as rings at forming septa and at the bases of emerging germ tubes and branches, and as punctate spots and filaments in the cytoplasm and at the cell cortex. In conidiophores, AspA-GFP and AspC-GFP localize as diffuse bands or rings at the bases of emerging layers and conidial chains and as discrete spots or bars in newly formed conidia. AspA-GFP forms abnormal structures in
ΔaspC
strains while AspC-GFP does not localize in
ΔaspA
strains. Our results suggest that AspA and AspC interact with each other and are important for normal development, especially for preventing the inappropriate emergence of germ tubes and branches. This is the first report of a septin limiting the emergence of new growth foci in any organism.
Evidence for Mucin-Like Glycoproteins That Tether Sporozoites of
Cryptosporidium parvum
to the Inner Surface of the Oocyst Wall
Authors: Chatterjee Anirban , Banerjee Sulagna , Steffen Martin , O'Connor Roberta M. , Ward Honorine D. , Robbins Phillips W. , Samuelson John
Published: December 2009

Cryptosporidium parvum
oocysts, which are spread by the fecal-oral route, have a single, multilayered wall that surrounds four sporozoites, the invasive form. The
C
.
parvum
oocyst wall is labeled by the
Maclura pomifera
agglutinin (MPA), which binds GalNAc, and the
C
.
parvum
wall contains at least two unique proteins (

C

ryptosporidium
o
ocyst
w
all protein 1 [COWP1] and COWP8) identified by monoclonal antibodies.
C
.
parvum
sporozoites have on their surface multiple mucin-like glycoproteins with Ser- and Thr-rich repeats (e.g., gp40 and gp900). Here we used ruthenium red staining and electron microscopy to demonstrate fibrils, which appear to attach or tether sporozoites to the inner surface of the
C
.
parvum
oocyst wall. When disconnected from the sporozoites, some of these fibrillar tethers appear to collapse into globules on the inner surface of oocyst walls. The most abundant proteins of purified oocyst walls, which are missing the tethers and outer veil, were COWP1, COWP6, and COWP8, while COWP2, COWP3, and COWP4 were present in trace amounts. In contrast, MPA affinity-purified glycoproteins from
C
.
parvum
oocysts, which are composed of walls and sporozoites, included previously identified mucin-like glycoproteins, a GalNAc-binding lectin, a Ser protease inhibitor, and several novel glycoproteins (
C
.
parvum
MPA affinity-purified glycoprotein 1 [CpMPA1] to CpMPA4). By immunoelectron microscopy (immuno-EM), we localized mucin-like glycoproteins (gp40 and gp900) to the ruthenium red-stained fibrils on the inner surface wall of oocysts, while antibodies to the
O
-linked GalNAc on glycoproteins were localized to the globules. These results suggest that mucin-like glycoproteins, which are associated with the sporozoite surface, may contribute to fibrils and/or globules that tether sporozoites to the inner surface of oocyst walls.
TheStreptococcuspneumoniaeCapsule Inhibits Complement Activity and Neutrophil Phagocytosis by Multiple Mechanisms
Authors: Hyams Catherine , Camberlein Emilie , Cohen Jonathan M. , Bax Katie , Brown Jeremy S.
Published: December 2009

TheStreptococcuspneumoniaecapsule is vital for virulence and may inhibit complement activity and phagocytosis. However, there are only limited data on the mechanisms by which the capsule affects complement and the consequences forS. pneumoniaeinteractions with phagocytes. Using unencapsulated serotype 2 and 4S. pneumoniaemutants, we have confirmed that the capsule has several effects on complement activity. The capsule impaired bacterial opsonization with C3b/iC3b by both the alternative and classical complement pathways and also inhibited conversion of C3b bound to the bacterial surface to iC3b. There was increased binding of the classical pathway mediators immunoglobulin G (IgG) and C-reactive protein (CRP) to unencapsulatedS. pneumoniae, indicating that the capsule could inhibit classical pathway complement activity by masking antibody recognition of subcapsular antigens, as well as by inhibiting CRP binding. Cleavage of serum IgG by the enzyme IdeS reduced C3b/iC3b deposition on all of the strains, but there were still marked increases in C3b/iC3b deposition on unencapsulated TIGR4 and D39 strains compared to encapsulated strains, suggesting that the capsule inhibits both IgG-mediated and IgG-independent complement activity againstS. pneumoniae. Unencapsulated strains were more susceptible to neutrophil phagocytosis after incubation in normal serum, normal serum treated with IdeS, complement-deficient serum, and complement-deficient serum treated with IdeS or in buffer alone, suggesting that the capsule inhibits phagocytosis mediated by Fcγ receptors, complement receptors, and nonopsonic receptors. Overall, these data show that theS. pneumoniaecapsule affects multiple aspects of complement- and neutrophil-mediated immunity, resulting in a profound inhibition of opsonophagocytosis.
Streptococcus pneumoniae
Resistance to Complement-Mediated Immunity Is Dependent on the Capsular Serotype
Authors: Hyams Catherine , Yuste Jose , Bax Katie , Camberlein Emilie , Weiser Jeffrey N. , Brown Jeremy S.
Published: December 2009

Streptococcus pneumoniae
strains vary considerably in the ability to cause invasive disease in humans, and this is partially associated with the capsular serotype. The
S. pneumoniae
capsule inhibits complement- and phagocyte-mediated immunity, and differences between serotypes in these effects on host immunity may cause some of the variation in virulence between strains. However, the considerable genetic differences between
S. pneumoniae
strains independent of the capsular serotype prevent an unambiguous assessment of the effects of the capsular serotype on immunity using clinical isolates. We have therefore used capsular serotype-switched TIGR4 mutant strains to investigate the effects of the capsular serotype on
S. pneumoniae
interactions with complement. Flow cytometry assays demonstrated large differences in C3b/iC3b deposition on opaque-phase variants of TIGR4(−)+4, +6A, +7F, and +23F strains even though the thicknesses of the capsule layers were similar. There was increased C3b/iC3b deposition on TIGR4(−)+6A and +23F strains compared to +7F and +4 strains, and these differences persisted even in serum depleted of immunoglobulin G. Neutrophil phagocytosis of the TIGR4(−)+6A and +23F strains was also increased, but only in the presence of complement, showing that the effects of the capsular serotype on C3b/iC3b deposition are functionally significant. In addition, the virulence of the TIGR4(−)+6A and +23F strains was reduced in a mouse model of sepsis. These data demonstrate that resistance to complement-mediated immunity can vary with the capsular serotype independently of antibody and of other genetic differences between strains. This might be one mechanism by which the capsular serotype can affect the relative invasiveness of different
S. pneumoniae
strains.
Red Light Activates the σB-Mediated General Stress Response ofBacillus subtilisvia the Energy Branch of the Upstream Signaling Cascade
Authors: Ávila-Pérez Marcela , van der Steen Jeroen B. , Kort Remco , Hellingwerf Klaas J.
Published: December 2009

The σB-dependent general stress response in the common soil bacteriumBacillus subtiliscan be elicited by a range of stress factors, such as starvation or an ethanol, salt, or heat shock, via a complex upstream signaling cascade. Additionally, σBcan be activated by blue light via the phototropin homologue YtvA, a component of the environmental branch of the signaling cascade. Here we use a reporter-gene fusion to show that σBcan also be activated by red light via the energy branch of its upstream signaling cascade. Deletion mutagenesis and homologous overproduction experiments indicate that the RsbP protein (composed of an N-terminal Per-ARNT-Sim [PAS] domain and a C-terminal PP2C-type phosphatase domain) is involved in the red light response. This second light input pathway functions complementarily to YtvA; it shows broader spectral sensitivity but requires higher light intensities. These results are confirmed by transcriptome analyses, which show that both light effects result in upregulation of the σBregulon, with minimal activation of other responses.
Control of Directionality in Bacteriophage mv4 Site-Specific Recombination: Functional Analysis of the Xis Factor
Authors: Coddeville Michèle , Ritzenthaler Paul
Published: December 2009

The integrase of the temperate bacteriophage mv4 catalyzes site-specific recombination between the phageattPsite and the hostattBsite duringLactobacillus delbrueckiilysogenization. The mv4 prophage is excised during the induction of lytic growth. Excisive site-specific recombination between theattRandattLsites is also catalyzed by the phage-encoded recombinase, but the directionality of the recombination is determined by a second phage-encoded protein, the recombination directionality factor (RDF). We have identified and functionally characterized the RDF involved in site-specific excision of the prophage genome. The mv4 RDF,mv4Xis, is encoded by the second gene of the early lytic operon. It is a basic protein of 56 amino acids. Electrophoretic mobility shift assays demonstrated thatmv4Xis binds specifically to theattPandattRsites via two DNA-binding sites, introducing a bend into the DNA.In vitroexperiments andin vivorecombination assays with plasmids inEscherichiacoliandLactobacillusplantarumdemonstrated thatmv4Xis is absolutely required for inter- or intramolecular recombination between theattRandattLsites. In contrast to the well-known phage site-specific recombination systems, the integrative recombination between theattPandattBsites seems not to be inhibited by the presence ofmv4Xis.
A Comprehensive Proteomics and Transcriptomics Analysis ofBacillus subtilisSalt Stress Adaptation
Authors: Hahne Hannes , Mäder Ulrike , Otto Andreas , Bonn Florian , Steil Leif , Bremer Erhard , Hecker Michael , Becher Dörte
Published: December 2009

In its natural habitats,Bacillus subtilisis exposed to changing osmolarity, necessitating adaptive stress responses. Transcriptomic and proteomic approaches can provide a picture of the dynamic changes occurring in salt-stressedB. subtiliscultures because these studies provide an unbiased view of cells coping with high salinity. We applied whole-genome microarray technology and metabolic labeling, combined with state-of-the-art proteomic techniques, to provide a global and time-resolved picture of the physiological response ofB. subtiliscells exposed to a severe and sudden osmotic upshift. This combined experimental approach provided quantitative data for 3,961 mRNA transcription profiles, 590 expression profiles of proteins detected in the cytosol, and 383 expression profiles of proteins detected in the membrane fraction. Our study uncovered a well-coordinated induction of gene expression subsequent to an osmotic upshift that involves large parts of the SigB, SigW, SigM, and SigX regulons. Additionally osmotic upregulation of a large number of genes that do not belong to these regulons was observed. In total, osmotic upregulation of about 500B. subtilisgenes was detected. Our data provide an unprecedented rich basis for further in-depth investigation of the physiological and genetic responses ofB. subtilisto hyperosmotic stress.
Identification of the
Salmonella enterica damX
Gene Product, an Inner Membrane Protein Involved in Bile Resistance
Authors: López-Garrido Javier , Cheng Nancy , García-Quintanilla Fátima , García-del Portillo Francisco , Casadesús Josep
Published: December 2009

The
damX
gene product of
Salmonella enterica
serovar Typhimurium is a protein located in the inner membrane. DamX migrates as a 70-kDa protein in SDS-PAGE even though the predicted protein size is 46 kDa. Synthesis of DamX protein occurs in both exponential- and stationary-phase cultures. Disruption of
damX
causes severe sensitivity to bile. Lack of the outer membrane protein AsmA suppresses bile sensitivity in
Salmonella damX
mutants.
Genome Sequence of the Fleming Strain of
Micrococcus luteus
, a Simple Free-Living Actinobacterium
Authors: Young Michael , Artsatbanov Vladislav , Beller Harry R. , Chandra Govind , Chater Keith F. , Dover Lynn G. , Goh Ee-Been , Kahan Tamar , Kaprelyants Arseny S. , Kyrpides Nikos , Lapidus Alla , Lowry Stephen R. , Lykidis Athanasios , Mahillon Jacques , Markowitz Victor , Mavromatis Konstantinos , Mukamolova Galina V. , Oren Aharon , Rokem J. Stefan , Smith Margaret C. M. , Young Danielle I. , Greenblatt Charles L.
Published: December 2009

Micrococcus luteus
(NCTC2665, “Fleming strain”) has one of the smallest genomes of free-living actinobacteria sequenced to date, comprising a single circular chromosome of 2,501,097 bp (G+C content, 73%) predicted to encode 2,403 proteins. The genome shows extensive synteny with that of the closely related organism,
Kocuria rhizophila
, from which it was taxonomically separated relatively recently. Despite its small size, the genome harbors 73 insertion sequence (IS) elements, almost all of which are closely related to elements found in other actinobacteria. An IS element is inserted into the
rrs
gene of one of only two
rrn
operons found in
M. luteus
. The genome encodes only four sigma factors and 14 response regulators, a finding indicative of adaptation to a rather strict ecological niche (mammalian skin). The high sensitivity of
M. luteus
to β-lactam antibiotics may result from the presence of a reduced set of penicillin-binding proteins and the absence of a
wblC
gene, which plays an important role in the antibiotic resistance in other actinobacteria. Consistent with the restricted range of compounds it can use as a sole source of carbon for energy and growth,
M. luteus
has a minimal complement of genes concerned with carbohydrate transport and metabolism and its inability to utilize glucose as a sole carbon source may be due to the apparent absence of a gene encoding glucokinase. Uniquely among characterized bacteria,
M. luteus
appears to be able to metabolize glycogen only via trehalose and to make trehalose only via glycogen. It has very few genes associated with secondary metabolism. In contrast to most other actinobacteria,
M. luteus
encodes only one resuscitation-promoting factor (Rpf) required for emergence from dormancy, and its complement of other dormancy-related proteins is also much reduced.
M. luteus
is capable of long-chain alkene biosynthesis, which is of interest for advanced biofuel production; a three-gene cluster essential for this metabolism has been identified in the genome.
A novel Dance Dance Revolution (DDR) system for in-home training of stepping ability: basic parameters of system use by older adults
Authors: Smith S T , Sherrington C , Studenski S , Schoene D , Lord S R
Published: December 2009

Objective

This series of studies was conducted to develop and establish characteristics of exercise videogame play in older adults. The videogame was a modified version of the popular Dance Dance Revolution (DDR; Konomi).

Methods

Participants aged ≥70 were asked to make simple step movements in response to vertically drifting arrows presented on a video screen. Step responses were detected by a modified USB DDR mat, and characteristics of stepping performance such as step timing, percentage of missed target steps and percentage of correct steps were recorded by purpose-built software. Drift speed and step rate of visual stimuli were modified to increase task difficulty.

Results

Significant linear relationships between stepping performance and stimulus characteristics were observed. Performance of older adults decreased as stimulus speed and step rate were increased. Optimal step performance occurred for a stimulus speed of 17° of visual angle per second and a step rate of one step every 2 s. At fast drift speeds (up to 35°/s), participants were more than 200 ms too slow in coordinating their steps with the visual stimulus. Younger adults were better able to perform the stepping task across a wider range of drift speeds than older adults.

Conclusion

The findings suggest that older adults are able to interact with video games based upon DDR but that stepping performance is determined by characteristics of game play such as arrow drift speed and step rate. These novel “exergames” suggest a low-cost method by which older adults can be engaged in exercises that challenge balance and which can be conducted in their own homes.
CT guided percutaneous calcaneoplasty: a case of metastatic intra-articular calcaneus fracture
Authors: Kamalian S , Hirsch A E , Growney M L , Raskin K A , Yoo A J , Krag K J , Hirsch J A
Published: December 2009

Symptomatic bone metastases to the calcaneus are rare.1 Minimally invasive percutaneous augmentation is an option in the palliative management of patients with metastatic bone disease, and has been commonly used in the setting of vertebral compression fractures.2 Calcaneal augmentation can potentially allow for earlier weight bearing and a shorter period of disability. A case report is presented of percutaneous intra-articular calcaneal fracture augmentation using polymethyl methacrylate.
Impact of WASID and Wingspan on the frequency of intracranial angioplasty and stenting at a high volume tertiary care hospital
Authors: Moskowitz S I , Kelly M E , Obuchowski N , Fiorella D
Published: December 2009

Background and Purpose

The impact of the Warfarin and Aspirin for Symptomatic Intracranial Disease (WASID) data is described in combination with the commercial release of the Wingspan stenting system on the frequency of neurointervention for the treatment of symptomatic intracranial atherosclerotic disease (ICAD) in a tertiary care center.

Methods

Endovascular case logs were reviewed from April 2004 to July 2007. The total number of intracranial neurointerventions and the number of neurointerventions (percutaneous transluminal angioplasty alone (PTA) or with stenting (PTAS)) performed for symptomatic ICAD were calculated. The time period evaluated was divided into two equal 19.5-month epochs representing the time periods before and after the availability of the Wingspan system.

Results

The frequency of neurointerventions for ICAD increased by 763%, from seven of 354 total cases (2%) to 56 of 367 total cases (15.3%) (p<0.001) after the Wingspan system became available. The increase in intracranial PTAS volume occurred immediately and was stable throughout the “Wingspan era”.

Conclusions

The publication of the WASID trial results combined with the availability of the Wingspan stent system led to a marked increase in the frequency of neurointervention for symptomatic ICAD at our institution. The adoption of this technology occurred without direct evidence that PTAS with Wingspan is superior to traditional medical therapy. These findings underscore the need for a randomized trial of stenting and medical therapy for the treatment of this disease process.
A novel rearrangement in the mitochondrial genome of tongue sole,Cynoglossus semilaevis: control region translocation and a tRNA gene inversion
Authors: Kong Xiaoyu , Dong Xiaoli , Zhang Yanchun , Shi Wei , Wang Zhongming , Yu Ziniu
Published: December 2009

The organization of fish mitochondrial genomes (mitogenomes) is quite conserved, usually with the heavy strand encoding 12 of 13 protein-coding genes and 14 of 22 tRNA genes, and the light strand encoding ND6 and the remaining 8 tRNA genes. Currently, there are only a few reports on gene reorganization of fish mitogenomes, with only two types of rearrangements (shuffling and translocation) observed. No gene inversion has been detected in approximately 420 complete fish mitogenomes available so far. Here we report a novel rearrangement in the mitogenome of Cynoglossus semilaevis (Cynoglossinae, Cynoglossidae, Pleuronectiformes). The genome is 16 371 bp in length and contains 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and 2 main noncoding regions, the putative control region and the light-strand replication origin. A striking finding of this study is that the tRNAGlngene is translocated from the light to the heavy strand (Q inversion). This is accompanied by shuffling of the tRNAIlegene and long-range translocation of the putative control region downstream to a site between ND1 and the tRNAGlngene. The remaining gene order is identical to that of typical fish mitogenomes. Additionally, unique characters of this mitogenome, including a high A+T content and length variations of 8 protein-coding genes, were found through comparison of the mitogenome sequence with those from other flatfishes. All the features detected and their relationships with the rearrangements, as well as a possible rearrangement pathway, are discussed. These data provide interesting information for better understanding the molecular mechanisms of gene reorganization in fish mitogenomes.
THE ESTIMATION OF THE STROKE VOLUME OF RATS FROM THE BODY WEIGHT AND HEART WEIGHT
Authors: Grad Bernard
Published: December 2009

The object of this study was to arrive at useful empirical equations for the estimation of the stroke volume of rats from the body weight and heart weight.A logarithm of the [Formula: see text]against body weight or rats weighing from 5 to 500 gm. gave a significant positive regression which permitted the estimation of the stroke volume from the body weight, assuming a certain constant value for the arterio–venous oxygen difference. A similar plot of the heart weight of rats against body weight gave a curve which was superimposable on the first one, allowing the estimation of the stroke volume from the heart weight.
TRAUMATIC RENAL FAILURE IN THE RAT
Authors: Morton Kenneth S. , Friedman Sydney M.
Published: December 2009

A systematic study of the variables connected with the production of renal damage following a crushing limb injury in the rat has been made. The state of hydration of the animal immediately prior to and following the crushing injury appears to be the most critical variable. Myoglobin, while it intensifies the renal dysfunction, is not necessary for its production. The area involved in the crush as well as the amount of renal reserve, are important factors. Various steroid hormones were ineffective in altering the course of the disease.
SEROLOGICAL EFFECTS OF ETHIONINE IN THE RABBIT
Authors: Rice Christine E. , Boulanger P. , Annau E.
Published: December 2009

In relation to body weight, rabbits were found to tolerate a much lower dosage of ethionine than guinea pigs. Serum complement titers fell, however, even in rabbits receiving small daily doses of ethionine, an indication of a basal disturbance in the processes of protein synthesis. Total serum protein values remained within normal limits but globulin tended to increase in relation to albumin, whereas in many of the ethionine-injected guinea pigs previously studied, serum albumin had increased in relation to globulin. In rabbits given 250 mgm. daily doses of ethionine, a definite increase in the nonprotein nitrogen content of the serum was recorded.
THE ANAEROBIC DISSIMILATION OFD-GLUCOSE-1-C14,D-ARABINOSE-1-C14, ANDL-ARABINOSE-1-C14BYAEROBACTER AEROGENES
Authors: Neish A. C. , Simpson F. J.
Published: December 2009

D-Glucose-1-C14, D-arabinose-1-C14, and L-arabinose-1-C14were dissimilated anaerobically by Aerobacter aerogenes. The major products (2,3-butanediol, ethanol, acetic acid, lactic acid, formic acid, and carbon dioxide) were isolated and the location of C14determined. The products from glucose were all labeled, mainly in the methyl groups, in agreement with the hypothesis that they were derived from methyl-labeled pyruvate formed by the reactions of the classical Embden–Meyerhof scheme for glycolysis. The products from both pentoses appeared to have been formed from pyruvate labeled in both the methyl and carboxyl groups with twice as much C14in the methyl group as in the carboxyl group. This result may be explained quantitatively by a hypothesis assuming complete conversion of pentose to triose via a heptulose.
THE METABOLISM OF DL-HYDROXYPROLINE-2-C14 IN THE RAT
Authors: Gianetto R. , Bouthillier L. P.
Published: December 2009

DL-Hydroxyproline-2-C14 was synthesized with an over-all yield of about three per cent as calculated on the amount of carbon dioxide employed as radioactive starting material. DL-Hydroxyproline-2-C14 given to three normal rats by intraperitoneal injection was shown to be slowly metabolized. A scheme is suggested for the conversion of this amino acid into glutamic and aspartic acids.
AUTORADIOGRAPHIC VISUALIZATION OF THE ENTRY AND TRANSIT OF S35 IN CARTILAGE, BONE, AND DENTINE OF YOUNG RATS AND THE EFFECT OF HYALURONIDASE IN VITRO
Authors: Bélanger Leonard F.
Published: December 2009

The fate of radiosulphate introduced in young suckling rats has been followed by autoradiography of formalin–ethanol fixed celloidin sections. In the cartilaginous epiphyses of long bones, the radioelement has been detected first in the cells, particularly at the hypertrophic level, then in cells and matrix, and finally in matrix exclusively. The incorporated radiosulphate was later displaced towards the ossification area by the growth process. During this migration, the relative intensity of the autoradiograph decreased progressively. This series of events was interpreted as visualization of chondroitin sulphate synthesis, secretion, and utilization or turnover. The S35 labelled substance was rapidly hydrolyzed by hyaluronidase except in areas of tendon insertions. In bone and dentine, an over-all initial exchange picture was replaced by a growth line of acid fast, hyaluronidase labile, sulphated material which became displaced along with bone salts by further growth. This material was metachromatic with toluidine blue. It is thought to be also chondroitin sulphate and its possible relationship to mineralization has been discussed.
THE BIOSYNTHESIS OF CELL WALL CARBOHYDRATES: GLUCOSE-C14 AS A CELLULOSE PRECURSOR IN WHEAT PLANTS
Authors: Brown Stewart A. , Neish A. C.
Published: December 2009

The formation of cellulose in wheat plants has been studied using as precursors glucose-1-C14, sorbitol-1-C14, and succinic acid-2,3-C14, which were administered to the plant aseptically by injection of sterile solutions into the hollow internodes. Glucose obtained by hydrolysis of the cellulose, isolated after the plants had grown for 24 to 48 hr. following the injection of labelled glucose, showed a concentration of at least 50% of the total C14 in carbon-1. When sorbitol was injected most of the carbon-14 was fairly evenly divided between C-1 and C-6, and when succinic acid was used evidence of a comparatively uniform labelling of the glucose was obtained. The results indicate that there is considerable conversion of glucose to cellulose without skeletal rearrangement, and that there is also appreciable cellulose formation from resynthesized glucose. Sorbitol is not an intermediate between glucose and cellulose; it is suggested that it is converted to glucose largely by way of three-carbon fragments.
ESTIMATION OF PENTOSE NUCLEIC ACID AND DESOXYPENTOSE NUCLEIC ACID IN THE LIVER AND BRAIN TISSUE OF MICE FOLLOWING THE FEEDING OF THE INSECTICIDE ALDRIN (HEXACHLOROHEXAHYDRODIMETHANONAPHTHALENE)
Authors: Annau E.
Published: December 2009

Aldrin fed to mice in sublethal concentrations induced, after a period of time, increase in DNA content of the liver tissue, while its PNA content remained essentially unaltered. A slight increase in the weight of the brain was noted, which did not affect the DNA or PNA content of the brain tissue.
THE FAILURE OF GROWTH HORMONE TO MODIFY HORMONAL HYPERTENSION IN THE RAT
Authors: Friedman Sydney M. , Nakashima Miyoshi , Friedman Constance L.
Published: December 2009

Growth hormone (Connaught) was given in 1 mgm. daily doses to rats either otherwise untreated or given an amount of DCA or Compound F sufficient to elicit a moderate hypertensive response. At the dose level used, growth hormone did not affect either blood pressure or kidney size in treated or untreated animals. The dose used was one-fifth the amount found to increase renal function in the dog and 10 times the dose required for trophic effects in the rat.
THE GLYCEROL DEHYDROGENASES OF PSEUDOMONAS SALINARIA, VIBRIO COSTICOLUS, AND ESCHERICHIA COLI IN RELATION TO BACTERIAL HALOPHILISM
Authors: Baxter R. M. , Gibbons N. E.
Published: December 2009

Glycerol dehydrogenases from the extremely halophilic Pseudomonas salinaria and the moderately halophilic Vibrio costicolus are described and compared with the corresponding enzyme from the nonhalophilic Escherichia coli. The properties of all three enzymes are similar except their responses to salt concentration. The enzymes from E. coli and V. costicolus are most active at sodium chloride concentrations of about 0.25 M and 0.5 M respectively; that from P. salinaria is not only most active in the presence of 1.5 M NaCl but is irreversibly inactivated in the absence of salt. All three enzymes are more active in the presence of potassium chloride than of sodium chloride at any given molar concentration. These results suggest that the extremely halophilic bacteria contain high concentrations of salt and that their enzymes function maximally at these high concentrations. In contrast the moderately halophilic organisms contain relatively little salt and their enzymes are more comparable with those of nonhalophiles.
FACTORS AFFECTING THE BIOASSAY OF 11-OXYCORTICOSTEROIDS IN ADRENALECTOMIZED RATS
Authors: Stephenson N. R.
Published: December 2009

Total sugar in a protein-free liver hydrolyzate, determined by copper reduction, provides an acceptable criterion for estimating the response to graded doses of 11-oxycorticosteroids. Total carbohydrate in the liver, measured by the anthrone reagent, may also be used for this purpose. The glycogenic response to adrenal corticoids of fasting adrenalectomized male rats of the Wistar strain, weighing 40 to 50 gm., is increased by the administration of glucose, but appears to be depressed by ethanol. Although the precision of the assay is not influenced unfavorably by using the rats on the day following adrenalectomy, the mortality rate of the adrenalectomized animals is reduced by this procedure.
PHOSPHORUS METABOLISM OF THE ADRENAL GLAND EFFECT OF HYPOPHYSECTOMY AND ADMINISTRATION OF ACTH ON INCORPORATION OF P32 INTO INORGANIC PHOSPHATE
Authors: Riedel B. E. , Logan J. E. , DeLuca H. A. , Rossiter R. J.
Published: December 2009

In confirmation of the work of others, the concentration of inorganic phosphate (P) in the plasma of hypophysectomized rats was found to be less than that in the plasma of control animals. Hypophysectomy caused no significant change in the concentration of inorganic P in the adrenal gland or liver. A single intraperitoneal injection of each of two preparations of ACTH failed to cause any significant change in the concentrations of inorganic P in plasma, adrenal, or liver.The specific activity of the inorganic P in the plasma of hypophysectomized rats after an intraperitoneal injection of inorganic P labelled with P32 was greater than that in the control animals. Hypophysectomy caused a decrease in the specific activity of the inorganic P of the adrenal gland relative to that of the inorganic P of the plasma. Each of the two preparations of ACTH, given to the hypophysectomized animals as a single intraperitoneal injection 20 hr. before killing, restored the relative specific activity of the inorganic P of the adrenals to normal values. When the ACTH was administered six hours before killing, one of the preparations (ACTH A) caused an increase in the relative specific activity of the inorganic P of the adrenals, but a second preparation (ACTH C) was without significant effect.The increase in the specific activity of the inorganic P of the plasma comes on slowly (quite small two days after hypophysectomy), whereas the decrease in the relative specific activity of the inorganic P of the adrenal gland comes on rapidly (maximal two days after hypophysectomy). For this reason, at longer time intervals after hypophysectomy (greater than six days) the absolute activity of the acid-soluble P of the adrenal, i.e. the activity not referred to that of the inorganic P of the plasma, was greater in hypophysectomized animals, and not less, as reported by other workers. The activity of this fraction is less in hypophysectomized animals only if the observations are made at short time intervals after removal of the pituitary. Evidence is presented for the view that the increase in the specific activity of the inorganic P of the plasma is the result of changes brought about by a deficiency of growth hormone, whereas the decrease in the relative specific activity of the adrenal is due to a deficiency of ACTH.
PHOSPHORUS METABOLISM OF THE ADRENAL GLAND: THE MECHANISM OF THE CHANGE AFTER HYPOPHYSECTOMY
Authors: Riedel B. E. , Rossiter R. J.
Published: December 2009

The specific activity of the intracellular inorganic phosphate (P) of the adrenal gland relative to that of the inorganic P of the plasma is significantly decreased in hypophysectomized rats at time intervals as long as 16 hr. after the injection of inorganic P labelled with P32. The specific activity of the intracellular inorganic P was determined (1) by measuring the specific activity of the easily-hydrolyzable acid-soluble P, which rapidly comes into isotope equilibrium with the intracellular inorganic P, (2) by calculation, from the specific activity of the inorganic P of the whole adrenal, assuming values for the specific activity and the concentration of the inorganic P of the extracellular fluid and the volume of the extracellular fluid compartment, and (3) by measuring the specific activities of the inorganic P of both the adrenal gland and the plasma at a series of time intervals after the injection of the P32. It is concluded that the decrease in the relative specific activity of the intracellular inorganic P of the adrenal is the result of a slowing in the passage of inorganic P32 across the cell membrane, i.e. from the extracellular to the intracellular fluid.
CARDIOVASCULAR RESPONSES IN DOGS TO LARGE INTRAVENOUS INFUSIONS
Authors: Gowdey C. W. , Hatcher J. D. , Sunahara F. A.
Published: December 2009

The effects of a continuous, slow, intravenous infusion of gum acacia solution have been measured in anesthetized dogs. When the volume of the circulation was increased and the hematocrit value reduced, the cardiac output, intracardiac pressures, and peripheral blood flows began to increase. In some experiments these changes continued until sudden cardiac decompensation occurred during which the arterial blood pressure, cardiac output, and peripheral blood flows were reduced while the mean right auricular and ventricular pressures increased markedly. Analysis of the results indicates that up to the time of the highest cardiac output there is a closer correlation between cardiac output and hematocrit value than between cardiac output and mean right auricular pressure.
NUTRITION OF ANIMAL CELLS IN TISSUE CULTURE: VII. USE OF REPLICATE CELL CULTURES IN THE EVALUATION OF SYNTHETIC MEDIA
Authors: Parker Raymond C. , Healy George M. , Fisher Dorothy C.
Published: December 2009

The replicate culture assay procedures of Earle and his associates have been adapted for use in evaluating the effectiveness of synthetic media. For this purpose, use has also been made of Earle's L strain mouse cells. Washed and continuously stirred suspensions of these or similar strains of cells may be dispensed, with reasonable assurance of uniformity, into a series of replicate cultures, the number depending on the volume of the suspension and the capacity and effectiveness of the stirring and dispensing unit. For use with synthetic media, the original procedures for the preparation and care of the replicate cultures and for their subsequent treatment for the counting of isolated, stained nuclei have been modified considerably. This paper describes the procedures that were finally adopted and also describes a relatively simple screening procedure in which washed cell suspensions may be used to advantage in making preliminary assays of synthetic media and in testing the relative toxicity or growth stimulating effects of substances added to, or derived from, natural media.
STUDIES ON THE PRESERVATION OF BLOOD: III. REVERSIBLE INHIBITION OF GLYCOLYSIS IN BLOOD BY OXALATE
Authors: Pappius Hanna M. , Denstedt Orville F.
Published: December 2009

Oxalate was found to inhibit glycolysis in red blood cells during storage at 5 °C. The inhibition of glycolysis in blood cells by oxalate is reversible. The glycolytic activity of the oxalated cells can be restored by washing the cells, or by precipitating the oxalate with an equivalent concentration of added calcium ions. However, with increased duration of storage prior to removal of oxalate the rate of glycolysis in the erythrocytes was found to be decreased on removal of the oxalate.
THE LACK OF EFFECT OF VITAMIN E ON THE BLOOD CLOTTING MECHANISM
Authors: Paul Rachel M. , Lewis J. A. , DeLuca H. A.
Published: December 2009

The oral administration of vitamin E to human subjects in dail doses of 600 mgm. for seven days failed to evoke any blood change in clotting times, prothrombin times, or plasma fibrinogen levels. There was a small increase in platelet level which was statistically of borderline significance.
THE LIPID AND WATER CONTENT OF CARCASS, SKELETAL MUSCLE, AND TESTICLE IN THE HOST COMPONENT OF THE ALBINO RAT – WALKER CARCINOMA 256 DUAL ORGANISM AT PROGRESSIVE STAGES OF TUMOR GROWTH
Authors: Boyd Eldon M. , Boyd Carl E. , Hill J. Gilbert , Ravinsky Ely
Published: December 2009

The objective of this investigation was to find at what stage in the growth of Walker carcinoma 256 appear the shifts in water and lipid levels of host carcass, skeletal muscle, and testicle, noted at or near death of the dual organism by Boyd, Connell, and McEwen (1952). Lipid and water estimations were made upon these tissues, at intervals of one, two, and three weeks of tumor growth, in 35 tumor-bearing and 34 littermate control albino rats. In host carcass, the decline in concentration, per 100 gm. dry weight, of total lipid, neutral fat, and total fatty acids appeared after two weeks of tumor growth, while at or about the same time a rise occurred in the levels of water, total cholesterol, free cholesterol, and phospholipid. In hind limb skeletal muscle of the host, the levels of total lipid, neutral fat, and total fatty acids were lowered, while those of water were elevated, after two and three weeks of tumor growth. In host testicle, the levels of water and lipids were essentially similar to those of the controls. The rise in concentration of water, phospholipid, total cholesterol, and free cholesterol of the host varied, in general, with increase in the T/RC coefficient. Maximal low levels of host total lipid, neutral fat, and total fatty acids were reached at T/RC coefficient values of 20 to 30. Maintenance of total body weight (tumor plus host) was due mainly to accumulation of water in both components. The host component lost dry weight, total lipid, neutral fat, and total fatty acids more rapidly than these accumulated in the tumor in total amount. The smaller loss of total amounts of water, phospholipid, total cholesterol, and free cholesterol in the host was offset by an approximately equal accumulation of these substances in the tumor.
THE EFFECT OF CORTISONE ACETATE ON THE PRODUCTION OF CIRCULATING HEMOLYTIC ANTIBODIES IN THE MOUSE
Authors: Newsom Shirley E. , Darrach Marvin
Published: December 2009

Quantitative immunochemical methods have been used to show that in the mouse cortisone acetate suppresses the formation of circulating hemolytic antibodies to sheep erythrocytes. The amount of suppression varies with the dose of both cortisone acetate and antigen. Experimental conditions are defined whereby an almost complete inhibition of circulating antibody occurs. The method serves as a quantitative procedure for comparing the effect of different steroids and hormones on antibody production as well as a means of studying possible cortisone antagonists.
CONTINUOUS INTRAVENOUS INFUSION IN THE RAT, AND THE EFFECT ON THE ISLETS OF LANGERHANS OF THE CONTINUOUS INFUSION OF GLUCOSE
Authors: Kinash B. , Haist R. E.
Published: December 2009

A method is described for the continuous intravenous infusion of fluids in the unanesthetized rat. When rats were infused continuously with glucose solutions for 6–14 days the total amount of islet tissue was greatly increased, as compared to that of saline-infused controls. This increase was evident also when considered in relation to pancreas weight or body weight.
MODIFICATION OF PARATHION'S TOXICITY FOR RATS BY PRETREATMENT WITH CHLORINATED HYDROCARBON INSECTICIDES
Authors: Ball W. L. , Sinclair J. W. , Crevier M. , Kay Kingsley
Published: December 2009

Oral pretreatment of rats with chlorinated hydrocarbon insecticides greatly increases their resistance to poisoning by organic phosphate insecticides. Previous work in this laboratory demonstrated that the pretreatment is followed by a rise in the serum aliesterase level. Aldridge has shown that A-aliesterase is capable of hydrolyzing paraoxon (an organic phosphate). It is suggested that an increase in the level of this organophosphate hydrolyzing enzyme plays a part in the protection observed.
ACUTE ESERINE POISONING IN THE MONKEY
Authors: Candole C. A. de
Published: December 2009

The changes in pulmonary ventilation which follow the intravenous injection of lethal and sublethal doses of eserine in the urethanized monkey have been studied. A reduction in minute volume is the outstanding change; but periods of overventilation may occur both before and after the phase of underventilation.When death takes place within ten minutes respiration fails before the heart, whereas when death is delayed, both fail together.Different elements of the respiratory mechanism differ in their susceptibility to eserine. Thus diaphragmatic and intercostal function are rarely lost together, and gasping may continue for long periods after eupnoeic breathing has failed.
STARCH SYNTHESIS IN CHLORELLA VULGARIS
Authors: Bailey J. M. , Neish A. C.
Published: December 2009

Chlorella vulgaris was found to deposit starch, in amounts up to 20% of the dry weight of the cells, when grown in a medium containing glucose. The cells did not contain cellulose or chitin. The starch was difficult to extract, being associated with an alkali-soluble, dextrorotatory, cell-wall polysaccharide. The starch, after extraction by a 26% solution of calcium chloride at 120 °C., had properties quite similar to starches from higher plants. It was composed of amylose (30–40%) and amylopectin. Glucose-1-C14 was incorporated into the starch, by growing cells, without much breakdown and resynthesis. Cell-free extracts, obtained from the alga, contained a phosphorylase and a branching enzyme similar to those of the potato. These brought about the synthesis of an amylopectin–glycogen type polysaccharide from glucose-1-phosphate. It is concluded that the mechanism of starch synthesis in Chlorella vulgaris is essentially the same as in higher plants.
THE HYDROLYSIS OF ADENOSINE TRIPHOSPHATE BY BRAIN SUSPENSIONS
Authors: Pappius Hanna M. , Elliott K. A. C.
Published: December 2009

Adenosine triphosphate (ATP) hydrolysis by unfractionated brain suspensions has been measured manometrically and chemically in bicarbonate buffer. Both magnesium and, less strongly, calcium accelerate ATP hydrolysis by brain suspensions. The effectiveness of low magnesium or calcium concentrations is decreased by relatively high ATP concentrations. The optimum pH for phosphate liberation is about 8. The ratio of carbon dioxide evolved to inorganic phosphate liberated depends on the pH. Evidence is given that the suspensions contain a Mg-activated ATPase which is strongly inhibited by calcium, a Ca-activated ATPase, and a Mg-activated PPase which is strongly inhibited by calcium but is different from the Mg-ATPase. These enzymes were not obtained in "latent" form nor activated by glutathione. Ethylenediamine tetraacetate can inhibit Ca-ATPase completely but is less effective on the Mg-ATPase and can release the latter from calcium inhibition. Other inhibitors have been tested.The ATPase activity of cerebral cortex of mouse, rat, cat, beef, and man has been measured by a standard procedure. The activity per unit weight decreases with increasing size of the animal parallel to the decrease in the oxygen uptake rate. Calculations show that the ATPase activity is high enough to release all high energy phosphate which could be produced by esterification coupled with respiration.
INCORPORATION OF RADIOACTIVE PHOSPHATE INTO NON-NUCLEOTIDE PROTEIN-BOUND PHOSPHORUS FRACTIONS OF RESPIRING CAT BRAIN SLICES
Authors: Findlay Muriel , Strickland K. P. , Rossiter R. J.
Published: December 2009

Slices of cat brain respiring in a Krebs–Ringer bicarbonate medium were found to incorporate inorganic phosphate labelled with P32 into two non-nucleotide protein-bound phosphorus fractions, which are referred to as the residue organic phosphorus (ROP) and the "phosphoprotein" (PP), respectively. The addition of glucose or mannose to the medium increased the incorporation into both fractions. The addition of fructose, galactose, pyruvate, lactate, succinate, or L-glutamate failed to increase the incorporation into either fraction.Anaerobic conditions, homogenization of the tissue, or the addition of a wide range of metabolic inhibitors (cyanide, azide, iodoacetate, fluoride, nembutal, malononitrile, chloretone) inhibited the incorporation. The incorporation was also inhibited by 2,4-dinitrophenol in concentrations that do not inhibit oxygen consumption.These results are compared and contrasted with previous findings on the incorporation of P32 into the lipid phosphorus and pentosenucleic acid of brain slices.