bluuebunny/crossref_metadata_2025 huggingface dataset:
Long-range frequency-difference source localization in the Philippine Sea
Authors: Geroski David J. , Dowling David R.
Published: December 2019

Matched field processing (MFP) refers to a variety of source localization schemes for known complicated environments and involves matching measured and calculated (replica) fields to identify source locations. MFP may fail for several reasons, most notably when the calculated fields are insufficiently accurate. This error commonly prevents MFP-based long-range (>100 km) source localization in the deep ocean (from 5 to 6 km depth) for signal frequencies of hundreds of Hz, even when extensive high-signal-to-noise ratio field measurements are available. Recently, below-band MFP utilizing the frequency-difference autoproduct [Worthmann, Song, and Dowling (2015). J. Acoust. Soc. Am, 138(6), 3549–3562] achieved some shallow-ocean localization success at a 3 km source-to-array range with signal frequencies in the tens of kHz. The performance of this technique, when extended to matching the measured frequency-difference autoproduct with a composite mode-ray replica, is described here for deep ocean source localization. The ocean propagation data come from the PhilSea10 experiment and involve source-to-array ranges from 129 to 379 km and nominal 100-Hz-bandwidth signals having center frequencies from 250 to 275 Hz. Based on an incoherent average of five signal samples, the frequency-difference technique was 90%–100% successful at four different source-to-array ranges using single-digit-Hz difference frequencies.
Noise robust bird call localisation using the generalised cross-correlation with phase transform in the wavelet domain
Authors: Ollivier Benjamin , Pepperell Alexander , Halstead Zachary , Hioka Yusuke
Published: December 2019

Bird localisation using passive acoustic methods is a non-intrusive solution for taking a census of bird species. Through the recordings from a microphone array, the generalised cross-correlation with phase transform (GCC-PHAT) has been adopted widely in the estimation of the direction-of-arrival (DOA) of audio signals especially speech in indoor environments, as it performs very well in a reverberant environment. However, the performance is degraded when the signal to noise ratio is low. This study investigates the performance of DOA estimation when the GCC-PHAT is applied in the wavelet domain. Three configurations are considered in this study and the performance of these configurations is assessed by numerical simulation under ideal setup and practical experiments using audio signals recorded in a typical native forest in New Zealand. The results suggest the configuration which applies GCC-PHAT and denoising in the wavelet domain and estimates DOA from the reconstructed cross-correlation can give more accurate estimation compared to the conventional GCC-PHAT.
Toxicity of Smoke to Epiphytic Ice Nucleation-Active Bacteria
Authors: Zagory Devon , Lindow Steven E. , Parmeter John R.
Published: December 2019

Wheat straw smoke aerosols and liquid smoke condensates reduced significantly both the viability and the ice-nucleating activity of
Pseudomonas syringae
pv.
syringae
and
Erwinia herbicola
in vitro and on leaf surfaces in vivo. Highly significant reductions in numbers of bacterial ice nuclei on the surface of both corn and almond were observed after exposure to smoke aerosols. At −5°C, frost injury to corn seedlings colonized by ice nucleation-active bacteria was reduced after exposure to smoke aerosols. Effects on −9°C ice nuclei, although significant, were less than on ice nuclei active at −5°C. These results suggest that smoke from wildfires or smudge pots may reduce plant frost susceptibility and sources of ice nuclei important in other natural processes under some conditions.
Age-Dependent Levels of Select Immunological Mediators in Sera of Healthy Children
Authors: Sack Ulrich , Burkhardt Ullrich , Borte Michael , Schädlich Hiltrud , Berg Kerstin , Emmrich Frank
Published: December 2019

Serum cytokine levels were measured in 275 healthy children of different ages (3 to 17 years). Interleukin-1 receptor antagonist (IL-1RA), soluble IL-2R (sIL-2R) (sCD25), IL-6, IL-8, tumor necrosis factor alpha (TNF-α), soluble TNF receptor type II (sTNF-RII) (sCD120b), gamma interferon (IFN-γ), soluble intercellular adhesion molecule 1 (sICAM-1) (sCD54), soluble E selectin (sE-selectin) (ELAM-1; sCD62E), sCD14, and neopterin were measured with commercial test kits. The mean levels of IL-1RA, sIL-2R, TNF-α, sICAM-1, sE-selectin, and sCD14 were higher than in healthy adults. In contrast, IFN-γ and IL-8 were hardly detectable in children and thereby significantly lower than in adults. In the case of TNF-α, sICAM-1, sE selectin, and sCD14, there was a high interindividual variability, apparently unrelated to disease. The profiles of some cytokines, i.e., IL-1RA, IL-6, and TNF-α, showed age-related increases that overlapped with known patterns of physical growth. Of note, sIL-2R and sE-selectin instead declined with time. Because of the remarkable age-dependent variability in healthy pediatric subjects, disease-related changes, as well as therapy-dependent alterations, should be considered with caution.
Antifungal Activity of a Human Antiglucuronoxylomannan Antibody
Authors: Zhong Zhaojing , Pirofski Liise-anne
Published: December 2019

The human immunoglobulin M (IgM) monoclonal antibody (MAb) 2E9 binds the glucuronoxylomannan (GXM) ofCryptococcus neoformansserotypes A, B, and D. This study was undertaken to determine the opsonic efficacy of 2E9 and its ability to promote the antifungal activity of human polymorphonuclear neutrophils (PMNs) againstC. neoformans. We incubated purified PMNs with fluorescein isothiocyanate-labeledC. neoformanscells that were treated with the GXM IgM 2E9, IgM antibodies that do not bind GXM, and rabbit and human factor-B-deficient serum as complement sources. PMN-associatedC. neoformanscells fluoresced and were detected with a fluorescence-activated cell sorter. The amount of phagocytosis was defined as the percent fluorescing PMNs, which was 37% for yeast cells opsonized with 2E9 plus rabbit serum and 57% for yeast cells opsonized with 2E9 plus factor-B-deficient serum. Phagocytosis was significantly greater for yeast cells that were treated with 2E9 plus a complement source than for yeast cells treated with the complement sources alone or treated with the control IgMs alone or with the complement sources. Fluorescence quenching and light and electron microscopy of the phagocytosis mixtures revealed that 2E9-opsonized yeast cells were internalized by PMNs. Maximal inhibition ofC. neoformansgrowth occurred when PMNs were cocultured with yeast cells that were opsonized with 2E9 plus a complement source. Our data demonstrate that the human GXM IgM 2E9 can mediate PMN phagocytosis andC. neoformansgrowth inhibition in vitro. These findings strongly suggest that antibody-mediated deposition of complement components on the cryptococcal capsule can augment PMN complement receptor-mediated antifungal activity. Antibody activation of complement-mediated effector cell antifungal mechanisms may play a role in host defense against cryptococcosis and represents a goal for the use of MAbs to treat or prevent humanC. neoformansinfections.
Induction of Fimbriated
Vibrio cholerae
O139
Authors: Ehara Masahiko , Iwami Mamoru , Ichinose Yoshio , Hirayama Toshiya , Albert M. John , Sack R. Bradley , Shimodori Shoichi
Published: December 2019

Several fimbriated phases of
Vibrio cholerae
O139 strains were selectively induced and compared immunologically and biochemically with those of
V. cholerae
O1. Fimbrial antigens were detected on the surfaces of vibrio cells colonizing the epithelial cells of a rabbit small intestine. Convalescent-phase sera from six individuals infected with
V. cholerae
O139 revealed the development of antibody against the fimbrillin. These findings suggest that the fimbriae of
V. cholerae
O1 and O139 are expressed in vivo during infection and that consideration must be given to the use of fimbrial antigens as components of vaccines against cholera.
Heterogeneity of Immunoglobulin G Response to
Helicobacter pylori
Measured by the Unweighted Pair Group Method with Averages
Authors: Mayo Kathryn , Pretolani Stefano , Gasbarrini Giovanni , Ghironzi Giancarlo , Megraud Francis
Published: December 2019

The heterogeneity of the immune response to
Helicobacter pylori
has always been noticed but has never been evaluated by obtaining a quantitative measure. For this purpose, sera were tested by enzyme-linked immunosorbent assay, and 207 positive serum specimens were subsequently tested by immunoblotting. The presence or absence of six specific bands was noted. The homology of the different profiles of bands was measured by calculating the Dice coefficient, and a dendrogram was constructed. Thirty-four profiles were found, with each profile containing from 1 to 43 serum specimens. At a level of 72% similarity, 115 of the serum specimens studied fell into eight profiles. At a level of 48% similarity, 186 of the serum specimens studied fell into 22 profiles. The difference in immunoblot profiles was probably linked to the host immune response, but infection with strains carrying different antigens cannot be ruled out.
Decline of Measles-Specific Immunoglobulin M Antibodies after Primary Measles, Mumps, and Rubella Vaccination
Authors: Helfand Rita F. , Gary Howard E. , Atkinson William L. , Nordin James D. , Keyserling Harry L. , Bellini William J.
Published: December 2019

Detection of measles-specific immunoglobulin M (IgM) has become the standard diagnostic method for laboratory confirmation of measles. In outbreaks, the interpretation of an IgM-positive result can be complicated when persons with suspected measles receive a dose of measles vaccine as part of outbreak control measures. This investigation evaluated the decay of measles-specific IgM antibodies 1 to 4 months after primary vaccination with measles, mumps, and rubella vaccine (MMRII). Serum samples were obtained from 536 infants vaccinated when they were 15 months old as part of a study to assess primary and secondary measles vaccine failure. Sixty serum specimens per week were selected from specimens collected between 4 and 9 weeks after MMRII vaccination; all 176 available serum specimens collected between 10 and ≥16 weeks were included. Specimens were tested for the presence of measles-specific IgM by an antibody-capture enzyme immunoassay. The proportion of IgM-positive specimens dropped from 73% at 4 weeks after vaccination to 52% at 5 weeks after vaccination and then declined to 7% by 8 weeks after vaccination. Less than 10% of children remained IgM positive between 9 and 11 weeks. An IgM-negative result helps rule out the diagnosis of measles in a person with suspected infection and a history of recent vaccination. The interpretation of a positive IgM result from a person with a clinically suspected case of measles and a recent history of measles vaccination (especially within 8 weeks) is problematic, and the diagnosis of measles should be based on epidemiologic linkage to a confirmed case or on detection of wild-type measles virus.
Diversity of Hemagglutination Phenotypes among P-Fimbriated Wild-Type Strains of
Escherichia coli
in Relation to
papG
Allele Repertoire
Authors: Johnson James R. , Brown Jennifer J. , Ahmed Parvia
Published: December 2019

Data regarding the hemagglutination (HA) patterns of the three variants (classes I, II, and III) of the
Escherichia coli
adhesin PapG are conflicting. These HA patterns usually have been assessed for each
papG
allele separately with recombinant strains in slide HA assays. We rigorously evaluated an alternative microtiter tray HA assay and then used it to assess the HA of four erythrocyte types (human A
1
P
1
and OP
1
, rabbit, and sheep erythrocytes) by multiple wild-type
E. coli
strains representing the four naturally occurring combinations of the
papG
alleles, i.e., class I plus III, class III only, class II plus III, and class II only. The microtiter tray HA assay displayed significantly better reproducibility of intraobserver (83%) and interobserver (86%) results than did slide HA assays (39 and 73%, respectively). Novel findings from the study of 32 wild-type P-fimbriated strains included reproducible determinations of phenotypic diversity among different
papG
categories, among strains within each
papG
category, and from day to day for individual strains. There was also substantial overlap of phenotypes between
papG
categories I plus III and III only and between II plus III and II only. A class III
papG
recombinant strain’s HA pattern differed significantly from that of the wild-type class III strains. These data demonstrate that HA phenotypes of wild-type P-fimbriated
E. coli
strains can be reproducibly assessed by a microtiter HA assay and that they correspond broadly to
papG
genotype but in a more complex and varied fashion than previously recognized.
Immunodepression Induced byTrypanosoma cruziand Mouse Hepatitis Virus Type 3 Is Associated with Thymus Apoptosis
Authors: Verinaud Liana , Da Cruz-Höfling Maria Alice , Sakurada Júlia K. , Rangel Humberto A. , Vassallo José , Wakelin Derek , Sewell Herb F. , Camargo Irineu J. B.
Published: December 2019

Trypanosoma cruzi-infected mice show disturbance in the peripheral immune system such as polyclonal lymphocyte activation, autoantibody production, and immunosuppression of T lymphocytes. Previous observations in our laboratory showed that some stocks ofT. cruzican be contaminated with mouse hepatitis virus type 3 (MHV-3). Literature has shown that MHV-3 infection induces immunologic disorders characterized by thymic involution with marked cell depletion. However, the effects of interactions between MHV-3 and the parasite on the immune system are not well understood. In the present study specific-pathogen-free CBA mice were inoculated with MHV-3, alone or associated with different stocks ofT. cruzi. Concurrent murine virus infection resulted in increased pathogenicity ofT. cruziinfection shown by profound thymic atrophy; loss of cortical thymocytes; depletion of Thy1.2+, CD4+, and CD8+cells; enhancement of in situ labeling of nuclear DNA fragmentation; and eventually, death of the animals. Such lines of evidence show that the mechanism underlying this thymic atrophy is associated with apoptosis. These results also suggest that MHV-3 can account for the increased immunosuppression observed during experimental infection with the parasite.
PCR Detection of Human Cytomegalovirus DNA in Clinical Specimens Using Novel UL37 Exon 3 and US3 Primers
Authors: Lesperance Marci M. , Contopoulos-Ioannidis Despina G. , Gutiérrez María del Pilar , Colberg-Poley Anamaris M.
Published: December 2019

The sensitivity and specificity of novel UL37 exon 3 (UL37x3) and US3 immediate-early (IE) gene PCR primers to detect human cytomegalovirus (HCMV) DNA in clinical specimens are comparable to those of HCMV DNA polymerase (UL54) primers. The use of these IE primers increases the diagnostic performance of HCMV PCR.
Interleukin-1β Expression after Inhibition of Protein Phosphatases in Endotoxin-Tolerant Cells
Authors: Yoza Barbara K. , Wells Jon D. , McCall Charles E.
Published: December 2019

Endotoxin (lipopolysaccharide [LPS]) is a potent activator of a number of inflammatory genes in blood leukocytes, including interleukin-1 (IL-1). Blood leukocytes isolated from patients with septic shock fail to produce IL-1 in response to LPS, a phenomenon known as endotoxin tolerance. To study the regulation of IL-1 expression in endotoxin-tolerant cells, the protein phosphatase inhibitor okadaic acid was used to examine the effects of protein phosphorylation on IL-1β gene expression. We found that endotoxin-tolerant cells produced normal levels of IL-1β when protein phosphatases were inhibited. In the human pro-monocytic cell line THP-1, okadaic acid increased mRNA accumulation and synthesis of IL-1β protein. Normal and endotoxin-tolerant THP-1 cells accumulated IL-1β mRNA and protein with similar delayed kinetics. Okadaic acid stabilization of IL-1β mRNA appears to be the primary mechanism through which endotoxin-tolerant cells accumulate IL-1β mRNA and protein. Endotoxin-tolerant cells were unable to activate transcription in response to okadaic acid. However, the transcription factor NF-κB, which is known to be involved in IL-1β expression, was translocated to the nucleus in both normal and endotoxin-tolerant cells after treatment with okadaic acid. These studies revealed that protein phosphorylation can affect gene expression on at least two distinct levels, transcription factor activation and mRNA stability. Endotoxin-tolerant cells have decreased transcription activation potential, while IL-1β mRNA stability remains responsive to protein phosphorylation.
Development and Evaluation of a Chromatographic Procedure for Partial Purification of Substance P with Quantitation by an Enzyme Immunoassay
Authors: Fehder William P. , Ho Wen-Zhe , Campbell Donald E. , Tourtellotte Wallace W. , Michaels Lisa , Cutilli Joann R. , Uvaydova Marina , Douglas Steven D.
Published: December 2019

We have developed a simple chromatographic procedure for the partial purification of substance P (SP) from acidified plasma and serum samples. We have evaluated a sensitive antigen competition enzyme immunoassay (EIA) for the quantitation of SP. The chromatographic procedure has recovery efficiencies ranging from 94.8 to 125%. The immunoreactivity of unknown amounts of purified SP subjected to the preparative procedure yielded a coefficient of variance of 9.4%. The EIA yielded reproducible standard curves having an interassay (n= 8) correlation coefficient of 0.984. The evaluation of normal adult control serum yielded a mean value of 51 pg/ml (range, 35 to 61 pg/ml). The evaluation of 3.33× concentrates of serum-derived partially purified SP provided uncorrected SP values of 117 to 201 pg/ml, which fell within the midpoint of the three-decalog standard curve. These studies indicate that both the preparative and quantitative procedures are required for the detection of SP in plasma or serum samples collected from patients with several clinical disorders.
Comparison of Immunodot and Western Blot Assays for Diagnosing Lyme Borreliosis
Authors: Fawcett Paul T. , Rosé Carlos D. , Gibney Kathleen M. , Doughty Robert A.
Published: December 2019

Two commercially available serologic tests for use in diagnosing Lyme borreliosis were evaluated by using a test panel comprised of sera from patients diagnosed with Lyme borreliosis, non-Lyme disease controls, and healthy subjects. The test methods examined were a Western blot assay and an immunodot assay. The study was initiated to determine how the immunodot assay, which contains purified and recombinant proteins to those borrelial antigens recommended for immunoglobulin M (IgM) detection in the Dearborn criteria, would compare with the Western blot assay as a confirmatory method for serologic diagnosis of Lyme borreliosis. Results obtained showed that the two test methods performed comparably for detecting IgG antibodies. For IgM antibody detection, the immunodot and Western blot assays had similar sensitivities; however, the immunodot assay was more specific and had greater positive predictive value than the Western blot assay. The results obtained indicate that the immunodot assay performs as well as or better than the Western blot assay for diagnosing Lyme borreliosis. Furthermore, because it uses a limited panel (
n
= 5) of antigens, the immunodot is easier to read and interpret than standard Western blots.
Comparison of an Amplified Enzyme-Linked Immunosorbent Assay with Procedures Based on Molecular Biology for Assessing Human Immunodeficiency Virus Type 1 Viral Load
Authors: Goldschmidt Pablo L. , Devillechabrolle Andrée , Ait-Arkoub Zaina , Aubin Jean-Thierry
Published: December 2019

The sensitivity of the enzyme-linked amplified sorbent test (ELAST) was compared with those of other classic enzyme-linked immunosorbent assays (ELISAs), with or without previous acidic immunocomplex dissociation (ICD), in a series of samples at different stages of human immunodeficiency virus type 1 (HIV-1) infection. The limit of viral detection of ELAST was assessed with fresh HIV-1 preparations quantified by reverse transcription-PCR and with the P24 antigen (Ag) Sanofi Pasteur Calibrator containing lyophilized virus. The P24 Ag detection capacity of ELAST was compared with that of NASBA in samples obtained from infected subjects with less than 250 CD4
+
cells. The results of the present study show that ELAST was the most sensitive method for detecting P24 Ag compared to classic ELISA and ICD plus ELISA. ELAST was able to detect 0.5 pg of P24 Ag per ml in a whole virus preparation and the equivalent of 330 to 1,000 RNA copies/ml of HIV. The rate of detection of P24 Ag was always higher in subjects with low levels of anti-P24 antibodies. The number of positive results was dramatically enhanced (from 37% to 94% for subjects with <250 CD4
+
cells) when the incubation period was prolonged from 1 to 16 h. In a third series of 84 samples (<250 CD4
+
cells) tested in parallel, NASBA yielded 83% of the positive results and ELAST yielded 79%. Considering the high sensitivity, low cost, simplicity of equipment (only a plate reader), and possibility for full automation, ELAST appears to be a promising new tool for measuring viral load, especially in areas with few resources, in which the procedures based on molecular biology techniques may be difficult to install.
Detection of HumanToxoplasma-Specific Immunoglobulins A, M, and G with a RecombinantToxoplasma gondiiRop2 Protein
Authors: Martin Valentina , Arcavi Miriam , Santillan Graciela , Amendoeira Maria Regina R. , De Souza Neves Elizabeth , Griemberg Gloria , Guarnera Eduardo , Garberi Juan C. , Angel Sergio O.
Published: December 2019

TheToxoplasma gondiirhoptry protein Rop2 was expressed inEscherichia colias a fusion protein containing 44 kDa of the 55-kDa mature Rop2, supplied with six histidyl residues at the N-terminal end (Rop2196–561). Humoral response duringToxoplasmainfection of humans was analyzed by immunoglobulin G (IgG), IgA, and IgM enzyme-linked immunosorbent assay with Rop2196–561as the antigen substrate. The analyzed sera were divided according toT. gondii-specific serological tests (IgG, IgA, or IgM indirect immunofluorescence and IgA or IgM immunosorbent agglutination assay) as group A (IgG+IgA−IgM−;n= 35), group B (IgG+IgA+IgM+;n= 21), group C (IgG+IgA+IgM−;n= 5), and group D (IgG+IgA−IgM+;n= 16). Twenty-sixT. gondii-seronegative sera from individuals with other infections were also included (group E). Anti-Rop2 IgG antibodies were detected in 82.8% of group A sera and in 97.6% of the sera with acute-phase marker immunoglobulins (groups B, C, and D). The percentage of IgA antibody reactivity against Rop2196–561was 17.1% in group A, 50% in group D, and 80.8% in groups B and C. The percentage of IgM antibody reactivity was 0% in groups A and C and 62% in groups B and D. Sera from group E failed to show IgA, IgM, or IgG antibody reactivity. SinceT. gondiiRop2 elicits a strong humoral response from an early stage of infection, it is suggested that recombinant Rop2196–561would be suitable for use in diagnostic systems, in combination with otherT. gondiiantigens, to detect specific IgG, IgA, and IgM antibodies.
Low Relative Frequencies of CD26
+
CD4
+
Cells in Long-Term Nonprogressing Human Immunodeficiency Virus Type 1-Infected Subjects
Authors: Broström Christina , Sönnerborg Anders , Lindbäck Stefan , Gaines Hans
Published: December 2019

A broad antibody panel was used for immunophenotyping of human immunodeficiency virus type 1 (HIV-1)-infected patients who were long-term nonprogressors (LTNP). The LTNP were compared with patients in the early phase of infection and patients who had progressed to advanced immunodeficiency. Changes in CD8
+
subset distribution were observed mainly at acquisition of HIV-1 infection, whereas CD4
+
subset changes appeared during progression of HIV-1 infection. The decreasing levels of CD4
+
cells were characterized by an increasing frequency of cells expressing the activation markers HLA-Dr and CD45RO but not the CD28 surface antigen. The LTNP exhibited significant changes compared to HIV-negative patients in almost all markers. Compared to patients in the early phase of infection, the only difference was a relatively lower frequency of CD4
+
cells expressing CD26 among the LTNP. The results show that HIV-1-infected persons who have no signs of immunodeficiency despite many years of infection have an immunophenotypic pattern that is substantially different from that of noninfected persons. Despite the long duration of infection, the LTNP exhibit a pattern similar to that of newly infected persons, with the exception of lower expression of CD26 on CD4
+
cells.
Role of Arachidonic Acid and Its Metabolites in the Priming of NADPH Oxidase in Human Polymorphonuclear Leukocytes by Peritoneal Dialysis Effluent
Authors: Daniels I. , Lindsay M. A. , Keany C. I. C. , Burden R. P. , Fletcher J. , Haynes A. P.
Published: December 2019

Peritoneal dialysis effluent (PDE) contains a low-molecular-weight solute that will activate and prime the NADPH oxidase of human neutrophils via a phospholipase A2(PLA2)-dependent mechanism. Since the products of PLA2are known to activate and prime the oxidase we have investigated their role in the dialysis effluent-mediated activation and priming of human neutrophils. NADPH oxidase activity of PDE-primed and -unprimed neutrophils was measured by lucigenin-enhanced chemiluminescence in the presence of known inhibitors of the arachidonic acid cascade. Incubation of neutrophils with the nonselective PLA2inhibitor quinacrine (0 to 100 μM) reduced oxidase activity in both primed and unprimed cells. Furthermore, primed cells were more sensitive to the action of quinacrine than were unprimed cells. We were unable to determine the relative roles of secretory PLA2(sPLA2) and cytosolic PLA2(cPLA2) since the selectivesPLA2inhibitor scalaradial (0 to 100 μM) inhibited oxidase activity in both groups of cells by similar degrees, while the specificcPLA2inhibitor AACO-CF3(0 to 50 μM) failed to affect activity in either group. Inhibition of platelet-activating factor (PAF), cycloxygenase, and 5-lipoxygenase-activating protein by hexanolamino-PAF (0 to 25 μM), flurbiprofen (0 to 25 μM), and MK886 (0 to 5 μM), respectively, had no effect upon oxidase activity. However, the direct inhibition of 5-lipoxygenase by caffeic acid or lipoxin A4resulted in a similar concentration-dependent attenuation of oxidase activity in both primed and unprimed cells. Leukotriene B4(LTB4) release from primed neutrophils was comparable to that from unprimed cells with the exception of phorbol myristate acetate-stimulated cells, which released fivefold more LTB4than control. Taken together, these results suggest that it is arachidonic acid per se, and not its metabolites, that is important in priming of the neutrophil NADPH oxidase by dialysis effluent.
Effects of the Nature of Adjuvant and Site of Parenteral Immunization on the Serum and Mucosal Immune Responses Induced by a Nasal Boost with a Vaccine Alone
Authors: Guy Bruno , Fourage Sophie , Hessler Catherine , Sanchez Violette , Millet Marie José Quentin
Published: December 2019

Outbred OF1 mice were immunized subcutaneously with flu vaccine, either in the neck or in the lumbar region (back), in combination with adjuvants inducing either a Th1- or a Th2-type response, referred to as adjuvants A1 and A2, respectively. After two parenteral immunizations, the mice were boosted intranasally with nonadjuvanted vaccine. The serum response was analyzed after each immunization by measuring specific immunoglobulin A (IgA), IgG1, and IgG2a antibody levels, while the local response (same isotypes) was measured in the salivary glands after the mucosal boost by ELISPOTs. We observed that systemic priming at any of the two sites with a Th2 rather than a Th1 adjuvant dramatically enhanced the mucosal IgG1 and IgA responses following a mucosal boost with unadjuvanted vaccine. In addition, as judged by the IgG2a/IgG1 ratios and serum IgA levels, immunization of mice in the back induced a rise in Th2 response compared to neck immunization with adjuvant A1. In contrast, such back immunization with adjuvant A2 reversed the Th1-Th2 balance in favor of the Th1 response compared to neck immunization. Similar differences were observed in mucosal antibody levels according to the site of priming with one given adjuvant; priming in the back with adjuvant A1 increased the mucosal IgA and IgG1 responses compared to neck priming, while the local IgG2a levels were decreased. The reverse was true for adjuvant A2. Back versus neck priming with this latter adjuvant decreased the mucosal IgG1 response, while local IgG2a levels were increased. The different lymphatic drainages of the two sites of parenteral immunization may explain these differences, due to the targeting of particular lymphoid inductive sites. Some of these sites may represent crossroads between systemic and mucosal immunity.
Increases in the Levels ofCoxiella burnetii-Specific Immunoglobulin G1 and G3 Antibodies in Acute Q Fever and Chronic Q Fever
Authors: Capo Christian , Iorgulescu Ioana , Mutillod Maryse , Mege Jean-Louis , Raoult Didier
Published: December 2019

A detailed analysis of the humoral response toCoxiella burnetiimay provide insight into the pathogenesis of Q fever, a zoonosis caused byC. burnetii. The subclasses ofC. burnetii-specific antibodies were determined by immunofluorescence in 20 patients with acute Q fever and 20 patients with chronic Q fever. Although immunoglobulin G1 (IgG1) and IgG3 antibodies were found in acute and chronic Q fever, neither IgG2 nor IgG4 was detected. The detection of IgG1 and IgG3 antibodies was not due to an increase of the IgG1 and IgG3 subclasses. Moreover, IgG1 and IgG3 antibodies were not correlated, suggesting that they may play different roles in Q fever.
Pulmonary Bovine-Type Tuberculosis in Rabbits: Bacillary Virulence, Inhaled Dose Effects, Tuberculin Sensitivity, andMycobacterium vaccaeImmunotherapy
Authors: Converse P. J. , Dannenberg A. M. , Shigenaga T. , McMurray D. N. , Phalen S. W. , Stanford J. L. , Rook G. A. W. , Koru-Sengul T. , Abbey Helen , Estep J. E. , Pitt M. L. M.
Published: December 2019

This report elucidates four aspects of the immunology of pulmonary tuberculosis produced in rabbits: (i) the virulence of bovine-type tubercle bacilli, strain Ravenel S, (ii) systemic factors influencing the generation of visible primary pulmonary tubercles, (iii) differences in tuberculin sensitivity of rabbits and humans, and (iv) the effect ofMycobacterium vaccaeimmunotherapy on cavitary tuberculosis. Laboratory strain Ravenel S (ATCC 35720) was not fully virulent. Fully virulent strains produce one visible primary pulmonary tubercle for each three bacillary units inhaled. Strain ATCC 35720 produced one such tubercle for each 18 to 107 bacillary units inhaled, indicating that its virulence was reduced by 6- to 36-fold. When a low dose of this Ravenel S strain was inhaled, the host resistance (measured by the number of inhaled bacilli needed to generate one visible primary pulmonary tubercle) was increased at least 3.5-fold compared to the host resistance when a high dose was inhaled. Rabbits and humans differ in the degree and in the maintenance of their dermal sensitivities to tuberculin. Compared to rabbits, humans are 100 times more sensitive to tuberculin. Also, at 33 weeks rabbits with well-controlled cavitary tuberculosis usually showed a decrease in their tuberculin reactions of about 50% from peak values, whereas humans with such well-controlled tuberculosis are thought to maintain strong reactions for many years. These species differences may be due to desensitization to group II mycobacterial antigens in the rabbits because they have a different diet and a different type of digestive tract.M. vaccaeimmunotherapy of rabbits with cavitary tuberculosis produced no statistically significant effects. Experiments with many more rabbits would be required to prove whether or not such immunotherapy is beneficial.
Streptococcal DNase B Is Immunologically Identical to Superantigen SpeF but Involves Separate Domains
Authors: Eriksson Anna , Eriksson Björn , Holm Stig E. , Norgren Mari
Published: December 2019

The previous suggestion that streptococcal superantigen SpeF might be identical to DNase B was confirmed in this study. Polyclonal SpeF-specific antisera were able to inhibit depolymerization of methyl-green DNA by DNase B. However, T-cell mitogenicity and nuclease activity appear to involve separate immune epitopes on SpeF, since sera with the capacity to neutralize the mitogenic activity of SpeF did not always inhibit the DNase activity.
Incidence of Human Immunodeficiency Virus Antibody in a Prenatal Population at a Community Hospital
Authors: Alexander Thomas S. , Lee Joanne , Yen-Lieberman Belinda
Published: December 2019

Prenatal human immunodeficiency virus (HIV) screening may reduce vertical HIV transmission. We screened 4,419 prenatal sera and found 38 repeatedly reactive specimens with an HIV-1–HIV-2 enzyme-linked immunosorbent assay. Western blot analysis confirmed four of these specimens as positive for HIV-1 antibodies. Screening detects previously unidentified HIV infections, but false-positive results may also occur.
Prevalence ofBartonella henselaeandBartonella clarridgeiaein an Urban Indonesian Cat Population
Authors: Marston Eric L. , Finkel Barbara , Regnery Russell L. , Winoto Imelda L. , Graham R. Ross , Wignal Steven , Simanjuntak Gindo , Olson James G.
Published: December 2019

We studied evidence ofBartonella henselaeandBartonella clarridgeiaeinfection in 54 cats living in Jakarta, Indonesia. By using an indirect immunofluorescence assay, we found immunoglobulin G antibody toB. henselaein 40 of 74 cats (54%). The blood of 14 feral cats was cultured on rabbit blood agar plates for 28 days.Bartonella-like colonies were identified asB. henselaeorB. clarridgeiaeby using restriction fragment length polymorphism analysis and direct sequencing of the PCR amplicons. Of the cats sampled in the study, 6 of 14 (43%; all feral) were culture positive forB. henselae; 3 of 14 (21%; 2 feral and 1 pet) culture positive forB. clarridgeiae. This is the first report that documentsB. henselaeandB. clarridgeiaeinfections in Indonesian cats.
Potential Value of Major Antigenic Protein 2 for Serological Diagnosis of Heartwater and Related Ehrlichial Infections
Authors: Bowie Michael V. , Reddy G. Roman , Semu Shalt M. , Mahan Suman M. , Barbet Anthony F.
Published: December 2019

Cowdria ruminantium
is the etiologic agent of heartwater, a disease causing major economic loss in ruminants in sub-Saharan Africa and the Caribbean. Development of a serodiagnostic test is essential for determining the carrier status of animals from regions where heartwater is endemic, but most available tests give false-positive reactions with sera against related
Erhlichia
species. Current approaches rely on molecular methods to define proteins and epitopes that may allow specific diagnosis. Two major antigenic proteins (MAPs), MAP1 and MAP2, have been examined for their use as antigens in the serodiagnosis of heartwater. The objectives of this study were (i) to determine if MAP2 is conserved among five geographically divergent strains of
C. ruminantium
and (ii) to determine if MAP2 homologs are present in
Ehrlichia canis
, the causative agent of canine ehrlichiosis, and
Ehrlichia chaffeensis
, the organism responsible for human monocytic ehrlichiosis. These two agents are closely related to
C. ruminantium
. The
map2
gene from four strains of
C. ruminantium
was cloned, sequenced, and compared with the previously reported
map2
gene from the Crystal Springs strain. Only 10 nucleic acid differences between the strains were identified, and they translate to only 3 amino acid changes, indicating that MAP2 is highly conserved. Genes encoding MAP2 homologs from
E. canis
and
E. chaffeensis
also were cloned and sequenced. Amino acid analysis of MAP2 homologs of
E. chaffeensis
and
E. canis
with MAP2 of
C. ruminantium
revealed 83.4 and 84.4% identities, respectively. Further analysis of MAP2 and its homologs revealed that the whole protein lacks specificity for heartwater diagnosis. The development of epitope-specific assays using this sequence information may produce diagnostic tests suitable for
C. ruminantium
and also other related rickettsiae.
Genetic and Serological Analysis of Lipoprotein LppA in
Mycoplasma mycoides
subsp.
mycoides
LC and
Mycoplasma mycoides
subsp.
capri
Authors: Monnerat Marie-Pierre , Thiaucourt François , Poveda Jose B. , Nicolet Jacques , Frey Joachim
Published: December 2019

The genes encoding the 62-kDa lipoproteins from the
Mycoplasma mycoides
subsp.
mycoides
large-colony type (LC) strain Y-goat and the
M. mycoides
subsp.
capri
strain PG3 were cloned and analyzed by sequencing. These two lipoproteins have been named LppA[MmymyLC] and LppA[Mmyca], and their corresponding genes have been named
lppA
[MmymyLC] and
lppA
[Mmyca], respectively. The nucleotide and deduced amino acid sequences of these two lipoproteins showed a very high degree of similarity between these two mycoplasmas. Given the sequence data, LppA seems to fulfill the same structural functions as the previously described major lipoproteins P72 of
M. mycoides
subsp.
mycoides
small-colony type and P67 of the
Mycoplasma
species bovine group 7. Based on
lppA
gene sequences of
M. mycoides
subsp.
mycoides
LC and
M. mycoides
subsp.
capri
type strains, a specific PCR assay was developed so that it amplified this gene in all field strains of the two species analyzed in this study but not in the other members of the
M. mycoides
cluster. Analysis of the PCR-amplified
lppA
genes with frequently cutting restriction enzymes showed a certain degree of genetic variability which, however, did not cluster the two subspecies. This PCR therefore allows a rapid identification of
M. mycoides
subsp.
mycoides
LC and
M. mycoides
subsp.
capri
but does not distinguish between these two closely related subspecies. LppA was expressed in
Escherichia coli
K-12 and used for the production of polyclonal mouse antiserum. Antibodies against recombinant LppA[MmymyLC] reacted with a 62-kDa protein in all
M. mycoides
subsp.
mycoides
LC and
M. mycoides
subsp.
capri
type strains and field strains tested but not with the other members of the
M. mycoides
cluster, thus showing the antigenic specificity of LppA and further supporting the concept that a close relationship exists between these two mycoplasmas.
Identification of a Peptide from Mammal Albumins Responsible for Enhanced Pigment Production by Group B Streptococci
Authors: Rosa-Fraile Manuel , Sampedro Antonio , Varela Javier , Garcia-Peña Marisa , Gimenez-Gallego Guillermo
Published: December 2019

The peptide from peptones responsible for enhanced pigment production by
Streptococcus agalactiae
in culture media has been isolated from a peptic digest of human albumin and has been identified as Ile-Ala-Arg-Arg-His-Pro-Tyr-Phe. The related heptapeptide lacking the N-terminal Ile also had pigment-enhancing activity. A sequence similarity search showed that these sequences are present only in mammal albumins.
Levels of Proinflammatory Cytokines in Plasma after Pneumoccoccal Immunization in Human Immunodeficiency Virus Type 1-Infected Patients
Authors: Valdez Hernan , Purvis Scott , Asaad Robert , Valerio Ian , Sha Beverly E. , Landay Alan L. , Lederman Michael M.
Published: December 2019

To ascertain if immunization with pneumococcal polysaccharide vaccine is associated with rises in the levels of proinflammatory cytokines in the plasma of human immunodeficiency virus type 1 (HIV-1)-infected patients, the levels of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) were measured serially after immunization. IL-6 levels rose an average of 2.2- and 2.1-fold 6 and 8 h after immunization, respectively, but TNF-α levels remained unchanged. The levels of these cytokines were stable in unimmunized controls. Immunization with pneumococcal polysaccharide vaccine induces increases in the levels of IL-6 in the plasma of persons with HIV-1 infection.
Depletion of Alveolar Macrophages by Treatment with 2-Chloroadenosine Aerosol
Authors: Kubota Yutaka , Iwasaki Yoshinobu , Harada Hidehiko , Yokomura Ichiro , Ueda Mikio , Hashimoto Shinichi , Nakagawa Masao
Published: December 2019

Alveolar macrophages (AMs) are localized in the alveoli and alveolar ducts of the lung and are the only macrophages living in an aerobic environment. Recent studies have demonstrated that AMs play a central role in lung diseases, such as pneumonia and acute respiratory distress syndrome. It has become important to find a simple, effective way to eliminate AMs in order to investigate the function of AMs in vivo. 2-Chloroadenosine (2-CA), a purine analog, is reported to be selectively cytotoxic to cultured macrophages, and we hypothesized that it would deplete the number of AMs in the bronchoalveolar lavage fluid (BALF) of mice without any effect on neutrophil or lymphocyte counts. After mice had inhaled 1 mM aerosolized 2-CA for 2 h, AMs were found to be significantly depleted at 0 h [(4.42 ± 0.16) × 10
4
/ml], 24 h [(4.17 ± 0.89) × 10
4
/ml], 48 h [(3.17 ± 0.21) × 10
4
/ml], and 72 h [(5.00 ± 0.64) × 10
4
/ml] compared with concentrations in untreated controls [(12.1 ± 0.21) × 10
4
/ml]. Neutrophil and lymphocyte counts in BALF did not change and histological changes in the lung were not observed after 2-CA treatment. The lung wet-to-dry weight ratio did not change at 0, 24, and 48 h after 2-CA aerosol application. The 2-CA aerosol had no effect on lung vascular permeability, as assessed by the intravenous administration of Evans blue, or on other phagocytes, as assessed by Kupffer cell counts. Our study demonstrates the efficacy of 2-CA in reducing AM numbers in vivo.
Diagnostic and Prognostic Potential of a Competitive Enzyme-Linked Immunosorbent Assay for Leishmaniasis in India
Authors: Chatterjee Mitali , Jaffe Charles L. , Sundar Shyam , Basu Debasis , Sen Sandeep , Mandal Chitra
Published: December 2019

ALeishmania donovanispecies-specific monoclonal antibody (monoclonal antibody D2) was evaluated for its diagnostic and prognostic potential by a competitive enzyme-linked immunosorbent assay (C-ELISA) in sera from Indian patients with visceral leishmaniasis (VL) and seven patients with post-kala-azar dermal leishmaniasis (PKDL). These results were compared with those obtained by microscopy with Giemsa-stained tissue smears and a direct enzyme-linked immunosorbent assay (direct ELISA) with crude parasite antigen. Of 121 patients with clinically diagnosed VL examined, 103 (85.1%) were positive and 11 (9.1%) were negative by all three methods. An additional 7 (5.8%) who were negative by microscopy were positive by both C-ELISA and direct ELISA. Seven PKDL patients were also examined and were found to be positive by all three methods. Analysis of the chemotherapeutic response to sodium antimony gluconate of these 110 serologically positive VL patients showed that 57 (51.8%) were drug responsive and 53 (48.2%) were drug resistant. The C-ELISA with sera from 20 longitudinally monitored VL patients before and after chemotherapy showed a significant decrease in percent inhibition of monoclonal antibody D2 in drug-responsive patients. However, in drug-unresponsive patients, the percent inhibition of D2 was unchanged or was slightly increased. Our results therefore indicate (i) the applicability ofL. donovanispecies-specific monoclonal antibody D2 for sensitive and specific serodiagnosis by C-ELISA, (ii) that the C-ELISA is more sensitive than microscopy, especially for early diagnosis, (iii) thatL. donovaniis still the main causative agent of VL, irrespective of the chemotherapeutic response, and (iv) that the C-ELISA can be used to evaluate the success of drug treatment.
Pentoxifylline Inhibits Superantigen-Induced Toxic Shock and Cytokine Release
Authors: Krakauer Teresa , Stiles Bradley G.
Published: December 2019

Tumor necrosis factor alpha (TNF-α) is a critical cytokine that mediates the toxic effects of bacterial superantigens like staphylococcal enterotoxin B (SEB) and toxic shock syndrome toxin 1 (TSST-1). Pentoxifylline, an anti-inflammatory agent that inhibits endotoxemia and lipopolysaccharide (LPS)-induced release of TNF-α, was tested for its ability to inhibit SEB- and TSST-1-induced activation of human peripheral blood mononuclear cells (PBMCs) in vitro and toxin-mediated shock in mice. Stimulation of PBMCs by SEB or TSST-1 was effectively blocked by pentoxifylline (10 mM), as evidenced by the inhibition of TNF-α, interleukin 1β (IL-1β), gamma interferon (IFN-γ), and T-cell proliferation. The levels of TNF-α, IL-1α, and IFN-γ in serum after an SEB or TSST-1 injection were significantly lower in mice given pentoxifylline (5.5 mg/animal) versus control mice. Additionally, pentoxifylline diminished the lethal effects and temperature fluctuations elicited by SEB and TSST-1. Thus, in addition to treating endotoxemias, the cumulative in vitro and in vivo data suggest that pentoxifylline may also be useful in abrogating the ill effects of staphylococcal enterotoxins and TSST-1.
Conservation of Expression and N-Terminal Sequences of the
Pasteurella haemolytica
31-Kilodalton and
Pasteurella trehalosi
29-Kilodalton Periplasmic Iron-Regulated Proteins
Authors: Tabatabai Louisa B. , Frank Glynn H.
Published: December 2019

This study examined the conservation of expression of a 31-kDa iron-regulated protein by serotypes of
Pasteurella haemolytica
and
Pasteurella trehalosi
associated with pasteurellosis of cattle and sheep. A polyclonal antibody prepared against the purified 31-kDa periplasmic iron-regulated protein from
P. haemolytica
serotype A1 showed that all
P. haemolytica
serotypes expressed similar 31-kDa proteins with identical N-terminal sequences, whereas
P. trehalosi
serotypes expressed immunologically different 29-kDa proteins with a different N-terminal sequence. Antibody to the 31-kDa iron-regulated protein was a useful tool to distinguish similarities and differences of the iron-regulated proteins of
P. haemolytica
and
P. trehalosi
.
Redesignation of a Purported P1.15 Subtype-Specific Meningococcal Monoclonal Antibody as a P1.19-Specific Reagent
Authors: Wedege E. , Caugant D. A. , Musacchio A. , Saunders N. B. , Zollinger W. D.
Published: December 2019

Two reference monoclonal antibodies against the meningococcal P1.15 subtype PorA, MN3C5C and 2-1-P1.15, showed only partial concordant recognition of meningococcal isolates. Cyanogen bromide cleavage of P1.19,15 PorA, peptide mapping, and sequencing of
porA
regions demonstrated that 2-1-P1.15 was specific for subtype P1.19, and henceforth it is to be redesignated as 2-1-P1.19.