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1.
J Appl Microbiol ; 123(5): 1168-1183, 2017 Nov.
Article in English | MEDLINE | ID: mdl-28853204

ABSTRACT

AIMS: The work aimed at developing and evaluating practically relevant methods for testing of disinfectants on contaminated personal protective equipment (PPE). METHODS AND RESULTS: Carriers were prepared from PPE fabrics and contaminated with Bacillus subtilis spores. Peracetic acid (PAA) was applied as a suitable disinfectant. In method 1, the contaminated carrier was submerged in PAA solution; in method 2, the contaminated area was covered with PAA; and in method 3, PAA, preferentially combined with a surfactant, was dispersed as a thin layer. In each method, 0·5-1% PAA reduced the viability of spores by a factor of ≥6 log10 within 3 min. The technique of the most realistic method 3 proved to be effective at low temperatures and also with a high organic load. Vaccinia virus and Adenovirus were inactivated with 0·05-0·1% PAA by up to ≥6 log10 within 1 min. The cytotoxicity of ricin was considerably reduced by 2% PAA within 15 min of exposure. CONCLUSIONS: PAA/detergent mixture enabled to cover hydrophobic PPE surfaces with a thin and yet effective disinfectant layer. SIGNIFICANCE AND IMPACT OF THE STUDY: The test methods are objective tools for estimating the biocidal efficacy of disinfectants on hydrophobic flexible surfaces.


Subject(s)
Disinfectants/pharmacology , Disinfection/methods , Peracetic Acid/pharmacology , Personal Protective Equipment/microbiology , Bacillus subtilis/drug effects , Bacillus subtilis/growth & development , Disinfection/instrumentation
2.
Analyst ; 139(22): 5885-92, 2014 Nov 21.
Article in English | MEDLINE | ID: mdl-25237676

ABSTRACT

Simultaneous detection of small and large molecules on microarray immunoassays is a challenge that limits some applications in multiplex analysis. This is the case for biosecurity, where fast, cheap and reliable simultaneous detection of proteotoxins and small toxins is needed. Two highly relevant proteotoxins, ricin (60 kDa) and bacterial toxin staphylococcal enterotoxin B (SEB, 30 kDa) and the small phycotoxin saxitoxin (STX, 0.3 kDa) are potential biological warfare agents and require an analytical tool for simultaneous detection. Proteotoxins are successfully detected by sandwich immunoassays, whereas competitive immunoassays are more suitable for small toxins (<1 kDa). Based on this need, this work provides a novel and efficient solution based on anti-idiotypic antibodies for small molecules to combine both assay principles on one microarray. The biotoxin measurements are performed on a flow-through chemiluminescence microarray platform MCR3 in 18 minutes. The chemiluminescence signal was amplified by using a poly-horseradish peroxidase complex (polyHRP), resulting in low detection limits: 2.9 ± 3.1 µg L(-1) for ricin, 0.1 ± 0.1 µg L(-1) for SEB and 2.3 ± 1.7 µg L(-1) for STX. The developed multiplex system for the three biotoxins is completely novel, relevant in the context of biosecurity and establishes the basis for research on anti-idiotypic antibodies for microarray immunoassays.


Subject(s)
Enterotoxins/analysis , Immunoassay/methods , Ricin/analysis , Saxitoxin/analysis , Calibration , Luminescence
3.
Analyst ; 138(20): 6154-62, 2013 Oct 21.
Article in English | MEDLINE | ID: mdl-23971072

ABSTRACT

This article describes a novel bioluminescence assay for detecting the proteolytic activity of Botulinum NeuroToxins (BoNT) in complex matrices. The assay is capable of detecting traces of BoNT in blood samples as well as in food drinks. The assay was responsive to BoNT/A subtypes 1 to 5, and serotype E3 in buffered solutions. It was responsive to filtered Clostridium botulinum supernatants and BoNT/A1 in complex with neurotoxin associated proteins in bouillon and milk (3.8% fat) down to 400 fM after 4 h RT incubation and in bouillon at concentrations down to 120 fM after 21 h RT incubation. In combination with an immunocapture/enrichment step it could detect BoNT/A1 in citrated plasma at concentrations down to 30 fM (1.2 mouse LD50 per mL). The simplicity of the assay, combined with a demonstrated ability to lyophilize the reagents, demonstrates its usefulness for detection of BoNT in non-specialised analytical laboratories.


Subject(s)
Botulinum Toxins, Type A/analysis , Botulinum Toxins, Type A/chemistry , Luminescent Measurements/methods , Animals , Clostridium botulinum/chemistry , Humans , Mice , Mice, Inbred BALB C , Protein Structure, Secondary
4.
J Med Primatol ; 42(4): 220-4, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23617545

ABSTRACT

Few lethal pathogens in wild-living primates have been described, and little is known about infectious diseases of the reproductive tract and their possible impact on health and reproduction. This report describes the pathology and isolation of an alpha-toxin producing strain of Clostridium septicum in a case of necrotizing endometritis in a wild sooty mangabey found dead in a tropical rainforest of West Africa.


Subject(s)
Bacterial Toxins/biosynthesis , Cercocebus atys , Clostridium Infections/veterinary , Clostridium septicum/metabolism , Endometritis/veterinary , Monkey Diseases/microbiology , Animals , Clostridium septicum/isolation & purification , Cote d'Ivoire , Endometritis/microbiology , Endometritis/pathology , Female , Necrosis
5.
Poult Sci ; 88(2): 281-90, 2009 Feb.
Article in English | MEDLINE | ID: mdl-19151341

ABSTRACT

One of the key benefits in using chickens for immunization is the high yield of antibodies obtainable. It is known that egg production decreases over time, while animal maintenance costs remain stable. It would, however, be desirable to keep hens as long as possible to obtain maximal amounts of antibodies. To identify a suitable length of time that animals can be kept and to optimize the cost:yield ratio, we monitored the number of eggs laid, the total amount of chicken IgY, and the specific antibody titer from individually prepared eggs over a 2-yr period. The plant toxin ricin and the Clostridium botulinum neurotoxins type A and B were used to immunize 4 chickens. The number of eggs laid in 2 yr was approximately 600 per hen (about 80% of the maximum egg number), yielding about 20 to 40 g of total IgY per hen. A stable antibody titer of 1:100,000 to 1:1,000,000, as measured by ELISA, was obtained following up to 11 injections of 10 to 20 microg of immobilized native toxin. Laying capacities were found to decrease, on average, from 7 eggs/wk at the point of first immunization to 2 eggs/wk after more than 2 yr. In parallel, the yield of total and specific IgY increased over time, so that the antibody recovery remained high, even after prolonged immunization times. Using purified IgY preparations, classical immunological assays such as ELISA and Western blotting were performed. Furthermore, the IgY showed neutralizing capacity when used to block the functional activity of the toxins both in vitro and in vivo. Analysis of the total IgY content over time demonstrated a complex biological oscillation (and the antigen-specific titer), with a shorter time period of around 7 d (circaseptan rhythm). In summary, we successfully immunized chickens with ricin and botulinum neurotoxins and monitored laying capacity, IgY concentration, and specific antibody titer over an extended period of 2 yr.


Subject(s)
Chickens/physiology , Eggs/analysis , Immunoglobulins/analysis , Immunotoxins/blood , Oviposition/immunology , Animals , Botulinum Toxins/immunology , Chickens/immunology , Female , Immunization/veterinary , Ricin/immunology
6.
Phys Rev E Stat Nonlin Soft Matter Phys ; 69(6 Pt 1): 061203, 2004 Jun.
Article in English | MEDLINE | ID: mdl-15244550

ABSTRACT

A high resolution (1.5 meV) inelastic neutron scattering experiment was carried out, aiming at an accurate investigation of the high frequency and low momentum dynamic response in heavy water. The experimental data confirm the existence of a dispersionless mode, besides the ordinary longitudinal collective dynamics. A simplified model, based on the interaction of two vibrational branches, is proposed to interpret the observed features of the dynamic spectra. The validity extent of this scheme is proved by applying it to room temperature neutron and x-ray data, to temperature and pressure dependent x-ray data, and to room temperature neutron data of vibrational density of states. The overall successfull results provided by this model, in conjunction with the combined analysis of the x-ray and neutron data on collective dynamics, enable a deeper insight into the complex mechanisms of the water dynamics and provide a simple phenomenological explanation for the transition from ordinary to fast sound.

7.
Bone Marrow Transplant ; 31(10): 927-30, 2003 May.
Article in English | MEDLINE | ID: mdl-12748671

ABSTRACT

The so-called pinch-off syndrome is observed in up to 1% of all central venous catheters (CVCs), and is a valuable warning prior to fragmentation, which occurs in approximately 40% of the respective cases. As long-term indwelling CVCs are used with increasing frequency, this paper describes the necessity of pinch-off monitoring following the experiences of a case study and a review of the current literature on this specific topic in order to point out preventive practice guidelines. Besides easy preventive practices such as a high level of suspicion and adequate X-ray controls, findings give strong evidence that the most important specific factor might be the adequate approach. In our hands, the supraclavicular technique has provided the best results with regards to percutaneous introduction of large bore CVCs.


Subject(s)
Catheterization, Central Venous/adverse effects , Equipment Failure , Adolescent , Bone Marrow Transplantation , Female , Humans , Monitoring, Physiologic , Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy , Radiography, Thoracic , Transplantation, Autologous
8.
Br J Cancer ; 88(4): 496-501, 2003 Feb 24.
Article in English | MEDLINE | ID: mdl-12592361

ABSTRACT

A total of 55 patients with histologically proven glioblastoma multiforme (total gross resection: n=24, subtotal resection: n=20, stereotactic biopsy: n=11) were treated with the combination of dacarbazine (D) (200 mg m(-2)) and fotemustine (F) (100 mg m(-2)) and concomitant radiotherapy (2 Gy day(-1), 5 days per week using limited fields up to 60 Gy) to assess efficacy and toxicity of this regimen. Survival (median survival, 12-, 18- and 24-month survival rates) and time to progression (median time to progression (TTP), 6-month progression-free survival) were analysed by Kaplan-Meier's method. A total of 268 (range 1-8, median: 5) cycles were administered. Median survival is 14.5+ (range: 0.5-40+) months, and the 12-, 18- and 24-month survival rates are 58, 29 and 23%, respectively. Median TTP from the start of D/F therapy is 9.5+ (range: 0.5-33+) months. The 6-month progression-free survival is 54%. Partial remissions were observed in 3.6%. Main toxicity was thrombocytopenia. Five patients were excluded from further D/F application, four patients because of prolonged thrombocytopenia NCI-CTC grades 3 and 4 and one patient because of whole body erythrodermia. One patient died because of septic fever during thrombocytopenia and leukopenia NCI-CTC grade 4 after the first cycle. No other toxicities of NCI-CTC grade 3 or 4 occurred. The treatment is feasible in a complete outpatient setting and the results of the D/F regimen justify further investigations with these compounds.


Subject(s)
Dacarbazine/adverse effects , Dacarbazine/therapeutic use , Glioblastoma/diagnosis , Glioblastoma/drug therapy , Nitrosourea Compounds/adverse effects , Nitrosourea Compounds/therapeutic use , Organophosphorus Compounds/adverse effects , Organophosphorus Compounds/therapeutic use , Adolescent , Adult , Aged , Disease Progression , Female , Humans , Male , Middle Aged , Survival Analysis , Survival Rate
9.
Eur Phys J E Soft Matter ; 12 Suppl 1: S21-4, 2003 Nov.
Article in English | MEDLINE | ID: mdl-15011008

ABSTRACT

The temperature evolution of the structure of NaNO(2) nanocomposite ferroelectric material in a porous glass with 7 nm pores was studied by neutron diffraction in temperature region from room temperature up to the melting, i.e. in the ferro- and paraelectric phases. It is demonstrated that in the ferroelectric phase the structure is consistent with the structure of the bulk, but above the ferroelectric phase transition (and up to approximately 513 K) a volume premelted state is formed, manifesting itself in a growth of amplitudes of ion thermal vibrations, a steep increase of elementary cell volume and "softening" of lattice. For the first time the temperature dependence of order parameter eta for confined sodium nitrite is determined. eta (T) follows a power law with T(C)=425.6+/- 2.1 K and beta= 0.31+/- 0.04, which is essentially different from that for bulk NaNO(2). Our obtained data are in a good agreement with the results of earlier dielectric and neutron diffraction measurements.

10.
Phys Rev Lett ; 87(21): 215504, 2001 Nov 19.
Article in English | MEDLINE | ID: mdl-11736350

ABSTRACT

The ion dynamics of liquid mercury was investigated by inelastic neutron scattering. By exploiting an optimized high-resolution ( approximately 1 meV) experimental configuration, the dynamic response function was accurately measured. Collective excitations extending up to 0.6 A(-1) were observed with an associated velocity of 2100+/-80 m/s. This value is notably greater than the sound velocity, but it is provided by a simple Bohm-Staver calculation. The latter finding emphasizes those electron-related features in the ion dynamics, which are common to systems as different as polyvalent and alkali metals.

11.
Onkologie ; 24(4): 376-9, 2001 Aug.
Article in German | MEDLINE | ID: mdl-11574766

ABSTRACT

Multimodal Therapy of High Grade Gliomas in Adults For chemotherapy of high-grade gliomas only few substances with proven efficacy are available. Today, even the newest substances are not proven to be superior to nitrosoureas. Although nitrosoureas are still considered as commonly accepted standard treatment, these results are based on studies in which response evaluation was not performed according to standardized criteria: prognostic criteria were not taken into account and statistical power of these studies is limited. However, a lot of questions remain to be dealt with and are currently under investigation in multicentric randomized studies. Therefore, patients should be encouraged to participate in prospectively randomized trials. With or after chemotherapy, supportive and palliative care including neurorehabilitation are important to improve patients' quality of life.


Subject(s)
Antineoplastic Agents/therapeutic use , Glioma/drug therapy , Antineoplastic Agents/adverse effects , Chemotherapy, Adjuvant , Combined Modality Therapy , Glioma/mortality , Glioma/radiotherapy , Humans , Radiotherapy, Adjuvant , Randomized Controlled Trials as Topic , Survival Rate
12.
Eur J Immunol ; 31(7): 2051-9, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11449358

ABSTRACT

The heat shock proteins (HSP) gp96, Hsp70 and Hsp60 activate professional antigen-presenting cells (APC) to secrete proinflammatory cytokines and to express costimulatory molecules. Here, we analyze the impact of Hsp60 as a hypothetical danger signal on the antigen-specific activation of T cells derived from DO11.10 TCR-transgenic mice. The release of IFN-gamma, induced by the antigenic OVA(323-339)-peptide, is increased and accelerated dramatically by the addition of Hsp60 to ex vivo purified populations of T cells and peritoneal macrophages (PEC), while the antigen-specific IL-2 production or proliferation of the T cells remain unchanged. In contrast, "effector" T cells, undergoing secondary stimulation, displayed almost unchanged activation kinetics in the presence of Hsp60. The presence of Hsp60 induces IFN-gamma and up-regulation of CD69 in T cell/PEC cocultures even in the absence of antigenic peptide and this induction of IFN-gamma is strictly dependent on the ability of the macrophages to produce IL-12. Taken together, our data strongly suggest that the presence of eukaryotic mitochondrial Hsp60 allows antigen-specific IFN-gamma secretion under conditions when an antigenic stimulus alone is not sufficient to activate T cells.


Subject(s)
Chaperonin 60/pharmacology , Interferon-gamma/biosynthesis , T-Lymphocytes/immunology , Animals , Antigens/immunology , Antigens, CD/metabolism , Antigens, Differentiation, T-Lymphocyte/metabolism , Cell Line , Cells, Cultured , Eukaryotic Cells/metabolism , Interleukin-2/biosynthesis , Kinetics , Lectins, C-Type , Macrophages, Peritoneal/immunology , Mice , Mice, Inbred BALB C , Mice, Transgenic , Ovalbumin/immunology , Peptide Fragments/immunology , Receptors, Antigen, T-Cell/genetics , Up-Regulation
13.
Article in English | MEDLINE | ID: mdl-11088861

ABSTRACT

The low momentum collective dynamics of heavy water has been investigated by means of neutron three-axis spectroscopy. Working at the resolution limit of this instrument, an energy resolution of 2.6 meV was achieved for a constant analyzer energy of 120 meV. This good resolution allowed us to establish definitely that the collective dynamics of water is dominated by the presence and interaction of opticlike and acousticlike branches, coupling at wave-vector transfers of 0.3 to 0.35 A(-1). The transition from slow to fast sound has been attributed to the interaction between opticlike modes and a solidlike fast acoustic mode.

14.
Clin Transplant ; 14(1): 8-10, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10693628

ABSTRACT

Infectious complications after renal transplantation remain a major cause of morbidity and mortality. Mucormycosis is a rare infection in renal transplant recipients; however, mortality is exceedingly high. Risk factors predisposing to this disease include prolonged neutropenia, diabetes, and patients who are immunosuppressed (Singh N, Gayowski T, Singh J, Yu LV. Invasive gastrointestinal zygomycosis in a liver transplant recipient: case report and review of zygomycosis in solid-organ transplant recipients, Clin Infect Dis 1995: 20: 617). Life-threatening infections can occur, as this fungus has the propensity to invade blood vessel endothelium, resulting in hematological dissemination. We report a case of cavitary Rhizopus lung infection, 2 months after renal transplantation, where the patient was treated successfully with Amphotericin B and surgical resection of the lesions with preservation of his allograft function. In this era of intensified immunosuppression, we may see an increased incidence of mucormycosis in transplant population. Invasive diagnostic work-up is mandatory in case of suspicion; Amphotericin B and, in selected cases, surgical resection are the mainstays of therapy.


Subject(s)
Diabetic Nephropathies/surgery , Kidney Transplantation , Lung Diseases, Fungal/diagnosis , Mucormycosis/diagnosis , Opportunistic Infections/diagnosis , Rhizopus , Humans , Lung Diseases, Fungal/drug therapy , Lung Diseases, Fungal/etiology , Male , Middle Aged , Mucormycosis/drug therapy , Mucormycosis/etiology , Opportunistic Infections/drug therapy , Opportunistic Infections/etiology
15.
Phys Rev Lett ; 85(25): 5352-5, 2000 Dec 18.
Article in English | MEDLINE | ID: mdl-11135994

ABSTRACT

The investigation of the ion dynamics in the liquid alloy K52Cs48, which has the same electron density as Rb, was carried out by neutron inelastic scattering. Well defined collective excitations were observed up to the maximum value of momentum transfer of the experiment. A scaling of the dispersion relation data was found to hold between K-Cs alloys, Rb and Cs. The suggested scaling points out the key role of the conduction electron density in the collective dynamics of alkali metals.

17.
J Am Diet Assoc ; 97(10 Suppl 2): S172-6, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9336583

ABSTRACT

Advance directives can promote and improve communication between the patient and the provider and ideally safeguard the resident's interests by directing medical care. Participation on the facility's ethics committee allows the dietitian to become involved in the legal and ethical issues regarding feeding and to promote the use of specific advance directives for nutrition and hydration. Consider the goals of patient care carefully and assure that they are individualized and patient-centered. Continually reassess nutritional status and collaborate with the health care team in recommending interventions and care plans for each case.


Subject(s)
Advance Directives , Dietetics/standards , Homes for the Aged/standards , Nursing Homes/standards , Patient Care Team/standards , Dietetics/legislation & jurisprudence , Homes for the Aged/legislation & jurisprudence , Humans , Nursing Homes/legislation & jurisprudence , Nutritional Physiological Phenomena , Patient Care Team/legislation & jurisprudence , Risk Assessment , United States , Withholding Treatment
19.
J Biol Chem ; 272(12): 7699-706, 1997 Mar 21.
Article in English | MEDLINE | ID: mdl-9065428

ABSTRACT

Neuropeptide Y (NPY) is a potent inhibitor of neurotransmitter release through the Y2 receptor subtype. Specific antagonists for the Y2 receptors have not yet been described. Based on the concept of template-assembled synthetic proteins we have used a cyclic template molecule containing two beta-turn mimetics for covalent attachment of four COOH-terminal fragments RQRYNH2 (NPY 33-36), termed T4-[NPY(33-36)]4. This structurally defined template-assembled synthetic protein has been tested for binding using SK-N-MC and LN319 cell lines that express the Y1 and Y2 receptor, respectively. T4-[NPY(33-36)]4 binds to the Y2 receptor with high affinity (IC50 = 67.2 nM) and has poor binding to the Y1 receptor. This peptidomimetic tested on LN319 cells at concentrations up to 10 microM shows no inhibitory effect on forskolin-stimulated cAMP levels (IC50 for NPY = 2.5 nM). Furthermore, we used confocal microscopy to examine the NPY-induced increase in intracellular calcium in single LN319 cells. Preincubation of the cells with T4-[NPY(33-36)]4 shifted to the right the dose-response curves for intracellular mobilization of calcium induced by NPY at concentrations ranging from 0.1 nM to 10 microM. Finally, we assessed the competitive antagonistic properties of T4-[NPY(33-36)]4 at presynaptic peptidergic Y2 receptors modulating noradrenaline release. the compound T4-[NPY(33-36)]4 caused a marked shift to the right of the concentration-response curve of NPY 13-36, a Y2-selective fragment, yielding a pA2 value of 8.48. Thus, to our best knowledge, T4-[NPY(33-36)]4 represents the first potent and selective Y2 antagonist.


Subject(s)
Neuropeptide Y/antagonists & inhibitors , Peptides, Cyclic/chemical synthesis , Peptides, Cyclic/pharmacology , Receptors, Neuropeptide Y/antagonists & inhibitors , Binding, Competitive , Calcium/metabolism , Cyclic AMP/metabolism , Humans , Norepinephrine/metabolism , Peptides, Cyclic/metabolism , Receptors, Neuropeptide Y/metabolism , Tumor Cells, Cultured
20.
J Biol Chem ; 272(13): 8817-23, 1997 Mar 28.
Article in English | MEDLINE | ID: mdl-9079718

ABSTRACT

Recently, we identified a novel putative human cytokine expressed by activated CD8(+) T cells, which was designated ATAC (activation-induced, T cell-derived, and chemokine-related; the same molecule has been identified independently as lymphotactin and single cysteine motif-1). In this report, we provide evidence that ATAC is a secreted 93-amino acid protein that is generated from its precursor by proteolytic cleavage between Gly21 and Val22. An estimated 60% of ATAC (Val22-Gly114) is secreted as an unmodified protein with a molecular mass of 10,271.72 Da (apparent molecular mass of 12 kDa in SDS-polyacrylamide gel electrophoresis) and in which Cys32 and Cys69 are linked by a disulfide bridge. Unmodified ATAC is a cationic protein with a pI of 11.35 and is capable of binding to heparin. Some 40% of ATAC is O-glycosylated within 25 min of synthesis, giving rise to the appearance of a homogeneous 15-kDa (minor fraction) and a heterogeneous, terminally sialylated 17-19-kDa (major fraction) protein species in SDS-polyacrylamide gel electrophoresis. The secretion of all ATAC protein variants is completed within 30-40 min of synthesis. In terms of function, various ATAC protein forms were consistently ineffective in chemotaxis assays. In contrast, both purified natural ATAC and a chemically synthesized aglycosyl analog induced locomotion (chemokinesis) in purified CD4(+) and CD8(+) T cell populations at 400 ng/ml.


Subject(s)
CD8-Positive T-Lymphocytes/chemistry , Carrier Proteins/isolation & purification , Cytokines/isolation & purification , Lymphocyte Activation , CD4-Positive T-Lymphocytes/cytology , CD8-Positive T-Lymphocytes/cytology , CD8-Positive T-Lymphocytes/metabolism , Carrier Proteins/chemistry , Chemotaxis, Leukocyte , Chromatography, High Pressure Liquid , Cytokines/chemistry , Glycoproteins/chemistry , Glycoproteins/isolation & purification , Glycosylation , Humans , Kinetics , Mass Spectrometry , Peptide Mapping
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