bluuebunny/crossref_metadata_2025 huggingface dataset:
Indoor Air as a Risk Factor for Tuberculosis
Authors: Príkazský V. , Havelková M. , Bencko V.
Published: July 2003

Tuberculosis is a communicable disease that is compulsorily notifiable in the Czech Republic. As part of this process regular publication of epidemiological data on tuberculosis started in 1960 at the Institute of Health Information and Statistics. Here patients' data were collected in the Register of Tuberculosis (RT). In the early 1980s another information system that recorded reports of bacillary tuberculosis (ISBT) was started on mainframe computers as a centrally operated information system. Nowadays the two systems co-operate in the notification and validation of all individual data on tuberculosis cases. Risk information is gathered if available together with data relevant to the disease itself. The second information system, ISBT, collects data on positive laboratory results. ISBT notified 9.2 cases per 100,000 of population in the year 2000. This figure has decreased in the last few years. The RT notified 14/100,000 population incident cases in 2000. Both figures have fluctuated around these levels during the last ten years. The incidence of tuberculosis in risk groups is higher than in general population. In prisoners as well as in migrants (officially recorded) the incidence is higher than in the general population. Age distribution in these risk groups is different from the general population and recalls similar patterns from developing countries. There are also several cases of tuberculosis notified among health professionals, clinical and laboratory staff. It is always supposed that these professionals are infected in connection with their profession. The use of modern molecular techniques in everyday epidemiological practice will improve things further by documenting more precisely the spread of tuberculosis bacilli through the community.
German Cockroaches (Blattella Germanica L.) as a Potential Source of Pathogens Causing Nosocomial Infections
Authors: Gliniewicz A. , Czajka E. , Laudy A. E. , Kochman M. , Grzegorzak K. , Ziółkowska K. , Sawicka B. , Stypulkowska-Misiurewicz H. , Pancer K.
Published: July 2003

German cockroaches collected in two hospitals were examined bacteriologically. Twenty strains of different taxons were isolated, all considered to be potentially pathogenic to humans. Some strains were resistant to antibacterial drugs widely used for treatment of patients: Gram-negative rods were resistant to amoxicillin/clavulanic acid, Pseudomonas spp. additionally resistant to co-trimoxasole. Strains of Staphylococcus equorum, S. hominis were methicillin-resistant, a strain of S. epidermidis showed MLSb mechanism of resistance. Determination of the minimum inhibitory concentration (MIC) for the disinfectant sodium dichloroisocyanurate showed it was in the range 500- 2000 mg. L−1. The MIC for glucoprotamine was from 1.453 to 500 mg. L−1. A strain of S. epidermidis was less sensitive (MIC = 125 mg. L−1) to glucoprotamine than other staphylococci, Serratia marcescens was resistant to that agent (MIC = 500 mg. L−1). Product containing glucoprotamine was ineffective against strains of S. epidermidis MLSb and Pseudomonas putida. Our data suggest that cockroaches might serve as a source of infection, by acting as carriers and reservoirs of bacterial strains in nosocomial outbreaks.
Problems in Deciding the Effect of the Airborne Particulate Load in Hospitals on Patient Health
Authors: Hoskins John A.
Published: July 2003

Probably the most important factor to be considered when considering the health effects resulting from a level of pollutant concentration is that some fraction of the population will be affected more than the rest. This fraction is usually held to comprise the very young, the elderly and those compromised through various medical conditions. The patient population in a hospital is largely drawn from just this group of people. In most cases they spend 100% of their time in an indoor environment. Although many studies have been published associating particulates in the air with excess mortality, it is difficult to quantitate the effect and probably impossible to set an experimentally based guideline for particulates in indoor air that would have any validity for a hospital environment. The main reason for the lack of such evidence is that the epidemiological methods which have revealed associations between current outdoor concentrations of particles and health outcomes cannot be applied to indoor exposures. Those epidemiological studies using time-series techniques have shown statistically significant associations between outdoor particulate levels and admissions to hospital for treatment of respiratory and cardiovascular diseases and daily deaths from these causes. However, the effect of continued exposure in hospital on people compromised through age or health status will be difficult to ascertain. It may be that the best we will be able to do in the absence of acquiring hard data will be to apply the precautionary principle and lower particle levels - just in case it helps.
Computational Fluid Dynamics Applications in Hospital Ventilation Design
Authors: Colquhoun John , Partridge Lester
Published: July 2003

An investigation into modelling the internal environment in important hospital air conditioning applications, using computational fluid dynamics techniques (CFD) to improve air quality and health outcomes has been conducted.

Aggressive infections are developing which medical science is hard pressed to control with antibiotics. In facilities such as operating theatres where patients undergo deep surgery or in ICU/transplant/oncology wards where immuno-compromised patients are accommodated, the control of air quality is essential to minimise patient infection. This reduces the risk of compromising patient health, reduces the cost of expensive drug treatments and, in this day and age, reduces the risk of litigation when patients are infected. CFD analysis techniques allow a study of the air flow and temperature distribution and the optimisation of air delivery to achieve higher sterility in risk-affected areas. In the case of operating theatres, air quality is affected by many factors and the principal objectives are to deliver sterile air to the surgical site, ensure that inappropriate air mixing does not occur, air velocities do not adversely affect the operation and room comfort conditions are maintained.

Computational fluid dynamics analysis allows a proposed operating room to be modelled and each of the variables to be tested to identify flaws in air distribution and make adjustments to achieve the optimum flows and temperature. Graphic displays show velocity distributions and temperature distribution, leading to specific placement of filters, diffusers and allow the effects of lights, staff and pendants to be identified.
Integrated Environmental Management for Hospitals
Authors: Saad Samia Galal
Published: July 2003

The hospital indoor environment includes several unwanted elements including a mix of chemical pollutants, biological contaminants and a high physical and mental stress for all the staff offering medical services. Infection risks are increased by direct contact with sick people in addition to needle and medical equipment punctures conveying blood-transmitted diseases. Although infection control measures are considered seriously they are often inadequate because of concomitant construction problems and ingress of outdoor pollutants. This is especially so in metropolitan areas where the hospitals are engulfed in an urban environment and surrounded by small scale industrial pollution and traffic air emissions.

Every country and indeed every hospital has its own problems and this paper describes the progress of an environmental integrated management scheme initiated for the indoor environment of hospitals to provide greater control to protect the hospital staff from occupational hazards impacting their health, their home contacts and their patients. The scheme started with the preparation of an environmental assessment for newly built hospitals. For those hospitals currently operating the scheme comprised the organisation of an environmental health group and the preparation of a checklist of the environmental sources of indoor pollutants, contaminants and sources of infection and occupational health hazards. An environmental audit was conducted with monitoring for sources of environmental deterioration. There was a follow up for the adoption of hazardous waste management and cleaning and disinfection protocols. A reporting system was set up within the scheme which included follow up actions. The problems of adopting the system are discussed with methodologies for their solutions.
Quality of the Air and Health Assessment of the Medical Staff Handling Disinfection Chemicals in Lithuanian Hospitals
Authors: Glumbakaitė Edita , Žymantienė Zita , Šeškauskas Viktoras , Smolianskienė Gražina , Jurkuvėnas Vytautas
Published: July 2003

The use of chemical disinfectants where ventilation is insufficient can produce morbidity and allergic responses. Although disinfection is the universal tool for prevention of infections, the chemicals used can irritate the eyes and skin and may have neurotoxic and sensitising effects. The objective in the present study was to evaluate such chemical hazards, the use of personal protective equipment and the health status of those who handle disinfection chemicals in Lithuanian hospitals. Nine hospitals in Vilnius were studied in 2000-2001. Some 334 hospital employees from surgery, anaesthesiology, intensive care unit, resuscitation, nursing and endoscopy departments in contact with disinfecting chemicals were interviewed by questionnaire.

Levels of several chemicals were monitored: glutaraldehyde, chlorine compounds and hydrogen peroxide were determined by a photometric method; isopropyl and ethyl alcohols were determined by gas chromatography. It was found that maximum levels of glutaraldehyde exceeded standards by 1.3 times, hydrogen peroxide by 8 times and isopropyl alcohol by 11 times. When working with the chemicals rubber latex gloves were used by 84.1% of personnel and protective face masks by 45.9%. Other personal protective equipment (goggles and respirators) were rarely used. Adverse reactíons included skin reddening and itching (57.6%; p<0.05), allergic conjunctivitis (39.5%), allergic rhinitis (36.0%), as well as frank diseases like chronic bronchitis (9.9%), allergic contact dermatitis (8.3%), allergic rhinitis (7.0%) and allergic conjunctivitis (6.4%). The study concluded that improvements were necessary in all aspects of work with chemical disinfectants containing these active components.
Comparison of the Environment in Operating Theatres in Two Hospitals
Authors: Holcátová Ivana , Benešová Vilma , Hartlová Darina
Published: July 2003

Hospitals and other health care facilities are complex environments that require specialised ventilation for the comfort of patients, personnel and visitors and to control hazardous emissions. Among the most highly specialised environments in hospitals are operating theatres. Health care personnel who work in surgical suites are potentially exposed to a number of airborne hazards and an environment that relies on ventilation to achieve a high air quality. We have tried to compare such environments in two very different hospitals with different exposures to different risk factors. The hospitals were the University Hospital in Prague which is located in the centre of the city in historical buildings and Homolka Hospital. The latter is one of the newest mono-block hospitals in Prague, well equipped; smaller than the University Hospital but specialised (with Leksell Gamma Knife facilities for neurosurgery). We compared the concentrations of anaesthetic gases and room temperature during operations in theatres at the two hospitals with or without air-conditioning. The positive effect of ventilation systems was confirmed provided the systems were properly operated and well maintained. However, it is best to identify and control at source airborne pollutants since this is the most effective way of reducing or preventing of indoor air quality problems, followed by other measures including ventilation.
Gas Chromatographic Determination of Halothane Levels in Hospital Operating Theatres
Authors: Culea Monica , Nicoarã Simona , Nica Nicoleta-Alina , Gherman Carmen
Published: July 2003

A simple, rapid, and sensitive method for the analysis of halothane in the air of operating theatres has been developed. The procedure involves the pre-concentration of halothane on active charcoal in an adsorption tube, then desorption in toluene, followed by capillary gas chromatography (GC) analysis using a flame ionisation detector (FID). The recovery from air samples after pre-concentration was 67%. The linearity was established with standards over a concentration range between 0 and 3 μL mL−1 of halothane in toluene with an excellent coefficient of correlation (0.99) and a limit of detection of 1 μL halothane per cubic metre of air. The time for analysis was approximately 10 min with an adsorption step of 60-90 min per sample collected on site. The procedure was used for occupational exposure assessment, by quantitation of halothane in two different hospital operating theatres, during a single general anaesthesia for a routine orthopaedic surgery and for a routine internal surgery, respectively.
Development and Application of a Protocol for the Assessment of Indoor Air Quality
Authors: Möhle G. , Crump D. , Brown V. , Hunter C. , Squire R. , Mann H. , Raw G. J.
Published: July 2003

The main exposure to air pollutants for most people occurs in indoor environments and this can have an important influence on their health and well-being and affect productivity at the workplace. BRE has prepared a protocol for the measurement of key parameters to undertake an indoor air quality (IAQ) assessment. The measurements required depend upon the objectives and therefore the design of an appropriate strategy is an important first stage in any IAQ assessment. This paper summarises the recommended protocol and reports its evaluation by a study of 10 homes and 10 office buildings. The IAQ protocol was generally well accepted by occupants. There are problems with evaluating the significance of the measurements of some parameters because of a lack of relevant air quality guidelines and data about concentrations that normally occur in buildings. The BRE protocol can be used as guidance by specialists undertaking an IAQ assessment and by building owners/managers concerned about IAQ.
Relationships between Indoor and Outdoor Air Quality in Delhi
Authors: Srivastava Arun , Jain V. K.
Published: July 2003

The relationship between indoor and outdoor airborne particles was investigated for 24 residential, sensitive, commercial and heavy traffic sites in Delhi, India. Sampling of suspended particulate matter (SPM) was first conducted simultaneously indoors and outdoors at the reference site during February to March 2000, and later at all the sites during February to July 2001. In order to evaluate the heavy metal composition of the SPM, analyses of Mg, Ca, Cu, Cd, Pb, Cr, Mn, Fe and Ni were done by atomic absorption spectrometry (AAS). It was found that the outdoor SPM concentrations do indeed affect the indoor SPM concentrations in varying degrees depending upon the nature of the site. In case of metals Cu, Cr, Cd and Ni, very good correlation between the indoor and outdoor concentrations was observed irrespective of the nature of the site. The correlation between indoor and outdoor for Mg, Fe, Mn and Pb depends upon the nature of the site. No correlation was observed between indoor and outdoor Ca at any of the chosen sites.
Species of Conditionally Pathogenic Micromycetes in the Air of Dwellings and Occupational Premises
Authors: Lugauskas Albinas , Krikštaponis Arūnas , Šeškauskas Viktoras
Published: July 2003

The spread of filamentous fungi in occupational premises and dwellings predetermines not only the pollution of the surroundings, but also influences the health of their occupants. Problems caused by micromycetes are of major importance in Lithuania where, because of the climatic conditions, people spend a large part of their time indoors. Over the years 1994-2000 the microbiological state of various dwellings and occupational premises in Vilnius city has been investigated. Some 359 fungal species ascribed to 100 genera, 20 families, 11 ranges, 2 classes and 4 divisions were isolated and identified. Mitosporic fungi (74 genera, 290 species) constituted by far the majority of the identified isolates. In occupational and residential premises fungal species ascribed to Penicillium, Aspergillus, Alternaria, Cladosporium, Mucor, Rhizopus, Rhizomucor, Ulocladium, Mortierella, Aureobasidium, Oidiodendron, Geotrichum, Trichoderma, Acremonium, Scytalidium, Scolecobasidium genera dominated. The distribution of different species in the investigated premises varied. It was confirmed that respiratory and allergic diseases in the investigated premises were closely related to raised concentrations of airborne fungi.
Emission and Evolution of Air-Borne Microflora in Slaughter Houses
Authors: Adeeb Farah , Shooter David
Published: July 2003

The indoor environment can potentially place human occupants at greater risk than the outside environment, because enclosed spaces help to confine aerosols and allow them to build up to potentially infectious levels. The concentration of air-borne microflora (bacteria and fungi) was determined over a 12-month period in four slaughter houses in the city of Lahore, Pakistan. The overall level of airborne microflora (i.e. 3.9-8.5 log CFU m−3) obtained in Lahore was higher compared to the level of indoor microflora (i.e. 3.7-6.43 log CFU m−3) reported for other livestock buildings surveyed in 4 European countries (England, Germany, Denmark and The Netherlands). Differences in concentration may be due to the generation rate of air-borne microflora, accumulation potential and clearance by various routes, mainly ventilation and mechanical coagulation (i.e. settlement).
Primary Prevention of Chemical Risk: Evaluation of the Efficiency of Fume Hoods in the Laboratories of Bologna University
Authors: Serra R. , Rossi S. , Ferroni F. , Berardi B. M. , Badiello R.
Published: July 2003

The Service Department for Safety and Prevention of Accidents of Bologna University has analysed about 400 chemical fume hoods located in the laboratories of the Chemical and other Departments (Biology, Veterinary Medicine, Engineering). The parameters chosen were the face velocity of air immission and the velocity of aspiration. The results are not particularly encouraging and show that the percentage with a sufficient aspiration is too low (about 50%). The hooded benches in the majority of teaching laboratories contribute in some way to this negative result. In some cases preventative maintenance has improved the efficiency of aspiration although this has not always been sufficient to ensure the minimum parameters required by the safety standards were reached. We include recommendations that the correct choice of the hood should be made with regard to the TLV of the substances handled.
Murine neutrophils require α1,3-fucosylation but not PSGL-1 for productive infection with Anaplasma phagocytophilum
Authors: Carlyon Jason A. , Akkoyunlu Mustafa , Xia Lijun , Yago Tadayuki , Wang Tian , Cummings Richard D. , McEver Rodger P. , Fikrig Erol
Published: July 2003

Anaplasma phagocytophilum causes human granulocytic ehrlichiosis, the second most common tick-borne disease in the United States. Mice are natural reservoirs for this bacterium and man is an inadvertent host. A phagocytophilum's tropism for human neutrophils is linked to neutrophil expression of P-selectin glycoprotein ligand-1 (PSGL-1), as well as sialylated and α1,3-fucosylated glycans. To determine whether A phagocytophilum uses similar molecular features to infect murine neutrophils, we assessed in vitro bacterial binding to neutrophils from and infection burden in wild-type mice; mice lacking α1,3-fucosyltransferases Fuc-TIV and Fuc-TVII; or mice lacking PSGL-1. Binding to Fuc-TIV-/-/Fuc-TVII-/- neutrophils and infection of Fuc-TIV-/-/Fuc-TVII-/- mice were significantly reduced relative to wild-type mice. A phagocytophilum binding to PSGL-1-/- neutrophils was modestly reduced, whereas sialidase treatment significantly decreased binding to both wild-type and PSGL-1-/- neutrophils. A phagocytophilum similarly infected PSGL-1-/- and wild-type mice in vivo. A phagocytophilum induced comparable levels of chemokines from wild-type and PSGL-1-/- neutrophils in vitro, while those induced from Fuc-TIV-/-/Fuc-TVII-/- neutrophils were appreciably reduced. Therefore, A phagocytophilum infection in mice, as in humans, requires sialylation and α1,3-fucosylation of neutrophils. However, murine infection does not require neutrophil PSGL-1 expression, which has important implications for understanding how A phagocytophilum binds and infects neutrophils.
Phenotypic and functional T-cell aging in rhesus macaques (Macaca mulatta): differential behavior of CD4 and CD8 subsets
Authors: Janković Vladimir , Messaoudi Ilhem , Nikolich-Žugich Janko
Published: July 2003

A prominent theory of immune senescence holds that repeated antigenic stimulation and decreased production of naive cells combine to progressively exhaust the reserve of lymphocytes available to fight new pathogens, culminating in an accumulation of lymphocytes that achieved replicative senescence. A well-defined primate model of immune senescence in vivo would greatly facilitate testing of this theory. Here, we investigated phenotypic and functional T-cell aging in the rhesus macaques (RMs), currently the dominant primate model of AIDS. Our results show that sharp differences exist between the CD8 and CD4 T-cell subsets in (1) cell-cycle programs (as assessed by both in vitro proliferation and in vivo turnover measurement); (2) CD28 regulation on cell-cycle entry; and (3) accumulation of immediate effector cells among the CD28– cells, believed to be close to or at replicative senescence. These results further suggest poor reliability of CD28 as a marker for senescence. We suggest that some of the T-cell aging phenomenology in RMs can be ascribed to accentuation over time of the inherent differences in activation programs in CD8 and CD4 T cells.
Scaffold attachment region–containing retrovirus vectors improve long-term proviral expression after transplantation of GFP-modified CD34+ baboon repopulating cells
Authors: Kurre Peter , Morris Julia , Thomasson Bobbie , Kohn Donald B. , Kiem Hans-Peter
Published: July 2003

Sustained high-level proviral expression is important for clinical applications of gene therapy. Genetic elements including the β-interferon scaffold attachment region (SAR) have been shown to improve transgene expression in hematopoietic cells. We hypothesized that SAR elements might improve expression and allow the preselection of successfully transduced cells. Thus, we transplanted green fluorescent protein (GFP)–selected cells, half of which had been transduced with either SAR or non–SAR-containing retrovirus vectors, into 3 animals. All animals showed delayed engraftment compared with historic controls (28 vs 15.5 days). GFP marking was seen at levels up to 8% but declined over the first 6 weeks. Importantly, fluorescence intensity was 2- to 9-fold increased in progeny of SAR versus non–SAR vector–modified cells in all hematopoietic lineages for the duration of follow-up (6-12 months). In conclusion, the use of SAR-containing vectors improved transgene expression in hematopoietic repopulating cells, which may obviate the need for multicopy integration to achieve high-level expression and reduce the risk for insertional mutagenesis.
The zebrafish spi1 promoter drives myeloid-specific expression in stable transgenic fish
Authors: Ward Alister C. , McPhee Dora O. , Condron Melanie M. , Varma Sony , Cody Stephen H. , Onnebo Sara M. N. , Paw Barry H. , Zon Leonard I. , Lieschke Graham J.
Published: July 2003

The spi1 (pu.1) gene has recently been identified as a useful marker of early myeloid cells in zebrafish. To enhance the versatility of this organism as a model for studying myeloid development, the promoter of this gene has been isolated and characterized. Transient transgenesis revealed that a 5.3 kilobase promoter fragment immediately upstream of the spi1 coding sequence was sufficient to drive expression of enhanced green fluorescent protein (EGFP) in injected embryos in a manner that largely recapitulated the native spi1 gene expression pattern. This fragment was successfully used to produce a germ line transgenic line of zebrafish with EGFP-expressing myeloid cells. These TG(spi1:EGFP)pA301 transgenic zebrafish represent a valuable tool for further studies of myeloid development and its perturbation.
Protease-activated receptors 1 and 4 mediate thrombin signaling in endothelial cells
Authors: Kataoka Hiroshi , Hamilton Justin R. , McKemy David D. , Camerer Eric , Zheng Yao-Wu , Cheng Abby , Griffin Courtney , Coughlin Shaun R.
Published: July 2003

Defining the relative importance of protease-activated receptors (PARs) for thrombin signaling in mouse endothelial cells is critical for a basic understanding of thrombin signaling in these cells and for the rational use of knockout mice to probe the roles of thrombin's actions on endothelial cells in vivo. We examined thrombin- and PAR agonist–induced increases in cytoplasmic calcium, phosphoinositide hydrolysis, extracellular signal-regulated kinase (ERK) phosphorylation, and gene expression in endothelial cells from wild-type and PAR-deficient mice. PAR1 and PAR4 agonists triggered responses in wild-type but not in Par1–/– and Par4–/– endothelial cells, respectively. Calcium imaging confirmed that a substantial fraction of individual endothelial cells responded to both agonists. Compared with wild-type cells, Par1–/– endothelial cells showed markedly decreased responses to low concentrations of thrombin, and cells that lacked both PAR1 and PAR4 showed no responses to even high concentrations of thrombin. Similar results were obtained when endothelial-dependent vasorelaxation of freshly isolated mouse aorta was used as an index of signaling in native endothelial cells. Thus PAR1 is the major thrombin receptor in mouse endothelial cells, but PAR4 also contributes. These receptors serve at least partially redundant roles in endothelial cells in vitro and in vivo and together are necessary for the thrombin responses measured.
The shared tumor-associated antigen cytochrome P450 1B1 is recognized by specific cytotoxic T cells
Authors: Maecker Britta , Sherr David H. , Vonderheide Robert H. , von Bergwelt-Baildon Michael S. , Hirano Naoto , Anderson Karen S. , Xia Zhinan , Butler Marcus O. , Wucherpfennig Kai W. , O'Hara Carl , Cole Geoffrey , Kwak Silvia S. , Ramstedt Urban , Tomlinson Andy J. , Chicz Roman M. , Nadler Lee M. , Schultze Joachim L.
Published: July 2003

Cytochrome P450 1B1 (CYP1B1), a drug-metabolizing extrahepatic enzyme, was recently shown to be overexpressed in multiple types of cancer. Such tumor-associated genes may be useful targets for anticancer therapy, particularly cancer immunotherapeutics. We identified HLA-A*0201–binding peptides and a naturally processed and presented T-cell epitope capable of inducing CYP1B1-specific cytotoxic T lymphocytes (CTLs) in HLA-A2 transgenic mice. Furthermore, the induction of CYP1B1-specific T cells was demonstrated in healthy donors and cancer patients. These T cells efficiently lysed target cells pulsed with the cognate peptide. More important, HLA-A2–matched tumor cell lines and primary malignant cells were also recognized by CYP1B1-specific CTLs. These findings form the basis of a phase 1 clinical trial exploring a DNA-based vector encoding CYP1B1 for widely applicable cancer immunotherapy conducted at the Dana-Farber Cancer Institute.
Redirection of antileukemic reactivity of peripheral T lymphocytes using gene transfer of minor histocompatibility antigen HA-2-specific T-cell receptor complexes expressing a conserved alpha joining region
Authors: Heemskerk Mirjam H. M. , Hoogeboom Manja , de Paus Roelof A. , Kester Michel G. D. , van der Hoorn Menno A. W. G. , Goulmy Els , Willemze Roel , Falkenburg J. H. Frederik
Published: July 2003

Donor-derived T lymphocytes directed against minor histocompatibility antigens (mHags) exclusively expressed on cells of the hematopoietic lineages can eliminate hematologic malignancies. Transfer of T-cell receptors (TCRs) directed against these mHags into T lymphocytes may provide a strategy to generate antileukemic T cells. To investigate the feasibility of this strategy the TCR usage of mHag HA-2-specific T-cell clones was characterized. Thirteen different types of HA-2-specific T-cell clones were detected, expressing TCRs with diversity in TCR α- and β-chain usage, however, containing in the TCR α chain a single conserved gene segment Jα42, indicating that Jα42 is involved in HA-2-specific recognition. We transferred various HA-2 TCRs into T lymphocytes from HLA-A2-positive HA-2-negative individuals resulting in T cells with redirected cytolytic activity against HA-2-expressing target cells. Transfer of chimeric TCRs demonstrated that the HA-2 specificity is not only determined by the Jα42 region but also by the N-region of the α chain and the CDR3 region of the β chain. Finally, when HA-2 TCRs were transferred into T cells from HLA-A2-negative donors, the HA-2 TCR-modified T cells exerted potent antileukemic reactivity without signs of anti-HLA-A2 alloreactivity. These results indicate that HA-2 TCR transfer may be used as an alternative strategy to generate HA-2-specific T cells to treat hematologic malignancies of HLA-A2-positive, HA-2-expressing patients that received transplants from HLA-A2-matched or -mismatched donors. (Blood. 2003;102:3530-3540)
Male DNA in female donor apheresis and CD34-enriched products
Authors: Adams Kristina M. , Lambert Nathalie C. , Heimfeld Shelly , Tylee Tracy S. , Pang Jennifer M. , Erickson Timothy D. , Nelson J. Lee
Published: July 2003

Increased risk of graft-versus-host disease (GVHD) has been described in recipients of hematopoietic stem cell transplantations when the donor is a parous woman. Cells from prior pregnancies are now known to persist in women and could contribute to GVHD. We asked whether male DNA (presumed fetal microchimerism) is present in apheresis products of female donors. A total of 50 samples were studied by using real-time quantitative polymerase chain reaction (PCR) for the Y chromosome-specific sequence DYS14. Among 29 growth factor-mobilized peripheral blood mononuclear cell (G-PBMC) products, 34% were positive for male DNA. Quantitative results, expressed as DNA genome equivalent of male cells per million host cells (gEq/mil), ranged from 0 to 35 gEq/mil. Among 21 CD34-enriched cell fractions, 48% were positive with a range of 0 to 357 gEq/mil. In summary, male DNA was frequently detected in G-PBMC and CD34-enriched products from female donors. Whether fetal microchimerism contributes to GVHD merits further investigation. (Blood. 2003;102: 3845-3847)
Transforming growth factor–β1 in supernatants from stored red blood cells inhibits neutrophil locomotion
Authors: Ghio Massimo , Ottonello Luciano , Contini Paola , Amelotti Massimo , Mazzei Clemente , Indiveri Francesco , Puppo Francesco , Dallegri Franco
Published: July 2003

Studies comparing transfusion and nontransfusion patients suggest an increased risk of postoperative infections in transfusion groups. Supernatants of blood components have been shown to affect the function of T lymphocytes and natural killer cells. Here, we found that supernatants from stored red blood cells (RBCs) inhibit human neutrophil migration in response to formyl peptides and stimulate neutrophil locomotion. These effects can be observed with high dilutions of RBC supernatants, such as 1:5 × 106 (vol/vol), able to trigger locomotion as well as desensitization of the cells to alternative chemoattractants. The phenomenon might be mediated by chemoattractants present in the supernatants. As RBC supernatants failed to mobilize intracellular free calcium, the chemoattractants should belong to the group of pure chemoattractants, that is, soluble Fas ligand (sFasL) and transforming growth factor–β1 (TGF-β1), known to act without increasing calcium levels. Recombinant TGF-β1, but not sFasL, was found to reproduce the ability of RBC supernatants to both inhibit neutrophil response to formyl peptides and stimulate neutrophil locomotion. Moreover, TGF-β1–immunodepleted supernatants did not display neutrophil-directed activities. Finally, RBC supernatants from RBCs stored after depletion of leukocytes were incapable of affecting neutrophil function. With neutrophils acting as a first-line antimicrobial defense, the ability, shown here, of high dilutions of RBC supernatants to inhibit neutrophil chemotaxis through TGF-β1 may be a relevant determinant of infections in the postoperative period for transfusion patients. Consistently, the neutrophil chemotactic response to formyl peptide was inhibited by the plasma obtained from 5 transfusion patients.
Reconstitution of the Epstein-Barr virus–specific cytotoxic T-lymphocyte response following T-cell–depleted myeloablative and nonmyeloablative allogeneic stem cell transplantation
Authors: Chakrabarti Suparno , Milligan Donald W. , Pillay Deenan , Mackinnon Stephen , Holder Kathleen , Kaur Narinder , McDonald Dorothy , Fegan Christopher D. , Waldmann Herman , Hale Geoff , Rickinson Alan , Steven Neil
Published: July 2003

The recovery of circulating antigen-specific T-cell immunity to Epstein-Barr virus (EBV) was determined in ELIspot assays following allogeneic myeloablative or nonmyeloablative stem cell transplantation (MST/NST). In 8 of 12 MST patients receiving an alemtuzumab-treated graft, the frequency of the EBV-specific reactivities was similar to or greater than that seen in the healthy controls. A response was detectable in 3 of 6 and 6 of 9 patients by 3 and 6 months, respectively, and in all patients by one year following MST. In contrast, only 1 of 9 (95% confidence interval [CI], 0-2.8) patients made a detectable EBV-specific response by 6 months following NST conditioned with fludarabine, melphalan, and alemtuzumab. Responses were detected in 7 of 10 patients by 1 year after NST. Parallel surveillance demonstrated that other virus infections occurred more frequently and earlier after transplantation in NST patients. The use of alemtuzumab in vivo in the nonmyeloablative conditioning might have resulted in the delay in EBV-specific T-cell recovery and increased virus infections.
Are Older French Patients As Willing As Older American Patients to Undertake Chemotherapy?
Authors: Extermann Martine , Albrand Gilles , Chen Hongbin , Zanetta Sylvie , Schonwetter Ronald , Zulian Gilbert B. , Cantor Alan , Droz Jean-Pierre
Published: July 2003

Purpose: A view often held in Europe is that older Europeans are less willing than older Americans to undertake chemotherapy. This study assesses whether this view is valid.

Patients and Methods: Three-hundred twenty outpatients aged 70 years and older were interviewed via anonymous questionnaires: French patients with and without cancer and American patients with and without cancer. The response rate was 61% (195 of 320 questionnaires). Ages ranged from 70 to 95 years (29% aged 80 years and older). Two scenarios were presented: a strong chemotherapy (platinum/taxane combination–like) and a milder chemotherapy (weekly vinorelbine–like). The options were to refuse chemotherapy or to accept for a threshold chance of cure, of life prolongation, or of symptom relief. Functional status, education, self-rated health, and depression were controlled for.

Results: French noncancer patients (34%) were less willing to accept the strong chemotherapy than French cancer patients (77.8%), American noncancer patients (73.8%), and American cancer patients (70.5%) (P < .001 for each pair). This was also true for the moderate chemotherapy (67.9% v 100%, 95.2%, and 88.5%, respectively; P < .001). Age and sex did not correlate with response, but self-rated health, cancer status, and nationality did. Thresholds varied from patient to patient.

Conclusion: Whereas older French people without cancer are more reluctant than older Americans to envision chemotherapy, older cancer patients in both countries have the same amenability to treatment. Chemotherapy options should be fully discussed with older cancer patients, given that most are willing to consider them.
Targeted Overexpression of Luteinizing Hormone Causes Ovary-Dependent Functional Adenomas Restricted to Cells of the Pit-1 Lineage
Authors: Mohammad Helai P. , Abbud Rula A. , Parlow Al F. , Lewin Jonathan S. , Nilson John H.
Published: July 2003

The majority of pituitary adenomas in humans are nonmetastasizing, monoclonal neoplasms that occur in approximately 20% of the general population. Their development has been linked to a combination of extrinsic factors and intrinsic defects. We now demonstrate with transgenic mice that targeted and chronic overexpression of LH causes ovarian hyperstimulation and subsequent hyperproliferation of Pit-1-positive cells that culminates in the appearance of functional pituitary adenomas ranging from focal to multifocal expansion of lactotropes, somatotropes, and thyrotropes. Tumors fail to develop in ovariectomized mice, indicating that contributions from the ovary are necessary for adenoma development. Although the link between chronic ovarian hyperstimulation and PRL-secreting adenomas was expected, the involvement of somatotropes and thyrotropes was surprising and suggests that multiple ovarian hormones may contribute to this unusual pathological consequence. In support of this idea, we have found that ovariectomy followed by estrogen replacement results in the expansion of lactotropes selectively in LH overexpressing mice, but not somatotropes and thyrotropes. Collectively, these data indicate that estrogen is sufficient for the formation of lactotrope adenomas only in animals with a hyperstimulated ovary, whereas the appearance of GH- and TSH-secreting adenomas depends on multiple ovarian hormones. Together, our data expand current models of pituitary tumorigenesis by suggesting that chronic ovarian hyperstimulation may underlie the formation of a subset of pituitary adenomas containing lactotropes, somatotropes, and thyrotropes.
The WT1 Wilms’ Tumor Suppressor Gene: A Novel Target for Insulin-Like Growth Factor-I Action
Authors: Bentov Itay , LeRoith Derek , Werner Haim
Published: July 2003

IGF-I stimulates cell division in numerous cell types after activation of the IGF-I receptor, a transmembrane heterotetramer linked to the ras-raf-MAPK and phosphatidylinositol 3-kinase signaling pathways. The WT1 Wilms’ tumor suppressor is a zinc finger-containing transcription factor that is involved in a number of developmental processes, as well as in the etiology of certain neoplasias. In the present study, we demonstrated that IGF-I reduced WT1 expression in osteosarcoma-derived Saos-2 cells in a time- and dose-dependent manner. This effect was mediated through the MAPK signaling pathway, as shown by the ability of the specific inhibitor UO126 to abrogate IGF-I action. Furthermore, the effect of IGF-I involved repression of transcription from the WT1 gene promoter, as demonstrated using transient transfection assays. Taken together, our results suggest that the WT1 gene is a novel downstream target for IGF-I action. Reduced levels of WT1 may facilitate IGF-I-stimulated cell cycle progression. Most importantly, inhibition of WT1 gene expression by IGF-I may have significant implications in terms of cancer initiation and/or progression.
Disrupted Function of Tumor Necrosis Factor-α-Stimulated Gene 6 Blocks Cumulus Cell-Oocyte Complex Expansion
Authors: Ochsner Scott A. , Day Anthony J. , Rugg Marilyn S. , Breyer Richard M. , Gomer Richard H. , Richards Joanne S.
Published: July 2003

During ovulation, the oocyte and surrounding somatic cumulus cells contained within a specialized, mucoid matrix are released from the ovary. One matrix component, TNF-α-stimulated gene 6 (TSG-6), is a hyaluronan binding protein induced in cumulus cells of preovulatory follicles by the LH surge and is decreased in cumulus cells of COX-2 and prostaglandin E2 (PGE2) receptor subtype EP2 null mice that exhibit impaired ovulation and cumulus expansion. To determine if TSG-6 was hormonally induced in cumulus cells in vitro and was functional during the formation of the expanded matrix, we established a cumulus cell-oocyte complex (COC) culture system. This system was used to analyze the effects of FSH, PGE2, EP2 receptor, and selected protein kinase inhibitors on TSG-6 production as well as specific antibodies to the TSG-6 link module on TSG-6 function. We document that TSG-6 message and protein are induced by cAMP/protein kinase A/MAPK signaling pathways and that blocking these cascades prevents expansion and the production of TSG-6. FSH but not PGE2 rescued expansion and production of TSG-6 in the EP2 null COCs, indicating that generation of a cAMP signal is essential. Furthermore, disruption of the functional interactions between TSG-6, inter-α trypsin inhibitor, and hyaluronan with specific antibodies severely altered matrix formation and cumulus expansion, as recorded by time-lapse imaging. Collectively, these results indicate that TSG-6 mRNA is induced in cumulus cells in culture by cAMP and that the secreted TSG-6 protein is a key structural component of the mouse COC matrix.
Stereospecific reduction of the original anticancer drug oracin in rat extrahepatic tissues
Authors: Szotáková Barbora , Skálová Lenka , Jílek Petr , Buchta Vladimír , Wsól Vladimír
Published: July 2003

The liver is the major site of drug metabolism in the body. However, many drugs undergo metabolism in extrahepatic sites and in the gut wall and lumen. In this study, the distribution and activity of reductases in rat that reduced potential cytostatic oracin to its principal metabolite 11-dihydrooracin (DHO) were investigated. The extension and stereospecificity of oracin reduction to DHO were tested in microsomal and cytosolic fractions from the liver, kidney, heart, lung and wall of small intestine, caecum and large intestine. Intestinal bacterial reduction of oracin was studied as well. The amount of DHO enantiomers was measured by HPLC with Chiralcel OD-R as chiral column. Reductive biotransformation of oracin was mostly stereospecific for (+)-DHO, but the enantiomeric ratio differed significantly among individual tissues and subcellular fractions (from 56% (+)-DHO in heart microsomes to 92% (+)-DHO in liver cytosol). Stereospecificity for (-)-DHO (60%) was observed in bacterial oracin reduction in the lumen of small intestine, caecum and large intestine. Shift of the (+)-DHO/(-)-DHO enantiomeric ratio from 90:10 (in liver subcellular fractions) to 60:40 (in-vivo) clearly demonstrated the importance of the contribution of extrahepatic metabolism to the total biotransformation of oracin to DHO.
Hypochlorite scavenging activity of hydroxycinnamic acids evaluated by a rapid microplate method based on the measurement of chloramines
Authors: Firuzi Omidreza , Giansanti Luisa , Vento Roberta , Seibert Cathrin , Petrucci Rita , Marrosu Giancarlo , Agostino Roberta , Saso Luciano
Published: July 2003

Scavengers of hypochlorite (XOCI) could have beneficial effects in diseases in which this oxidant plays a pathogenic role. It has been reported that ferulic acid and chlorogenic acid, the quinic ester of caffeic acid, are good hypochlorite scavengers, but a systematic evaluation of the naturally occurring hydroxycinnamic acids (HCAs), which these substances belong to, has not been performed yet. Thus, in this work we studied, by two different in-vitro methods, the antioxidant activity of five HCAs: p-coumaric acid, ferulic acid, sinapinic acid, caffeic acid and chlorogenic acid. The methods applied in this study were based on the oxidation of human serum albumin (HSA) by XOCI, a new microplate method based on the measurement of chloramines and a previously described carbonyl assay. Firstly, lysine-derived chloramines, in the presence or absence of the HCAs, were detected using 5-thio-2-nitrobenzoic acid (TNB), measuring the absorbance at 415nm by a microplate reader. To remove excess XOCI, Trolox, a known XOCI scavenger, was added before TNB. Secondly, lysine-derived carbonyls, in the presence or absence of the HCAs, were detected by using 2,4-dinitro-phenylhydrazine. Hydroxycinnamic acids appeared active (caffeic≥sinapinic>chlorogenic≅ferulic>p-coumaric acid) by both methods, suggesting possible pharmacological applications for these compounds, which are present at high concentrations in the plant kingdom.
A new animal model for evaluation of long-term growth rate over one month by rhGH/PLGA microcapsule formulations
Authors: Takada Shigeyuki , Kurokawa Tomofumi , Misaki Masafumi , Taira Keiko , Yamagata Yutaka
Published: July 2003

A new animal model to evaluate the long-term growth rate produced by a sustained-release formulation of recombinant human growth hormone (rhGH) over one month was developed and the usefulness of our microcapsule formulations was demonstrated in this model. Long-term pharmacological effects by subcutaneous injection of microcapsules for sustained release of rhGH were evaluated in hypophysectomized (Hpx) rats treated with immunosuppressive agent along with hormone supplement. Copoly(dl-lactic/glycolic)acid (PLGA) microcapsules for sustained release of rhGH, a two-week sustained-release formulation (rhGH-SR-2W) and a one-month sustained-release formulation (rhGH-SR-1M), were prepared by a solid-in-oil-in-water emulsion solvent evaporation technique. Body-weight gain, body-length gain and serum levels of rat insulin-like growth factor-l (rlGF-l) induced by subcutaneous injection of rhGH-SR were compared with those by daily injections of rhGH solution in Hpx rats for 35 days. Serum IGF-l levels in Hpx rats after the injection of rhGH-SR-2W microcapsules were higher than those after daily injections of rhGH solution. Body-length gain, a new parameter, after single injection of rhGH-SR-1M microcapsules demonstrated the higher growth rate than that after daily injections of rhGH solution for 35 days. Thus, single injection of rhGH-SR microcapsules demonstrated long-term pharmacological effects greater than those by daily injections of rhGH solution in a newly developed model, immunosuppressed Hpx rats.
Controlled release of anti-cocaine catalytic antibody from biodegradable polymer microspheres
Authors: Homayoun P , Mandal T , Landry D , Komiskey H
Published: July 2003

Recent reports have shown that anti-cocaine catalytic monoclonal antibody 15A10 reduces the toxic effect of cocaine by increasing its breakdown to systemically inert products ecgonine methylester and benzoic acid. This study reports the microencapsulation of antibody 15A10 using biodegradable poly (lactic-glycolic) acid (PLGA) by double emulsion technique. Formulation parameters such as protein loading, polymer molecular weight and the presence of zinc carbonate were studied for their effects on in-vitro release of antibody from microspheres. The initial burst release was decreased by the reduction of the protein (as % of total ingredients) in the formulation. Although changing the polymer molecular weight did not cause a reduction in initial burst release, it was effective in improving the release rate. The inclusion of zinc carbonate in microsphere preparation resulted in increase in initial burst release. An in-vivo study in mice revealed the presence of antibody in blood up to ten days following subcutaneous injections. These data demonstrate a potential for a sustained-release formulation of monoclonal antibody 15A10 for treatment of cocaine addiction.