U. P. Kodavanti, R. H. Jaskot, W. Y. Su, D. L. Costa, A. J. Ghio and K. L. Dreher
Experimental Toxicology Division, United States Environmental Protection Agency, Research Triangle Park, North ...Carolina 27711, USA.
Occupational exposure to anthropogenic particles is associated with lung
injury in humans. We hypothesized that residual oil fly ash (ROFA), an
emission source particulate, may induce acute lung injury and fibrosis in
sensitive rat strains and that fibronectin (Fn) gene expression will
correspond to the development of fibrosis. Male Sprague-Dawley (SD), Wistar
(WIS), and Fischer 344 (F-344) rats (60 days old) were exposed to saline or
ROFA (8.3 mg/kg) by intratracheal instillation and examined for up to 12
wk. Histology indicated focal areas of lung damage showing inflammatory
cell infiltration as well as alveolar, airway, and interstitial thickening
in all three rat strains during 1-7 days postexposure. Trichrome staining
of the lung sections indicated a sporadic incidence of focal alveolar
fibrosis at 1, 3, and 12 wk in SD rats, whereas WIS and F-344 rats showed
only a modest increase in trichrome staining in the septal areas. Of all Fn
mRNA isoforms examined by polymerase chain reaction, only EIIIA(+) was
upregulated during 6 h-1 wk in ROFA-exposed SD and WIS rats but not in
F-344 rats. In situ hybridization analysis in SD rats revealed Fn mRNA
expression by macrophage and alveolar and airway epithelium and within
fibrotic areas. Immunohistochemical analysis revealed increased presence of
Fn EIIIA(+) protein in the areas of fibrotic injury and basally to the
airway epithelium. In summary, Fn EIIIA(+) increases early in the course of
particle-induced lung injury and remodeling, which may or may not result in
discernible alveolar fibrosis. There is a rat strain variation in
ROFA-induced fibrosis and associated Fn EIIIA(+) expression.
Transcribed sequences in the human genome can be identified with confidence only by alignment with sequences derived from cDNAs synthesized from naturally occurring mRNAs. We constructed a set of ...250,000 cDNAs that represent partial expressed gene sequences and that are biased toward the central coding regions of the resulting transcripts. They are termed ORF expressed sequence tags (ORESTES). The 250,000 ORESTES were assembled into 81,429 contigs. Of these, 1,181 (1.45%) were found to match sequences in chromosome 22 with at least one ORESTES contig for 162 (65.6%) of the 247 known genes, for 67 (44.6%) of the 150 related genes, and for 45 of the 148 (30.4%) EST-predicted genes on this chromosomes. Using a set of stringent criteria to validate our sequences, we identified a further 219 previously unannotated transcribed sequences on chromosome 22. Of these, 171 were in fact also defined by EST or full length cDNA sequences available in GenBank but not utilized in the initial annotation of the first human chromosome sequence. Thus despite representing less than 15% of all expressed human sequences in the public databases at the time of the present analysis, ORESTES sequences defined 48 transcribed sequences on chromosome 22 not defined by other sequences. All of the transcribed sequences defined by ORESTES coincided with DNA regions predicted as encoding exons by GENSCAN. (http://genes.mit.edu/GENSCAN.html).
Bronchoalveolar lavage (BAL) was used to sample lung cells and biochemical components in the lung air spaces at various times from 1 to 91 d after intrapulmonary instillation of 2.6 microm-diameter ...iron oxide particles in human subjects. The instillation of particles induced transient acute inflammation during the first day post instillation (PI), characterized by increased numbers of neutrophils and alveolar macrophages as well as increased amounts of protein, lactate dehydrogenase, and interleukin-8 in BAL fluids. This response was subclinical and was resolved within 4 d PI. A similar dose-dependent response was seen in rats 1 d after intratracheal instillation of the same particles. The particles contained small amounts of soluble iron (240 ng/mg) and possessed the capacity to catalyze oxidant generation in vitro. Our findings indicate that the acute inflammation after particle exposure may, at least partially, be the result of oxidant generation catalyzed by the presence of residual amounts of ferric ion, ferric hydroxides, or oxyhydroxides associated with the particles. These findings may have relevance to the acute health effects associated with increased levels of ambient particulate air pollutants.
ELISA for early diagnosis of histoplasmosis Guimaraes, Allan Jefferson; Pizzini, Claudia Vera; de Matos Guedes, Herbert Leonel ...
Journal of medical microbiology,
06/2004, Letnik:
53, Številka:
6
Journal Article
Recenzirano
Odprti dostop
1 Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil 2 Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 3 St John's ...Institute of Dermatology, Guy's Hospital, King's College, London, UK
Correspondence Rosely Maria Zancopé-Oliveira zancope{at}ipec.fiocruz.br
Received September 24, 2003
Accepted January 23, 2004
An ELISA was developed and evaluated as a method for detecting antibodies against glycosylated and deglycosylated histoplasmin (HMIN). Sera from patients with histoplasmosis, paracoccidioidomycosis, sporotrichosis, coccidioidomycosis, aspergillosis, cryptococcosis and healthy donors were tested by ELISA against purified, deglycosylated histoplasmin (ptHMIN) and compared with purified, native (i.e. glycosylated) histoplasmin (pHMIN). Although cross-reactivity was not abolished when ptHMIN was used in the test, it was reduced (pHMIN ELISA 93 % versus ptHMIN ELISA 96 %). However, there were statistically significant differences between the sensitivities of these two methods for the detection of antibodies (pHMIN ELISA 57 % versus ptHMIN ELISA 92 %; P < 0.001) and between the efficiency of the methods (pHMIN ELISA 83 % versus ptHMIN ELISA 95 %; P < 0.001). These parameters compare better than previously published data relating to the use of treated HMIN in diagnostic ELISAs. Some of the reactivities of serum samples were compared by immunoblotting using deglycosylated HMIN and by immunodiffusion using the crude antigen. The results demonstrated that cross-reactions with heterologous sera in both ELISAs could also be observed in immunoblotting and arose from shared protein epitopes. These data suggest that ELISA using deglycosylated HMIN is a very sensitive diagnostic method and, by using commercially available antigen, it can be easily standardized and performed faster than previous Western blot-based tests using the same antigen. It provides a useful adjunct to existing methods of diagnosis that could be applied even in situations where laboratory facilities were relatively limited.
Abbreviations: CF, complement fixation; HMIN, histoplasmin; ID, immunodiffusion; pHMIN, purified histoplasmin; ptHMIN, purified, treated histoplasmin.
This study was conducted to evaluate the ocular hypotensive efficacy, safety, and side effects of latanoprost 0.005% administered as adjunctive therapy in patients with Sturge-Weber syndrome (SWS) ...and glaucoma.
Commercially available latanoprost 0.005% was added as a single drop once daily to other antiglaucoma medications. Intraocular pressure (IOP) was measured at 1, 3, and 6 months of treatment. A successful response was defined as a reduction of at least 20% in IOP at the final follow-up evaluation without additional medical or surgical therapy and no adverse events related to latanoprost.
18 eyes of 18 patients with SWS and glaucoma were enrolled from 9 clinical centers. Mean baseline IOP was 28.4 +/- 7.1 mmHg (range, 17-42 mmHg). Using Kaplan-Meier analysis, a successful response to latanoprost was observed in 3 of 18 (16.7%) patients at the 6-month interval. Seven (38.9%) patients required surgery; three (16.7%) patients required additional medical therapy, seven (38.9%) patients had no change in therapy. One (5.6%) patient discontinued latanoprost treatment because of intolerable conjunctival hyperemia. Two successfully treated patients had significantly greater episcleral vessel engorgement after initiation of latanoprost therapy.
Patients with SWS and glaucoma respond poorly to adjunctive latanoprost therapy and often require additional medical or surgical intervention. Increased episcleral vascular engorgement might result in greater operative risks should filtration surgery become necessary in these patients.
Abstract only Background: Refractory angina(Ang) effects 150 –250,000 pts in the US alone. These individuals have exhausted options for revasc and continue to have lifestyle limiting ang despite ...optimal medical Rx. Pre-clinical studies provided evidence that human CD34+ cells(CD34) can stimulate new blood vessel formation in ischemic myocardium(Myo), improving perfusion and function. We performed a phase 2 study of intraMyo injection of autologous CD34 in pts with CCS Class 3 and 4 Ang to obtain evidence for feasibility, safety and bioactivity. Methods: A phase 2 randomized, double-blind, placebo-controlled clinical trial was performed at 26 centers in the United States. There were 3 treatment groups: placebo, low dose (1×10^5 CD34/kg body wt) and high dose (5×10^5 CD34/kg). All pts underwent mobilization with GCSF 5 mcg/kg/day SC for 5 d, followed by apheresis on d5 to collect mononuclear cells. On day 6 CD34 were purified from the collected cells using the Isolex 300i device. After lot release testing, CD34 were injected endocardially at 10 locations in the ischemic Myo using the Myostar catheter following NOGA mapping. Placebo injections consisted of identical volumes of the diluent only. Results: A total of 167 pts were randomized and completed the injection procedure and 162 completed the 6 mo evaluation. The mean age of subjects was 61.0±8.9, there were 22 female and 145 male subjects. Trop was elevated in 26 pts at baseline and in 71 at some point following GCSF or injection. At 6 mo CD34 treated pts showed more reduction in ang frequency (−8.6 placebo vs 14.2 low dose) and improved ETT time (+69 sec placebo vs. +138 low dose). At 12m CD34 treated continue to show more reduction in angina (−8.2 placebo vs. −15.0 low dose) and improved ETT (+58 sec placebo vs. + 139 sec low dose). For key endpoints the high dose group results were inferior to low dose. The 12m data set has been locked and is being analyzed and the results of the completed analysis will be presented. Conclusions: Autologous CD34 cell therapy was associated with improved exercise tolerance and reduced angina in no-option pts with intractable angina. Ongoing analysis will determine endpoints, sample size and suitability of this therapy for a phase III study in no-option pts with refractory angina.
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