Space shuttle main engine fault detection using neural networks

A method for on-line Space Shuttle Main Engine (SSME) anomaly detection and fault typing using a feedback neural network is described. The method involves the computation of features representing time-variance of SSME sensor parameters, using historical test case data. The network is trained, using backpropagation, to recognize a set of fault cases. The network is then able to diagnose new fault cases correctly. An essential element of the training technique is the inclusion of randomly generated data along with the real data, in order to span the entire input space of potential non-nominal data

New candidate biomarkers in the female genital tract to evaluate microbicide toxicity.

Vaginal microbicides hold great promise for the prevention of viral diseases like HIV, but the failure of several microbicide candidates in clinical trials has raised important questions regarding the parameters to be evaluated to determine in vivo efficacy in humans. Clinical trials of the candidate microbicides nonoxynol-9 (N9) and cellulose sulfate revealed an increase in HIV infection, vaginal inflammation, and recruitment of HIV susceptible lymphocytes, highlighting the need to identify biomarkers that can accurately predict microbicide toxicity early in preclinical development and in human trials. We used quantitative proteomics and RT-PCR approaches in mice and rabbits to identify protein changes in vaginal fluid and tissue in response to treatment with N9 or benzalkonium chloride (BZK). We compared changes generated with N9 and BZK treatment to the changes generated in response to tenofovir gel, a candidate microbicide that holds promise as a safe and effective microbicide. Both compounds down regulated mucin 5 subtype B, and peptidoglycan recognition protein 1 in vaginal tissue; however, mucosal brush samples also showed upregulation of plasma proteins fibrinogen, plasminogen, apolipoprotein A-1, and apolipoprotein C-1, which may be a response to the erosive nature of N9 and BZK. Additional proteins down-regulated in vaginal tissue by N9 or BZK treatment include CD166 antigen, olfactomedin-4, and anterior gradient protein 2 homolog. We also observed increases in the expression of C-C chemokines CCL3, CCL5, and CCL7 in response to treatment. There was concordance in expression level changes for several of these proteins using both the mouse and rabbit models. Using a human vaginal epithelial cell line, the expression of mucin 5 subtype B and olfactomedin-4 were down-regulated in response to N9, suggesting these markers could apply to humans. These data identifies new proteins that after further validation could become part of a panel of biomarkers to effectively evaluate microbicide toxicity

ELISA analysis of rabbit proteins in vaginal brush samples increased in response to treatment with N9 or BZK.

<p>Statistical analysis were performed using a two-tailed Student's T-test of N9 and BZK samples compared to vehicle controls (** = p≤0.01, * = p≤0.05, NS = not significant).</p

Summary of proteins whose levels significantly altered upon N9 or BZK treatment determined by proteomic analysis in mouse vaginal brush and tissue samples.

<p>*Average signal compared to vehicle controls which are set to a default value of 1,</p>†<p>number of studies where protein was identified (for both N9 and BZK), ND = not detected,</p>††<p>proteins shown in bold are identified in both brush and tissue samples.</p><p>Summary of proteins whose levels significantly altered upon N9 or BZK treatment determined by proteomic analysis in mouse vaginal brush and tissue samples.</p

RT-PCR analysis of mouse CCL3, CCL5, and CCL7 gene expression.

<p>Relative expression levels are normalized to Glyceraldehyde 3-phosphate dehydrogenase (GADPH). Statistical analysis was performed using a two-tailed Student's T-test of N9, BZK, and Tenofovir samples (3 replicates) compared to vehicle controls (*** = p≤0.001, ** = p≤0.01,* = p≤0.05, NS = not significant).</p

Quantitative proteomic analysis of mouse brush samples.

<p>(A) LC-MS/MS analysis of mouse proteins in vaginal brush samples that are affected by treatment with N9 (open, black circles) or BZK (closed squares). Shown are proteins that are increased compared to vehicle controls. The following proteins are depicted in the graph: fibrinogen alpha polypeptide isoform 2 (FGA), plasminogen (PLG), apolipoprotein A-1 (Apo A-1), and apolipoprotein C-1 (Apo C-1). Each data point represents the average of three replicates per individual experimental study. Only proteins shown to change in four studies are represented. (B) ELISA analysis of mouse proteins in vaginal brush samples that are increased in response to treatment with 8% N9 or 2% BZK. Statistical analysis was performed using a two-tailed Student's T-test of N9 and BZK samples compared to vehicle controls (**** = p≤0.0001, ** = p≤0.01). (C) ELISA analysis of mouse proteins in vaginal brush samples that are increased in response to treatment with vehicle, 1% N9, 4% N9, or 8% N9. Statistical analysis was performed using a two-tailed student's t-test of N9 and BZK samples compared to vehicle controls (**** = p≤0.0001, *** = p≤0.001, ** = p≤0.01, * = p≤0.05, NS = not significant).</p

mRNA expression of cyclooxygenase-2 (cox2), Interleukin 8 (IL-8), mucin 5B, and olfactomedin-4 (OLFM-4) in Vκ2 cells treated with N9 for 6, 24, or 48 hours.

<p>Shown is the average of two replicate experiments, each sample performed in duplicate for each time point. Also shown is the relative expression levels of N9 treated cells compared to untreated cells, which was set to an expression level of 1 (dashed line). Statistical analysis was performed using a two-tailed Student's T-test of N9 samples compared to untreated controls at each time point (*** = p≤0.001, ** = p≤0.01, * = p≤0.05, NS = not significant).</p

Model for the protective roles of mucin 5B, olfactomedin-4 (OLFM-4), and peptidoglycan recognition protein 1 (PGLYRP-1) in vaginal epithelium.

<p>In normal epithelium (top) these proteins maintain a balanced microflora and suppress inflammation. However, when exposed to toxic microbicides such as N9 (bottom) the epithelium becomes inflamed causing bleeding and release of serum proteins into the vaginal cavity. In addition, the microflora is altered leading to dysbiosis which could boost the inflammation response. The result is an influx and expansion of leukocytes that may be susceptible to HIV.</p

Quantitative proteomic and RT-PCR analysis of mouse tissue samples.

<p>(A) LC MS/MS proteomic analysis of mouse vaginal tissue samples down-regulated in response to treatment with N9 (open, black circles) or BZK (black squares). The following proteins are depicted in the graphs: peptidoglycan recognition protein 1 (PGLYRP-1), CD166 antigen (CD166), mucin 5 subtype B (mucin 5B), olfactomedin-4 (OLFM-4), and anterior gradient protein 2 homolog (AGR2). Each data point represents the average of three replicates per individual experimental study. Only proteins shown to change in a minimum of two studies are represented. (B) RT-PCR analysis of mouse mRNA expression. Relative expression levels are normalized to Glyceraldehyde 3-phosphate dehydrogenase (GADPH). Statistical analysis was performed using a two-tailed Student's T-test of N9, BZK, and tenofovir samples compared to vehicle controls (*** = p≤0.001, * = p≤0.05, NS = not significant). (C) Immunoblot of mouse CD166 antigen. Three mice per condition were analyzed. The CD166 signal was normalized to the β-actin signal for quantitation. Statistical analysis was performed using a two-tailed student's T-test of N9 and BZK samples compared to vehicle controls (*** = p≤0.001, ** = p≤0.01).</p

LC MS/MS proteomic analysis of mouse proteins in vaginal brush samples that are down-regulated in response to treatment with N9 (open, black circles) or BZK (closed squares).

<p>Shown are proteins that were decreased relative to vehicle controls. The following proteins are shown: peptidoglycan recognition protein 1 (PGLYRP-1), mucin 5 subtype B (mucin 5B), destrin, and carbonyl reductase 3 (CAR3). Each data point represents the average of three replicates per individual experimental study. Only proteins shown to change in a minimum of two studies are represented.</p