Douglas Landsittel

Gene Expression Alterations in Prostate Cancer Predicting Tumor Aggression and Preceding Development of Malignancy

PURPOSE The incidence of prostate cancer is frequent, occurring in almost one-third of men older than 45 years. Only a fraction of the cases reach the stages displaying clinical significance. Despite the advances in our understanding of prostate carcinogenesis and disease progression, our knowledge of this disease is still fragmented. Identification of the genes and patterns of gene expression will provide a more cohesive picture of prostate cancer biology. PATIENTS AND METHODS In this study, we performed a comprehensive gene expression analysis on 152 human samples including prostate cancer tissues, prostate tissues adjacent to tumor, and organ donor prostate tissues, obtained from men of various ages, using the Affymetrix (Santa Clara, CA) U95a, U95b, and U95c chip sets (37,777 genes and expression sequence tags). RESULTS Our results confirm an alteration of gene expression in prostate cancer when comparing with nontumor adjacent prostate tissues. However, our study also indicates that the gene expression pattern in tissues adjacent to cancer is so substantially altered that it resembles a cancer field effect. CONCLUSION We also found that gene expression patterns can be used to predict the aggressiveness of prostate cancer using a novel model.

Multiplexed Immunobead-Based Cytokine Profiling for Early Detection of Ovarian Cancer

Early detection of ovarian cancer might improve clinical outcome. Some studies have shown the role of cytokines as a new group of tumor markers for ovarian cancer. We hypothesized that a panel comprised of multiple cytokines, which individually may not show strong correlation with the disease, might provide higher diagnostic power. To evaluate the diagnostic utility of cytokine panel, we used a novel multianalyte LabMAP profiling technology that allows simultaneous measurement of multiple markers. Concentrations of 24 cytokines (cytokines/chemokines, growth, and angiogenic factors) in combination with cancer antigen-125 (CA-125), were measured in sera of 44 patients with early-stage ovarian cancer, 45 healthy women, and 37 patients with benign pelvic tumors. Six markers, i.e., interleukin (IL)-6, IL-8, epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (MCP-1), and CA-125, showed significant differences in serum concentrations between ovarian cancer and control groups. Out of this group, IL-6, IL-8, VEGF, EGF, and CA-125, were used in a classification tree analysis that resulted in 84% sensitivity at 95% specificity. The receiver operator characteristic curve created using the combination of markers produced sensitivities between 90% and 100% in the area of 80% to 90% specificity, whereas the receiver operator characteristic curve for CA-125 alone resulted in sensitivities of 70% to 80%. The classification tree analysis for discrimination of benign condition from ovarian cancer used CA-125, granulocyte colony-stimulating factor (G-CSF), IL-6, EGF, and VEGF resulting in 86.5% sensitivity and 93.0% specificity. The presented data show that simultaneous testing of a panel of serum cytokines and CA-125 using LabMAP technology may present a promising approach for ovarian cancer detection.

Kidney Volume and Functional Outcomes in Autosomal Dominant Polycystic Kidney Disease

BACKGROUND AND OBJECTIVES Autosomal dominant polycystic kidney disease (ADPKD) is characterized by increased total kidney volume (TKV) and renal failure. This study aimed to determine if height-adjusted TKV (htTKV) predicts the onset of renal insufficiency. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS This prospective, observational, longitudinal, multicenter study included 241 adults with ADPKD and preserved renal function. Magnetic resonance imaging and iothalamate clearance were used to measure htTKV and GFR, respectively. The association between baseline htTKV and the attainment of stage 3 CKD (GFR <60 ml/min per 1.73 m(2)) during follow-up was determined. RESULTS After a mean follow-up of 7.9 years, stage 3 CKD was attained in 30.7% of the enrollees. Using baseline htTKV, negative correlations with GFR increased from -0.22 at baseline to -0.65 at year 8. In multivariable analysis, a baseline htTKV increase of 100 cc/m significantly predicted the development of CKD within 8 years with an odds ratio of 1.48 (95% confidence interval: 1.29, 1.70). In receiver operator characteristic curve analysis, baseline htTKV of 600 cc/m most accurately defined the risk of developing stage 3 CKD within 8 years with an area under the curve of 0.84 (95% confidence interval: 0.79, 0.90). htTKV was a better predictor than baseline age, serum creatinine, BUN, urinary albumin, or monocyte chemotactic protein-1 excretion (P<0.05). CONCLUSIONS Baseline htTKV ≥600 cc/m predicted the risk of developing renal insufficiency in ADPKD patients at high risk for renal disease progression within 8 years of follow-up, qualifying htTKV as a prognostic biomarker in ADPKD.

Quality of Life, Functional Outcome, and Costs of Early Glottic Cancer†

Objective To analyze quality of life, functional outcome, and hidden costs by primary treatment with surgery or radiation therapy in patients with early glottic cancer.

Detection of Prostate Cancer with a Blood-Based Assay for Early Prostate Cancer Antigen

Prostate-specific antigen lacks specificity for prostate cancer, so the identification and characterization of a unique blood-based marker for the disease would provide for a more accurate diagnosis, reducing both unnecessary biopsies and patient uncertainty. We previously identified a novel biomarker for prostate cancer, early prostate cancer antigen (EPCA). EPCA antibodies positively stained the negative biopsies of men who, as much as 5 years later, were diagnosed with prostate cancer. The goal of this study was to determine whether EPCA antibodies could be used in a clinically applicable plasma-based immunoassay to specifically detect prostate cancer. Using an EPCA-based ELISA, the protein was measured in the plasma of 46 individuals, including prostate cancer patients, healthy individuals, other cancer patients, spinal cord injury victims, and patients with prostatitis. With a predetermined cutoff value of 1.7 absorbance at 450 nm, only the prostate cancer population, as a whole, expressed plasma-EPCA levels above the cutoff. Statistical analysis showed a significant difference in EPCA levels between the prostate cancer population and each of the other groups, specifically the healthy donors (P < 0.0001), bladder cancer patients (P = 0.03), and spinal cord injury patients (P = 0.001). Sensitivity of the EPCA assay for prostate cancer patients was 92% whereas the overall specificity was 94%. Specificity for the healthy donors was 100%. Although larger trials are required, this initial study shows the potential of EPCA to serve as a highly specific blood-based marker for prostate cancer. EPCA, when coupled with prostate-specific antigen, may help reduce the number of both unnecessary biopsies and undetected prostate tumors.

Corticospinal tract lesion load: An imaging biomarker for stroke motor outcomes

The aim of this work was to investigate whether an imaging measure of corticospinal tract (CST) injury in the acute phase can predict motor outcome at 3 months in comparison to clinical assessment of initial motor impairment.

Sentinel Lymph Node Micrometastasis as a Predictor of Axillary Tumor Burden

Abstract:   The sentinel lymph node biopsy (SLNB) procedure is an alternative method for assessing the axillary lymph node (ALN) status in patients with breast cancer. The SLNB carries the risk of a false‐negative result, with patients harboring positive ALNs in the face of a negative SLNB examination. In addition, the significance of a SLNB with cells identified only with keratin or with deposits less than 0.2 mm remains unresolved. We analyzed our SLNB data over the past 5 years in order to determine the relationship between SLN tumor burden and ALN tumor burden. Pathology files for the past 5 years at Magee‐Womens Hospital were searched for all SLNB cases that had an axillary lymph node dissection (ALND). Each SLNB case was reviewed and tabulated for breast tumor size, SLN tumor size, and largest tumor size in the ALND. Correlation and frequency distribution were performed for the status of all SLNs and ALNDs. Patterns of lymph node metastasis were recorded and the sizes of the SLN metastases were reported according to the recent Philadelphia Consensus Conference on Sentinel Lymph Nodes and the revised American Joint Committee on Cancer (AJCC) staging. SLN metastases were classified as immunohistochemistry (IHC) positive if only single keratin‐positive cells or clusters were present and were not observed with standard tissue stains, as submicrometastatic (SMM) if tumors were less than 0.2 mm (excluding IHC positive), as micrometastatic if tumors were larger than 0.2 mm but ≤2 mm, or as macrometastatic if tumors were larger than 2 mm. A total of 445 patients had both SLNB and ALND. Fifty percent (224/445) of cases were SLN positive, including 58 SLN positive/ALN positive cases and 166 SLN positive/ALN negative cases. Of the 221 patients in the SLN‐negative group, 4 were ALN positive (false‐negative SLN). The incidence of SLN metastases increased with tumor stage, with the percentage of SLN positives as follows: T1a, 2.1%; T1b, 10.9%; T1c, 51.7%; and T2, 35.3%. There were 4 of 41 patients (10%) with SLNs that were IHC positive that had macrometastases in a solitary ALN. Three of 22 patients (13.6%) that were SMM positive had ALN macrometastasis in a solitary ALN. Four of 49 patients (8.1%) with micrometastatic SLNs had a solitary positive ALN, 3 of which were macrometastases (6.1%). Overall a total of 10 of 112 patients (9.0%) with traditionally defined SLN micrometastases of 2.0 mm or less had a solitary ALN macrometastasis. The vast majority (90%) of these macrometastases were found with T1c and T2 breast tumors. There was a significant difference in the means of SLN tumor sizes for the SLN‐positive/ALND‐negative (4.5 mm) versus SLN‐positive/ALND‐positive (10.1 mm) patients, although the range of SLN tumor sizes within each group were similar. There is an increasing incidence of SLN‐positive and ALN‐positive cases with increasing T stage. Overall in this series, 9% of patients with SLN metastases ≤2 mm had a solitary axillary macrometastasis. Ninety percent of these metastases occurred with T1c/T2 breast tumors, indicating the important codependence of T stage. Overall there is a subset of patients who are IHC positive, SMM positive, or micrometastatic positive with ALNs that are macrometastatic who are at risk of harboring axillary macrometastases. Keratin IHC of breast SLNs is useful for defining these subsets. 

Facial Anthropometric Differences among Gender, Ethnicity, and Age Groups

OBJECTIVES The impact of race/ethnicity upon facial anthropometric data in the US workforce, on the development of personal protective equipment, has not been investigated to any significant degree. The proliferation of minority populations in the US workforce has increased the need to investigate differences in facial dimensions among these workers. The objective of this study was to determine the face shape and size differences among race and age groups from the National Institute for Occupational Safety and Health survey of 3997 US civilian workers. METHODS Survey participants were divided into two gender groups, four racial/ethnic groups, and three age groups. Measurements of height, weight, neck circumference, and 18 facial dimensions were collected using traditional anthropometric techniques. A multivariate analysis of the data was performed using Principal Component Analysis. An exploratory analysis to determine the effect of different demographic factors had on anthropometric features was assessed via a linear model. The 21 anthropometric measurements, body mass index, and the first and second principal component scores were dependent variables, while gender, ethnicity, age, occupation, weight, and height served as independent variables. RESULTS Gender significantly contributes to size for 19 of 24 dependent variables. African-Americans have statistically shorter, wider, and shallower noses than Caucasians. Hispanic workers have 14 facial features that are significantly larger than Caucasians, while their nose protrusion, height, and head length are significantly shorter. The other ethnic group was composed primarily of Asian subjects and has statistically different dimensions from Caucasians for 16 anthropometric values. Nineteen anthropometric values for subjects at least 45 years of age are statistically different from those measured for subjects between 18 and 29 years of age. Workers employed in manufacturing, fire fighting, healthcare, law enforcement, and other occupational groups have facial features that differ significantly than those in construction. CONCLUSIONS Statistically significant differences in facial anthropometric dimensions (P < 0.05) were noted between males and females, all racial/ethnic groups, and the subjects who were at least 45 years old when compared to workers between 18 and 29 years of age. These findings could be important to the design and manufacture of respirators, as well as employers responsible for supplying respiratory protective equipment to their employees.

TIME COURSE OF PULMONARY RESPONSE OF RATS TO INHALATION OF CRYSTALLINE SILICA: NF-kappa B ACTIVATION, INFLAMMATION, CYTOKINE PRODUCTION, AND DAMAGE

In vitro studies suggest that silica-induced lung disease may be linked to processes regulated by nuclear factor- κ B (NF- κ B) activation, but this has not been examined in vivo. Rats were exposed to a silica aerosol of 15 mg/m 3 (6 h/day, 5 days/wk) for 116 days, and bronchoalveolar lavage (BAL) was conducted at various times during the exposure. Silica-induced pulmonary inflammation and damage were determined by measuring BAL cell differentials and first BAL fluid lactate dehydrogenase (LDH) activity and serum albumin concentrations, respectively. NF- κ B activation and production of tumor necrosis factor-α (TNF-α) and interleukin-1 (IL-1) by BAL cells were also measured. The results demonstrate that NF- κ B activation occurred after 5 days exposure, and continued to increase thereafter. BAL cell production of IL-1 and TNF-α had increased incrementally by 10 and 30 days of exposure, respectively. This elevation continued through 79 days of exposure before further increasing at 116 days of exposure. Pulmonary inflammation and damage in silica-exposed rats were also significantly elevated at 5 days of exposure, further increased at a slow rate through 41 days of exposure, and dramatically increased thereafter. Taken together, the results indicate that the initial molecular response of NF- κ B activation in BAL cells occurs in response to low levels of silica deposition in the lung and increases more rapidly versus exposure duration than silica-induced pulmonary inflammation, cellular damage, and cytokine production by BAL cells. This suggests that NF- κ B activation in BAL cells may play an important role in the initiation and progression of silica-induced pulmonary inflammation, cellular damage, and fibrosis.

Polymorphisms of the IL-1 gene complex in coal miners with silicosis.

BACKGROUND Silicosis is characterized by fibrosing nodular lesions that eventually develop into progressive pulmonary fibrosis. Pro-inflammatory cytokines, such as interleukin-1 (IL-1), play a key role in the development of silicosis by regulating mediators which are responsible for lung injury, inflammation, and potentially fibrosis. To study whether functional single nucleotide polymorphisms (SNPs) located in the regulatory elements of genes coding for the IL-1alpha, IL-1beta, and IL-1 receptor antagonist (RA) cytokines are associated with silicosis, we examined 318 Caucasian cases confirmed histopathologically with pulmonary silicosis and 163 controls without any apparent inflammation or other pulmonary disease. METHODS Genotyping was carried out by polymerase chain reaction-restriction fragment length polymorphism technique. RESULTS The proportion of the IL-1RA (+ 2018) allele 2 genotype was increased in miners with silicosis (0.27) compared to controls (0.16). The odds of being a case were 2.15 (CI = 1.4-3.3) times higher for subjects with at least one copy of allele 2. No statistically significant differences in the allelic frequencies or genotype distributions for IL-1alpha (+ 4845) or IL-1beta (+ 3953) were found between the control and disease groups. CONCLUSIONS This is the first report showing an association between the IL-1RA (+ 2018) polymorphism and silicosis, and suggests that this polymorphism may confer increased risk for the development of the disease.