Evaluation of symmetric dimethylarginine and creatinine in dogs with primary hypoadrenocorticism receiving long-term mineralocorticoid replacement therapy.

OBJECTIVES: To investigate kidney function by determining serum symmetric dimethylarginine (sSDMA) and serum creatinine (sCr) concentrations in dogs with primary hypoadrenocorticism (PH) receiving long-term mineralocorticoid replacement therapy. METHODS: Dogs with PH receiving a minimum of 12 months of either desoxycorticosterone pivalate or fludrocortisone acetate were included in the study provided that banked frozen serum samples were available for sSDMA analysis. sCr concentrations were retrieved from the medical records. In dogs still alive and presented for regular re-evaluations and in newly diagnosed patients, blood was prospectively collected for sSDMA and sCr determination. RESULTS: Thirty-two dogs met the inclusion criteria. The treatment time ranged from 12 to 146 months after initial diagnosis (median, 55.5 months). The majority of dogs had normal sSDMA and sCr concentrations throughout the hormone replacement treatment. Both sSDMA and sCr concentrations were persistently elevated in three of 32 dogs. Further workup confirmed chronic kidney disease (CKD) in all three dogs. CONCLUSIONS: Based on these data, the prevalence of CKD could be higher in dogs with PH receiving long-term mineralocorticoid replacement treatment than in the general dog population. However, additional studies with a larger number of dogs are needed to confirm it.

Sonographic findings for the common extensor tendon of the elbow in the general population.

OBJECTIVE: To describe structural characteristics and sonographic alterations of the common extensor tendon (CET). METHODS: In 240 patients, we determined the body mass index; sonographic CET thickness, morphologic characteristics, and alterations; and color Doppler measurements of the epicondylar artery width and peak systolic velocity. Age, sex, epicondylalgia history, activities (work, sports, and hobbies), and dominant and nondominant elbows were noted. RESULTS: The CET was thicker in the dominant elbow (4.77 versus 4.61 mm [P = .023]), male patients (dominant, 5.09 versus 4.46 mm [P < .001]; nondominant, 5.00 versus 4.21 mm [P < .001]), patients involved in risk activities (dominant, 5.21 versus 4.70 mm [P < .001]; nondominant, 5.12 versus 4.53 mm [P < .001]), and those with a history of epicondylalgia (right, 5.27 versus 4.70 mm [P < .001]; left, 4.86 versus 4.60 mm [P = .316]). Thickness correlated weakly with age (dominant, r = 0.284; nondominant, r = 0.215) and moderately with weight (dominant, r = 0.492; nondominant, r = 0.502). The mean epicondylar artery diameter was 1.35 mm (SD, 0.96 mm); mean peak velocity, 13.01 cm/s (SD, 4.90 cm/s). Morphologic abnormalities were found in 79.5% of patients with a history of epicondylalgia; 7.7% with no history had abnormalities; and 55.9% with abnormalities but no history were older than 55 years. Bone spurs (49.2% versus 16.4% [P < .001]), tendon calcifications (21.5% versus 3.9% [P < .001]), and bone cortex abnormalities (12.3% versus 1.7% [P < .001]) were found more often in non-normal elbows; 79.4% of bone spurs in normal elbows occurred in patients older than 55 years. CONCLUSIONS: Mild sonographic alterations in the CET and bone spurs should be considered with caution when assessing epicondylar pain, especially in patients older than 55 years and those with a history of epicondylalgia.

Tissue sample collection for proteomics analysis.

Successful collection of tissue samples for molecular analysis requires critical considerations. We describe here our procedure for tissue specimen collection for proteomic purposes with emphasis on the most important steps, including timing issues and the procedures for immediate freezing, storage, and microdissection of the cells of interest or "tissue targets" and the lysates for protein isolation for SELDI, MALDI, and 2DGE applications. The pathologist is at the cornerstone of this process and is an invaluable collaborator. In most institutions, pathologists are responsible for "tissue custody," and they closely supervise the tissue bank. In addition, they are optimally trained in histopathology in order to they assist investigators to correlate tissue morphology with molecular findings. In recent years, the advent of the laser capture microscope, a tool ideally designed for pathologists, has tremendously facilitated the efficiency of collecting tissue targets for molecular analysis.

MALDI/SELDI protein profiling of serum for the identification of cancer biomarkers.

The ability to visualize the full depth of the serum proteome in a high-throughput manner is a major goal of clinical proteomics. Methodologies, which combine higher throughput with the ability to observe differential protein expression levels, have been applied to this goal. An example of such a system is the coupling of robotic sample processing to matrix-assisted laser desorption time of flight mass spectrometry (MALDI-TOF-MS). Within this paradigm is a modification of MALDI-TOF termed surface-enhanced laser desorption/ionization-TOF (SELDI-TOF). Both conventional MALDI and SELDI have been used to generate protein expression profiles reflective of potential peptide changes in serum. This information can be used to identify proteins, which may enable new diagnostic and therapeutic strategies.

Computer modeling technology to assess extracapsular tissue coverage of whole mount sections after retropubic and laparoscopic radical prostatectomy.

PURPOSE: The introduction of new surgical approaches to radical prostatectomy requires methodologies that permit valid comparison that are more expedient than long-term outcomes of biochemical local and distant failure and survival. We used a computer modeling program to assess the percent of extracapsular tissue coverage of prostate glands removed by the open retropubic and laparoscopic approaches. MATERIALS AND METHODS: Specimens were available for 15 and 17 patients who underwent open and laparoscopic radical prostatectomy, respectively. Serial whole mount sections were taken at 5 mm intervals. A genitourinary pathologist drew the contours of the prostate capsule on each tissue section. The whole mount was scanned to produce digital images. A software program was used to create a file with capsule information and a file with extraprostatic fibroadipose tissue information. Two separate point cloud files were generated to represent the capsule and extraprostatic models, and software algorithms were used to generate differences in the point clouds to quantify the extent of extracapsular tissue coverage. RESULTS: When separated into sides dissected by a nerve or nonnerve sparing technique, the overall percent of gland surface coverage by extracapsular fibroadipose tissue was statistically greater with laparoscopic dissection than with the open approach. When a segmental analysis of gland coverage was evaluated, a statistically greater percent of fibroadipose coverage was associated with laparoscopic dissection in the apical and inferolateral segments with nonnerve sparing, and in the apical segment with nerve sparing. CONCLUSIONS: This small radical prostatectomy series, analyzed by computer reconstruction as described, provides information suggesting that overall extracapsular tissue coverage is at least equal if not superior using the laparoscopic vs the open approach. This was specifically the case in areas of inferolateral and apical dissection with nonnerve sparing procedures and in areas of the apical dissection with nerve sparing procedures.

Overexpression of alpha-defensin is associated with bladder cancer invasiveness.

Alpha-defensin (alpha-defensin) has been identified as a potential marker for bladder cancer in urine by surface enhanced laser desorption ionization studies, and confirmed using both immunoabsorption and immunodepletion studies. The objective of this study was to investigate the role of alpha-defensin in bladder cancer. Immunohistochemical analysis of tissue sections showed that alpha-defensin peptides are frequently expressed in bladder cancer cells. It is noteworthy that expression of alpha-defensins increased with tumor invasiveness. Surface enhanced laser desorption ionization analysis showed the presence of alpha-defensin in the T24 and A498 cancer cell lines. These cell lines show higher classically aggressive in vitro characteristics compared with the J82 cells that did not express alpha-defensin. Exogenously added alpha-defensin increased the proliferation and motility/invasiveness of these cell lines using respective assays. It is interesting that alpha-defensin peptides increased intracellular calcium ions (Ca2+). These data are consistent with a role for alpha-defensin in bladder cancer via modulation of cell motility and invasiveness using common intracellular signals, such as Ca2+. We propose that autocrine tumor expression of alpha-defensins may play an important role in facilitating the invasive phenotype of bladder cancer in patients.

Laparoscopic radical prostatectomy: evaluation of specimen pathologic features to critically assess and modify surgical technique.

OBJECTIVES: To evaluate laparoscopic radical prostatectomy (LRP) specimens for pathologic stage and the malignant and benign surgical margin status to determine areas for improvement in surgical technique. LRP is gaining significant attention as a viable alternative to retropubic radical prostatectomy. METHODS: A total of 110 consecutive LRP specimens were retrospectively reviewed. A single experienced genitourinary pathologist examined all slides. In addition, exposure of benign glands at the inked margin was reported. Oncologic data were assessed by standard pathologic examination for stage, grade, and margin status. The pathologic data from year 1 (group 1, 52 patients) were compared with the data from subsequent years (group 2, 58 patients). RESULTS: The overall positive surgical margin rate was 18.2% (13.5% for pT2 disease). Benign parenchymal exposure was identified in 16 patients (14.5%). Six of these patients had concomitant negative surgical margins for cancer. The positive margin rate for patients with Stage pT2 disease was 22.2% in group 1 and 5.9% in group 2. The positive margin rate was 34.8%, 13.6%, and 13.8% for bilateral, unilateral, and non-nerve-sparing procedures, respectively. CONCLUSIONS: LRP offers comparable surgical margins to those of reported open prostatectomy series. Careful pathologic review provides feedback so that technique modifications can be implemented and the positive margin rates improved further. A low, but significant, positive margin or benign parenchymal exposure rate in pT2 disease indicates room for improvement in the surgical technique. The presence of benign glands at the inked margin is not routinely reported but does provide an additional parameter of surgical adequacy.

Serum levels of an isoform of apolipoprotein A-II as a potential marker for prostate cancer.

PURPOSE: We recently showed that protein expression profiling of serum using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) has potential as a diagnostic approach for detection of prostate cancer. As a parallel effort, we have been pursuing the identification of the protein(s) comprising the individual discriminatory "peaks" and evaluating their utility as potential biomarkers for prostate disease. EXPERIMENTAL DESIGN: We employed liquid chromatography, gel electrophoresis and tandem mass spectroscopy to isolate and identify a protein that correlates with observed SELDI-TOF MS mass/charge (m/z) values. Immunodepletion, immunoassay, and Western analysis were used to verify that the identified protein generated the observed SELDI peak. Subsequent immunohistochemistry was used to examine the expression of the proteins in prostate tumors. RESULTS: An 8,946 m/z SELDI-TOF MS peak was found to retain discriminatory value in each of two separate data sets with an increased expression in the diseased state. Sequence identification by liquid chromatography-MS/MS and subsequent immunoassays verified that an isoform of apolipoprotein A-II (ApoA-II) is the observed 8,946 m/z SELDI peak. Immunohistochemistry revealed that ApoA-II is overexpressed in prostate tumors. SELDI-based immunoassay revealed that an 8.9-kDa isoform of ApoA-II is specifically overexpressed in serum from individuals with prostate cancer. ApoA-II was also overexpressed in the serum of individuals with prostate cancer who have normal prostate-specific antigen (0-4.0 ng/mL). CONCLUSIONS: We have identified an isoform of ApoA-II giving rise to an 8.9K m/z SELDI "peak" that is specifically overexpressed in prostate disease. The ability of ApoA-II to detect disease in patients with normal prostate-specific antigen suggests potential utility of the marker in identifying indolent disease.

Selective capture of prostatic basal cells and secretory epithelial cells for proteomic and genomic analysis.

Basal cells play an undefined role in signaling the growth and differentiation of normal secretory epithelial cells in the human prostate. Because basal cells disappear during malignant transformation, we hypothesize that loss of basal cell function may have a permissive role in progression of prostate intraepithelial neoplasia into invasive carcinoma. We describe an immuno-laser capture microdissection approach to selectively capture basal cells. Using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, we identified several protein candidates selectively expressed in microdissected basal cells. We also demonstrate that the RNA derived form this technique is an excellent source for gene-array studies. Thus, we provide evidence that proteomic and microgenomic techniques can be successfully applied to investigate the expression profiles of basal and secretory cells after immuno-capture.

Laparoscopic radical prostatectomy: is intact organ removal attainable? Study of margin status.

PURPOSE: To determine the initial oncologic results (pathology) of specimens removed by laparoscopic radical prostatectomy (LRP) by examining the surgical margins. PATIENTS AND METHODS: The 70 consecutive LRP procedures performed for clinically localized prostate cancer at Eastern Virginia Medical School from April 2001 to November 2002 were reviewed for preoperative and important intraoperative variables. The initial histopathology report and a prospective review by a single genitourinary pathologist for margin status as well parenchymal exposure of benign glands were assessed. Postoperative prostate specific antigen (PSA) levels were noted. RESULTS: The clinical stage distribution was as follows: T1c = 59, T2a = 10, and T2b = 1. The preoperative median PSA value was 6.96 ng/mL. The mean operative time was 307 minutes. The mean estimated blood loss was 298 mL. The prospective pathologic review results were as follows: 1 pT0 (1.4%), 60 pT2 (85.7%), and 9 pT3 (12.8%). The overall positive surgical-margin rate was 15.7%. Of those patients with pT2 disease; 8 specimens (13.3%) had a positive margin, whereas 33% of patients with pT3 disease had a positive surgical margin. Parenchymal exposure of benign glands on the inked surface was recognized in 8 patients (13.3%). Data from PSA assays 1 month postoperatively were available in 69 patients. Serum PSA was undetectable (< or =0.1 ng/mL) in 67 men (97%). CONCLUSION: An LRP can offer surgical margins comparable to those of open procedures series reported in the literature. Long-term progression and survival outcome data are necessary before this procedure should be offered as a replacement for open prostatectomy.

Multilocular prostatic cystadenoma with high-grade prostatic intraepithelial neoplasia.

Multilocular prostatic cystadenoma is a rarely encountered neoplasm located in the midline between the bladder and rectum that is either attached to the prostate by a pedicle or separate from the prostate entirely. Histologically and immunohistochemically these lesions resemble benign prostate tissue. We report the first case of this entity for which multifocal high-grade prostatic intraepithelial neoplasia (PIN) is identified. Conceptually, the finding of high-grade PIN in multilocular prostatic cystadenomas provides further evidence that these lesions are fully analogous to the prostate gland not only in their morphology and immunohistochemistry but also in their predilection for the same diseases.

Incidencia, etiología y manifestaciones clínicas de anemia ferropénica: estudio en un área de salud / Incidence, etiology and clinical manifestation of iron deficiency anemia: study in a health area

Se diagnostican durante un período de 10 meses, en la consulta externa de un área de salud predominantemente urbana, de 18 194 habitantes, 76 pacientes afectos de anemia ferropénica, por presentar síntomas y signos, o por estudios hematológicos realizados por otras causas. Se les realiza a todos los casos una encuesta, en busca de datos de interés relacionados con la etiología y las manifestaciones clínicas. Se informa que la anemia ferropénica fue más frecuente en pacientes del sexo femenino, de la raza blanca y con edades inferiores a los 50 años. Predominaron las formas ligeras y moderadas de anemia y las principales causas fueron los sangramientos por causas ginecológicas o por alteraciones del tracto digestivo

Incidencia, etiología y manifestaciones clínicas de anemia ferropénica: estudio en un área de salud / Incidence, etiology and clinical anifestation of iron deficiency anemia: study in a health area

Se diagnostican durante un período de 10 meses, en la consulta externa de un área de salud predominantemente urbana, de 18 194 habitantes, 76 pacientes afectos de anemia ferropénica, por presentar síntomas y signos, o por estudios hematológicos realizados por otras causas. Se les realiza a todos los casos una encuesta, en busca de datos de interés relacionados con la etiología y las manifestaciones clínicas. Se informa que la anemia ferropénica fue más frecuente en pacientes del sexo femenino, de la raza blanca y con edades inferiores a los 50 años. Predominaron las formas ligeras y moderadas de anemia y las principales causas fueron los sangramientos por causas ginecológicas o por alteraciones del tracto digestivo (AU)