Comparison of Dako HercepTest and Ventana PATHWAY Anti-HER2 (4B5) Tests and Their Correlation With Fluorescent In Situ Hybridization in Breast Carcinoma.

OBJECTIVES: We compared the performance of two Food and Drug Administration-approved HER2 immunohistochemistry (IHC) tests: HercepTest (Dako) and PATHWAY anti-HER2 (4B5) (Ventana). MATERIALS AND METHODS: In total, 180 invasive breast carcinomas previously tested by both HercepTest and fluorescent in situ hybridization (FISH) were retested with 4B5. Three pathologists scored the HER2 IHC using the 2013 American Society of Clinical Oncology/College of American Pathologists guidelines. The HER2 IHC results were correlated with FISH. RESULTS: Among 135 equivocal cases by HercepTest, 100 (74.1%) were negative by 4B5. Among 45 positive HercepTest cases 9 (20%) were equivocal by 4B5. Among 135 equivocal HercepTest results, 100 (74.1%) were nonamplified, 18 (13.3%) equivocal, and 17 (12.6%) amplified by FISH. Among the 45 positive results with HercepTest, 2 (4.5%) were nonamplified and 1 (2.2%) was equivocal by FISH. All 37 positive and 3 negative by 4B5 cases were amplified by FISH. The absolute interobserver agreement was high for both tests (Fleiss kappa=0.838 for HercepTest and 0.771 for 4B5). CONCLUSIONS: PATHWAY anti-HER2 (4B5) significantly reduced the number of equivocal results that require additional testing. Although HercepTest was positive in a small number of HER2 nonamplified cases, 4B5 failed to detect 3 cases that were interpreted as positive by FISH, all with nonclassic or low levels of amplification.

Acute Myeloid Leukemia With a Rare t(7;14)(q21;q32) and Trisomy 4 With Poor Clinical Outcome: A Case Report.

OBJECTIVES: Recurrent cytogenetic abnormalities and/or molecular aberrations play an important role in the diagnosis and prognostification of acute myeloid leukemia (AML). We describe a case of a 40 year old woman diagnosed with de novo AML with a novel t(7;14)(q21,q32) and trisomy 4 with poor clinical outcome. Methods: Morphologic, flow cytometry and cytogenetic results of the patient's peripheral blood and bone marrow samples were analyzed. RESULTS: The diagnostic bone marrow was hypercellular for age (>95%) with increased blasts (62%) that by flow cytometry exhibited myeloid differentiation with a few T/NK lineage markers. Cytogenetics showed a t(7;14)(q21,q32) and trisomy 4. The patient had extremely poor response to two rounds of induction chemotherapy with persistent leukemia following therapy. CONCLUSION: To the best of our knowledge, the t(7;14) is a novel cytogenetic abnormality that has not been reported previously in acute myeloid leukemia, and is important to report as it appears to be associated with poor prognosis.

Pathologic Findings of Breast Lesions Detected on Magnetic Resonance Imaging.

CONTEXT: - Breast magnetic resonance imaging (MRI) is now used routinely for high-risk screening and in the evaluation of the extent of disease in newly diagnosed breast cancer patients. Morphologic characteristics and the kinetic pattern largely determine how suspicious a breast lesion is on MRI. Because of its high sensitivity, MRI identifies a large number of suspicious lesions. However, the low to moderate specificity and the additional cost have raised questions regarding its frequent use. OBJECTIVES: - To identify the pathologic entities that frequently present as suspicious enhancing lesions and to identify specific MRI characteristics that may be predictive of malignancy. DESIGN: - One hundred seventy-seven MRI-guided biopsies from 152 patients were included in the study. The indication for MRI, MRI features, pathologic findings, and patient demographics were recorded. The MRI findings and the pathology slides were reviewed by a dedicated breast radiologist and breast pathologists. RESULTS: - Seventy-one percent (126 of 177) of MRI-guided breast biopsies were benign, 11% (20 of 177) showed epithelial atypia, and 18% (31 of 177) showed malignancy. The vast majority (84%; 62 of 74) of MRI lesions with persistent kinetics were benign. However, 57% (17 of 30) of lesions with washout kinetics and 65% (62 of 95) of mass lesions were also benign. CONCLUSIONS: - Magnetic resonance imaging detects malignancies undetected by other imaging modalities but also detects a wide variety of benign lesions. Benign and malignant lesions identified by MRI share similar morphologic and kinetic features, necessitating biopsy for histologic confirmation.

Visually-driven ocular growth in mice requires functional rod photoreceptors.

PURPOSE: Proper refractive eye growth depends on several features of the visual image and requisite retinal pathways. In this study, we determined the contribution of rod pathways to normal refractive development and form deprivation (FD) myopia by testing Gnat1(-/-) mice, which lack functional rods due to a mutation in rod transducin-α. METHODS: Refractive development was measured in Gnat1(-/-) (n = 30-36) and wild-type (WT) mice (n = 5-9) from 4 to 12 weeks of age. FD was induced monocularly from 4 weeks of age using head-mounted diffuser goggles (Gnat1(-/-), n = 9-10; WT, n = 7-8). Refractive state and ocular biometry were obtained weekly using a photorefractor, 1310 nm optical coherence tomography, and partial coherence interferometry. We measured retinal dopamine and its metabolite, DOPAC, using HPLC. RESULTS: During normal development, the refractions of WT mice started at 5.36 ± 0.68 diopters (D) and became more hyperopic before plateauing at 7.78 ± 0.64 D. In contrast, refractions in Gnat1(-/-) mice were stable at 7.39 ± 1.22 D across all ages. Three weeks of FD induced a 2.54 ± 0.77 D myopic shift in WT mice, while Gnat1(-/-) mice did not respond to FD at any age. Axial lengths of Gnat1(-/-) and WT mice increased with age, but differences between genotypes or with goggling did not reach statistical significance and fell within the precision of the instruments. The DOPAC levels were significantly lower in Gnat1(-/-) mice from 2 to 12 weeks of age with DOPAC/dopamine ratio peaking earlier in Gnat1(-/-) compared to WT mice. No differences in dopamine were seen in response to FD or between genotypes. CONCLUSIONS: Functional rod photoreceptors are critical to normal refractive development and the response to FD in mice. Dopamine levels may not directly modulate the refractive state of the mouse eye, but tonic levels of dopamine during development may determine susceptibility to myopia.

Fatal caffeine overdose: a case report and review of literature.

Caffeine is a central nervous system stimulant that is consumed by large numbers of people on a routine basis, usually in the form of coffee or tea. However, if consumed in high doses, this xanthine alkaloid is profoundly toxic and can result in death. Increasingly being sold as a dietary supplement, many people, particularly those in the health and fitness community, where it is touted as a fitness and muscle building aid, are consuming caffeine anhydrous on a daily basis. We report a case of fatal caffeine overdose in a 39-year-old man resulting from the self-administered ingestion of approximately 12 g of pure caffeine anhydrous. Autopsy blood caffeine levels were 350 mg/L. We recommend mandated labeling of pure caffeine anhydrous, highlighting the toxicity risk of ingesting this chemical; and we recommend ensuring that caffeine levels are included in the comprehensive forensic toxicology panel performed on all cases.

Retinal degeneration increases susceptibility to myopia in mice.

PURPOSE: Retinal diseases are often associated with refractive errors, suggesting the importance of normal retinal signaling during emmetropization. For instance, retinitis pigmentosa, a disease characterized by severe photoreceptor degeneration, is associated with myopia; however, the underlying link between these conditions is not known. This study examines the influence of photoreceptor degeneration on refractive development by testing two mouse models of retinitis pigmentosa under normal and form deprivation visual conditions. Dopamine, a potential stop signal for refractive eye growth, was assessed as a potential underlying mechanism. METHODS: Refractive eye growth in mice that were homozygous for a mutation in Pde6b, Pde6b(rd1/rd1) (rd1), or Pde6b(rd10/rd10) (rd10) was measured weekly from 4 to 12 weeks of age and compared to age-matched wild-type (WT) mice. Refractive error was measured using an eccentric infrared photorefractor, and axial length was measured with partial coherence interferometry or spectral domain ocular coherence tomography. A cohort of mice received head-mounted diffuser goggles to induce form deprivation from 4 to 6 weeks of age. Dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC) levels were measured with high-performance liquid chromatography in each strain after exposure to normal or form deprivation conditions. RESULTS: The rd1 and rd10 mice had significantly greater hyperopia relative to the WT controls throughout normal development; however, axial length became significantly longer only in WT mice starting at 7 weeks of age. After 2 weeks of form deprivation, the rd1 and rd10 mice demonstrated a faster and larger myopic shift (-6.14±0.62 and -7.38±1.46 diopter, respectively) compared to the WT mice (-2.41±0.47 diopter). Under normal visual conditions, the DOPAC levels and DOPAC/dopamine ratios, a measure of dopamine turnover, were significantly lower in the rd1 and rd10 mice compared to the WT mice, while the dopamine levels were similar or higher than WT in the rd10 mice. Lower basal levels of DOPAC were highly correlated with increasing myopic shifts. CONCLUSIONS: Refractive development under normal visual conditions was disrupted toward greater hyperopia from 4 to 12 weeks of age in these photoreceptor degeneration models, despite significantly lower DOPAC levels. However, the retinal degeneration models with low basal levels of DOPAC had increased susceptibility to form deprivation myopia. These results indicate that photoreceptor degeneration may alter dopamine metabolism, leading to increased susceptibility to myopia with an environmental visual challenge.

Assessment of axial length measurements in mouse eyes.

PURPOSE: To compare measurements of murine ocular axial lengths (ALs) made with 780 nm partial coherence interferometry (PCI) and 1310 nm spectral domain-optical coherence tomography (SD-OCT). METHODS: AL was measured at postnatal day (P) 58 in C57BL/6J mice. Repeated AL measurements were taken using a custom-made 780 nm PCI and a commercial 1310 nm SD-OCT. Intra- and interuser variability was assessed along the central optical axis and 2-degree off-axes angles with the SD-OCT. Data were collected and analyzed using Cronbach alpha (α), Bland-Altman coefficient of repeatability, agreement plots, and intraclass correlation coefficients (ICC). RESULTS: AL measurements agreed well between the two instruments (3.262 ± 0.042 mm for PCI; 3.264 ± 0.047 mm for SD-OCT; n = 20 eyes). The ICC for PCI compared with SD-OCT was 0.92, confirming high agreement between the two instruments. Intrauser ICC for the PCI and SD-OCT were 0.814 and 0.995, respectively. Similarly, interuser ICC for PCI and SD-OCT were 0.970 and 0.943, respectively. Using SD-OCT, a 2-degree misalignment of the eye along the horizontal meridian produced mean differences in AL of -0.002 ± 0.017 mm relative to the centrally aligned images, whereas similar misalignment along the vertical meridian created 0.005 ± 0.018 mm differences in AL measurements. CONCLUSIONS: AL measurements from the 780 nm PCI and 1310 nm SD-OCT correlate well. Multiple statistical indices indicate that both instruments have good precision and agreement for measuring murine ocular AL in vivo. Although the vertical meridian had the greater variability in AL in the small mouse eye; 2-degree off-axes differences were within the SD of centrally aligned AL.

Retinal function and structure in Ant1-deficient mice.

PURPOSE: Mutations in ANT, a mitochondrial ATP transporter, are typically associated with myopathy. Because of the high metabolic demands of the retina, the authors examined whether elimination of the Ant1 isoform in a transgenic mouse affects retinal function or morphology. METHODS: RT-PCR was used to confirm Ant1 expression in retinas of wild-type (WT) or Ant1(-/-) mice. Full-field ERGs were used to test retinal function under dark- and light-adapted conditions and the recovery of the photoresponse to a bright flash. Using histologic methods, the authors assessed the retinal location of ANT and ANT1-ß-gal reporter protein, mitochondrial activity with cytochrome c oxidase (COX) and succinate dehydrogenase (SDH) staining, retinal layer thickness, and bipolar cell types using Chx10 and recoverin. RESULTS: Ant1(-/-) mice had supernormal ERG b-waves under both dark- and light-adapted conditions. X-Gal staining was detected in a subset of cells within the inner retina. The following characteristics were normal in Ant1(-/-) mice compared with age-matched WT mice: recovery of the photoresponse, COX and SDH activity, retinal morphology, and bipolar cell morphology. CONCLUSIONS: The presence of ANT1 in a subset of inner retinal cells accompanied by supernormal ERG responses suggests that ANT1 may be localized to hyperpolarizing bipolar cells. However, the immunohistochemical techniques used here did not show any differences in bipolar cells. Moderate functional changes coupled with a lack of detectable morphologic changes suggest that ANT1 is not essential for ATP transport in the retina.