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Clinical, histopathologic, and biologic features of pleomorphic lobular (ductal-lobular) carcinoma in situ of the breast: a report of 24 cases.

We reviewed 10 cases of pleomorphic lobular (ductal lobular) carcinoma in situ (PL/DLCIS) of the breast and compared them with 14 cases of pleomorphic lobular carcinoma in situ (PLCIS) found in association with invasive pleomorphic lobular carcinoma. The histologic features; immunohistochemical staining for estrogen receptors (ERs), p53, Ki67, E-cadherin, and gross cystic disease fluid protein-15 (GCDFP-15); and results of fluorescence in situ hybridization for HER-2/neu gene amplification were evaluated in all 24 cases. Histologically, PL/DLCIS cells were similar to those of PLCIS with invasion in that they were discohesive and medium to large in size with moderate to marked nuclear pleomorphism, small to prominent nucleoli, and moderate to abundant eosinophilic or vacuolated cytoplasm. In both groups, central necrosis was present in a small number of cases, and classic LCIS coexisted with the in situ lesion in less than half of the cases; in situ carcinomas were positive for ERs in 23 (100%) of 23 cases, p53 in 6 (25%) of 24 cases, and GCDFP-15 in 14 (74%) of 19 cases. The percentage of Ki67-positive tumor nuclei indicated moderate to high (more than 20%) proliferative activity in 8 (47%) of 17 cases. Immunostaining for E-cadherin was negative in all 24 cases. HER-2/neu gene amplification was observed in 1 (4%) of 23 cases. In cases with associated invasion, PLCIS had cytologic features and immunostaining patterns similar to those of the invasive pleomorphic component. Seven of the 10 patients who had PL/DLCIS without invasion underwent lumpectomy or simple mastectomy. Six of these patients had no evidence of disease in follow-up periods ranging from 4 to 32 months; the seventh patient developed recurrent disease 12 months after undergoing lumpectomy. We conclude that the cytologic features and biomarker expression profile of PL/DLCIS are similar to those of PLCIS with invasion but somewhat different from those of classic LCIS and ductal carcinoma in situ. Long-term follow-up studies are needed to further define the natural history of PL/DLCIS and its optimal management.

Adult↗

Benign CD10-positive T cells in reactive lymphoid proliferations and B-cell lymphomas.

Recent reports have indicated that the neoplastic T cells of angioimmunoblastic T-cell lymphoma express CD10. It has been suggested that the demonstration of a CD10+ T-cell population may assist in establishing a diagnosis of angioimmunoblastic T-cell lymphoma and in distinguishing angioimmunoblastic T-cell lymphoma from other peripheral T-cell lymphomas. It has been unclear, however, whether this phenotypically unusual T-cell population might be present in other settings as well. In this report, we have retrospectively examined 64 cases of lymph node and solid tissue biopsies for the presence of CD10+ T cells using multicolor flow cytometry. Discrete populations of CD10+ T cells were found in 5 of 28 cases (18%) of reactive lymphoid hyperplasia, 4 of 17 cases (23%) of follicular lymphoma, and 9 of 19 cases (47%) of marginal zone B-cell lymphomas. The CD10+ T cells constituted 1-6% of total cells analyzed and </=14% of the total T-cell population. Using two-color immunohistochemical stains, many of the CD10+ PAX5-negative presumptive T cells were found to be located within germinal centers. These findings indicate that a normal small subset of CD10+ peripheral T cells exists and, at least when present in small numbers, should not be considered an indication of a T-cell neoplasm.

Biomarkers, Tumor↗

Evaluating nipple discharge.

Nipple discharge is the third most common breast complaint after breast pain and breast mass. It is most often a benign process. Up to 50% women in their reproductive years can express one or more drops of fluid from the breast. Nipple discharge can be of several types, including milky, multicolored and sticky, purulent, clear and watery, yellow or serous, pink or serosanguinous, bloody or sanguinous. The characteristics of the nipple discharge help in the early diagnosis and management of breast disease. The most common cause of pathologic nipple discharge is a benign papilloma followed by ductal ectasia, and the least likely is carcinoma. Most nipple discharges are the result of a clinically insignificant benign process; therefore, less invasive, nonsurgical diagnostic modalities have been explored to reduce the need for surgical intervention. The evaluation and diagnosis of nipple discharge is important for the early detection of carcinoma, when present; and, in the case of benign disease, it is necessary to stop the incommodious discharge.

Breast Diseases↗

Multiparameter immunofluorescence on paraffin-embedded tissue sections.

Immunohistochemical techniques have gained increasing importance in diagnostics and research. While formalin-fixed, paraffin-embedded human tissue retains excellent morphology, the detection of antigens by immunofluorescence in its sections and especially the demonstration of multiple simultaneous antibodies have limitations. Double immunofluorescence labeling of routinely processed paraffin sections has been described previously. The signal intensity observed after triple labeling has been reported to be significantly inferior to that obtained by application of double fluorochromes. The authors show multicolor labeling of three and four primary antibodies in routinely processed paraffin-embedded tissue sections using a standardized immunofluorescence technique. In addition, procedures to reduce background staining and to avoid nonspecific double staining are described.

Antibodies↗

Venous malformations of skeletal muscle.

Intramuscular venous malformations are often mistaken for tumors because of a similar presentation and improper nomenclature. This is a review of 176 patients with venous malformations localized to skeletal muscle compiled from the Vascular Anomalies Center at Children's Hospital from 1980 through 1999. The female-to-male ratio was 2:1. Two-thirds of skeletal muscle venous malformations were noted at birth; the remainder manifested in childhood and adolescence. Venous malformations occurred in every muscle group, most often in the head and neck and extremities. Pain and swelling were the usual presenting complaints. Skeletal problems, such as fracture, deformation, or growth abnormalities, were rare. Hormonal exacerbation and intralesional bleeding were infrequent. Magnetic resonance imaging showed the lesions to be isointense to surrounding muscle on T1-weighted sequences and hyperintense on T2-weighted images. Characteristic tubular or serpentine components were oriented along the muscular long axis. Thrombi were hyperintense on T1-weighted and hypointense on T2-weighted sequences; phleboliths were seen as signal voids on all sequences. Gross examination of resected specimens revealed multicolored tissue with dilated vascular channels, frequently containing phleboliths. Light microscopy showed aggregates of primarily medium-sized, thin-walled vascular channels with flat endothelium and variable smooth muscle, most closely resembling dysplastic veins. Three lesions had a different histologic appearance consisting predominantly of small vessels with capillary structure and proliferative activity admixed with large feeding and draining vessels, similar to a lesion called intramuscular capillary hemangioma in the literature. The endothelium in these three lesions was negative for glucose transporter-1 by immunostaining. Eight percent of the patients, who had minor or no symptoms, were not treated. Twenty-four percent of the patients were managed conservatively (with aspirin and compressive garments); for 17 of these patients (10 percent of 176), noninvasive therapy was not successful, and they proceeded to sclerotherapy, excision, or both. A total of 31 percent of the patients had sclerotherapy, 20 percent had excision, and 27 percent had combined sclerotherapy and excision. Sclerotherapy was used for diffuse lesions, except for those with multiple intralesional thromboses, neurologic impairment, or compressive signs and symptoms. Resection was preferred for venous malformations well localized to a single muscle or muscle group, particularly if the muscles are expendable. Therapeutic outcomes were recorded in the charts or obtained by telephone interview in 122 of the patients (69 percent). Of these, compression garment and aspirin, resection, sclerotherapy, or combined excision and sclerotherapy improved symptoms in 121 patients (92 percent); no change was noted in 10 patients (8 percent). Only one patient was worse (self-reported) after intervention.

Adolescent↗

Tumor infiltrating T lymphocytes in colorectal cancer: Tumor-selective activation and cytotoxic activity in situ.

OBJECTIVE: To examine whether tumor-selective infiltration, activation, and cytotoxic activity of tumor infiltrating T lymphocytes (TIL) can be demonstrated in situ in colorectal cancer samples. SUMMARY BACKGROUND DATA: Recent studies indicated a correlation between the presence of TIL and an improved prognosis in colorectal cancer. However, tumor-selective activation and cytotoxic activity of CD8 TIL in situ in colorectal cancer patients have not yet been examined. METHODS: Tumor samples from 49 patients and corresponding normal mucosa samples from 23 patients with colorectal cancer (UICC stages II-IV) were examined for TIL. Two-color fluorescence immunohistochemistry and multicolor flowcytometric (FACS) analysis were used for quantification of CD8 T cells and measurement of their activation status (CD69-expression) and cytotoxic activity (CD107a-expression) in situ. Presence of tumor antigen-reactive T cells in tumor, blood, and bone marrow was evaluated by IFN-gamma Elispot analysis. RESULTS: While absolute numbers of CD8 T cells were similar, CD4 T helper cells were significantly increased in tumor tissue compared with normal mucosa. There was a significantly higher proportion of activated and cytotoxically active CD8 TIL in colorectal cancer compared with normal mucosa. Increased activation, cytotoxic activity, and functional reactivity of TIL were correlated with the presence of functional tumor antigen-reactive T cells in the blood and bone marrow. The proportion of activated TIL decreased significantly with higher tumor stage. CONCLUSIONS: Tumor-selective activation and cytotoxic activity of CD8 TIL and tumor-selective migration of CD4 T helper cells were demonstrated in colorectal cancer for the first time. Our data support the immunogenicity of colorectal cancer and suggest clinical significance of tumor-specific immune responses.

Adenocarcinoma↗

An improved multiple-locus variable number of tandem repeats analysis for Leptospira interrogans serovar Australis: a comparison with fluorescent amplified fragment length polymorphism analysis and its use to redefine the molecular epidemiology of this serovar in Queensland, Australia.

In this study, an improved multiple-locus variable number of tandem repeats analysis (MLVA) method based upon a previously published method is described. Improvements to the method included redesigned primers and PCR conditions, combined with pooled capillary electrophoresis using multicolored dyes. Allele sizes were converted into an allele string, and each unique allele string was assigned a numerical MLVA type (MVT). The improved MLVA method was then applied to 96 previously characterized Leptospira interrogans serovar Australis isolates from human and animal sources. The improved MLVA was found to have between six and 13 alleles at each locus, compared with three to eight in the original. The mean Hunter-Gaston diversity index (HGDI) for the improved MLVA method was 0.654, compared with 0.599 in the original; this increase in diversity was largely due to changes in the analysis of the variable number of tandem repeat (VNTR) data. When the improved MLVA method was compared with the fluorescent amplified fragment length polymorphism (FAFLP) method, there was a high level of concordance between the profiles; however, the MLVA method produced an additional four unique profiles amongst the subset of 30 isolates tested. Given that the improved MLVA method was found to be superior to the original MLVA method, it was subsequently used to redefine the molecular epidemiology of L. interrogans serovar Australis in Queensland, Australia. Using cluster analysis, the authors were able to demonstrate clonal links amongst rodent isolates, rodent and human isolates, and rodent and canine isolates. These results highlight the role of rodents in the disease, and also the potential role of MLVA in defining the molecular epidemiology of L. interrogans.

Animals↗

The large-scale organization of the centromeric region in Beta species.

In higher eukaryotes, the DNA composition of centromeres displays a high degree of variation, even between chromosomes of a single species. However, the long-range organization of centromeric DNA apparently follows similar structural rules. In our study, a comparative analysis of the DNA at centromeric regions of Beta species, including cultivated and wild beets, was performed using a set of repetitive DNA sequences. Our results show that these regions in Beta genomes have a complex structure and consist of variable repetitive sequences, including satellite DNA, Ty3-gypsy-like retrotransposons, and microsatellites. Based on their molecular characterization and chromosomal distribution determined by fluorescent in situ hybridization (FISH), centromeric repeated DNA sequences were grouped into three classes. By high-resolution multicolor-FISH on pachytene chromosomes and extended DNA fibers we analyzed the long-range organization of centromeric DNA sequences, leading to a structural model of a centromeric region of the wild beet species Beta procumbens. The chromosomal mutants PRO1 and PAT2 contain a single wild beet minichromosome with centromere activity and provide, together with cloned centromeric DNA sequences, an experimental system toward the molecular isolation of individual plant centromeres. In particular, FISH to extended DNA fibers of the PRO1 minichromosome and pulsed-field gel electrophoresis of large restriction fragments enabled estimations of the array size, interspersion patterns, and higher order organization of these centromere-associated satellite families. Regarding the overall structure, Beta centromeric regions show similarities to their counterparts in the few animal and plant species in which centromeres have been analyzed in detail.

Amino Acid Sequence↗

DNA mapping using microfluidic stretching and single-molecule detection of fluorescent site-specific tags.

We have developed a rapid molecular mapping technology--Direct Linear Analysis (DLA)--on the basis of the analysis of individual DNA molecules bound with sequence-specific fluorescent tags. The apparatus includes a microfluidic device for stretching DNA molecules in elongational flow that is coupled to a multicolor detection system capable of single-fluorophore sensitivity. Double-stranded DNA molecules were tagged at sequence-specific motif sites with fluorescent bisPNA (Peptide Nucleic Acid) tags. The DNA molecules were then stretched in the microfluidic device and driven in a flow stream past confocal fluorescence detectors. DLA provided the spatial locations of multiple specific sequence motifs along individual DNA molecules, and thousands of individual molecules could be analyzed per minute. We validated this technology using the 48.5 kb lambda phage genome with different 8-base and 7-base sequence motif tags. The distance between the sequence motifs was determined with an accuracy of +/-0.8 kb, and these tags could be localized on the DNA with an accuracy of +/-2 kb. Thus, DLA is a rapid mapping technology, suitable for analysis of long DNA molecules.

Bacteriophage lambda↗

Approximate symmetry laws for percolation in complex systems: Percolation in polydisperse composites.

The concept of so-called global symmetry of percolation models is discussed and extended to multicolored models. An integral equation is obtained, which determines the partial percolation probabilities P(a) for sites of color a. This equation is applied to a polydisperse particulate composite: a mixture of conducting (of relative fraction x(m)) and nonconducting spheres with distributions of sizes n(m)(R) and n(i)(R), respectively. We find the probability P(R) for a conducting particle of radius R to belong to the percolation cluster as a function of x(m) and a functional of n(m)(R') and n(i)(R'). The percolation threshold x is shown to decrease with increasing dispersion Delta of particle sizes. A simple law x=1/(3[1+(Delta/4)]) is obtained in the range of moderate dispersions. The theory is applicable also to a mixture of electronic and ionic conductors.

Journal Article↗

Cnoidal wave patterns in quadratic nonlinear media.

We report the existence of whole families of stationary cnoidal, periodic wave patterns in quadratic nonlinear media. We study the main physical features of the multicolored light patterns, including their shape, contrast, multifrequency energy sharing, asymptotics in the cascading limit, and excitation. Our numerical simulations predict that the cnoidal waves with high and even with moderate contrasts are robust enough against modulational instabilities to be experimentally observable.

Journal Article↗

Metastability of dark snoidal-type waves in quadratic nonlinear media.

We report the existence and basic properties of dark snoidal-type waves self-sustained in quadratic nonlinear media. Using a stability analysis technique, we reveal that they are almost completely stable, or metastable, in suitable ranges of input energy flows and material parameters. This opens the way to the experimental observation of dark-type multicolor periodic wave patterns supported by quadratic nonlinearities.

Journal Article↗

Quantum information processing with semiconductor macroatoms.

An all optical implementation of quantum information processing with semiconductor macroatoms is proposed. Our quantum hardware consists of an array of quantum dots and the computational degrees of freedom are energy-selected interband optical transitions. The quantum-computing strategy exploits exciton-exciton interactions driven by ultrafast multicolor laser pulses. Contrary to existing proposals based on charge excitations, our approach does not require time-dependent electric fields, thus allowing for a subpicosecond, decoherence-free, operation time scale in realistic semiconductor nanostructures.

Journal Article↗

Direct production of tripartite pump-signal-idler entanglement in the above-threshold optical parametric oscillator.

We calculate the quantum correlations existing among the three output fields (pump, signal, and idler) of a triply resonant nondegenerate optical parametric oscillator operating above threshold. By applying the standard criteria [P. van Loock and A. Furusawa, Phys. Rev. A 67, 052315 (2003)], we show that strong tripartite continuous-variable entanglement is present in this well-known and simple system. Furthermore, since the entanglement is generated directly from a nonlinear process, the three entangled fields can have very different frequencies, opening the way for multicolored quantum information networks.

Journal Article↗

Cytogenetic and molecular analysis of a family with three brothers afflicted with germ-cell cancer.

A thorough cytogenetic investigation and an analysis of detailed questionnaires were performed in a family with three brothers afflicted with germ-cell tumors (GCTs), in an attempt to detect a congenital factor related either to a hereditary genetic background or an environmental/lifestyle influence. One brother had an intracranial tumor in the pineal region and the two others had testicular tumors. Peripheral blood was studied by traditional karyotyping, multicolor-FISH, high-resolution comparative genomic hybridization (HR-CGH), and molecular analysis of selected loci on sex chromosomes (Yq11 region, TSPY, and the androgen receptor gene); however, no abnormalities were detected. The HR-CGH analysis of microdissected histological components of the overt tumors and the adjacent carcinoma in situ demonstrated a pattern of genomic imbalances characteristic for sporadic GCTs, including gain of 12p. The questionnaire and interview revealed a history of different cancers in the extended family, and a possible in utero and/or infantile exposure of the three brothers with GCTs to compounds suspected of endocrine-disrupting properties. Although no genetic aberration was detected in this family, we suspect the presence of a recessive hereditary factor pre-disposing to cancer, which probably was manifested as GCTs in the three brothers because of an adverse effect of an environmental factor on the early germ-cell differentiation.

Adult↗

Gain of chromosome 3q is an early and consistent genetic aberration in carcinomas of the vulva.

The aim was to determine whether specific gains of chromosome 3q and laminin-5gamma2-chain expression can improve early detection of invasive capacity in precancerous and squamous cell carcinoma of the vulva (VSCC). Six VSCC and three precancerous lesions were studied. Multicolor fluorescence in situ hybridization (FISH) probe sets were applied to nuclei suspensions prepared from archival material using the Hedley method. The probe panel consists of the centromers of chromosome 7, chromosome 3, and the TERC gene residing on the long arm of chromosome 3. Laminin-5gamma2-chain immunohistochemical analysis was performed on corresponding specimens and was expressed only in the VSCC. The genome-specific FISH analysis revealed 3q amplification in 43% of the nuclei analyzed for the VSCC and 22% of the nuclei for the precancerous lesions. Low-level 3q amplifications were found in precancerous lesions with an average fold increase of 1.15 for 3q. The invasive lesions showed higher average fold increases for 3q, averaging 1.32. Laminin-5gamma2-chain protein was expressed only in VSCC, whereas 3q gains were observed both in precancerous lesions and in VSCC, indicating that gain of chromosome 3q is an early and consistent event during carcinogenesis of VSCC.

Adult↗

Biochemical key to eight species of adult Egyptian mosquitoes.

1. An electrophoretic key is described, based on enzyme relative mobility, to distinguish eight species of Egyptian mosquito adults. 2. Using horizontal starch gel electrophoresis, five different enzyme assays unambiguously separated Culex pipiens L., Cx antennatus (Becker), Cx pusillus Macquart, Aedes caspius (Pallas), Culiseta longiareolata (Macquart), Uranotaenia unguiculata Edwards, Anopheles multicolor Cambouliu and An.pharoensis Theobald. 3. Diagnostic loci between species were: Ak-2, G6pd, alpha-Gpd, Gpi and 6-Pgd; the probability of correct diagnosis in each case was at least 0.989.

Aedes↗

A patient with Edwards syndrome caused by a rare pseudodicentric chromosome 18 of paternal origin.

We present an unusual case of trisomy 18 due to a pseudodicentric chromosome 18 of paternal origin. The karyotype was: 46,XY, -18, +psu dic(18)(qter-->cen-->p11.31::p11.31-->psucen-->qter). The origin of the abnormal chromosome was verified by FISH with a painting probe from chromosome 18. Absence of short-arm telomeres was shown by multicolor FISH, and the results of DNA analysis showed monosomy for loci D18S59 and D18S170 as well as paternal inheritance of the aberrant chromosome. The child's phenotype was characteristic of trisomy 18.

Centromere↗