Preparation of mango peel-waste pectin-based nanofibers by solution blow spinning (SBS).

An essential prerequisite for successful solution blow spinning (SBS) is the presence of effective molecular entanglements of polymers in the solution. However, the fabrication of biopolymer fibers is not as straightforward as synthetic polymers. Particularly for biopolymers such as pectin, molecular entanglements are essential but insufficient for successful spinning through the SBS production method. Such a challenge is due to the biopolymer's complex nature. However, incorporating an easily spinnable polymer precursor, such as polyacrylonitrile (PAN), to pectin effectively enabled the production of fibers from the SBS process. In this process, PAN-assisted pectin nanofibers are produced with average diameters ranging from 410.75 ± 3.73 to 477.09 ± 6.60 nm using a feed flow rate of 5 ml h-1, air pressure of 3 bars, syringe tip to collector distance at 30 cm, and spinning time of 10 min. PAN in DMSO solvent at different volume ratios (i.e. 35%-55% v/v) was critical in assisting pectin to produce nanofibers. The addition of a high molecular weight polymer, PAN, to pectin also improved the viscoelasticity of the solution, eventually contributing to its successful SBS process. Furthermore, the composite SBS-spun fibers obtained suggest that its formation is concentration-dependent.

Measures, Uncertainties, and Significance Test in Operational ROC Analysis.

In receiver operating characteristic (ROC) analysis, the sampling variability can result in uncertainties of performance measures. Thus, while evaluating and comparing the performances of algorithms, the measurement uncertainties must be taken into account. The key issue is how to calculate the uncertainties of performance measures in ROC analysis. Our ultimate goal is to perform the significance test in evaluation and comparison using the standard errors computed. From the operational perspective, based on fingerprint-image matching algorithms on large datasets, the measures and their uncertainties are investigated in the three scenarios: 1) the true accept rate (TAR) of genuine scores at a specified false accept rate (FAR) of impostor scores, 2) the TAR and FAR at a given threshold, and 3) the equal error rate. The uncertainties of measures are calculated using the nonparametric two-sample bootstrap based on our extensive studies of bootstrap variability on large datasets. The significance test is carried out to determine whether the difference between the performance of one algorithm and a hypothesized value, or the difference between the performances of two algorithms where the correlation is taken into account is statistically significant. Examples are provided.

IL-4 production by CD8+ lymphomatoid papulosis, type C, attracts background eosinophils.

There are two subsets of CD8+ T cells: Tc1 and Tc2. INF-gamma production by Tc1 cells causes granulomatous inflammation. IL-4 production by Tc2 cells attracts eosinophils. A 76-year-Caucasian female presented with CD8+ lymphomatoid papulosis (LyP), type C. We hypothesized that the LyP cells belonged to the Tc2 subset because of abundant background eosinophils. Hematoxylin and eosin and immunohistochemical stains were carried out on tissue sections from a skin punch biopsy. Antibodies for immunohistochemical stains included CD3, CD4, CD5, CD7, CD8, CD30, CD56, ALK-1, clusterin and IL-4. There was involvement of the dermis by a dense lymphoid infiltrate composed of large atypical cells and numerous eosinophils. The LyP cells expressed CD5, CD8, CD30 and IL-4. Keratinocytes showed a membranous pattern of immunoreactivity for IL-4. IL-4 production by CD8+ LyP, type C indicates that it belongs to the Tc2 subset. The cytokine milieu produced by the LyP cells attracted eosinophils. The IL-13R complex on keratinocytes bound IL-4 and produced a membranous staining pattern. Although CD8+ LyP is rare, we believe that this CD30+ lymphoproliferative disorder should be included in the World Health Organization-European Organization for Research and Treatment of Cancer classification of cutaneous T-cell lymphomas.

Biotin-rich intranuclear inclusions: have you checked your immunohistochemical controls?

Intranuclear biotin interacts with avidin-biotin-complex used for viral immunoperoxidase detection giving false-positive results.

Monte Carlo studies of bootstrap variability in ROC analysis with data dependency.

ROC analysis involving two large datasets is an important method for analyzing statistics of interest for decision making of a classifier in many disciplines. And data dependency due to multiple use of the same subjects exists ubiquitously in order to generate more samples because of limited resources. Hence, a two-layer data structure is constructed and the nonparametric two-sample two-layer bootstrap is employed to estimate standard errors of statistics of interest derived from two sets of data, such as a weighted sum of two probabilities. In this article, to reduce the bootstrap variance and ensure the accuracy of computation, Monte Carlo studies of bootstrap variability were carried out to determine the appropriate number of bootstrap replications in ROC analysis with data dependency. It is suggested that with a tolerance 0.02 of the coefficient of variation, 2,000 bootstrap replications be appropriate under such circumstances.

Development of cystic glandular hyperplasia of the endometrium in Mullerian inhibitory substance type II receptor-pituitary tumor transforming gene transgenic mice.

The pituitary tumor transforming gene (PTTG)/securin is an oncogene that is involved in cell cycle regulation and sister chromatid separation. PTTG is highly expressed in various tumors including ovarian tumors, suggesting that PTTG may play a role in ovarian tumorigenesis. Overexpression of PTTG resulted in induction of cellular transformation in vitro and tumor formation in nude mice. To ascertain PTTG function in ovarian tumorigenesis, we generated a transgenic mouse model of PTTG by cloning PTTG cDNA downstream of Mullerian inhibitory substance type II receptor gene promoter (MISIIR) in order to target the ovarian surface epithelium. By screening of transgenic animals, we identified five founders (four males and one female). Using the four male founders, we developed four transgenic lines. PTTG expression was increased in ovarian surface epithelium, ovarian granulosa cells, as well as in the pituitary gland. Transgenic females did not develop any visible ovarian tumors at 8-10 months of age; however, there was an overall increase in the corpus luteum mass in transgenic ovary, suggesting increased luteinization. These changes were associated with an increase in serum LH and testosterone levels. In addition, there was a generalized hypertrophy of the myometrium of MISIIR-PTTG transgenic uteri with cystic glandular and hyperplasia of the endometrium. Based on these results, we conclude that the overexpression of PTTG may be required to initiate precancerous conditions but is not sufficient to induce ovarian tumorigenesis and may require another partner to initiate cellular transformation.

Pathologic evaluation of cryoprobe-assisted lumpectomy for breast cancer.

Cryoprobe-assisted lumpectomy is a relatively new technique that converts nonpalpable carcinomas into well-defined, palpable ones by creating an ice ball under ultrasonographic guidance, thus eliminating the need for preoperative needle localization. We evaluated the effect of cryoprobe-induced freezing on tumor tissue, peritumoral tissue, and margin status in 6 cases of cryoprobe-assisted lumpectomy performed for infiltrating ductal carcinoma. Immunohistochemical stains for estrogen and progesterone receptors and the proliferation marker Ki-67 were performed on 4 cases and results compared with those of the pretreatment biopsy specimens. Although it was possible to recognize the tumor as infiltrating carcinoma in all cases, the alteration in tumor morphology interfered with tumor grading, distinguishing in situ and invasive components, and assessment of mitoses and lymphovascular invasion. The expression of estrogen and progesterone receptors was greatly reduced, whereas the Ki-67 staining was not significantly different when compared with pretreatment biopsy specimens. The "cryoprobe effect" did not interfere with evaluation of the margins and surrounding breast tissue.

Tartrate-resistant acid phosphatase as an immunohistochemical marker for inflammatory macrophages.

Human serum contains 2 isoforms of type-5 tartrate-resistant acid phosphatase (TRACP): 5a and 5b. TRACP-5b is osteoclastic. Our goal was to determine if serum TRACP-5a could originate from inflammatory macrophages (MPhi). We stained 246 paraffin-embedded tissue samples for TRACP using monoclonal antibody 9C5 (mab9C5) to isoforms 5a and 5b and a novel mab220 specific to isoform 5a. CD68 and lysozyme were also stained. MPhi of chronic and granulomatous inflammation and in tissues that undergo strong antigenic stimulation were strongly positive for TRACP, more so with mab220 than with mab9C5. Noninflammatory MPhi in lymph node sinuses or germinal centers and red pulp MPhi of spleen were weak or negative for TRACP. Marginal zone lymphocytes and sebaceous glands of skin were weakly positive for TRACP. Tissue mast cells displayed strong TRACP staining. Neuroendocrine cells of gastrointestinal tissues were strongly immunoreactive with mab9C5 but negative with mab220. Restricted expression of TRACP primarily in inflammatory MPhi supports our hypothesis that circulating TRACP-5a could be a biomarker of chronic inflammatory disease activity.

Performance generalization in biometric authentication using joint user-specific and sample bootstraps.

Biometric authentication performance is often depicted by a detection error trade-off (DET) curve. We show that this curve is dependent on the choice of samples available, the demographic composition and the number of users specific to a database. We propose a two-step bootstrap procedure to take into account the three mentioned sources of variability. This is an extension to the Bolle et al.'s bootstrap subset technique. Preliminary experiments on the NIST2005 and XM2VTS benchmark databases are encouraging, e.g., the average result across all 24 systems evaluated on NIST2005 indicates that one can predict, with more than 75 percent of DET coverage, an unseen DET curve with eight times more users. Furthermore, our finding suggests that with more data available, the confidence intervals become smaller and, hence, more useful.

The Impact of Data Dependence on Speaker Recognition Evaluation.

The data dependency due to multiple use of the same subjects has impact on the standard error (SE) of the detection cost function (DCF) in speaker recognition evaluation. The DCF is defined as a weighted sum of the probabilities of type I and type II errors at a given threshold. A two-layer data structure is constructed: target scores are grouped into target sets based on the dependency, and likewise for non-target scores. On account of the needed equal probabilities for scores being selected when resampling, target sets must contain the same number of target scores, and so must non-target sets. In addition to the bootstrap method with i.i.d. assumption, the nonparametric two-sample one-layer and two-layer bootstrap methods are carried out based on whether the resampling takes place only on sets, or subsequently on scores within the sets. Due to the stochastic nature of the bootstrap, the distributions of the SEs of the DCF estimated using the three different bootstrap methods are created and compared. After performing hypothesis testing, it is found that data dependency increases not only the SE but also the variation of the SE, and the two-layer bootstrap is more conservative than the one-layer bootstrap. The rationale regarding the different impacts of the three bootstrap methods on the estimated SEs is investigated.

Validation of Nonparametric Two-Sample Bootstrap in ROC Analysis on Large Datasets.

The nonparametric two-sample bootstrap is applied to computing uncertainties of measures in ROC analysis on large datasets in areas such as biometrics, speaker recognition, etc., when the analytical method cannot be used. Its validation was studied by computing the SE of the area under ROC curve using the well-established analytical Mann-Whitney-statistic method and also using the bootstrap. The analytical result is unique. The bootstrap results are expressed as a probability distribution due to its stochastic nature. The comparisons were carried out using relative errors and hypothesis testing. They match very well. This validation provides a sound foundation for such computations.

Positive pregnancy tests in a nongravid, premenopausal woman due to hCG beta-chain production by multiple myeloma.

Positive pregnancy test results occurred in a nongravid, premenopausal woman while she was receiving chemotherapy for multiple myeloma. We tested 2 hypotheses to account for this finding: (1) Heterophil antibodies caused positive interference in the immunoassays. (2) Genuine human chorionic gonadotropin (hCG) originated from a nonsyncytiotrophoblastic source. Paraprotein was eliminated as a source of positive interference because 3 different instruments with unique capture and signal antibodies gave similar results (83, 90, and 97 mIU/mL [83, 90, and 97 IU/L]). Human antimouse antibodies (HAMAs) were unlikely to cause positive interference because immunoreactivity was maintained after serum was treated to neutralize heterophil antibodies. Immunoassays performed after gel filtration of serum indicated that immunoreactivity was due to genuine hCG. The high-molecular-weight fraction (heterophil antibody) had 6 mIU/mL (6 IU/L) of hCG. The low-molecular-weight fraction (hCG) had 86 mIU/mL (86 IU/L) of hCG. Immunohistochemical stains revealed that myeloma cells expressed immunoreactive hCG. Hence, multiple myeloma caused positive pregnancy test results in a nongravid woman.

Differential expression of cdk inhibitors p16, p21cip1, p27kip1, and cyclin E in cervical cytological smears prepared by the ThinPrep method.

Our objective was to correlate p16, p21cip1, p27kip1, and cyclin E protein expression with the degree of dysplasia on ThinPrep Papanicolaou (Pap) smears using a modified immunoperoxidase staining. Smears read as normal, atypical squamous cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), or high-grade SIL (HSIL) were identified and tested for high-risk human papillomavirus (HR-HPV). Additional smears were processed for immunoperoxidase for p16, p21cip1, p27kip1, and cyclin E. Thirty-four smears were satisfactory for study. The p16 was positive in all nine HSIL, in four of nine LSIL, and in one of seven ASC-US. The p27kip1 was positive in all nine HSIL, in eight of nine LSIL, and in one of seven ASC-US. The p21cip1 was positive in all nine HSIL, in one of nine LSIL, and in one of seven ASC-US. Cyclin E was positive in seven of nine HSIL and in one of nine LSIL and in none of the ASC-US smears. Normal smears were negative for all the antigens. There was poor correlation of protein expression and HR-HPV infection. We concluded that p16, p21cip1, p27kip1, and cyclin E can be demonstrated on Pap smears and they are expressed differentially in dysplastic cells, with highest expression in HSIL. The p21cip1 and cyclin E showed the greatest correlation with HSIL.

CD5+ follicular lymphoma: a clinicopathologic study of three cases.

Follicular lymphoma (FL) is a low-grade lymphoma that typically lacks CD5 antigen expression. We report 3 cases of FL with unusual expression of CD5. All cases showed histologic features of FL, including effaced nodal architecture, follicular growth pattern, and a spectrum of grades from 1 to 3 using World Health Organization criteria. In flow cytometric studies, all 3 cases showed a light chain-restricted, CD19+, CD20+ B-cell population coexpressing CD10 and low-level CD5. Immunohistochemical studies demonstrated an identical B-cell immunophenotype with weak expression of CD5 and coexpression of bcl-2 protein and the germinal center-associated markers, CD10 and bcl-6 protein. None of the cases showed expression of CD43, cyclin D1, or IgD. By molecular analysis, immunoglobulin heavy chain gene rearrangements were demonstrated in all 3 cases, and 2 of 3 cases had a t(14;18). These cases highlight the difficulty classifying these lymphomas by flow cytometric studies alone and emphasize the importance of recognizing FL in the differential diagnosis of CD5+ B-cell lymphomas.

Plasma cell neoplasm in conjunction with glioblastoma of the conus medullaris.

We report a plasma cell neoplasm in conjunction with a glioblastoma multiforme (GBM) of the conus medullaris in a 42-year-old man. Glioblastoma is a World Health Organization (WHO) grade IV neoplasm that requires surgical intervention, radiation, and possibly chemotherapy. Astrocytomas of the spinal cord are rare neoplasms, with intramedullary glioblastomas comprising only 1% to 3%. Plasma cell neoplasms result from monoclonal proliferation of mature B cells; they have been reported as a primary malignancy with gliomas arising after treatment. Secondary plasma cell neoplasms arising within glioblastomas have not previously been described. However, there have been reports of glioblastomas related to other plasma cell and hematopoietic diseases such as Waldenstrom's macroglobulinemia and myeloid sarcomas.

Expression of p16 in sinonasal undifferentiated carcinoma (SNUC) without associated human papillomavirus (HPV).

Sinonasal undifferentiated carcinoma (SNUC) is an uncommon and highly aggressive neoplasm of the paranasal sinuses and nasal cavity. Its undifferentiated histologic appearance often requires immunohistochemical studies to distinguish it from other high-grade neoplasms. Due to the rarity of SNUC, its immunohistochemical staining profile has been incompletely characterized, and little work has been done on its expression of the markers for human papillomavirus (HPV). Our objective is to expand our knowledge of its immunophenotype and its association with HPV in order to define markers with mechanistic potential in the disease process, or of possible therapeutic importance. A total of five patients (one woman and four men) with SNUC, ranging in age from 26 to 75 years (mean 56.8 years) were compared to five patients (five men) with poorly differentiated squamous cell carcinoma (PDSCC), ranging in age from 53 to 75 years (mean 62.2 years). PDSCC was chosen as a control, given its well-reported immunohistochemical profile and negativity for HPV markers. The immunohistochemical panel included: CK7, CK19, EMA, NSE, chromogranin, p53, CK5/6, p63, CK14, S100, HMB-45, desmin, muscle specific actin, and CD45. Additionally, tests for p16, EBV, and HPV (subtypes 6, 11 16, 18) were performed. The diagnosis of SNUC was confirmed in all cases by histology and immunohistochemical stains. An interesting finding of strong diffuse positivity for p16 was noted in all SNUC cases, compared to only two of five PDSCC that were positive for p16. HPV DNA was not detected in any SNUC cases or any cases of PDSCC. All SNUC cases demonstrated over expression of p16 in the absence of HPV DNA expression. This may represent residual epithelial p16 staining, which is normally present in the sinonasal tract. Due to the rarity of SNUC, more cases will need to be evaluated to confirm the absence of HPV DNA.

Lymphomas involving the breast: a study of 106 cases comparing localized and disseminated neoplasms.

Lymphomas involving the breast account for approximately 2% of extranodal and <1% of all non-Hodgkin lymphomas. Our aim in this study was to classify breast lymphomas using the World Health Organization classification and then compare this classification with clinical, histologic, and radiologic findings as well as survival. The study group included 106 patients with breast lymphoma (105 women and 1 man). The neoplasms were divided into 2 groups based on extent of disease at initial diagnosis: localized disease (n=50) and disseminated disease (n=56). The follow-up period ranged from 4 to 252 months (median, 49 mo). Almost all (97%) patients presented with a palpable breast mass or masses. In the localized group, diffuse large B-cell lymphoma (DLBCL) was most frequent (n=32, 64%). In the disseminated group, follicular lymphoma was most frequent and exclusive to this group (P=0.0004). Mucosa-associated lymphoid tissue lymphomas occurred in both groups without a significant difference in frequency. A variety of other types of B-cell and T-cell non-Hodgkin lymphomas and classical Hodgkin lymphoma involved the breast at much lower frequency; most of these neoplasms involved the breast as part of disseminated disease. The clinical presentation correlated with radiologic findings: localized lymphomas presented as solitary masses, whereas disseminated lymphomas commonly presented as multifocal masses. There was a significant difference in the disease-free survival between patients with localized and disseminated DLBCL (P=0.003). In the disseminated group, patients with DLBCL had a worse disease-free survival compared with patients with mucosa-associated lymphoid tissue lymphoma or follicular lymphoma (P=0.01).

CD10 expression in cutaneous adnexal neoplasms and a potential role for differentiating cutaneous metastatic renal cell carcinoma.

CONTEXT: Recent investigations have demonstrated the utility of CD10 as a marker for renal cell carcinoma (RCC). Cutaneous metastases occur in up to 11% of patients with RCC and may be the presenting sign of widespread disease. The differential diagnosis in histopathologic evaluation of these cases includes cutaneous adnexal neoplasms, and describing the expression of CD10 in these tumors may be helpful in delineating the differential diagnosis. OBJECTIVE: To determine CD10 expression in a variety of adnexal lesions and to determine the diagnostic utility of CD10 in an immunohistochemical panel differentiating metastatic cutaneous renal cell carcinoma from cutaneous adnexal neoplasms. DESIGN: We studied 57 primary adnexal neoplasms of eccrine (n = 31), apocrine (n = 16), and sebaceous (n = 10) differentiation as well as normal skin (n = 3) and RCC metastatic to the skin (n = 4). A CD10 monoclonal antibody was applied to formalin-fixed, paraffin-embedded tissue. Specimens were randomized and categorized as immunopositive or immunonegative by a pathologist with expertise in immunohistochemistry who was blinded to the diagnoses. RESULTS: Two (6.5%) of 31 eccrine, 1 (6%) of 16 apocrine, and 4 (40%) of 10 sebaceous neoplasms demonstrated CD10 immunopositivity. Four (100%) of 4 RCC were CD10 immunopositive. CD10 expression was significant for eccrine and apocrine neoplasms (P < .001) compared to metastatic RCC, but not for sebaceous neoplasms (P = .08). CONCLUSION: Based on these results, CD10 is a useful additional immunostain for the discrimination of RCC metastatic to the skin and cutaneous adnexal neoplasms with eccrine and apocrine differentiation, but not with sebaceous differentiation.