Immunohistochemical and Molecular Diagnosis of Mucocutaneous and Mucosal Leishmaniasis

2019 ◽  
Vol 28 (2) ◽  
pp. 138-145 ◽  
Author(s):  
Celeste Sánchez-Romero ◽  
Hercílio Martelli Júnior ◽  
Vania Lúcia Ribeiro da Matta ◽  
Larisse Miranda Freitas ◽  
Carina de Mattos Soares ◽  
...  
Keyword(s):  
Low Cost ◽  
Parasitic Infection ◽  
Positive Staining ◽  
Upper Aerodigestive Tract ◽  
Tissue Samples ◽  
Infected Female ◽  
Molecular Features ◽  
Mucocutaneous Leishmaniasis ◽  
Mucosal Involvement ◽  
Mucosal Leishmaniasis

Leishmaniasis is a parasitic infection transmitted by the bite of infected female sandflies. It principally affects the skin, and the frequency of mucosal involvement is about 5% to 20%. Considering the rarity of leishmaniasis affecting the upper aerodigestive tract mucosa, we evaluated the characteristics of mucocutaneous leishmaniasis and mucosal leishmaniasis and the diagnostic difficulty when the parasites are scarce in tissue samples. The clinical, histopathological, histochemical, immunohistochemical, and molecular features of 17 cases of mucocutaneous leishmaniasis and mucosal leishmaniasis were assessed. Mucosal disease was principally found in the soft palate, oropharynx, and nose, manifesting mainly as a solitary ulcer. In hematoxylin and eosin–stained sections, 10 cases revealed abundant amastigotes within the macrophages. Giemsa staining was not shown to be helpful to confirm the diagnosis in 6 cases with scarce or nondetectable amastigotes. Immunohistochemistry (IHC) showed high sensitivity by positive staining in 14 out of 17 cases (82.3%). Polymerase chain reaction was shown to be more sensitive than IHC with 13 out of 14 (92.8%) positive cases, including the 3 IHC negative cases; however, this technique is not available in many endemic regions. In summary, we suggest that the IHC is a simple technique with rapid results and relatively low cost, when compared with other laboratorial procedures; thus, IHC is a helpful tool that should be implemented in the routine diagnosis of leishmania.

scholarly journals Immunohistochemical expression of β-catenin, Ki67, CD3 and CD18 in canine colorectal adenomas and adenocarcinomas

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Kristin M. V. Herstad ◽  
Gjermund Gunnes ◽  
Runa Rørtveit ◽  
Øyvor Kolbjørnsen ◽  
Linh Tran ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
T Cells ◽  
Colorectal Adenomas ◽  
Colorectal Carcinogenesis ◽  
Positive Staining ◽  
Colonic Tissue ◽  
Retrospective Case ◽  
Tissue Samples ◽  
Colonic Cells ◽  
Control Samples

Abstract Background Inflammation is believed to influence human colorectal carcinogenesis and may have an impact on prognosis and survival. The mucosal immunophenotype in dogs with colorectal cancer is poorly described. The aim of this study was to investigate whether the density, distribution and grade of tumor-infiltrating immune cells (TIIs) are different in normal colonic tissue vs benign stages (adenomas) and malignant stages (adenocarcinomas) of canine colorectal carcinogenesis, and thus, whether they can be considered as prognostic factors in dogs. This retrospective case-control study was performed on formalin-fixed, paraffin-embedded tissue samples from dogs with histologically confirmed colorectal adenoma (n = 18) and adenocarcinoma (n = 13) collected from archived samples. The samples had been collected by colonoscopy, surgery or during postmortem examination. Healthy colonic tissue obtained post mortem from dogs euthanized for reasons not involving the gastrointestinal tract served as control tissue (n = 9). Results The tumor samples had significantly lower numbers of CD3+ T-cells in the epithelium compared to controls (adenocarcinoma vs control, Kruskal-Wallis test, p = 0.0004, and adenoma vs control, p = 0.002). Adenomas had a significantly lower number of CD18+ cells in the lamina propria, compared to control samples (Kruskal-Wallis test, p = 0.008). Colonic samples from control dogs had uniform staining of β-catenin along the cell membrane of epithelial cells. Compared to normal colonic cells, the expression levels of cytoplasmic β-catenin were significantly higher in adenomas and adenocarcinomas (adenoma vs control Kruskal-Wallis test, p = 0.004, and adenocarcinoma vs control, p = 0.002). None of the control samples showed positive staining of β-catenin in the nucleus of colonic cells. In contrast, adenocarcinomas and adenomas showed moderate to strong staining of the cell nucleus. The nuclear β-catenin expression (signal strength and distribution) was significantly higher in adenomas compared to adenocarcinomas (Kruskal-Wallis test, p < 0.05). Conclusions β-catenin and Ki67 were not useful markers for demonstrating tumor progression from adenomas to adenocarcinomas. The lower presence of CD18 and CD3+ cells in colorectal tumors compared to controls indicates a reduced presence of histiocytes and T-cells, which may have implications for the pathogenesis and progression of colorectal cancer in dogs.

Sarcomatoid (srcRCC) versus clear cell (ccRCC) renal cell carcinoma: A comparative comprehensive genomic profiling (CGP) study.

2021 ◽  
Vol 39 (6_suppl) ◽  
pp. 349-349
Author(s):  
Gennady Bratslavsky ◽  
Joseph M Jacob ◽  
Andrea Necchi ◽  
Philippe E. Spiess ◽  
Petros Grivas ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Clinical Course ◽  
Neuroendocrine Differentiation ◽  
Suppressor Gene ◽  
Rapid Progression ◽  
Positive Staining ◽  
Tumor Type ◽  
Tissue Samples ◽  
Comprehensive Genomic Profiling ◽  
Tumor Types

349 Background: srcRCC is a well-described histologic entity often featuring rapid progression and aggressive clinical course when compared with classic ccRCC. We queried whether CGP would uncover opportunities for targeted and immunotherapy (IO) for srcRCC patients that could individualize their treatment and entry into clinical trials. Methods: Using a hybrid capture-based CGP assay to evaluate all classes of genomic alterations (GA), 160 cases of srcRCC and 1,664 cases of ccRCC were sequenced from FFPE tissue samples. Tumor mutational burden (TMB) was determined on up to 1.1 Mbp of sequenced DNA and microsatellite instability (MSI) was determined on up to 114 loci. PD-L1 expression was determined by IHC (Dako 22C3) with low tumor cell positive staining set at 1-49% and high staining >50% expression. Results: Gender and age distributions for both tumor types were similar. srcRCC featured significantly higher GA/tumor than ccRCC (P < .0001). CGP revealed major differences with ccRCC associated more frequently with tumor suppressor gene (TSG) losses in VHL, PBRM1, TSC2 and SETD2 (all P < .0001). In contrast, srcRCC is associated with cell proliferation with increased inactivation of cell cycle regulatory genes including TP53, CDKN2A/B, MDM2 and TERT (all P < .0001). RB1 GA in srcRCC may reflect neuroendocrine differentiation occasionally found in these tumors. NF2 GA were more frequent in srcRCC (P < .0001). Conclusions: CGP reveals striking differences between srcRCC and ccRCC which may in part explain the differing histologic appearances and typical clinical course of these 2 aggressive malignancies. ccRCC is driven more by TSG loss and srcRCC is driven more by cell cycle dysregulation. Targeted therapy opportunities were uncommon for both tumor types although each featured biomarkers potentially predictive of mTOR inhibitor responses ( TSC2 in ccRCC and NF2 in srcRCC). Although the higher PBRM1 GA frequency in ccRCC may explain the IO benefit well-known for this tumor type, the srcRCC group features significantly increased TMB, CD274 amplification and PD-L1 staining which may also create IO opportunities for srcRCC patients. [Table: see text]

A New Cadmium Reduction Device for the Microplate Determination of Nitrate in Water, Soil, Plant Tissue, and Physiological Fluids

2011 ◽  
Vol 94 (6) ◽  
pp. 1896-1905 ◽  
Author(s):  
James D Crutchfield ◽  
John H Grove
Keyword(s):  
Plant Tissue ◽  
Low Cost ◽  
Statistical Validation ◽  
Soil Science Society ◽  
Widespread Application ◽  
Tissue Samples ◽  
New Device ◽  
Low Concentrations ◽  
Groundwater Samples ◽  
Interlaboratory Validation

Abstract A reusable catalytic reductor consisting of 96 copperized-cadmium pins attached to a microplate lid was developed to simultaneously reduce nitrate (NO3–) to nitrite (NO2–) in all wells of a standard microplate. The resulting NO2– is analyzed colorimetrically by the Griess reaction using a microplate reader. Nitrate data from groundwater samples analyzed using the new device correlated well with data obtained by ion chromatography (r2 = 0.9959). Soil and plant tissue samples previously analyzed for NO3– in an interlaboratory validation study sponsored by the Soil Science Society of America were also analyzed using the new technique. For the soil sample set, the data are shown to correlate well with the other methods used (r2 = 0.9976). Plant data correlated less well, especially for samples containing low concentrations of NO3–. Reasons for these discrepancies are discussed, and new techniques to increase the accuracy of the analysis are explored. In addition, a method is presented for analyzing NO3– in physiological fluids (blood serum and urine) after matrix modification with Somogyi's reagent. A protocol for statistical validation of data when analyzing samples with complex matrixes is also established. The simplicity, adaptability, and low cost of the device indicate its potential for widespread application.

scholarly journals Evaluation of an Immunochromatographic Assay as a Canine Rabies Surveillance Tool in Goa, India

Viruses ◽  
2019 ◽  
Vol 11 (7) ◽  
pp. 649 ◽  
Author(s):  
Yale ◽  
Gibson ◽  
Mani ◽  
P.K. ◽  
Costa ◽  
...  
Keyword(s):  
Brain Tissue ◽  
Low Cost ◽  
Antibody Test ◽  
Immunochromatographic Assay ◽  
Human Rabies ◽  
Post Mortem ◽  
Laboratory Equipment ◽  
Tissue Samples ◽  
Test Kit ◽  
The Government

Rabies is a fatal zoonotic disease transmitted by the bite of a rabid animal. More than 95% of the human rabies cases in India are attributed to exposure to rabid dogs. This study evaluated the utility of a lateral flow immunochromatographic assay (LFA) (Anigen Rapid Rabies Ag Test Kit, Bionote, Hwaseong-si, Korea) for rapid post mortem diagnosis of rabies in dogs. Brain tissue was collected from 202 animals that were screened through the Government of Goa rabies surveillance system. The brain tissue samples were obtained from 188 dogs, nine cats, three bovines, one jackal and one monkey. In addition, 10 dogs that died due to trauma from road accidents were included as negative controls for the study. The diagnostic performance of LFA was evaluated using results from direct fluorescence antibody test (dFT); the current gold standard post mortem test for rabies infection. Three samples were removed from the analysis as they were autolysed and not fit for testing by dFT. Of the 209 samples tested, 117 tested positive by LFA and 92 tested negative, while 121 tested positive by dFT and 88 tested negative. Estimates of LFA sensitivity and specificity were 0.96 (95% CI 0.91–0.99) and 0.99 (95% CI 0.94–1.00), respectively. The LFA is a simple and low-cost assay that aids in the rapid diagnosis of rabies in the field without the need for expensive laboratory equipment or technical expertise. This study found that Bionote LFA has potential as a screening tool in rabies endemic countries.

scholarly journals Gonadal malignant germ cell tumors express immunoreactive inhibin/activin subunits

2001 ◽  
pp. 779-784 ◽  
Author(s):  
L Cobellis ◽  
P Cataldi ◽  
FM Reis ◽  
G De Palo ◽  
F Raspagliesi ◽  
...  
Keyword(s):  
Germ Cell ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Ovarian Tissue ◽  
Germ Cell Tumors ◽  
Yolk Sac ◽  
Yolk Sac Tumor ◽  
Positive Staining ◽  
Tissue Samples ◽  
Malignant Germ Cell Tumors

OBJECTIVE: Inhibin and activin are proteins produced by ovarian granulosa cells and testicular Sertoli cells and are members of the transforming growth factor-beta superfamily. Since increased circulating levels of immunoreactive inhibin were detected in women with malignant ovarian tumors, they were proposed as tumor markers for ovarian carcinoma. Immunohistochemical studies later confirmed the presence of inhibin and activin subunits in granulosa cell tumors and epithelial ovarian cancer, as well as in Sertoli and Leydig cell testicular cancer. However, there is discrepant information on the detection of inhibin and activin in malignant germ cell tumors (MGCT). The aim of the present study was to evaluate the immunohistochemical expression of the inhibin/activin alpha, betaA and betaB subunits in ovarian and testicular MGCT specimens using polyclonal antisera. METHODS: The ovarian tissue samples were composed of 19 MGCT, including dysgerminoma (n=18) and yolk sac tumor (n=1). The testis specimens included classic seminomas (n=20), embryonal carcinomas (n=7), choriocarcinomas (n=2), and yolk sac tumor (n=1). RESULTS: Ovarian and testicular malignant germ cell tumors expressed positive staining for inhibin/activin alpha, betaA and betaB subunits, with some variations between and within individual tumors: while ovarian dysgerminomas were diffusely positive for alpha, betaA and betaB, testicular tumors expressed alpha and betaB subunits, whereas betaA staining was weak. CONCLUSIONS: The present results show positive staining for inhibin/activin subunits in ovarian and testicular MGCT, suggesting a possible role in tumorigenesis with the resultant clinical implication.

Polycythemia and Inappropriate Erythropoietin Concentrations in Two Dogs with Renal T-cell Lymphoma

2011 ◽  
Vol 47 (2) ◽  
pp. 122-128 ◽  
Author(s):  
Alexandra S. Durno ◽  
Jinelle A. Webb ◽  
Meredith J. Gauthier ◽  
Dorothee Bienzle
Keyword(s):  
T Cell ◽  
Cell Lymphoma ◽  
Abdominal Ultrasound ◽  
T Cell Lymphoma ◽  
Positive Staining ◽  
Renal Cells ◽  
Tissue Samples ◽  
Immunohistochemistry Staining ◽  
Postmortem Examinations ◽  
Serum Erythropoietin

Two dogs presented with suspected renal disease and polycythemia. Abdominal ultrasound examinations performed on both dogs revealed coalescing masses causing bilateral renomegaly. Serum erythropoietin (EPO) concentrations were physiologically inappropriate. Postmortem examinations revealed renal T-cell lymphoma in both dogs. One of the two dogs also had involvement of the liver and mesentery. EPO-immunohistochemistry on tissue samples demonstrated positive staining in tumor cells and occasional normal renal cells. This report illustrates that paraneoplastic EPO production may induce polycythemia. The pattern of EPO-immunohistochemistry staining suggested that the mechanism of production was due to tumor production of EPO and local hypoxia-induced EPO production from compression of normal renal cells and vasculature.

scholarly journals Histopathological Findings in Immunohistological Staining of the Granulomatous Tissue Reaction Associated with Tuberculosis

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Shirin Karimi ◽  
Masoud Shamaei ◽  
Mihan Pourabdollah ◽  
Makan Sadr ◽  
Mehrdad Karbasi ◽  
...  
Keyword(s):  
Immunohistochemical Staining ◽  
Plasma Cells ◽  
Tissue Reaction ◽  
Positive Staining ◽  
National Research Institute ◽  
Diagnostic Challenge ◽  
Tissue Samples ◽  
Granulomatous Tissue ◽  
Immunohistological Staining ◽  
Mycobacterial Antigens

Purpose. The histological diagnosis of Mycobacterium tuberculosis (MTB) remains a diagnostic challenge despite different methods. Immunohistochemistry (IHC) not only could confirm granulomatous tissue involvement but also can demonstrate MTB antigen immunolocalization. This study tries to clarify the details of immunohistochemical staining for MTB with pAbBCG.Materials/Methods. Twenty-three confirmed TB granulomatous tissue samples were studied by Ziehl-Neelsen and immunohistochemistry (IHC) staining with pAbBCG. Samples were selected from the archive of the Department of Pathology, National Research Institute of Tuberculosis and Lung Disease, Tehran, Iran.Results. IHC staining was positive in all samples, whereas Ziehl-Neelsen was positive in 9 cases out of 23 (39.1%). Tissue types used were pleural tissue, lymph nodes, and lung tissue. IHC showed positive coarse granular cytoplasmic and round, fragmented bacillary staining. In this study, epithelioid cells clearly showed more positive staining at the periphery of the granuloma rather than the center of granuloma. There is also positive staining in endothelial cells, fibroblasts, plasma cells, lymphocytes, and macrophages outside the granuloma.Conclusion. Considering the criteria of positive immunohistochemical staining of TB granulomatous reactions, this stain not only highlights the presence of mycobacterial antigens for tissue diagnosis, but also could morphologically localize its distribution in different cells.

scholarly journals Variation among intact tissue samples reveals the core transcriptional features of human CNS cell classes

2018 ◽  
Author(s):  
Kevin W. Kelley ◽  
Hiromi Inoue ◽  
Anna V. Molofsky ◽  
Michael C. Oldham
Keyword(s):  
Ectopic Expression ◽  
Brain Regions ◽  
Cellular Composition ◽  
Tissue Samples ◽  
Intact Tissue ◽  
The Core ◽  
Molecular Features ◽  
Online Resource ◽  
Cell Class

AbstractIt is widely assumed that cells must be physically isolated to study their molecular profiles. However, intact tissue samples naturally exhibit variation in cellular composition, which drives covariation of cell-class-specific molecular features. By analyzing transcriptional covariation in 7221 intact CNS samples from 840 individuals representing billions of cells, we reveal the core transcriptional identities of major CNS cell classes in humans. By modeling intact CNS transcriptomes as a function of variation in cellular composition, we identify cell-class-specific transcriptional differences in Alzheimer’s disease, among brain regions, and between species. Among these, we show that PMP2 is expressed by human but not mouse astrocytes and significantly increases mouse astrocyte size upon ectopic expression in vivo, causing them to more closely resemble their human counterparts. Our work is available as an online resource (http://oldhamlab.ctec.ucsf.edu) and provides a generalizable strategy for determining the core molecular features of cellular identity in intact biological systems.

Serum c-erbB-2 ectodomain (ECD) correlates with immunohistochemical c-erbB-2 expression and predicts an aggressive clinical outcome in hormone-independent prostate cancer (HIPC) patients (pts) treated with docetaxel (D)

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 4632-4632
Author(s):  
J. Domingo-Domenech ◽  
E. Gallardo ◽  
X. Filella ◽  
E. Fernandez ◽  
P. Fernandez ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Clinical Outcome ◽  
Median Time ◽  
Serum Levels ◽  
Tissue Expression ◽  
High Serum ◽  
Positive Staining ◽  
Tissue Samples ◽  
Staining Score ◽  
Psa Progression

4632 Background: The overexpression of c-erbB-2 has been described as a common event associated to the hormone-independent progression in prostate cancer. The clinical implications of c-erbB-2 expression in HIPC pts are not fully established. In this study we investigated the clinical significance of c-erbB-2 serum levels in HIPC pts treated with D and the correlation between serum c-erbB-2 ECD and c-erbB-2 tissue expression. Methods: Pts with HIPC treated with D were tested for serum c-erbB-2 ECD levels by immunoassay before chemotherapy. In pts with available biopsy specimens of hormone-independent tumor, c-erbB-2expression was determined by immunohistochemistry in paraffin-embedded tissue sections. PSA response to D, time to PSA progression and survival were analyzed. Results: Forty-one pts were included in the study. Median follow-up time was 23.3 (3.7–43.1) months. Median time to D progression was 4.1 (0,4–15.1) months and median survival was 11.3 (1,6–43.1). Serum c-erbB-2 correlated with PSA (r=0.351, p=0.025), inversely with time to PSA progression (r = −0.338, p = 0.033) and inversely with survival (r = −0.305, p = 0.05). Median time to PSA progression in pts with high serum c-erbB-2 (≥15 ng/ml) was 1.5 months compared to 4.6 months in pts with low levels (p = 0.0003). Survival in pts with high c-erbB-2 was 11 months and in pts with low c-erbB-2 was 15 months (p = 0.053). HIPC tissue samples from 10 pts were assessed for c-erbB-2 expression. Immunohistochemical c-erbB-2 positive staining (score +2 and +3) was detected in 4 (40%). C-erbB-2 staining showed correlation with serum c-erbB2 levels. Pts with positive staining had 19.1 ± 1.78 ng/ml and negative pts 8.8 ± 2.6 ng/ml (p = 0.001).The 4 pts with tissue c-erbB-2 overexpression had serum c-erbB-2 levels >15 ng/ml, while pts with no c-erbB-2 overexpression in tissue had also low c-erbB-2 levels in serum. Conclusions: High c-erbB-2 ECD in serum is associated with a worse clinical outcome of HIPC. Our data suggest that the determination of c-erbB-2 ECD in serum translates the c-erbB-2 status in the hormone-independent tumor and may be useful to select pts for c-erbB-2-targeted therapy. No significant financial relationships to disclose.

Biomarker analyses from JAVELIN Renal 101: Avelumab + axitinib (A+Ax) versus sunitinib (S) in advanced renal cell carcinoma (aRCC).

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. 101-101 ◽  
Author(s):  
Toni K. Choueiri ◽  
Laurence Albiges ◽  
John B. A. G. Haanen ◽  
James M.G. Larkin ◽  
Motohide Uemura ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Activation ◽  
De Novo ◽  
Nk Cell ◽  
Objective Response ◽  
Progression Free Survival ◽  
Significant Treatment ◽  
Tissue Samples ◽  
Molecular Features ◽  
Tumor Mutational Burden

101 Background: The phase 3 JAVELIN Renal 101 trial in previously untreated patients (pts) with aRCC demonstrated a progression-free survival (PFS) benefit and higher objective response rate with A+Ax vs S (Motzer, ESMO 2018; LBA6_PR). Here, we report outcomes from biomarker analyses of baseline tumor samples. Methods: We correlated efficacy with the results of molecular analyses of tissue samples from all 886 pts enrolled in JAVELIN Renal 101. Nephrectomy or tumor samples were characterized by immunohistochemistry (CD8 and PD-L1), whole-exome sequencing (WES), and RNAseq. WES and RNAseq were used to examine somatic mutations and analyze relevant gene expression signatures (GES) in relation to clinical outcomes. GES analyses included published and de novo signatures: effector T cell (Teff), angiogenesis (angio),T cell-inflamed (Tinf), and a novel immune-related signature incorporating pathway indicators for T- and NK-cell activation and IFNγ signaling, among others. Results: PD-L1 expression (≥1% immune cells) was associated with the longest PFS in the A+Ax arm and the shortest in the S arm (HR, 0.63; 95% CI, 0.49, 0.81). Significant treatment arm–specific differences in PFS were observed relative to wildtype when mutations in genes such as CD1631L, PTEN, or DNMT1 were present. Tumor mutational burden did not distinguish pts with respect to PFS. High-angio GES was associated with significantly improved PFS in the S arm but did not differentiate PFS in the A+Ax arm. In the low-angio subset, A+Ax improved PFS vs S. Pts with high Teff and Tinf in the A+Ax arm had longer PFS vs the S arm. In the A+Ax arm, PFS was enhanced in patients with immune GES–positive tumors vs those in the negative group (HR, 0.63; 95% CI, 0.46, 0.86; 2-sided p = 0.004), as well as in an independent dataset (JAVELIN Renal 100; Choueiri, Lancet Oncol, 2018) (HR, 0.46; 95% CI, 0.20, 1.05; 2-sided p = 0.064). Updated efficacy, including overall survival, will be presented. Conclusions: These findings define molecular features that differentiate therapy-specific outcomes in first-line aRCC and may inform personalized therapy strategies for pts with aRCC. Funding: Pfizer and Merck KGaA. Clinical trial information: NCT02684006.

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