Valeria Denninghoff

Helicobacter pylori associated with glossitis and halitosis.

Background.  Helicobacter pylori is a curved microaerophilic Gram‐negative bacterium considered as a risk factor for gastric cancer. The aim of this study was to find an association between burning sensations, acid taste, halitosis, and lingual hyperplasia with the effect of H. pylori on the mouth.

Helicobacter pylori and oral pathology: Relationship with the gastric infection

Helicobacter pylori (H. pylori) has been found in the oral cavity and stomach, and its infection is one of the most frequent worldwide. We reviewed the literature and conducted a Topic Highlight, which identified studies reporting an association between H. pylori-infection in the oral cavity and H. pylori-positive stomach bacterium. This work was designed to determine whether H. pylori is the etiologic agent in periodontal disease, recurrent aphthous stomatitis (RAS), squamous cell carcinoma, burning and halitosis. Record selection focused on the highest quality studies and meta-analyses. We selected 48 articles reporting on the association between saliva and plaque and H. pylori-infection. In order to assess periodontal disease data, we included 12 clinical trials and 1 meta-analysis. We evaluated 13 published articles that addressed the potential association with RAS, and 6 with squamous cell carcinoma. Fourteen publications focused on our questions on burning and halitosis. There is a close relation between H. pylori infection in the oral cavity and the stomach. The mouth is the first extra-gastric reservoir. Regarding the role of H. pylori in the etiology of squamous cell carcinoma, no evidence is still available.

Sentinel Lymph Node

AbstractIntroduction: Lymph node status in patients with cutaneous malignant melanoma is the most important prognostic factor. Patients with clinically positive nodes (stage III) should undergo therapeutic lymphadenectomy; however, the surgical approach to the regional disease in patients with negative clinical examination (stage I and II) is still controversial. Selective lymphadenectomy consists of the intraoperative identification of the first node in the nodal basin, the sentinel lymph node (SLN). Routine examination, serial sectioning, and immunohistochemistry may underestimate the presence of tumor cells. PCR is a molecular biology technique that may be useful for the detection of malignant melanoma nodal metastases in the SLN. Aim: The aim of this study was to use tyrosinase messenger RNA (mRNA) amplification for the detection of micrometastases in fresh frozen SLNs. Methods: 46 hematoxylin-eosin (HE)-negative sentinel node samples from 42 patients with malignant melanoma were included in this study. Formalin-fixed paraffin-embedded sections were immunostained with S-100 protein and HMB-45. A central portion of the node was submitted for PCR. This method was accomplished with a combination of reverse transcription and amplification of the tyrosinase complementary DNA and double-round PCR (nested reverse transcriptase [RT]-PCR). Results: In 1 of the 42 SLN-negative patients, immunohistochemistry stains allowed the detection of micrometastases. With molecular biology, 14 of the 42 SLN patients were positive (33%); in another 12 (29%), only the nested RT-PCR was positive. Of the 42 patients, 24 were put into 3 groups and followed for a 5-year period with 1, 7, and 16 patients, respectively, in the groups. The first group involved 1 patient who had provided 2 SLN samples that were found to be SLN-positive using both techniques, immunohistochemistry stains and nested RT-PCR (he had hepatic metastasis and died 24 months after diagnosis). The second group, with only nested RT-PCR positive SLN samples, included 7 of 12 patients who were followed and had a median survival of 37 months; 4 died of widespread metastatic disease, the other 3 patients had event-free survival, but 1 consented to undergo a therapeutic lymphadenectomy as a result of a positive test. The last group consisting of 16 of 32 patients, with complete 5-year survival, who were SLN-negative with both techniques, immunohistochemistry stains and nested RT-PCR. Fourteen of the 16 (88%) were event-free survival during the follow-up, and 2 had local relapse. Conclusion: Tyrosinase mRNA amplification may be a negative prognostic factor for the detection of micrometastases in fresh frozen SLNs using molecular biology techniques.

Sentinel node in melanoma patients: triple negativity with routine techniques and PCR as positive prognostic factor for survival.

Lymph node mapping and sentinel lymph node biopsy are currently used to stage patients with cutaneous malignant melanoma. Immunohistochemical stains contribute to the detection of micrometastases; however, molecular biology techniques are associated with better diagnostic sensitivity. Sixty sentinel lymph nodes were included in this study. The primary lesions were malignant melanoma stage I or II, with a follow-up of longer than 2 years. Sentinel lymph nodes were studied with hematoxylin–eosin, immunohistochemistry for S-100 and HMB-45, and molecular biology techniques (reverse transcription (RT)-PCR) for the detection of tyrosinase messenger RNA. In 15 of 60 cases (25%), tyrosinase was detected by RT-PCR; three of these cases were also positive by immunohistochemistry. The population was divided into three groups: (i) hematoxylin–eosin−/immunohistochemistry+/molecular biology techniques+ (3 cases); (ii) hematoxylin–eosin−/immunohistochemistry−/molecular biology techniques+ (12 cases); (iii) hematoxylin–eosin−/immunohistochemistry−/molecular biology techniques− (45 cases). Correlation of the groups with overall survival showed the following: (i) 2 of 3 patients died (67%); (ii) 5 of 12 died (42%), and (iii) all 45 patients are alive, with no lymphadenectomy and a median follow-up of 84 months. The inclusion of molecular biology techniques appears to be of great value for the detection of sentinel lymph node micrometastases in patients with cutaneous malignant melanoma. In our series, those patients who showed negativity with all the three methods had a null recurrence rate. Therefore, this triple negativity could be a positive prognostic factor for overall survival. Our findings suggest the possibility of molecular oncological staging, which would allow the selection of patients with submicroscopic metastases for a complete treatment.

Immunohistochemical characterization of neoplastic cells of breast origin.

BackgroundAfter skin cancer, breast cancer is the most common malignancy in women. Tumors of unknown origin account for 5-15% of malignant neoplasms, with 1.5% being breast cancer. An immunohistochemical panel with conventional and newer markers, such as mammaglobin, was selected for the detection of neoplastic cells of breast origin. The specific objectives are: 1) to determine the sensitivity and specificity of the panel, with a special emphasis on the inclusion of the mammaglobin marker, and 2) to compare immunohistochemistry performed on whole tissue sections and on Tissue Micro-Array.MethodsTwenty-nine metastatic breast tumors were included and assumed as tumors of unknown origin. Other 48 biopsies of diverse tissues were selected and assumed as negative controls. Tissue Micro-Array was performed. Immunohistochemistry for mammaglobin, gross cystic disease fluid protein-15, estrogen receptor, progesterone receptor and cytokeratin 7 was done.ResultsMammaglobin positive staining was observed in 10/29 cases, in 13/29 cases for gross cystic disease fluid protein-15, in 20/29 cases for estrogen receptor, in 9/29 cases for progesterone receptor, and in 25/29 cases for cytokeratin 7. Among the negative controls, mammaglobin was positive in 2/48, and gross cystic disease fluid protein-15 in 4/48.ConclusionsThe inclusion of MAG antibody in the immunohistochemical panel for the detection of tumors of unknown origin contributed to the detection of metastasis of breast cancer. The diagnostic strategy with the highest positive predictive value (88%) included hormone receptors and mammaglobin in serial manner.Virtual slidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1366310812718988

Sentinel lymph node: detection of micrometastases of melanoma in a molecular study.

INTRODUCTION Lymph node status in patients with cutaneous malignant melanoma is the most important prognostic factor. Patients with clinically positive nodes (stage III) should undergo therapeutic lymphadenectomy; however, the surgical approach to the regional disease in patients with negative clinical examination (stage I and II) is still controversial. Selective lymphadenectomy consists of the intraoperative identification of the first node in the nodal basin, the sentinel lymph node (SLN). Routine examination, serial sectioning, and immunohistochemistry may underestimate the presence of tumor cells. PCR is a molecular biology technique that may be useful for the detection of malignant melanoma nodal metastases in the SLN. AIM The aim of this study was to use tyrosinase messenger RNA (mRNA) amplification for the detection of micrometastases in fresh frozen SLNs. METHODS 46 hematoxylin-eosin (HE)-negative sentinel node samples from 42 patients with malignant melanoma were included in this study. Formalin-fixed paraffin-embedded sections were immunostained with S-100 protein and HMB-45. A central portion of the node was submitted for PCR. This method was accomplished with a combination of reverse transcription and amplification of the tyrosinase complementary DNA and double- round PCR (nested reverse transcriptase [RT]-PCR). RESULTS In 1 of the 42 SLN-negative patients, immunohistochemistry stains allowed the detection of micrometastases. With molecular biology, 14 of the 42 SLN patients were positive (33%); in another 12 (29%), only the nested RT-PCR was positive. Of the 42 patients, 24 were put into 3 groups and followed for a 5-year period with 1, 7, and 16 patients, respectively, in the groups. The first group involved 1 patient who had provided 2 SLN samples that were found to be SLN-positive using both techniques, immunohistochemistry stains and nested RT-PCR (he had hepatic metastasis and died 24 months after diagnosis). The second group, with only nested RT-PCR positive SLN samples, included 7 of 12 patients who were followed and had a median survival of 37 months; 4 died of widespread metastatic disease, the other 3 patients had event-free survival, but 1 consented to undergo a therapeutic lymphadenectomy as a result of a positive test. The last group consisting of 16 of 32 patients, with complete 5-year survival, who were SLN-negative with both techniques, immunohistochemistry stains and nested RT-PCR. Fourteen of the 16 (88%) were event-free survival during the follow-up, and 2 had local relapse. CONCLUSION Tyrosinase mRNA amplification may be a negative prognostic factor for the detection of micrometastases in fresh frozen SLNs using molecular biology techniques.

Sentinel Lymph Nodes Study: How to Do it Right? The Argentinean Experience

To the Editor: We would like to express our opinion about the article by Treseler et al. (1). We congratulate them on the exhaustive review of the most recent and relevant information about sentinel lymph nodes (SLN) evaluation and its clinical implications. In this letter we would like to stress some points regarding molecular studies. Even though most published data have shown a decrease in overall and disease-free survival in patients with micrometastases detected by hematoxylineosin (HE) or immunohistochemistry (IHC), the latter is not routinely recommended for clinical practice in most countries (1). However, retrospective studies of serial sections with HE have revealed metastases in approximately 30% of cases (2). To date, molecular-techniques for detection of specific markers by Retro Transcription-Polymerase Chain Reaction (RT-PCR) are only used for experimental purposes. In the last few years, several molecular markers has been proposed in the literature, such as CEA, PIP, CK19, MUC1, PSE, mammaglobin (MAG) A, and MAG-B (3). Recently, worldwide literature has demonstrated that MAG is the most sensitive and specific (3–6). Over the last 2 years, our research group has pioneered the study of MAG (both isoforms A and B) in Argentine. The purpose of our study was to analyze the presence of messenger RNA of MAG A and B in breast SLN by RT-PCR, and to correlate these findings with routine techniques (HE and IHC). To date, this experimental protocol includes 40 patients, all of them with primary tumor size £3 cm, and no palpable axillary lymphadenopathies. Conventional techniques (HE) showed metastases in 4 ⁄ 40 cases. We found 5 ⁄ 40 cases with IHC (Cytokeratin AE1-AE3), one of them negative with HE. Molecular biology techniques revealed 12 ⁄ 40 positive SLN with MAG A and B by RT-PCR, which comprised the five above-mentioned cases (four cases were positive with MAG-A, three with MAG-B, and five with both). Our study is one of the first designs using an RT-PCR multiplex, which includes both isoforms of MAG (A and B), in an attempt to increase the detection rate. Our results with molecular biology (12 ⁄ 40) are similar to other published data (3,7,8). Multiplex RT-PCR technique for MAG A and B proves to be specific and sensitive. To date, the only patient with HE(+), IHC(+), MGBA(+), and MGB-B(+) had disease relapse at 23 months of follow-up. All the other patients appear to be disease-free. However, we believe that they should be followed up for a longer period of time to determine whether there is a significant impact of RT-PCR positive findings. Our study has shown the Argentinean experience with a known molecular marker but with a novel approach to RT-PCR technique. Finally, we hope to encourage the development of new research protocols that include the use of molecular techniques in SLN evaluation. Based on current knowledge, we think other experimental protocols including more patients and a longer follow-up should be further developed, in order to assess their real prognostic value.

Short-term menhaden oil rich diet changes renal lipid profile in acute kidney injury.

Weanling male Wistar rats fed a choline-deficient diet develop acute kidney injury. Menhaden oil, which is a very important source of omega-3 fatty acids, has a notorious protective effect. The mechanism of this protection is unknown; one possibility could be that menhaden oil changes renal lipid profile, with an impact on the functions of biological membranes. The aim of this work was to study the renal lipid profile in rats fed a choline-deficient diet with menhaden oil or vegetable oil as lipids. Rats were divided into 4 groups and fed four different diets for 7 days: choline-deficient or choline-supplemented diets with corn and hydrogenated oils or menhaden oil. Serum homocysteine, vitamin B12, and folic acid were analyzed. Renal lipid profile, as well as the fatty acid composition of the three oils, was measured. Choline-deficient rats fed vegetable oils showed renal cortical necrosis. Renal omega-6 fatty acids were higher in rats fed a cholinedeficient diet and a choline-supplemented diet with vegetable oils, while renal omega-3 fatty acids were higher in rats fed a choline-deficient diet and a choline-supplemented diet with menhaden oil. Rats fed menhaden oil diets had higher levels of renal eicosapentaenoic and docosahexaenoic acids. Renal myristic acid was increased in rats fed menhaden oil. The lipid renal profile varied quickly according to the type of oil present in the diet.