Recent developments in data acquisition, treatment and analysis with ion mobility-mass spectrometry for lipidomics.

Lipids are involved in many biological processes and their study is constantly increasing. To identify a lipid among thousand requires of reliable methods and techniques. Ion Mobility (IM) can be coupled with Mass Spectrometry (MS) to increase analytical selectivity in lipid analysis of lipids. IM-MS has experienced an enormous development in several aspects, including instrumentation, sensitivity, amount of information collected and lipid identification capabilities. This review summarizes the latest developments in IM-MS analyses for lipidomics and focuses on the current acquisition modes in IM-MS, the approaches for the pre-treatment of the acquired data and the subsequent data analysis. Methods and tools for the calculation of Collision Cross Section (CCS) values of analytes are also reviewed. CCS values are commonly studied to support the identification of lipids, providing a quasi-orthogonal property that increases the confidence level in the annotation of compounds and can be matched in CCS databases. The information contained in this review might be of help to new users of IM-MS to decide the adequate instrumentation and software to perform IM-MS experiments for lipid analyses, but also for other experienced researchers that can reconsider their routines and protocols.

Secondary node analysis as an indicator for axillary lymphadenectomy in breast cancer patients. / Ganglio secundario como indicador de linfadenectomía axilar en pacientes afectas de cáncer de mama.

INTRODUCTION: Currently, there is no agreement regarding if it would be necessary to perform an axillary lymph node dissection (ALND) in patients who have macrometastases in the sentinel lymph node (SLN). We studied the utility of the secondary node analysis (SN), defined as the following node after the SLN in an anatomical and lymphatic pathway, as a sign of malignant axillary involvement. METHODS: An observational, retrospective and multicentre study was designed to assess the utility of the SN as a sign of axillary involvement. Among 2273 patients with breast cancer, a valid sample of 283 was obtained representing those who had the SN studied. Main endpoints of our study were: the SLN, the SN and the ALND histological pattern. Sensitivity, specificity and precision of the test were also calculated. RESULTS: SN test, in cases with positive SLN, has a sensitivity of 61.1%, a specificity of 78.7%, a positive predictive value of 45.8% and a negative predictive value of 87.3% with a precision of 74.7%. CONCLUSION: The study of the SN together with the technique of the SLN allows a more precise staging of the axillary involvement, in patients with breast cancer, than just the SLN technique.

[Use of platelet gel in breast reconstruction after breast-conserving cancer surgery]. / Reconstrucción de la mama con gel de plaquetas en la cirugía conservadora del cáncer.

INTRODUCTION: With the aim of decreasing breast defects after conservative cancer surgery, we present a new breast reconstruction technique using breast volume restitution with platelet gel. PATIENTS AND METHOD: A pilot study was conducted on 20 breast cancer patients undergoing tumorectomy with placement a gel obtained by plateletpheresis of a healthy allogeneic donor in the surgical cavity. Patients had a clinical, photographic and histological follow-up, as well as an assessment of the aesthetic outcome. RESULTS: The mean age was 50.5±8.6 years (range 42-70 years) and with a mean Charlson comorbidity index of 1.1±1.2 (range 0-5). The mean tumour volume was 63.1±31.1 ml (range 30-160 ml) and the mean restitution volume with platelet gel was 111.5±60.9 ml (range 40-250 ml). After a mean follow-up of 17 months, 80% of the patients maintained the breast volume and no further operations were needed due to surgical margin involvement. No recurrences were observed in any patient. CONCLUSIONS: Platelet gel allows restitution of the breast volume adjusted to the tumorectomy volume, minimising the usual retractions and deformities after conservative surgery. It enables wide resections and safety margins.

Prognostic value of hematogenous dissemination and biological profile of the tumor in early breast cancer patients: a prospective observational study.

BACKGROUND: The aim of this study was to investigate the incidence and prognostic value of disseminated tumor cells in bone marrow of breast carcinoma patients with early disease, and to analyze this finding in relation to lymph node involvement, determined by sentinel lymph node (SLN) biopsy analysis, and to prognostic factors of interest. METHODS: 104 patients with operable (T<3 cm) breast cancer and clinically- and sonographically-negative axillary lymph nodes were scheduled for SLN biopsy. Bone marrow aspirates were collected before the start of surgery from both iliac crests, and mononuclear cell layers were separated by density centrifugation (Lymphoprep). Slide preparations were then examined for the presence of disseminated tumor cells by immunocytochemistry with anti-cytokeratin antibodies (A45-B/B3). Lymphoscintigraphy was performed 2 hours after intratumor administration of 2 mCi (74 MBq) of 99mTc colloidal albumin. The SLN was evaluated for the presence of tumor cells by hematoxylin-eosin staining and, when negative, by immunocytochemistry using anti-cytokeratin antibody (CAM 5.2). Survival analyses and comparative analyses were performed on the results of bone marrow determinations, SLN biopsy, and known prognostic factors, including breast cancer subtypes according to the simplified classification based on ER, PR and HER2. RESULTS: Lymph node and hematogenous dissemination occur in one-third of patients with early-stage breast cancer, although not necessarily simultaneously. In our study, disseminated tumor cells were identified in 22% of bone marrow aspirates, whereas 28% of patients had axillary lymph node involvement. Simultaneous lymph node and bone marrow involvement was found in only 5 patients (nonsignificant). In the survival study (60 months), a higher, although nonsignificant rate of disease-related events (13%) was seen in patients with disseminated tumor cells in bone marrow, and a significant association of events was documented with the known, more aggressive tumor subtypes: triple negative receptor status (21%) and positive ERBB2 status (29%). CONCLUSIONS: Tumor cell detection in bone marrow can be considered a valid prognostic parameter in patients with early disease. However, the classic prognostic factors remain highly relevant, and the newer breast cancer subtypes are also useful for this purpose.

Unravelling the Encapsulation of DNA and Other Biomolecules in HAp Microcalcifications of Human Breast Cancer Tissues by Raman Imaging.

Microcalcifications are detected through mammography screening and, depending on their morphology and distribution (BI-RADS classification), they can be considered one of the first indicators of suspicious cancer lesions. However, the formation of hydroxyapatite (HAp) calcifications and their relationship with malignancy remains unknown. In this work, we report the most detailed three-dimensional biochemical analysis of breast cancer microcalcifications to date, combining 3D Raman spectroscopy imaging and advanced multivariate analysis in order to investigate in depth the molecular composition of HAp calcifications found in 26 breast cancer tissue biopsies. We demonstrate that DNA has been naturally adsorbed and encapsulated inside HAp microcalcifications. Furthermore, we also show the encapsulation of other relevant biomolecules in HAp calcifications, such as lipids, proteins, cytochrome C and polysaccharides. The demonstration of natural DNA biomineralization, particularly in the tumor microenvironment, represents an unprecedented advance in the field, as it can pave the way to understanding the role of HAp in malignant tissues.

Is it possible to refine the indication for sentinel node biopsy in high-risk ductal carcinoma in situ?

BACKGROUND: The indication for sentinel node biopsy (SNB) has not been fully established yet for patients with ductal carcinoma in situ (DCIS). AIM: To relate the conversion rate to invasive carcinoma with sentinel node positivity in high risk DCIS, and to refine the clinical presentation analysis in order to better select patients for SNB. For this purpose, a risk score was devised. METHODS: From 1998 to 2005, 151 high-risk DCIS patients from six clinical centres were included in a prospective sentinel node database. The conversion rate to invasive carcinoma was 39%. Ten of 142 (7%) successful SNBs showed a positive sentinel node (eight micrometastatic). The sentinel node was positive in 1% of pure DCIS, in 5.5% of DCIS with micro-invasion, and in 19.5% of invasive carcinoma. RESULTS: Both clinical presentation and corresponding risk score were closely related to conversion to invasive carcinoma. The association of risk score and sentinel node positivity approached but did not reach statistical significance (P=0.06); therefore a subset of further selected higher risk patients could not be defined. CONCLUSION: The relevance of SNB positivity cannot be overlooked in high-risk DCIS patients, however, because SNB is not free from morbidity and cost, more studies are needed to refine its final indication.

Localized renal cystic disease: imaging findings, pathologic correlation, and management approach.

A case of unilateral renal cystic disease of the kidney is reported, in which the cysts were located to the lower pole of the right kidney. The radiologic and pathologic findings are described and their differential diagnosis is discussed. Conservative treatment and imaging follow-up is proposed for this condition.

Detection of disseminated tumor cells in locally advanced breast cancer patients before primary systemic therapy.

AIM: To assess the prevalence and prognostic power of disseminated tumor cells (DTC) in patients with locally advanced breast cancer (LABC) before primary systemic therapy (PST). MATERIALS AND METHODS: LABC patients attending our Breast Unit were studied between 2002 and 2012, all of them being considered for PST. To determine the presence of DTC, posterior iliac crest aspirates were obtained and marrow samples were processed by gradient separation with Ficoll (Lymphoprep(®)) and immunohistochemical staining using the antiCK A45-B/B3 (EPIMET) antibody. Clinicopathologic variables were recorded before and after PST to assess response. Disease-free survival (DFS) and overall survival (OS) were determined after follow-up. The presence of DTC as a predictor of response to PST and as a prognostic tool for OS and DSF was evaluated. RESULTS: DTC were observed in 26% of 47 patients included in the study. PST consisted of chemotherapy in 94% and hormone therapy in 6%. Breast-conserving therapy was attained in 33%. Mean follow-up was 68 months. Complete clinical response (CR) after PST was seen in 26%, disease recurrence in 38%, and cancer-related death in 8%; tumor size and negative estrogen receptors were significant predictors of CR and mastectomy was associated with DFS. Persistent axillary disease after PST and previous recurrence were predictive of OS. DTC were detected more often in patients who did not achieve CR and those who presented recurrence. DTC detection was a significant prognostic factor for a worse OS (OR = 7.62; CI95%: 1.46-39.61; p = 0.009) and a decreased survival time (62 versus 82 months, p = 0.004). CONCLUSION: Presence of DTC before PST was found in a significant number of patients with LABC. DTC were found to be a significant prognostic factor for cancer-related death. DTC could be a surrogate predictor of response to PST and also of disease recurrence in LABC patients.

[Radio-guided occult lesion localization combined with sentinel node biopsy in women with breast cancer]. / Radiolocalización de lesiones mamarias no palpables combinada con la biopsia del ganglio centinela en mujeres con cáncer de mama.

INTRODUCTION: The practice of intratumoral radiocolloid injection for sentinel node biopsy (SNB) allows localization of nonpalpable breast tumors in the same surgical intervention, using the technique known as radio-guided occult lesion localization (ROLL). PATIENTS AND METHOD: We present our experience with the combined use of both techniques (SNB and ROLL). A total of 233 patients were retrospectively analyzed: 65 patients underwent ROLL and 168 patients underwent wire-guided localization (control group) as the excision modality. Both groups had comparable pathologic and radiologic features. RESULTS: Complete excision of the lesion was achieved at the first attempt in only 169 patients. There were no significant differences among groups in surgical margin status (margins were adequate in 80% of ROLL patients vs 69.9% of non-ROLL patients) or in maximum specimen diameter (6.85 cm vs 6.52 cm, respectively). CONCLUSION: We conclude that ROLL is an acceptable alternative to wire-guided localization in patients with nonpalpable breast tumors undergoing SNB, leading to easier lesion excision and greater patient comfort.

Replacement lipomatosis related to renal tuberculosis: imaging findings in one case.

Replacement lipomatosis of the kidney in a case of long-standing renal tuberculosis is reported. The radiologic and pathologic findings are described and the differential diagnosis is discussed. A hypothesis is given to explain the association of renal tuberculosis and replacement lipomatosis of the kidney.

Prognostic value of CT in the early assessment of patients with acute pancreatitis.

OBJECTIVE: This study investigates the prognostic value of early CT in acute pancreatitis, the role of pancreatic necrosis as a indicator of prognosis, and the need for the routine use of IV iodinated contrast material in early CT to assess prognosis in these patients. MATERIALS AND METHODS: We conducted a retrospective review of 148 patients who underwent unenhanced and contrast-enhanced helical CT within 72 hr after onset of symptoms of a first episode of acute pancreatitis. Patients were classified by CT grade and grouped into two categories (mild: grades A, B, C; and severe: grades D and E) that were correlated with complications and death. In the grades including patients with pancreatic necrosis, it was also correlated with complications and death. RESULTS: All complications (n = 15) and deaths (n = 4) occurred in patients with a CT grade of severe disease; differences as compared with mild grade were significant (p < 0.001 and p < 0.03, respectively). CT grade had a sensitivity and specificity of 100% and 61.6%, respectively, for predicting morbidity and 100% and 56.9% for predicting mortality. The 13 patients with necrosis were all in the severe group (p < 0.001). Necrosis detection on early CT had a sensitivity and specificity of 53.3% and 90.2%, respectively, for predicting morbidity and 75% and 83.8% for mortality. CONCLUSION: Early unenhanced CT alone was a good indicator of severity of acute pancreatitis in our selected population. CT grade was sensitive for predicting outcome in acute pancreatitis. Pancreatic necrosis, estimated on early, contrast-enhanced CT and seen only in patients having severe disease, was a specific predictor of morbidity and mortality. These findings lead us to suggest that the use of iodinated contrast material to assess necrosis can be reserved for only those patients classified as having severe disease on unenhanced CT.

Internal mammary sentinel node metastases in an otherwise lymph-node negative breast cancer patient.

A 35 year old woman with biopsy proved breast cancer was submitted for sentinel node (SN) biopsy. Preoperative lymphoscintigraphy displayed both axillary and internal mammary (IM) uptake foci consistent with SNs. Full axillary dissection was completed because of a greater-than 2 cm primary lesion. Two axillary SNs were excised. An IM SN was also excised through the second intercostal space, with the aid of the gamma probe. Fourteen axillary nodes, including SNs, were negative, whereas the IM SN harbored several metastatic implants. Implications for staging, prognosis and further therapy of such IM-only positive sentinel nodes are discussed.

Garantía de calidad de los alimentos fortificados: manual para inspectores / Garanty of quality for strengthen foods: Manual for inspectors

El propósito de este manual es codayuvar en la formación y las funciones que cumplen los inspectores del control de la calidad de los alimentos, sistematizando las principales tareas del control de calidad y los aspectos relevantes de la fortificación