Relevance of BRAF(V600E) Mutation Testing Versus RAS Point Mutations and RET/PTC Rearrangements Evaluation in the Diagnosis of Thyroid Cancer

A molecular profile including BRAF and RAS mutations as well as RET/PTC rearrangement evaluation has been proposed to provide an accurate presurgical assessment of thyroid nodules and to reduce the number of unnecessary diagnostic surgeries, sparing patients' health and saving healthcare resources. However, the application of such molecular analyses may provide different results among different centers and populations in real-life settings. Our aims were to evaluate the diagnostic utility of assessing the presence of BRAF and RAS mutations and RET/PTC1 and RET/PTC3 rearrangements in all cytological categories in an Italian group of thyroid nodule patients assessed prospectively, and to understand whether and which mutation testing might be helpful in cytologically indeterminate nodules. A total of 911 patients were submitted to ultrasound and fine-needle aspiration biopsy examination. Cytological evaluation was performed in parallel with molecular testing and compared to pathological results in 940 thyroid nodules, including 140 indeterminate lesions. BRAF mutation testing provided the best contribution to cancer diagnosis, allowing the disease to be detected at an early stage, and identifying indeterminate nodules in which diagnostic lobectomy could be spared. On the contrary, RAS and RET/PTC analysis did not further increase diagnostic sensitivity for thyroid cancer. In addition, we found RET/PTC rearrangements in benign lesions, indicating that this molecular marker might not be useful for the detection of thyroid cancer. BRAF(V600E) mutation analysis is superior to RAS point mutations and evaluation of RET/PTC rearrangements in the diagnosis of thyroid cancer, even in indeterminate lesions

Magmas, a gene newly identified as overexpressed in human and mouse ACTH-secreting pituitary adenomas, protects pituitary cells from apoptotic stimuli

Abstract Pituitary tumors are mostly benign, being locally invasive in 5-35% of cases. Deregulation of several genes has been suggested as a possible alteration underlying the development and progression of pituitary tumors. We here report the identification of a cDNA, corresponding to Magmas gene (mitochondria-associated protein involved in granulocyte-macrophage colony-stimulating factor signal transduction), which is highly expressed in two different ACTH-secreting mouse pituitary adenoma cell lines as compared with normal pituitary as well as in two thirds of 64 examined pituitary adenomas as compared with human normal pituitary. Tim 16, the mitochondrial protein encoded by Magmas, was indeed expressed in a mouse ACTH-secreting pituitary adenoma cell line, AtT-20 D16v-F2 cells, in a subcellular compartment likely corresponding to mitochondria. Magmas silencing determined a reduced rate of DNA synthesis, an accumulation in G1 phase, and a concomitant decrease in S phase in At-T20 D16v-F2 cells. Moreover, Magmas-silenced cells displayed basal caspase 3/7 activity and DNA fragmentation levels similar to control cells, which both increased under proapoptotic stimuli. Our data demonstrate that Magmas is overexpressed in mouse and human ACTH-secreting pituitary adenomas. Moreover, our results show that Magmas protects pituitary cells from apoptosis, suggesting its possible involvement in neoplastic transformation

Proposal for a novel management of indeterminate thyroid nodules on the basis of cytopathological subclasses

Indeterminate thyroid nodules include heterogeneous lesions that could benefit from a differential management. Our aim is to better define the management of the Bethesda System for Reporting Thyroid Cytopathology class III and IV nodules, by identifying cytological subcategories among Bethesda System for Reporting Thyroid Cytopathology class III associated with different clinical risk, by means of ultrasound, repeated FNAB, and BRAFV600E molecular analysis. We also evaluated the outcome of nodules not operated, over a 5-year follow-up. Out of 460 nodules (269 Bethesda System for Reporting Thyroid Cytopathology class III and 191 Bethesda System for Reporting Thyroid Cytopathology class IV), 344 were operated on surgical group and 116 followed-up conservatively (follow-up group). Bethesda System for Reporting Thyroid Cytopathology class III was divided into four subcategories on the basis of cytomorphological features (III-1, III-2, III-3, III-4). Clinical risk was defined on the basis of histological, cytological, and ultrasound data. Malignancy was higher in Bethesda System for Reporting Thyroid Cytopathology class III vs. Bethesda System for Reporting Thyroid Cytopathology class IV (34.4 vs. 26.2 %; p < 0.01). Papillary thyroid carcinoma was the most frequent cancer in each Bethesda System for Reporting Thyroid Cytopathology class (35 %). BRAFV600E diagnostic accuracy was 87 %. Repeated FNAB reclassified as benign nearly 40 % of nodules, selecting patients where surgery could be spared. Significant nodule growth occurred in 13.7 % of nodules, belonging mostly to Bethesda System for Reporting Thyroid Cytopathology class III-2 and Bethesda System for Reporting Thyroid Cytopathology class IV. Overall clinical risk was higher in Bethesda System for Reporting Thyroid Cytopathology III-1, III-4, and IV classes. We propose a differential management of Bethesda System for Reporting Thyroid Cytopathology III and IV classes and related subcategories: surgery may be indicated in Bethesda System for Reporting Thyroid Cytopathology class III-1, III-4, and IV; a conservative follow-up avoiding repeated FNAB may be appropriated in class III-3, while repeated FNAB may be useful in class III-2

An Evaluation of Hospital Cleaning Regimes&mdash;Microbiological Evaluation and LCA Analysis after Traditional and Sustainable/Green Procedures

The development of sustainable processes has a significant role in balancing social productivity demands, environmental protection, and natural resources. The control of microbial contamination has so far been concentrated on the use of chemical-based sanitation procedures, which may have various limitations, as testified by the persistence of contamination itself, by the growing antimicrobial resistance (AMR) of hospital microbes, and by chemical-related pollution. This communication aims to present a comparative analysis between the use of traditional and green sanitation products and processes in hospital environments. The sampling campaign was conducted in a day hospital ward of a general hospital (Imola, Santa Maria della Scaletta Hospital). Each sample comes from a specific surface, furniture or sanitary, and was taken using RODAC contact plates and swabs with a neutralizing agent in order to standardize the result of the microbiological evaluation. Sampling occurred before and after traditional and green cleaning procedures. The green experimental protocol using 100% natural and biodegradable products in sanitization procedures of hospital areas at a medium-high health risk was found to be a technique of relevant interest. From CAM requirements, the green protocol must give equal or better results than the traditional protocol. It can be concluded that the green experimental system meets this criterion and has shown better antimicrobial activity performance than the traditional system; all findings are in an acceptable state of sanitation, with no evidence of pathogenic micro-organisms specified in the guideline

An Evaluation of Hospital Cleaning Regimes—Microbiological Evaluation and LCA Analysis after Traditional and Sustainable/Green Procedures

The development of sustainable processes has a significant role in balancing social productivity demands, environmental protection, and natural resources. The control of microbial contamination has so far been concentrated on the use of chemical-based sanitation procedures, which may have various limitations, as testified by the persistence of contamination itself, by the growing antimicrobial resistance (AMR) of hospital microbes, and by chemical-related pollution. This communication aims to present a comparative analysis between the use of traditional and green sanitation products and processes in hospital environments. The sampling campaign was conducted in a day hospital ward of a general hospital (Imola, Santa Maria della Scaletta Hospital). Each sample comes from a specific surface, furniture or sanitary, and was taken using RODAC contact plates and swabs with a neutralizing agent in order to standardize the result of the microbiological evaluation. Sampling occurred before and after traditional and green cleaning procedures. The green experimental protocol using 100% natural and biodegradable products in sanitization procedures of hospital areas at a medium-high health risk was found to be a technique of relevant interest. From CAM requirements, the green protocol must give equal or better results than the traditional protocol. It can be concluded that the green experimental system meets this criterion and has shown better antimicrobial activity performance than the traditional system; all findings are in an acceptable state of sanitation, with no evidence of pathogenic micro-organisms specified in the guideline

Control of Erwinia amylovora Growth by Moringa oleifera Leaf Extracts: In Vitro and in Planta Effects

Abstract: Erwinia amylovora (EA) is a phytopathogenic bacterium, the causative agent of bacterial fire blight, a disease that affects Rosaceaes. In order to replace antibiotics and copper, the antimi-crobial activity of three extracts of Moringa oleifera Lam., methanolic (MeOH-MOE), hydroalco-holic (HA-MOE) and hydroalcoholic with maltodextrins (HAMD-MOE), was tested on eleven strains of EA isolated from apple trees by the Emilia-Romagna Phytosanitary Department. MIC and MBC have been evaluated; biofilm formation, swarming motility and amylovoran produc-tion were performed with the crystalviolet, soft-agar assay and the amylovoran method. All ex-tracts demonstrated bacteriostatic activity at a concentration of 1 mg/mL, resulting in a 80% re-duction in biofilm formation. HAMD-MOE, MeOH-MOE and HA-MOE caused an inhibition of motility of 60%, 65% and 30% after 6 days and a decrease in amylovoran synthesis of 84%, 63% and 93%, respectively. In planta results showed how the compounds were able to inhibit EA vir-ulence on apple trees, mainly if they were applied as a preventive treatment, although the treat-ment showed a significant reduction in fire blight symptoms progression. The antibacterial activ-ity of the extracts is mainly due to the high concentration of polyphenolic compounds detected in the extracts that was able to alter the permeability of bacterial membrane, resulting in slowing the synthesis of ATP and consequently of all ATP-dependent functions, such as motility and less se-lectivity towards harmful compounds, which can, thus, enter the cytoplasm and inhibit enzymes involved in replication and quorum sensing. The efficacy, eco-compatibility and low cost make such extracts a potential tool for the control of bacterial fire blight

Analysis of Civil Environments Cleaning Services&mdash;Microbiological and LCA Analysis after Traditional and Sustainable Procedures

The development of sustainable processes has an important role to play in balancing social productivity requirements, protecting the environment and natural resources. Up to date, efforts to combat microbial contamination have focused on the use of chemical-based sanitation procedures, which may have various limitations, as testified by the persistence of contamination itself, by the growing antimicrobial resistance of microbes and by the chemical related pollution. The purpose of this paper is to present a comparative analysis of the use of conventional and sustainable cleaning products and processes in civilian environments. The sampling campaign is conducted in a sports hall in Turin, Italy. Each sample comes from a specific environment, surface, furniture and sanitary and is taken using RODAC contact plates and swabs with a neutralizing agent in order to standardize the result of the microbiological evaluation. Sampling occurred before and after traditional and sustainable cleaning procedures. The sustainable experimental design using ecological products in the cleaning procedures of the analyzed areas proved to be a relevant technique. From CAM (minimal environmental criteria) requirements, the sustainable protocol must give equal or better results than the traditional protocol from both microbiological and environmental (LCA) point of view. It can be concluded that the ecological experimental system meets this criterion and has demonstrated better performance both in antimicrobial activity and in environmental impact than the conventional system: all findings are in an acceptable state of sanitation, with no evidence of pathogenic micro-organisms specified in the guideline

Effects of Moringa oleifera Leaf Extracts on Xanthomonas campestris pv. campestris

Abstract: Xanthomonas campestris pv. campestris (Xcc) is a Gram-negative bacterium belonging to the Xanthomonodaceae family, causing black rot in crucifers. To control this pathogen, the study investigated the effect of different leaves extracts of Moringa oleifera Lam., a tropical plant, well known for its food properties and with countless applications in many different fields, from nutraceutical (hypoglycemic) to the cosmetic (sunscreen) properties. Nevertheless, several studies pointed to its antibacterial action against both Gram-negative and Gram-positive bacteria. Many bioactive compounds, including flavonoids, phenolic acids, alkaloids, isothiocyanates, tannins and saponins, contained in these extracts, are responsible for its countless activities. The analyses carried out in this study show that the methanolic, hydroalcoholic and hydroalcoholic malto-dextrin extracts have both bacteriostatic and bactericidal effects at concentrations of 0.5, 0.5L and 0.1 mg/mL respectively. In particular, the study shows how all extracts can alter membrane permeability, to adversely affect swarming motility, and to alter biofilm formation in Xcc. The in planta experiments showed a reduction of the necrosis area in the infected radishes, although the ability of the extracts to be absorbed by root systems is yet to be understood, in order to reach the target point

Role of piuitary tumour transforming gene 1 in medullary thyroid carcinoma

Pituitary tumour transforming gene 1 (PTTG1) is over-expressed in a variety of endocrine-related tumors. We investigated PTTG1 expression in human C-cell hyperplasia (CCH), human medullary thyroid carcinoma (MTC) and in the human MTC cell line, TT. PTTG1 expression was significantly higher (P<0.01) in CCH (threefold), in papillary thyroid cancer and in MTC (fivefold) than in normal thyroid, and in MTC lymph-node metastases as compared to primary lesions (approximately twofold; P<0.05). PTTG1 mRNA and protein correlated with tumor diameter and TNM status (P<0.05). In TT cells, PTTG1 silencing did not completely block DNA synthesis, but significantly reduced [3H]Thymidine incorporation (~50%; P<0.01) for up to 3 days. Furthermore, PTTG1 overexpression significantly decreased cell proliferation, lasting for at least 3 days (30–60%; P<0.01). Therefore, PTTG1 levels correlate with tumor aggressiveness. PTTG1 silencing and overexpression cause reduced MTC cell proliferation, supporting the hypothesis that PTTG1 is an important determinant of C-cell neoplastic proliferation