LDOC-1 and PARP-1 mRNA expression in leukocytes of father and son with cutaneous malignant melanoma

Abstract Apoptosis is central to the biology of cutaneous malignant melanoma (CMM). The leucine zipper, down regulated in cancer 1 (LDOC-1) gene, is known to be a regulator of the nuclear factor kappa B (NF-kB) through inhibition of the same NF-kB. The poly (ADP-ribose) polymerase-1 (PARP1) gene plays an important role for the efficient maintenance of genome integrity. PARP-1 protein is required for the apoptosis-inducing factor (AIF) translocation from the mitochondria to the nucleus. We report here two interesting cases of family melanoma, a father and son 84 and 40 years old, respectively. The histological evaluation of the lesions of both men revealed diffused superficial melanoma with epithelioid cells. We evaluated the differential expression of LDOC-1 and PARP-1 mRNA in peripheral blood leukocytes of both the father and son. We found that both LDOC-1 and PARP-1 genes were down-regulated in both patients compared with those of controls. These data suggest that low levels of expression of LDOC-1 and PARP-1 mRNA may be associated with familial melanoma

The Rapid Identification of Anoplophora chinensis (Coleoptera: Cerambycidae) From Adult, Larval, and Frass Samples Using TaqMan Probe Assay

A molecular diagnostic method using TaqMan probe qPCR is presented for the identification of Anoplophora chinensis (Förster) (Coleoptera: Cerambycidae) from whole body insects (adults and larvae) and frass samples stored under different conditions. The results showed a perfect amplification of DNA from all samples; the repeatability and reproducibility of the protocol were very good, with standard deviations of inter-run and intrarun variability less than or equal to 0.5. The assay allowed to discern all A. chinensis samples from those of the other non-target wood-borer species, with 100% correspondence to the homologous sequences. No amplification or cross reactions were observed with A. glabripennis (Motschulsky) (Coleoptera: Cerambycidae), which is the most related species among those tested. The protocol was validated by an internal blind panel test which showed a good correspondence between the results obtained by different operators in the same lab. The analytical sensitivity for the lab frass with the Probe qPCR, namely the lowest amount of A. chinensis DNA that can be detected (LoD), was 0.64 pg/μl with a Cq of 34.87. The use of indirect evidence for the identification of a pest is an important feature of the method, which could be crucial to detect the presence of wood-boring insects. This diagnostic tool can help prevent the introduction of A. chinensis into new environments or delimit existing outbreak areas thanks to indirect frass diagnosis

First record of Aleurocanthus camelliae Kanmiya & Kasai, 2011 (Hemiptera, Aleyrodidae) from Italy, on ornamental Camellia spp. plants

This paper provides a first report of Aleurocanthus camelliae, the Camellia spiny whitefly, from Italy. The pest was found on plants of Camellia spp. grown in the nursery. Brief morphological and biological information is provided on this whitefly, as well as some considerations on the phytosanitary measures to be adopted to reduce the potential risk of its spread on ornamental plants in Europe and the EPPO region

Rapid Detection of Pityophthorus juglandis (Blackman) (Coleoptera, Curculionidae) with the Loop-Mediated Isothermal Amplification (LAMP) Method

The walnut twig beetle Pityophthorus juglandis is a phloem-boring bark beetle responsible, in association with the ascomycete Geosmithia morbida, for the Thousand Cankers Disease (TCD) of walnut trees. The recent finding of TCD in Europe prompted the development of effective diagnostic protocols for the early detection of members of this insect/fungus complex. Here we report the development of a highly efficient, low-cost, and rapid method for detecting the beetle, or even just its biological traces, from environmental samples: the loop-mediated isothermal amplification (LAMP) assay. The method, designed on the 28S ribosomal RNA gene, showed high specificity and sensitivity, with no cross reactivity to other bark beetles and wood-boring insects. The test was successful even with very small amounts of the target insect’s nucleic acid, with limit values of 0.64 pg/µL and 3.2 pg/µL for WTB adults and frass, respectively. A comparison of the method (both in real time and visual) with conventional PCR did not display significant differences in terms of LoD. This LAMP protocol will enable quick, low-cost, and early detection of P. juglandis in areas with new infestations and for phytosanitary inspections at vulnerable sites (e.g., seaports, airports, loading stations, storage facilities, and wood processing companies)

MMP-9 as a Candidate Marker of Response to BRAF Inhibitors in Melanoma Patients With BRAFV600E Mutation Detected in Circulating-Free DNA

The BRAFV600E mutation is associated with melanoma development and its detection in circulating-free DNA cannot be observed in all melanoma patients harboring this mutation in tumor specimens. Beside the circulating-free DNA BRAFV600E mutation, other markers of therapeutic response should be identified. Matrix metalloproteinase-9 (MMP-9) could be one of them as its role as indicator of invasiveness in melanoma have been explored. In this study, MMP-9 was evaluated in melanoma cells after treatment with dabrafenib. In vitro data were validated in 26 melanoma patients, of which 14 treated with BRAF inhibitor alone and 12 treated with both BRAF and MEK inhibitors, by ELISA assay and droplet digital PCR for measuring MMP-9 serum levels and circulating-free DNA BRAFV600E mutation, respectively. Statistical analyses were performed to evaluate the prognostic significance of MMP-9, progression-free survival (PFS) and overall survival (OS) according to the BRAFV600E mutation and MMP-9 levels. The performed analyses showed that MMP-9 and pEKR1-2 were statistically down-regulated in melanoma cells after treatment with dabrafenib. Circulating-free DNA BRAFV600E mutation was detected in 11 out of 26 melanoma patients showing higher levels of MMP-9 compared to those with undetectable BRAFV600E mutation. Furthermore, higher levels of MMP-9 and circulating-free DNA BRAFV600E mutation were associated with lower PFS and OS. Finally, the monitoring of therapy showed that MMP-9 significantly decreased at T1 and T2, but not at T-last, for the patients with detectable circulating-free DNA BRAFV600E mutation. In conclusion, high levels of MMP-9 and circulating-free DNA BRAFV600E mutation are associated with poor PFS and OS. MMP-9 may represent a promising indicator of response to BRAF inhibitors in combination with the detection of BRAFV600E mutation

An Italian Multicenter Perspective Harmonization Trial for the Assessment of MET Exon 14 Skipping Mutations in Standard Reference Samples

Lung cancer remains the leading cause of cancer deaths worldwide. International societies have promoted the molecular analysis of MET proto-oncogene, receptor tyrosine kinase (MET) exon 14 skipping for the clinical stratification of non-small cell lung cancer (NSCLC) patients. Different technical approaches are available to detect MET exon 14 skipping in routine practice. Here, the technical performance and reproducibility of testing strategies for MET exon 14 skipping carried out in various centers were evaluated. In this retrospective study, each institution received a set (n = 10) of a customized artificial formalin-fixed paraffin-embedded (FFPE) cell line (Custom METex14 skipping FFPE block) that harbored the MET exon 14 skipping mutation (Seracare Life Sciences, Milford, MA, USA), which was previously validated by the Predictive Molecular Pathology Laboratory at the University of Naples Federico II. Each participating institution managed the reference slides according to their internal routine workflow. MET exon 14 skipping was successfully detected by all participating institutions. Molecular analysis highlighted a median Cq cut off of 29.3 (ranging from 27.1 to 30.7) and 2514 (ranging from 160 to 7526) read counts for real-time polymerase chain reaction (RT-PCR) and NGS-based analyses, respectively. Artificial reference slides were a valid tool to harmonize technical workflows in the evaluation of MET exon 14 skipping molecular alterations in routine practice

Neuronavigated-biopsy in the diagniosis of demyelinating pseudotumoral brain lesions. Case report.

Introduction: Pseudotumoral-demyelinating lesions are typically characterized by severe difficulties in differential diagnosis from gliomas, metastatic tumors and abscesses. Marburg’s disease (MD) is an extremely rare and aggressive form of Mutiple Sclerosis (MS) with acute onset, rapid neurological deterioration, and poor prognosis. MRI images often show pseudotumoral lesions characterized by mild and focal or, sometimes, ring enhancement. Perhaps, radiological aspect can vary, making necessary hystopathological confirmation. Objectives: There is increasing interest in the role of surgical biopsy for a correct and early diagnosis of pseudotumoral demyelinating lesions. We report an histopathologically confirmed case of MD. Thanks to the analysis of actual literature, our purpose is to ameliorate diagnostic criteria, the timing of treatment and, subsequently, the prognosis of patients affected. Matherials and methods: A 24 years-old woman presented with a sudden onset of motor subaphasia and numbness to the right hemiface, followed, in a few hours by rapid deterioration with mild pyramidal right hemiparesis and complete aphasia. Patient underwent to a CT brain examination which showed three unspecific hypodense lesions: the bigger one, in the left subcortical frontal white matter, and two smaller lesions in the deep parietal and in the right paratrigonal white matter. For better characterization, a brain MRI was performed. The lesions in the right paratrigonal and left parietal white matter where appreciable as highly hyper-intense on TSE-T2 and FLAIR-T2 sequences, hypo-intense on TSE-T1, and responsible for a moderate mass effect on the surrounding structures; the left parietal white matter lesion also showed an incomplete and irregular ring-enhancement. The left subcortical frontal lesion was only mild hyper-intense on TSE-T2 and FLAIR-T2 but strongly hyper-intense on DWI, and a subsequent MRI scan showed a pseudotumoral development with an atypical patchy enhancement. Total body CT scan, auto-antibodies and viral serology on blood and CSF were negative. Isoelectric focusing in agarose gel detected the presence of CSF oligoclonal bands. Steroids were administrated without significant improvement. On the basis of the obtained data, and in consideration of the poor responsiveness to steroids, we performed a neuronavigator-aided biopsy of the main lesion through an open microsurgical approach. Preoperative planning was performed the day before by using contrast agent-enhanced brain CT scan, after positioning markers on the scalp. The exam was then transferred on a data storage device to insert CT images on the neuronavigator. The principal landmarks were used to realize a temporal craniotomy and a small corticectomy centred on the lesion. Finally, we detected, at a depth of about 2 cm, a pathological curdy and whitish tissue, ablated in small fragments and sent for hystological examination. A post-operative CT scan excluded any complications and the patient had a regular course. At demission, neurological exam was slightly better with partial regression of hemiparesis and partial amelioration of speech function. Hystopathological report confirmed the hypothesis of Marburg’s variant of MS. Results and Conclusions: Initial therapeutic approach to MS is pharmacological. However, in some atypical cases, CSF analisys, MRI images and/or clinical course after sterioids administration are not sufficiently diagnostic to completely rule out tumors or abscesses. So, early biopsy is an unavoidable necessity for the differential diagnosis of pseudotumoral-demyelinating lesions, to prevent inappropriate surgical or irradiative treatments and ameliorate the prognosis

Transcranial magnetic resonance-guided focused ultrasound thalamotomy as a safe treatment option in multiple sclerosis patients with essential tremor

Transcranial magnetic resonance-guided focused ultrasound is a recently introduced incisionless treating option for essential tremor and tremor-dominant idiopathic Parkinson disease. There is preliminary evidence that it may result in a promising effective treatment option for other movement disorders too. Here, we report on two patients with multiple sclerosis with medication refractory debilitating essential tremor comorbidity who successfully underwent unilateral Vim tcMRgFUS thalamotomy for tremor control. Patients' clinical condition and expanded disability status scale scores showed no changes during the 1-year follow-up period with no evidence of multiple sclerosis activity or progression