GDNA qPCR is statistically more reliable than mRNA analysis in detecting leukemic cells to monitor CML

Chronic Myeloid Leukemia (CML) is a stem cell cancer that arises when t(9;22) translocation occurs in a hematopoietic stem cells. This event results in the expression of the BCR-ABL1 fusion gene, which codes for a constitutively active tyrosine kinase that is responsible for the transformation of a HSC into a CML stem cell, which then gives rise to a clonal myeloproliferative disease. The introduction of Tyrosine Kinase Inhibitors (TKIs) has revolutionized the management of the disease. However, these drugs do not seem to be able to eradicate the malignancy. Indeed, discontinuation trials (STIM; TWISER; DADI) for those patients who achieved a profound molecular response showed 50% relapsing within 12 months. We performed a comparative analysis on 15 CML patients and one B-ALL patient, between the standard quantitative reverse-transcriptase PCR (qRT-PCR) and our genomic DNA patient-specific quantitative PCR assay (gDNA qPCR). Here we demonstrate that gDNA qPCR is better than standard qRT-PCR in disease monitoring after an average follow-up period of 200 days. Specifically, we statistically demonstrated that DNA negativity is more reliable than RNA negativity in indicating when TKIs therapy can be safely stopped

Peroneal tendoscopy

Peroneal tendoscopy is an innovative technique that allows visualization of the tendons from the myotendinous junction to the peroneal tubercle, together with adjacent anatomic structures such as the recently unveiled vincula. Through a minimally invasive approach, it is possible to diagnose and treat several disorders, such as common tenosynovitis, accessory muscles, hypertrophic bony prominences, and thickened vincula, that can cause pain and tendon catching. Surgical morbidity and postoperative pain are significantly reduced when compared with open procedures. In this paper, the main indications for peroneal tendoscopy are discussed, the available literature is reviewed, and the surgical technique is described. Advantages of this procedure and current limitations are also presented. Anatomic and histological studies were also performed in order to verify: 1) the feasibility of peroneal tendoscopy for evaluation of peroneal tendons, using cadaver specimens; 2) the presence of nervous tissue in cadaver peroneal vincula as well as in tendoscopic vincula biopsies from patients undergoing surgery for chronic lateral ankle pain. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s12178-012-9123-1) contains supplementary material, which is available to authorized users

OTX1 and OTX2 as possible molecular markers of sinonasal carcinomas and olfactory neuroblastomas

OTX Homeobox genes are involved in embryonic morphogenesis and in the development of olfactory epithelium in adult. Mutations occurring in the OTX genes are reported to be associated to tumorigenisis in human. No reports correlate the expression of OTX genes and neoplasms of the nasal cavity. Thus, through immunohistochemical and Real-time PCR analysis we investigated OTX1 and OTX2 expression in the more frequent types of nasal and sinonasal tumours. Variable expression of both genes were found in normal sinonasal mucosa and in tumours. Interestingly, no expression of both OTX genes were detected in sinonasal intestinal-type adenocarcinomas; only OTX1 was found in non-intestinal-type adenocarcinomas and OTX2 was selectively expressed in olfactory neuroblastomas. In conclusion, OTX1 and OTX2 genes might have a role in the pathogenesis of different types of sinonasal neoplasms.</p

Nitric oxide regulates homeoprotein OTX1 and OTX2 expression in the rat myenteric plexus after intestinal ischemia/reperfusion injury

Neuronal and inducible NO synthase (nNOS and iNOS) play a protective and damaging role, respectively, on the intestinal neuromuscular function after ischemia and reperfusion (I/R) injury. To uncover the molecular pathways underlying this dichotomy we investigated their possible correlation with orthodenticle homeobox proteins OTX1 and OTX2 in the rat small intestine myenteric plexus after in vivo I/R. Homeobox genes are fundamental for the regulation of the gut wall homeostasis both during development and in pathological conditions (inflammation, cancer). I/R injury was induced by temporary clamping the superior mesenteric artery under anaesthesia, followed by 24 and 48 hours of reperfusion. At 48hr I/R intestinal transit decreased and was further reduced by NPLA, nNOS selective inhibitor. By contrast this parameter was restored to control values by 1400W, iNOS selective inhibitor. In longitudinal muscle myenteric plexus (LMMP) preparations, iNOS, OTX1 and OTX2 mRNA and protein levels increased at 24hr and 48hr I/R. At both time periods, the number of iNOS and OTX immunopositive myenteric neurons increased. nNOS mRNA, protein levels and neurons were unchanged. In LMMPs, OTX1 and OTX2 mRNA and protein up-regulation was reduced by 1400W and NPLA, respectively. In myenteric ganglia OTX1 and OTX2 staining was superimposed with that of iNOS and nNOS, respectively. Thus in myenteric ganglia iNOS and nNOS-derived NO may promote OTX1 and OTX2 up-regulation, respectively. We hypothesize that the neurodamaging and neuroprotective roles of iNOS and nNOS during I/R injury in the gut may involve corresponding activation of molecular pathways downstream of OTX1 and OTX2

gDNA qPCR is statistically more reliable than mRNA analysis in detecting leukemic cells to monitor CML

Chronic Myeloid Leukemia (CML) is a stem cell cancer that arises when t(9; 22) translocation occurs in a hematopoietic stem cells. This event results in the expression of the BCR-ABL1 fusion gene, which codes for a constitutively active tyrosine kinase that is responsible for the transformation of a HSC into a CML stem cell, which then gives rise to a clonal myeloproliferative disease. The introduction of Tyrosine Kinase Inhibitors (TKIs) has revolutionized the management of the disease. However, these drugs do not seem to be able to eradicate the malignancy. Indeed, discontinuation trials (STIM; TWISER; DADI) for those patients who achieved a profound molecular response showed 50% relapsing within 12 months. We performed a comparative analysis on 15 CML patients and one B-ALL patient, between the standard quantitative reverse-transcriptase PCR (qRT-PCR) and our genomic DNA patient-specific quantitative PCR assay (gDNA qPCR). Here we demonstrate that gDNA qPCR is better than standard qRT-PCR in disease monitoring after an average follow-up period of 200 days. Specifically, we statistically demonstrated that DNA negativity is more reliable than RNA negativity in indicating when TKIs therapy can be safely stopped

Anatomical MRI and SPECT of Patient’s GN Brain.

<p>(A) T1-weighted MRI transversal and coronal images showing atrophy in the left temporal lobe (white circles). (B) SPECT transversal and coronal images showing hypoperfusion in the left tempo-parietal region (white circles).</p

Areas Activated in Patient GN for HFF (unknown faces wrongly recognized as familiar > unknown faces correctly recognized as unfamiliar).

<p>Areas Activated in Patient GN for HFF (unknown faces wrongly recognized as familiar > unknown faces correctly recognized as unfamiliar).</p

Sulcal Depth of Patient’s GN Brain.

<p>(A). Three-dimensional reconstruction of the sulcal depth in GN’s brain, from surface and shallow depth values (green/yellow) to deep values (orange/red). Note the deepening and widening of the temporal sulci in the left compared to right hemisphere as outlined by expanded red areas in lateral sulcus and in the STS.</p

Recognition of Face Familiarity in Patient GN and in the Twelve Age-, Gender- and Education-matched Controls.

<p>Recognition of Face Familiarity in Patient GN and in the Twelve Age-, Gender- and Education-matched Controls.</p