Circulating cell-free DNA: a powerful biomarker for tumor management and a possible monitor tool in other pathological conditions.

\u2018Liquid biopsy\u2019, i.e. the analysis of cfDNA in blood or body fluids, can give a live, \u2018total\u2019 image representing the entire heterogeneity of the system. The application of high throughput analytical procedures, such NGS or ddPCR, are necessary to obtain reliable data on such a small amount of starting material. Only few applications have achieved a clinical validation, due to the great variability in preanalytical procedures. In our work we evaluated cfDNA with three different aims. \u2022 Presence of mutation in a panel of 16 genes of the HR pathway in genomic and cfDNA in patients affected by breast cancer. We observed mutations in 3 out of 6 samples; in one case variant fraction in cfDNA was higer than in genomic DNA, probably due to limited ability to detect clonal heterogeneity in tissue. \u2022 Monitoring tool for determining septic risk in patients undergoing dialysis. We detected in a sample, in accordance with the emoculture, a Staphylococcus strain together with Propionibacterium and Streptococcus strains. The detection of Burkholderia multivorans in another sample raised the possibility to identify those bacteria that take more than the canonical 5 days of emoculture to growth, or that completely do not grow in emoculture conditions. \u2022 Detection of donor-derived cfDNA in transplanted patients. We identified more than 50% of donor derived polymorphisms just after reperfusion, falling down to 10% one day after surgery and then disappearing. The possibility to cross our data with clinical parameters will help us to better describe the pertinence of our results to the effective status of patients. In all the three settings, we are collecting other samples to have a broader amount of data that will allow us to perform statistical analysis to effectively validate our procedures

Is First Person Vision Challenging for Object Tracking?

Understanding human-object interactions is fundamental in First Person Vision (FPV). Tracking algorithms which follow the objects manipulated by the camera wearer can provide useful cues to effectively model such interactions. Visual tracking solutions available in the computer vision literature have significantly improved their performance in the last years for a large variety of target objects and tracking scenarios. However, despite a few previous attempts to exploit trackers in FPV applications, a methodical analysis of the performance of state-of-the-art trackers in this domain is still missing. In this paper, we fill the gap by presenting the first systematic study of object tracking in FPV. Our study extensively analyses the performance of recent visual trackers and baseline FPV trackers with respect to different aspects and considering a new performance measure. This is achieved through TREK-150, a novel benchmark dataset composed of 150 densely annotated video sequences. Our results show that object tracking in FPV is challenging, which suggests that more research efforts should be devoted to this problem so that tracking could benefit FPV tasks.Comment: IEEE/CVF International Conference on Computer Vision (ICCV) 2021, Visual Object Tracking Challenge VOT2021 workshop. arXiv admin note: text overlap with arXiv:2011.1226

Implant sonication versus intraoperative tissue sample cultures for periprosthetic joint infection (PJI) of shoulder arthroplasty

Introduction: Periprosthetic joint infection (PJI) is the most problematic complications after shoulder arthroplasty. Many diagnostic tools have been identified to find infection, such as hystopatologic examination of tissue sections or cultures of intraoperative tissue. Implant sonication fluid culture showed good results in order to enhance diagnostic accuracy, but literature results are still controversial. Aim of our study is to compare the results of sonication with intraoperative tissue sample cultures. Patients and Methods: From February 2016 to January 2018 we performed 102 revisions of Total Shoulder Arthroplasty (TSA) for suspected PJI. Sixty-five patients respected the criteria for admission to the study and were enrolled. In each case periprostethic specimens were collected and explanted prosthesis were put inside sterile fluid, sonicated and then placed under culture. Results: Among the sixty-five patients, 36 were considered as possible, probable or certain infection. Tissue cultures were positive for infection in thirty-four cases (52,3%) and in nineteen cases was found the positivity for Cutibacterium acnes. Sonication fluid cultures were positive in forty cases (61,5%), with a positivity for Cutibacterium acnes in twenty-seven cases. The sensitivities of sonication and tissue cultures for the diagnosis of shoulder PJI were 83.3% and 88,9% (P = 0,08); the specificities were 65.5% and 93,1% (P < 0.01) respectively. Conclusion: Our results suggest that sonication technique had not shown a clear advantage in postoperative shoulder PJI diagnosis, but it’s a real aid to detect Cutibacterium acnes. In any case, sensitivity and mostly specificity were higher with tissue cultures. (www.actabiomedica.it)

OTX Genes in Adult Tissues

OTX homeobox genes have been extensively studied for their role in development, especially in neuroectoderm formation. recently, their expression has also been reported in adult physiological and pathological tissues, including retina, mammary and pituitary glands, sinonasal mucosa, in several types of cancer, and in response to inflammatory, ischemic, and hypoxic stimuli. reactivation of OTX genes in adult tissues supports the notion of the evolutionary amplification of functions of genes by varying their temporal expression, with the selection of homeobox genes from the "toolbox" to drive or contribute to different processes at different stages of life. OTX involvement in pathologies points toward these genes as potential diagnostic and/or prognostic markers as well as possible therapeutic targets

A complication following ACL reconstruction using bioabsorbable cross-pins

This is a case of a proximal pin migration after ACL reconstruction in medial soft tissue with pain, inflammatory reaction and functional reduction. 33-year-old male presented at our clinic with a complete ACL rupture. Reconstruction with autogenous gracilis and semitendinosus hamstring tendons was performed and graft fixed in the femoral canal with two PLLA bioabsorbable pins (RIGIDFIX\uae Cross Pin System). Two months postoperatively the patient presented swelling and pain on the medial side of the knee, full range of motion and negative results at the Lachman and Pivot shift tests. MRI examination showed the superior femoral tunnel crossing both the lateral and medial cortex lodging the pin in the knee\u2019s medial soft tissue corresponding to the swelling area reported by the patient. The tendon graft was properly positioned. After surgical removal of the pin through a small skin incision, the pain and swelling promptly subsided allowing the patient return to normal activities in few weeks without any pain. In our opinion the painful swelling of the knee was due to a displacement of the pin that had been accidentally lodged in the soft tissues instead of the bone causing a foreign-body reaction resulting in granuloma formation with local inflammation. This dislodgement could have been due to an inappropriately long femoral tunnel

Donor Cell Acute Myeloid Leukemia after Hematopoietic Stem Cell Transplantation for Chronic Granulomatous Disease: A Case Report and Literature Review

The patient reported here underwent hematopoietic stem cell transplantation (HSCT) due to chronic granulomatous disease (CGD) caused by biallelic mutations of the NCF1 gene. Two years later, he developed AML, which was unexpected and was recognized via sex-mismatched chromosomes as deriving from the donor cells; the patient was male, and the donor was his sister. Donor cell leukemia (DCL) is very rare, and it had never been reported in patients with CGD after HSCT. In the subsequent ten years, the AML relapsed three times and the patient underwent chemotherapy and three further HSCTs; donors were the same sister from the first HSCT, an unrelated donor, and his mother. The patient died during the third relapse. The DCL was characterized since onset by an acquired translocation between chromosomes 9 and 11, with a molecular rearrangement between the MLL and MLLT3 genes-a quite frequent cause of AML. In all of the relapses, the malignant clone had XX sex chromosomes and this rearrangement, thus indicating that it was always the original clone derived from the transplanted sister's cells. It exhibited the ability to remain quiescent in the BM during repeated chemotherapy courses, remission periods and HSCT. The leukemic clone then acquired different additional anomalies during the ten years of follow-up, with cytogenetic results characterized both by anomalies frequent in AML and by different, non-recurrent changes. This type of cytogenetic course is uncommon in AML

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

Evidence of Infection by H5N2 Highly Pathogenic Avian Influenza Viruses in Healthy Wild Waterfowl

The potential existence of a wild bird reservoir for highly pathogenic avian influenza (HPAI) has been recently questioned by the spread and the persisting circulation of H5N1 HPAI viruses, responsible for concurrent outbreaks in migratory and domestic birds over Asia, Europe, and Africa. During a large-scale surveillance programme over Eastern Europe, the Middle East, and Africa, we detected avian influenza viruses of H5N2 subtype with a highly pathogenic (HP) viral genotype in healthy birds of two wild waterfowl species sampled in Nigeria. We monitored the survival and regional movements of one of the infected birds through satellite telemetry, providing a rare evidence of a non-lethal natural infection by an HP viral genotype in wild birds. Phylogenetic analysis of the H5N2 viruses revealed close genetic relationships with H5 viruses of low pathogenicity circulating in Eurasian wild and domestic ducks. In addition, genetic analysis did not reveal known gallinaceous poultry adaptive mutations, suggesting that the emergence of HP strains could have taken place in either wild or domestic ducks or in non-gallinaceous species. The presence of coexisting but genetically distinguishable avian influenza viruses with an HP viral genotype in two cohabiting species of wild waterfowl, with evidence of non-lethal infection at least in one species and without evidence of prior extensive circulation of the virus in domestic poultry, suggest that some strains with a potential high pathogenicity for poultry could be maintained in a community of wild waterfowl

Circulating cell-free DNA: a powerful biomarker for tumor management and a possible monitor tool in other pathological conditions.

‘Liquid biopsy’, i.e. the analysis of cfDNA in blood or body fluids, can give a live, ‘total’ image representing the entire heterogeneity of the system. The application of high throughput analytical procedures, such NGS or ddPCR, are necessary to obtain reliable data on such a small amount of starting material. Only few applications have achieved a clinical validation, due to the great variability in preanalytical procedures. In our work we evaluated cfDNA with three different aims. • Presence of mutation in a panel of 16 genes of the HR pathway in genomic and cfDNA in patients affected by breast cancer. We observed mutations in 3 out of 6 samples; in one case variant fraction in cfDNA was higer than in genomic DNA, probably due to limited ability to detect clonal heterogeneity in tissue. • Monitoring tool for determining septic risk in patients undergoing dialysis. We detected in a sample, in accordance with the emoculture, a Staphylococcus strain together with Propionibacterium and Streptococcus strains. The detection of Burkholderia multivorans in another sample raised the possibility to identify those bacteria that take more than the canonical 5 days of emoculture to growth, or that completely do not grow in emoculture conditions. • Detection of donor-derived cfDNA in transplanted patients. We identified more than 50% of donor derived polymorphisms just after reperfusion, falling down to 10% one day after surgery and then disappearing. The possibility to cross our data with clinical parameters will help us to better describe the pertinence of our results to the effective status of patients. In all the three settings, we are collecting other samples to have a broader amount of data that will allow us to perform statistical analysis to effectively validate our procedures