Miscarriage is one of the main complications occurring in pregnancy. The association between adverse pregnancy outcomes and silent bacterial infections has been poorly investigated. Ureaplasma parvum ...and urealiticum, Mycoplasma genitalium and hominis and Chlamydia trachomatis DNA sequences have been investigated by polymerase chain reaction (PCR) methods in chorionic villi tissues and peripheral blood mononuclear cells (PBMCs) from females with spontaneous abortion (SA, n = 100) and females who underwent voluntary interruption of pregnancy (VI, n = 100). U. parvum DNA was detected in 14% and 15% of SA and VI, respectively, with a mean of bacterial DNA load of 1.3 × 10−1 copy/cell in SA and 2.8 × 10
−3 copy/cell in VI; U. urealiticum DNA was detected in 3% and 2% of SA and VI specimens, respectively, with a mean DNA load of 3.3 × 10−3 copy/cell in SA and 1.6 × 10−3 copy/cell in VI; M. hominis DNA was detected in 5% of SA specimens with a DNA load of 1.3 × 10−4 copy/cell and in 6% of VI specimens with a DNA load of 1.4 × 10−4 copy/cell; C. trachomatis DNA was detected in 3% of SA specimens with a DNA load of 1.5 × 10−4 copy/cell and in 4% of VI specimens with a mean DNA load of 1.4 × 10−4 copy/cell. In PBMCs from the SA and VI groups, Ureaplasma spp, Mycoplasma spp and C. trachomatis DNAs were detected with a prevalence of 1%–3%. Bacteria were investigated, for the first time, by quantitative real‐time PCR (qPCR) in chorionic villi tissues and PBMCs from women affected by SA and VI. These data may help to understand the role and our knowledge of the silent infections in SA.
Miscarriage is one of the main complications occurring in pregnancy. Silent bacterial infections in miscarriage have been poorly investigated. Our data may help to understand the role of silent infections in miscarriage.
Treatment of high-grade osteosarcoma, the most common malignant tumor of bone, is largely based on administration of cisplatin and other DNA damaging drugs. Altered DNA repair mechanisms may thus ...significantly impact on either response or resistance to chemotherapy. In this study, by using a panel of human osteosarcoma cell lines, either sensitive or resistant to cisplatin, we assessed the value as candidate therapeutic targets of DNA repair-related factors belonging to the nucleotide excision repair (NER) or base excision repair (BER) pathways, as well as of a group of 18 kinases, which expression was higher in cisplatin-resistant variants compared to their parental cell lines and may be indirectly involved in DNA repair. The causal involvement of these factors in cisplatin resistance of human osteosarcoma cells was validated through gene silencing approaches and
reversal of CDDP resistance. This approach highlighted a subgroup of genes, which value as promising candidate therapeutic targets was further confirmed by protein expression analyses. The
activity of 15 inhibitor drugs against either these genes or their pathways was then analyzed, in order to identify the most active ones in terms of inherent activity and ability to overcome cisplatin resistance. NSC130813 (NERI02; F06) and triptolide, both targeting NER factors, proved to be the two most active agents, without evidence of cross-resistance with cisplatin. Combined
treatments showed that NSC130813 and triptolide, when administered together with cisplatin, were able to improve its efficacy in both drug-sensitive and resistant osteosarcoma cells. This evidence may indicate an interesting therapeutic future option for treatment of osteosarcoma patients who present reduced responsiveness to cisplatin, even if possible effects of additive collateral toxicities must be carefully considered. Moreover, our study also showed that targeting protein kinases belonging to the mitogen-activated protein kinase (MAPK) or fibroblast growth factor receptor (FGFR) pathways might indicate new promising therapeutic perspectives in osteosarcoma, demanding for additional investigation.
Viral infections are considered to be risk factors for spontaneous abortion (SA). Conflicting results have been reported on the association between Human Papillomavirus (HPV) and SA. HPV DNA was ...investigated in matched chorionic villi tissues and peripheral blood mononuclear cells (PBMCs) from women who experienced SA (
= 80, cases) and women who underwent a voluntary interruption of pregnancy (VI;
= 80, controls) by qualitative PCR and quantitative droplet digital PCR (ddPCR). Viral genotyping was performed using real-time PCR in HPV-positive samples. Specific IgG antibodies against HPV16 were investigated in sera from SA (
= 80) and VI (
= 80) females using indirect ELISA assays. None of the DNA samples from SA subjects was HPV-positive (0/80), whilst HPV DNA was detected in 2.5% of VI women (
> 0.05), with a mean viral DNA load of 7.12 copy/cell. VI samples (
= 2) were found to be positive for the HPV45 genotype. The ddPCR assay revealed a higher number of HPV-positive samples. HPV DNA was detected in 3.7% and 5% of SA and VI chorionic tissues, respectively, with mean viral DNA loads of 0.13 copy/cell in SA and 1.79 copy/cell in VI (
>0.05) samples. All DNA samples from the PBMCs of SA and VI females tested HPV-negative by both PCR and ddPCR. The overall prevalence of serum anti-HPV16 IgG antibodies was 37.5% in SA and 30% in VI (
> 0.05) women. For the first time, HPV DNA was detected and quantitatively analyzed using ddPCR in chorionic villi tissues and PBMCs from SA and VI women. Circulating IgG antibodies against HPV16 were detected in sera from SA and VI females. Our results suggest that HPV infection in chorionic villi may be a rare event. Accordingly, it is likely that HPV has no significant role in SA.
Introduction: Current treatment of conventional and non-conventional high-grade
osteosarcoma (HGOS) is based on the surgical removal of primary tumor and, when possible, of
metastases and local ...reccurrence, together with systemic pre- and post-operative chemotherapy
with drugs that have been used since decades.
Areas covered: This review is intended to summarize the new agents and
therapeutic strategies that are under clinical evaluation in HGOS, with the aim to increase the
cure probability of this highly malignant bone tumor, which has not significantly improved
during the last 30-40 years. The list of drugs, compounds and treatment modalities
presented and discussed here has been generated by considering only those that are included in
presently ongoing and recruiting clinical trials, or which have been completed in the last 2
years with reported results, on the basis of the information obtained from different and
continuously updated databases.
Expert opinion: Despite HGOS is a rare tumor, several clinical trials are
presently evaluating different treatment strategies, which may hopefully positively impact on
the outcome of patients who experience unfavorable prognosis when treated with conventional
therapies.
Trial registration:
ClinicalTrials.gov identifier: NCT01459484.
Trial registration:
ClinicalTrials.gov identifier: NCT01669369.
Trial registration:
ClinicalTrials.gov identifier: NCT00134030.
Trial registration:
ClinicalTrials.gov identifier: NCT00180908.
Trial registration:
ClinicalTrials.gov identifier: NCT00470223.
Trial registration:
ClinicalTrials.gov identifier: NCT01532687.
Trial registration:
ClinicalTrials.gov identifier: NCT02357810.
Trial registration:
ClinicalTrials.gov identifier: NCT03163381.
Trial registration:
ClinicalTrials.gov identifier: NCT02432274.
Trial registration:
ClinicalTrials.gov identifier: NCT02048371.
Trial registration:
ClinicalTrials.gov identifier: NCT02389244.
Trial registration:
ClinicalTrials.gov identifier: NCT02243605.
Trial registration:
ClinicalTrials.gov identifier: NCT02867592.
Trial registration:
ClinicalTrials.gov identifier: NCT03210714.
Trial registration:
ClinicalTrials.gov identifier: NCT03526250.
Trial registration:
ClinicalTrials.gov identifier: NCT03213678.
Trial registration:
ClinicalTrials.gov identifier: NCT03718091.
Trial registration:
ClinicalTrials.gov identifier: NCT03233204.
Trial registration:
ClinicalTrials.gov identifier: NCT03698994.
Trial registration:
ClinicalTrials.gov identifier: NCT03220035.
Trial registration:
ClinicalTrials.gov identifier: NCT03220035.
Trial registration:
ClinicalTrials.gov identifier: NCT02689336.
Trial registration:
ClinicalTrials.gov identifier: NCT03678883.
Trial registration:
ClinicalTrials.gov identifier: NCT01962103.
Trial registration:
ClinicalTrials.gov identifier: NCT02945800.
Trial registration:
ClinicalTrials.gov identifier: NCT01669369.
Trial registration:
ClinicalTrials.gov identifier: NCT03598595.
Trial registration:
ClinicalTrials.gov identifier: NCT02644460.
Trial registration:
ClinicalTrials.gov identifier: NCT02517918.
Trial registration:
ClinicalTrials.gov identifier: NCT03002805.
Trial registration:
ClinicalTrials.gov identifier: NCT02013336.
Trial registration:
ClinicalTrials.gov identifier: NCT02536183.
Trial registration:
ClinicalTrials.gov identifier: NCT02557854.
Trial registration:
ClinicalTrials.gov identifier: NCT02390843.
Trial registration:
ClinicalTrials.gov identifier: NCT03643133.
Trial registration:
ClinicalTrials.gov identifier: NCT03006848.
Trial registration:
ClinicalTrials.gov identifier: NCT03676985.
Trial registration:
ClinicalTrials.gov identifier: NCT03277924.
Trial registration:
ClinicalTrials.gov identifier: NCT03190174.
Trial registration:
ClinicalTrials.gov identifier: NCT02304458.
Trial registration:
ClinicalTrials.gov identifier: NCT02406781.
Trial registration:
ClinicalTrials.gov identifier: NCT02502786.
Trial registration:
ClinicalTrials.gov identifier: NCT03860207.
Trial registration:
ClinicalTrials.gov identifier: NCT03320330.
Trial registration:
ClinicalTrials.gov identifier: NCT03610490.
Trial registration:
ClinicalTrials.gov identifier: NCT02100891.
Trial registration:
ClinicalTrials.gov identifier: NCT02508038.
Trial registration:
ClinicalTrials.gov identifier: NCT03356782.
Trial registration:
ClinicalTrials.gov identifier: NCT01953900.
Trial registration:
ClinicalTrials.gov identifier: NCT03618381.
Trial registration:
ClinicalTrials.gov identifier: NCT03462316.
Trial registration:
ClinicalTrials.gov identifier: NCT02487979.
Droplet‐digital polymerase chain reaction (ddPCR) technique was set up to detect/quantify Merkel cell polyomavirus (MCPyV) DNA in clinical specimens, including chorionic villi and peripheral blood ...mononuclear cells (PBMCs) from spontaneous abortion (SA)‐affected females. This ddPCR assay showed high accuracy, sensitivity, and specificity in detecting MCPyV DNA cloned in a recombinant plasmid vector, the control. ddPCR was extended to MCPyV DNA to investigate/quantify its sequences in clinical samples. Overall, 400 samples were analyzed, that is, 100 chorionic villi and 100 PBMCs, from SA females (n = 100), the cases, and 100 chorionic villi and 100 PBMCs from females who underwent voluntary pregnancy interruption (VI, n = 100), the control. MCPyV DNA was detected in 4/100 (4%) and 5/100 (5%) of SA and VI chorionic villi, respectively. The mean viral DNA load was 1.99 ( ± 0.94 standard mean deviation SD) copy/104 cells in SA and 3.02 ( ± 1.86 SD) copy/104 cells in VI. In PBMCs, MCPyV DNA was revealed in 9/100 (9%) and 14/100 (14%) of SA and VI, with a mean of 2.09 ( ± 1.17 SD) copy/104 cells and 4.09 ( ± 4.26 SD) copy/104 cells in SA and VI, respectively. MCPyV gene expression analysis by quantitative PCR for the large T antigen (LT) and viral capsid protein 1 (VP1) showed their mRNAs in 2/4 (50%) SA‐ and 2/5 (40%) VI‐MCPyV‐positive samples. MCPyV DNA was detected/quantified using the ddPCR technique, in chorionic villi and PBMCs from SA and VI. In our experimental conditions, ddPCR provided a powerful tool to detect/quantify MCPyV DNA sequences in clinical samples.
Droplet‐digital polymerase chain reaction (ddPCR) technique was set up to detect/quantify Merkel cell polyomavirus (MCPyV) DNA in clinical specimens, including chorionic villi and peripheral blood mononuclear cells (PBMCs) from spontaneous abortion (SA)‐affected females. This ddPCR assay showed high accuracy, sensitivity and specificity in detecting MCPyV DNA in clinical samples. MCPyV gene expression analysis by quantitative PCR for the large T antigen (LT) and viral capsid protein 1 (VP1) showed their mRNAs in 2/4 (50%) SA‐ and 2/5 (40%)
Introduction: DNA damaging drugs are widely used for the chemotherapeutic treatment of high-grade osteosarcoma (HGOS). In HGOS patients, several germline polymorphisms have been reported to impact on ...the development of adverse toxic events related to DNA damaging drugs treatment. Some of these polymorphisms, when present in tumor cells, may also influence treatment response and prognosis of HGOS patients.
Area covered: In this review, the authors have focused on pharmacogenetic markers (mainly germline polymorphisms) described in patients with HGOS, which have proved or indicated to be related to the susceptibility to adverse toxic reactions and/or to influence response to DNA damaging drugs. The concordant and discordant results reported in different studies have also been discussed.
Expert opinion: Response and toxicity predisposition to DNA damaging drugs are influenced by genes encoding proteins involved in their uptake, efflux, activation, inactivation, and in DNA repair, activity of which may vary according to specific gene variations. In HGOS, there is a substantial medical need for biomarkers predictive for individual response and toxicity predisposition to DNA-targeting drugs, which may be used to tailor therapy in order to decrease the occurrence of adverse side effects and increase treatment efficacy and safety.
Introduction: Genetic characterization of osteosarcoma has evolved during the last decade, thanks to the integrated application of conventional and new candidate-driven and genome-wide technologies.
...Areas covered: This review provides an overview of the state of art in genetic testing applied to osteosarcoma, with particular regard to novel candidate genetic biomarkers that can be analyzed in tumor tissue and blood samples, which might be used to predict toxicity and prognosis, detect disease relapse, and improve patients' selection criteria for tailoring treatment.
Expert commentary: Genetic testing based on modern technologies is expected to indicate new osteosarcoma-related prognostic markers and driver genes, which may highlight novel therapeutic targets and patients stratification biomarkers. The definition of tailored or targeted treatment approaches may improve outcome of patients with localized tumors and, even more, of those with metastatic disease, for whom progress in cure probability is highly warranted.
The loss of SMARCB1/INI1 protein has been recently described in poorly differentiated chordoma, an aggressive and rare disease variant typically arising from the skull base.
Retrospective study aimed ...at 1) examining the differential immunohistochemical expression of SMARCB1/INI1 in conventional skull base chordomas, including the chondroid subtype; 2) evaluating SMARCB1 gene deletions/copy number gain; and 3) analyzing the association of SMARCB1/INI1 expression with clinicopathological parameters and patient survival.
65 patients (35 men and 30 women) affected by conventional skull base chordoma, 15 with chondroid subtype, followed for >48 months after surgery were collected. Median age at surgery was 50 years old (range 9-79). Mean tumor size was 3.6 cm (range 2-9.5). At immunohistochemical evaluation, a partial loss of SMARCB1/INI1 (>10% of neoplastic examined cells) was observed in 21 (32.3%) cases; the remaining 43 showed a strong nuclear expression. Fluorescence
hybridization (FISH) analysis was performed in 15/21 (71.4%) cases of the chordomas with partial SMARCB1/INI1 loss of expression. Heterozygous deletion of SMARCB1 was identified in 9/15 (60%) cases and was associated to copy number gain in one case; no deletion was found in the other 6 (40%) cases, 3 of which presenting with a copy number gain. No correlations were found between partial loss of SMARCB1/INI1 and the clinicopathological parameters evaluated (i.e., age, tumor size, gender, tumor size and histotype). Overall 5-year survival and 5-year disease-free rates were 82% and 59%, respectively. According to log-rank test analysis the various clinico-pathological parameters and SMARCB1/INI1 expression did not impact on overall and disease free-survival.
Partial loss of SMARCB1/INI1, secondary to heterozygous deletion and/or copy number gain of SMARCB1, is not peculiar of aggressive forms, but can be identified by immunohistochemistry in a significant portion of conventional skull base chordomas, including the chondroid subtype. The variable protein expression does not appear to correlate with clinicopathological parameters, nor survival outcomes, but still, it could have therapeutic implications.
A BCL6 corepressor (BCOR) gene alteration is a genetic signature of rare subsets of sarcomas. The identification of this alteration has recently contributed to the definition of new entities in the ...current WHO (2020) classification of soft tissue and bone tumours. We retrospectively examined cases of BCOR-rearranged sarcoma (BRS) to assess the reliability of the BCOR FISH analysis using an IVD (in vitro diagnostic) probe.
We investigated and compared the molecular diagnostic strategies and features by collecting 17 data from patients with a BCOR gene rearrangement detected using quantitative-Reverse Transcription-Polymerase Chain Reaction (qRTPCR), Next-Generation Sequencing (NGS) and Fluorescence in situ hybridization (FISH).
We describe fourteen BCOR::CCNB3 sarcomas, one spindle cell sarcoma with a novel BCOR::MAML1 fusion, one spindle cell sarcoma with a novel BCOR::AHR fusion, and one ossifying fibromyxoid tumour with a BCOR::ZC3H7B fusion. FISH analysis of all, except one, BCOR::CCNB3 sarcoma, showed a FISH break-apart pattern with mild signal separation. The MAML1::BCOR sarcoma showed large-space split signals, while in the two patients with AHR::BCOR and ZC3H7B::BCOR fusions, no BCOR rearrangement was observed using FISH.
Our study indicates that BCOR FISH analysis using an IVD probe, may be useful to detect the presence of a BCOR rearrangement, including both translocations and inversions; however, negative results, in some cases, can occur.
The standard treatment for young patients with untreated PTCLs is based on anthracycline containing-regimens followed by high-dose-chemotherapy and stem-cell-transplantation (HDT + SCT), but only 40% ...of them can be cured. Romidepsin, a histone-deacetylase inhibitor, showed promising activity in relapsed PTCLs; in first line, Romidepsin was added with CHOP. We designed a study combining romidepsin and CHOEP as induction before HDT + auto-SCT in untreated PTCLs (PTCL-NOS, AITL/THF, ALK-ALCL), aged 18-65 years. A phase Ib/II trial was conducted to define the maximum tolerated dose (MTD) of Ro-CHOEP, and to assess efficacy and safety of 6 Ro-CHOEP as induction before HDT. The study hypothesis was to achieve a 18-month PFS of 70%. Twenty-one patients were enrolled into phase Ib; 7 dose-limiting toxicities were observed, that led to define the MTD at 14 mg/ms. Eighty-six patients were included in the phase II. At a median follow-up of 28 months, the 18-month PFS was 46.2% (95%CI:35.0-56.7), and the 18-month overall survival was 73.1% (95%CI:61.6-81.7). The overall response after induction was 71%, with 62% CRs. No unexpected toxicities were reported. The primary endpoint was not met; therefore, the enrollment was stopped at a planned interim analysis. The addition of romidepsin to CHOEP did not improve the PFS of untreated PTCL patients.
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