Abstract
BACKGROUND:
Since precise diagnostic criteria for atypical and malignant meningiomas (AMMs) were provided for the first time in the 2000 World Health Organization (WHO) criteria, there is ...only sparse information about possible prognostic factors in the group of AMMs.
OBJECTIVE:
To evaluate the prognostic significance of various histological and clinical parameters in AMMs, with an emphasis on location, mitotic count, brain invasion, and Ki67 labeling index.
METHODS:
We analyzed 86 primary AMMs, 76 of which were atypical and 10 of which were malignant, diagnosed according to the 2000 WHO classification. Multivariate Cox survival analyses were performed.
RESULTS:
High mitotic count, brain invasion, and the parasagittal-falcine location of the tumor were significantly associated with decreased recurrence-free survival in multivariate analysis. Brain invasion was present in 25 of 37 cases in which brain tissue was identified in the tumor specimens. When brain invasion was not included in the analysis because of the limited number of cases in which it could be assessed, high mitotic count, Ki67 index >4%, the presence of macronucleoli, and parasagittal-falcine location were significant predictors of shorter recurrence-free survival.
CONCLUSION:
AMMs, as defined by 2000 WHO, are biologically heterogeneous. Recurrence-free survival can be further stratified by location and histological parameters, especially mitotic count, brain invasion, and Ki67 labeling index. Not only brain invasion, but also the presence or absence of brain tissue in surgical specimens should be reported, because the absence of brain invasion, when brain tissue is identified, provides very important positive prognostic information.
The number of prosthetic joint infection (PJI) cases is increasing along with total joint arthroplasties. There is currently no diagnostic test available with 100% sensitivity to identify PJI. The ...aim of the study was to assess and compare two different bacteriophage K‐based methods with standard microbiological culturing methods to detect staphylococci. Samples were retrieved from 104 patients undergoing revision surgery due to suspected PJI. Implants were subjected to sonication and sonicate fluid (SF) was assessed with the methods of qPCR detection of bacteriophage K DNA and adenosine triphosphate (ATP) detection after bacteriophage K lysis. The results were compared with the results of standard microbiological culturing methods. PJI was confirmed in 33 cases according to the PJI definition. Using the methods of ATP and bacteriophage K DNA detection 100% specificity and predictive value were achieved. The sensitivity of qPCR detection was higher (81.25%) than the sensitivity of ATP detection (62.50%) when analyzing SF directly. The sensitivity of the methods significantly improved (to 94.12%) with SF pre‐cultivation. Importantly, both methods provided results in 3–4 h when analyzing SF directly, while results from pre‐cultivated SF were obtained 19–20 h after sample collection. Our results suggest that bacteriophage‐based methods are specific and sensitive and importantly, faster than standard culturing methods. The addition of new bacteriophages to expand the bacterial detection spectrum could lead to the development of a faster, more sensitive, specific, and also economical, and handy method for PJI diagnosis.
Infection with high-risk human papillomavirus (HPV) strains is one of the risk factors for the development of oral squamous cell carcinoma (OSCC). Some patients with HPV-positive OSCC have a better ...prognosis and respond better to various treatment modalities, including radiotherapy or immunotherapy. However, since HPV can only infect human cells, there are only a few immunocompetent mouse models available that enable immunological studies. Therefore, the aim of our study was to develop a transplantable immunocompetent mouse model of HPV-positive OSCC and characterize it in vitro and in vivo.
Two monoclonal HPV-positive OSCC mouse cell lines were established by inducing the expression of HPV-16 oncogenes E6 and E7 in the MOC1 OSCC cell line using retroviral transduction. After confirming stable expression of HPV-16 E6 and E7 with quantitative real-time PCR and immunofluorescence staining, the cell lines were further characterized in vitro using proliferation assay, wound healing assay, clonogenic assay and RNA sequencing. In addition, tumor models were characterized in vivo in C57Bl/6NCrl mice in terms of their histological properties, tumor growth kinetics, and radiosensitivity. Furthermore, immunofluorescence staining of blood vessels, hypoxic areas, proliferating cells and immune cells was performed to characterize the tumor microenvironment of all three tumor models.
Characterization of the resulting MOC1-HPV cell lines and tumor models confirmed stable expression of HPV-16 oncogenes and differences in cell morphology, in vitro migration capacity, and tumor microenvironment characteristics. Although the cell lines did not differ in their intrinsic radiosensitivity, one of the HPV-positive tumor models, MOC1-HPV K1, showed a significantly longer growth delay after irradiation with a single dose of 15 Gy compared to parental MOC1 tumors. Consistent with this, MOC1-HPV K1 tumors had a lower percentage of hypoxic tumor area and a higher percentage of proliferating cells. Characteristics of the newly developed HPV-positive OSCC tumor models correlate with the transcriptomic profile of MOC1-HPV cell lines.
In conclusion, we developed and characterized a novel immunocompetent mouse model of HPV-positive OSCC that exhibits increased radiosensitivity and enables studies of immune-based treatment approaches in HPV-positive OSCC.
Prosthetic joint infections (PJIs) are commonly diagnosed via culture-based methods, which may miss hard-to-grow pathogens. This study contrasts amplicon metagenomic sequencing (16S AS) with ...traditional culture techniques for enhanced clinical decision-making. We analyzed sonicate fluid from 27 patients undergoing revision arthroplasty using both methods, emphasizing the distinction between contaminants and true positives. Our findings show moderate agreement between the two methods, with a Cohen's kappa of 0.490, varying across bacterial genera (Cohen's kappa -0.059 to 1). The sensitivity of 16S AS compared to culture was 81% (95% CI, 68% to 94%). Sequencing revealed greater microbial diversity, including anaerobic genera like Anaerococcus and Citrobacter. Interestingly, several culture-negative PJI samples showed diverse bacteria via 16S AS. Despite rigorous controls and algorithms to eliminate contaminants, confirming bacteria presence with 16S AS remains a challenge. This highlights the need for improved PJI diagnostic methods, while also pointing out the limitations of next-generation sequencing (NGS) as a clinical diagnostic tool.
Background and purpose - The correct diagnosis of prosthetic joint infection (PJI) can be difficult because bacteria form a biofilm on the surface of the implant. The sensitivity of culture from ...sonication fluid is better than that from periprosthetic tissue, but no comparison studies using molecular methods on a large scale have been performed. We assessed whether periprosthetic tissue or sonication fluid should be used for molecular analysis. Patients and methods - Implant and tissue samples were retrieved from 87 patients who underwent revision operation of total knee or total hip arthroplasty. Both sample types were analyzed using broad-range (BR-) PCR targeting the 16S rRNA gene. The results were evaluated based on the definition of periprosthetic joint infection from the Workgroup of the Musculoskeletal Infection Society. Results - PJI was diagnosed in 29 patients, whereas aseptic failure was diagnosed in 58 patients. Analysis of sonication fluid using BR-PCR detected bacteria in 27 patients, whereas analysis of periprosthetic tissue by BR-PCR detected bacteria in 22 patients. In 6 of 7 patients in whom BR-PCR analysis of periprosthetic tissue was negative, low-virulence bacteria were present. The sensitivity and specificity values for periprosthetic tissue were 76% and 93%, respectively, and the sensitivity and specificity values for sonication fluid were 95% and 97%. Interpretation - Our results suggest that sonication fluid may be a more appropriate sample than periprosthetic tissue for BR-PCR analysis in patients with PJI. However, further investigation is required to improve detection of bacteria in patients with so-called aseptic failure.
Introduction
The main aim was to analyse the series of 29 collected cemented Charnley–Muller Alivium retrievals with the meantime in situ of 27 years. In addition, the revision rate of 1425 Alivium ...prostheses implanted at our institution between 1977 and 1992 was calculated.
Materials and methods
The revision percentage of the Alivium cohort was calculated up to 45 years of follow-up and compared to that of all total hip arthroplasties (THAs) implanted in the same period (No. 5535). Metal and polyethylene retrieved components were inspected in 29 cases for wear damage and roughness. Wear particles were retrieved from periprosthetic tissue using digestion protocols and their composition, morphology, and size distribution were investigated. Periprosthetic tissue was analysed histologically.
Results
The revision percentage of the Alivium cohort was 16% at 45 years of follow-up. It was comparable to all the THAs implanted at the same time (18%). The shape of polyethylene particles isolated from periprosthetic tissue corresponded to the wear pattern on polyethylene cups. Polyethylene particles were the main wear product, with the majority (68%) of particles smaller than 0.1 µm. Metal particles were rare with two types: CoCr and Cr based. Histological analysis showed that in 14 out of 18 specimens, the metal particles were graded + 1, reflecting that the metal loading in the periprosthetic tissue was low.
Conclusions
Our study represents valuable data not reported previously on the survival rate of Charnley–Muller prostheses at 45 years of follow-up and a unique insight into the collected retrievals from the materials’ point of view.
Bacteriophages, prokaryotic viruses, hold great potential in genetic engineering to open up new avenues for vaccine development. Our study aimed to establish engineered M13 bacteriophages expressing ...MAGE-A1 tumor peptides as a vaccine for melanoma treatment. Through in vivo experiments, we sought to assess their ability to induce robust immune responses. Using phage display technology, we engineered two M13 bacteriophages expressing MAGE-A1 peptides as fusion proteins with either pVIII or pIIII coat proteins. Mice were intraperitoneally vaccinated three times, two weeks apart, using two different engineered bacteriophages; control groups received a wild-type bacteriophage. Serum samples taken seven days after each vaccination were analyzed by ELISA assay, while splenocytes harvested seven days following the second boost were evaluated by ex vivo cytotoxicity assay. Fusion proteins were confirmed by Western blot and nano-LC-MS/MS. The application of bacteriophages was safe, with no adverse effects on mice. Engineered bacteriophages effectively triggered immune responses, leading to increased levels of anti-MAGE-A1 antibodies in proportion to the administered bacteriophage dosage. Anti-MAGE-A1 antibodies also exhibited a binding capability to B16F10 tumor cells in vitro, as opposed to control samples. Splenocytes demonstrated enhanced CTL cytotoxicity against B16F10 cells. We have demonstrated the immunogenic capabilities of engineered M13 bacteriophages, emphasizing their potential for melanoma immunotherapy.
Prosthetic joint infections are frequently associated with biofilm formation and the presence of viable but non-culturable (VBNC) bacteria. Conventional sample culturing remains the gold standard for ...microbiological diagnosis. However, VBNC bacteria lack the ability to grow on routine culture medium, leading to culture-negative results. Bacteriophages are viruses that specifically recognize and infect bacteria. In this study, we wanted to determine if bacteriophages could be used to detect VBNC bacteria. Four staphylococcal strains were cultured for biofilm formation and transferred to low-nutrient media with different gentamycin concentrations for VBNC state induction. VBNC bacteria were confirmed with the BacLight
viability kit staining. Suspensions of live, dead, and VBNC bacteria were incubated with bacteriophage K and assessed in a qPCR for their detection. The VBNC state was successfully induced 8 to 19 days after incubation under stressful conditions. In total, 6.1 to 23.9% of bacteria were confirmed alive while not growing on conventional culturing media. During the qPCR assay, live bacterial suspensions showed a substantial increase in phage DNA. No detection was observed in dead bacteria or phage non-susceptible
suspensions. However, a reduction in phage DNA in VBNC bacterial suspensions was observed, which confirmed the detection was successful based on the adsorption of phages.
Amplification of the epidermal growth factor receptor gene on double minutes is recurrently observed in cells of advanced gliomas, but the structure of these extrachromosomal circular DNA molecules ...and the mechanisms responsible for their formation are still poorly understood. By using quantitative PCR and chromosome walking, we investigated the genetic content and the organization of the repeats in the double minutes of seven gliomas. It was established that all of the amplicons of a given tumor derive from a single founding extrachromosomal DNA molecule. In each of these gliomas, the founding molecule was generated by a simple event that circularizes a chromosome fragment overlapping the epidermal growth factor receptor gene. In all cases, the fusion of the two ends of this initial amplicon resulted from microhomologybased nonhomologous end-joining. Furthermore, the corresponding chromosomal loci were not rearranged, which strongly suggests that a postreplicative event was responsible for the formation of each of these initial amplicons.
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