Cell State of Origin Impacts Development of Distinct Endometriosis-Related Ovarian Carcinoma Histotypes.

Clear cell ovarian carcinoma (CCOC) and endometrioid ovarian carcinoma (ENOC) are ovarian carcinoma histotypes, which are both thought to arise from ectopic endometrial (or endometrial-like) cells through an endometriosis intermediate. How the same cell type of origin gives rise to two morphologically and biologically different histotypes has been perplexing, particularly given that recurrent genetic mutations are common to both and present in nonmalignant precursors. We used RNA transcription analysis to show that the expression profiles of CCOC and ENOC resemble those of normal endometrium at secretory and proliferative phases of the menstrual cycle, respectively. DNA methylation at the promoter of the estrogen receptor (ER) gene (ESR1) was enriched in CCOC, which could potentially lock the cells in the secretory state. Compared with normal secretory-type endometrium, CCOC was further defined by increased expression of cysteine and glutathione synthesis pathway genes and downregulation of the iron antiporter, suggesting iron addiction and highlighting ferroptosis as a potential therapeutic target. Overall, these findings suggest that while CCOC and ENOC arise from the same cell type, these histotypes likely originate from different cell states. This "cell state of origin" model may help to explain the presence of histologic and molecular cancer subtypes arising in other organs. SIGNIFICANCE: Two cancer histotypes diverge from a common cell of origin epigenetically locked in different cell states, highlighting the importance of considering cell state to better understand the cell of origin of cancer.

Laser-cutting: A novel alternative approach for point-of-care manufacturing of bespoke tablets.

A novel subtractive manufacturing method to produce bespoke tablets with immediate and extended drug release is presented. This is the first report on applying fusion laser cutting to produce bespoke furosemide solid dosage forms based on pharmaceutical-grade polymeric carriers. Cylindric tablets of different sizes were produced by controlling the two-dimensional design of circles of the corresponding diameter. Immediate and extended drug release patterns were achieved by modifying the composition of the polymeric matrix. Thermal analysis and XRD indicated that furosemide was present in an amorphous form. The laser-cut tablets demonstrated no significant drug degradation (<2%) nor the formation of impurities were identified. Multi-linear regression was used to quantify the influences of laser-cutting process parameters (laser energy levels, scan speeds, and the number of laser applications) on the depth of the laser cut. The utility of this approach was exemplified by manufacturing tablets of accurate doses of furosemide. Unlike additive or formative manufacturing, the reported approach of subtractive manufacturing avoids the modification of the structure, e.g., the physical form of the drug or matrix density of the tablet during the production process. Hence, fusion laser cutting is less likely to modify critical quality attributes such as release patterns or drug contents. In a point-of-care manufacturing scenario, laser cutting offers a significant advantage of simplifying quality control and a real-time release of laser-cut products such as solid dosage forms and implants.

The use of near-infrared as process analytical technology (PAT) during 3D printing tablets at the point-of-care.

Fused deposition modelling (FDM) is one of the most researched 3D printing technologies that holds great potential for low-cost manufacturing of personalised medicine. To achieve real-time release, timely quality control is a major challenge for applying 3D printing technologies as a point-of-care (PoC) manufacturing approach. This work proposes the use of a low-cost and compact near-infrared (NIR) spectroscopy modality as a process analytical technology (PAT) to monitor a critical quality attribute (drug content) during and after FDM 3D printing process. 3D printed caffeine tablets were used to manifest the feasibility of the NIR model as a quantitative analytical procedure and dose verification method. Caffeine tablets (0-40 % w/w) were fabricated using polyvinyl alcohol and FDM 3D printing. The predictive performance of the NIR model was demonstrated in linearity (correlation coefficient, R2) and accuracy (root mean square error of prediction, RMSEP). The actual drug content values were determined using the reference high-performance liquid chromatography (HPLC) method. The model of full-completion caffeine tablets demonstrated linearity (R2 = 0.985) and accuracy (RMSEP = 1.4 %), indicated to be an alternative dose quantitation method for 3D printed products. The ability of the models to assess caffeine contents during the 3D printing process could not be accurately achieved using the model built with complete tablets. Instead, by building a predictive model for each completion stage of 20 %, 40 %, 60 % and 80 %, the model of different completion caffeine tablets displayed linearity (R2 of 0.991, 0.99, 0.987, and 0.983) and accuracy (RMSEP of 2.22 %, 1.65 %, 1.41 %, 0.83 %), respectively. Overall, this study demonstrated the feasibility of a low-cost NIR model as a non-destructive, compact, and rapid analysis dose verification method enabling the real-time release to facilitate 3D printing medicine production in the clinic.

Molecular analysis suggests oligoclonality and metastasis of endometriosis lesions across anatomically defined subtypes.

OBJECTIVE: To investigate the heterogeneity of somatic cancer-driver mutations within patients and across endometriosis types. DESIGN: A single-center cohort, retrospective study. SETTING: Tertiary specialist-care center at a university hospital. PATIENT(S): Patients with surgically and histologically confirmed endometriosis of at least 2 anatomically distinct types (ovarian, deep infiltrating, and superficial). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Specimens were analyzed for the presence or absence of somatic cancer-driver mutations using targeted panel sequencing with orthogonal validation using droplet digital polymerase chain reaction and mutation-surrogate immunohistochemistry. RESULT(S): It was found that 13 of 27 patients had informative somatic driver mutations in endometriosis lesions; of these 13 patients, 9 had identical mutations across distinct lesions. Endometriomas showed a higher mutational complexity, with functionally redundant driver mutations in the same gene and within the same lesions. CONCLUSION(S): Our data are consistent with clonality across endometriosis lesions, regardless of subtype. Further, the finding of redundancy in mutations within the same gene and lesions is consistent with endometriosis representing an oligoclonal disease with dissemination likely to consist of multiple epithelial clones traveling together. This suggests that the current anatomically defined classification of endometriosis does not fully recognize the etiology of the disease. A novel classification should consider genomic and other molecular features to promote personalized endometriosis diagnosis and care.

A critical appraisal of the circulating levels of differentially expressed microRNA in endometriosis†.

Endometriosis is a common gynecological condition characterized by estrogen dependence, chronic pelvic pain, infertility, and diagnostic delay of between 5.4 and 12 years. Despite extensive study, no biomarker, either alone or in combination with other markers, has proven superior to laparoscopy for the diagnosis of endometriosis. Recent studies report that circulating levels of differentially expressed microRNA (miRNA) in women with endometriosis compared with controls are potential diagnostic tools. However, the lack of replication and absence of validated differential expression in novel study populations have led some to question the diagnostic value of miRNA. To elucidate potential reasons for the lack of replication of study results and explore future directions to enhance replicability of circulating miRNA results, we carried out an electronic search of the miRNA literature published between 2000 and 2020. Eighteen studies were identified in which 63 different miRNAs were differentially expressed in the circulation of women with endometriosis compared with controls. However, the differential expressions of only 14 miRNAs were duplicated in one or more studies. While individual miRNAs lacked diagnostic value, miRNA panels yielded sensitivity and specificity equal to or better than laparoscopy in five studies. Important differences in study design, sample processing, and analytical methods were identified rendering direct comparisons across studies problematic and could account for the lack of reproducibility of study results. We conclude that while the results of miRNA studies to date are encouraging, refinements to study design and analytical methods should enhance the reliability of circulating miRNA for the diagnosis of endometriosis.

3D and 4D printing in dentistry and maxillofacial surgery: Printing techniques, materials, and applications.

3D and 4D printing are cutting-edge technologies for precise and expedited manufacturing of objects ranging from plastic to metal. Recent advances in 3D and 4D printing technologies in dentistry and maxillofacial surgery enable dentists to custom design and print surgical drill guides, temporary and permanent crowns and bridges, orthodontic appliances and orthotics, implants, mouthguards for drug delivery. In the present review, different 3D printing technologies available for use in dentistry are highlighted together with a critique on the materials available for printing. Recent reports of the application of these printed platformed are highlighted to enable readers appreciate the progress in 3D/4D printing in dentistry.

Spatial frequency processing in inferred PC- and MC-pathways.

The goal of this study was to investigate the role of inferred parvocellular (PC) and magnocellular (MC) pathways in spatial contrast sensitivity. Localized, spatially narrow-band patterns (sixth derivatives of Gaussians, D6s) were presented at various peak spatial frequencies. When the D6 appeared on a pulsed luminance pedestal (Pulsed-Pedestal Paradigm), the spatial contrast sensitivity showed a band-pass shape with good contrast sensitivity at medium spatial frequencies. When the D6 appeared on a steady luminance pedestal (Steady-Pedestal Paradigm), the spatial contrast sensitivity showed a low-pass shape with decreased sensitivity at high spatial frequencies. The band-pass CSF was interpreted as reflecting PC-pathway mediation; the lower spatial frequency region of the low-pass CSF as reflecting MC-pathway mediation.