Estimating price elasticities of demand for pain relief drugs: evidence from Medicare Part D.

Overdose deaths from prescription opioids remain elevated, and policymakers seek solutions to curb opioid misuse. Recent proposals call for price-based solutions, such as opioid taxes and removal of opioids from insurance formularies. However, there is limited evidence on how opioid consumption responds to price stimuli. This study addresses that gap by estimating the effects of prices on the utilization of opioids, as well as other prescription painkillers. I use nationally representative individual-level data on prescription drug purchases and exploit the introduction of Medicare Part D in 2006 as an exogenous change in out-of-pocket drug prices. I find that new users have a relatively high price elasticity of demand for prescription opioids, and that consumers treat over-the-counter painkillers as substitutes for prescription painkillers. My results suggest that increasing out-of-pocket prices of opioids, through formulary design or taxes, may be effective in reducing new opioid use.

Medicare Advantage Prior Authorization Requirements for Otolaryngologic Procedures in 2021.

Little is known about the extent of prior authorization requirements in otolaryngology. We performed a secondary analysis of data comparing prior authorization (PA) policies across 5 major Medicare Advantage insurers to estimate the counterfactual proportion of 2021 Medicare Part B fee-for-service spending and utilization for commonly performed otolaryngologic procedures that would have required PA. The counterfactual proportion of spending (range: 20.4%-27.6%) and utilization (range: 1.8%-4.5%) requiring PA was relatively consistent across insurers and largely attributable to rhinologic procedures. However, PA requirements for specific services varied widely among insurers. Among the 70 (of 196; 35.7%) services subject to PA by any insurer, nearly half were subject to PA by a single insurer (n = 34; 48.6%). Only 10 (14.3%) services were subject to PA by 4 (n = 6; 8.6%) or 5 (n = 4; 5.7%) insurers. These discrepancies illustrate the challenges of navigating discordant insurer policies for otolaryngologists and raise concerns about the validity of certain PA requirements.

Resected lung adenocarcinoma with lymph node metastasis: is ground glass opacity component a prognostic factor?

Background: Ground glass opacity (GGO)-featured lung adenocarcinoma generally has excellent prognosis, and here is rarely the occurrence of lymph node metastasis. We conducted a retrospective cohort study to explore the prognostic impact of GGO component in node-positive lung adenocarcinomas. Methods: A total of 669 patients with pathologic N1/N2 lung adenocarcinoma receiving R0 resection and systemic lymph node dissection from 2008 to 2015 were reviewed, including 635 solid and 34 part-solid lesions. Propensity score matching (PSM) was performed to compare survival outcomes of solid and part-solid lesions, in order to determine the prognostic value of GGO component. Cox proportional hazard model was performed to identify significant prognostic factors for resected node positive lung adenocarcinoma. Results: About 5.1% (34 of 669) of resected node-positive lung adenocarcinoma presented as part-solid nodules on computed tomography (CT) images in this cohort. The median nodule size on CT of the 34 part-solid lesions was 31 mm (range, 15-68 mm), median solid component size on CT was 24 mm (range, 12-62 mm), and median consolidation/tumor ratio was 0.8 (range, 0.64-0.95). After 1:4 PSM, 136 patients and 34 patients were matched from the solid and part-solid groups. No significant difference in either recurrence-free survival (RFS) (P=0.71) or overall survival (OS) (P=0.82) was found between the solid and part-solid groups. Multivariable Cox regression showed that pN stage was the strongest prognostic factor for RFS and OS. GGO component was not an independent prognostic factor toward for RFS [P=0.75; hazard ratio (HR) =0.93; 95% confidence interval (CI): 0.59-1.46] or OS (P=0.53; HR =1.19; 95% CI: 0.69-2.05). Conclusions: A minority of resected node-positive lung adenocarcinoma presents as GGO component on CT. The presence of GGO component does not predict better prognosis in node-positive lung adenocarcinoma.

Healthcare expenditures for people with substance use disorders in drug courts compared to their peers in traditional courts.

Individuals within the criminal justice system are at greater risk of substance use-related morbidity and mortality and have substantial healthcare needs. In this quasi-experimental study, we assessed utilization patterns of Massachusetts Medicaid Program (MassHealth) services and associated expenditures among drug court probationers compared to a propensity score-matched sample of traditional court probationers. Risk of reoffending, employment status, age, and living arrangement data were used to calculate propensity scores and match probationers between the two court types, producing a final sample of 271 in each court (N=542). Utilization of services and associated expenditures were analyzed using a two-part model to address the skewed distribution of the data and to control for residual differences after matching from the perspective of the payer (i.e., MassHealth). The largest categories of MassHealth spending were prescription pharmaceuticals, hospital inpatient visits, and physician visits. In the unadjusted analysis, drug court probationers exhibited greater MassHealth services utilization and expenditures than traditional court probationers. However, drug courts enrolled more females, more people at higher risk of reoffending, and more people with opioid use disorders. After controlling for differences between the two court types, the difference in MassHealth services utilization and associated expenditures did not reach statistical significance. Drug court probationers were more likely to engage with healthcare services but did not incur significantly greater expenditures than traditional court probationers after controlling for differences between the samples.

Development and Manufacturing of a Fibre Reinforced Thermoplastic Composite Spar Produced by Oven Vacuum Bagging.

The limited recyclability of fibre-reinforced thermoset composites has fostered the development of alternative thermoplastic-based composites and their manufacturing processes. The most common thermoplastic-based composites are often costly due to their availability in the form of prepreg materials and to the high pressure and temperatures required for their manufacturing. Yet, the manufacturing of economic and recyclable composites, made of semi-preg composite materials using traditional composite manufacturing technologies, has only been proved at a laboratory scale through the manufacturing of flat plates. This work reports the manufacturing of a real structural part, a wing spar section with complex geometry, made of commingled polyamide 12 (PA12) fibres and carbon fibres (CFs) semi-preg and by oven vacuum bagging (OVB). The composite layup was studied using finite element analysis, and processing simulation assisted in the determination of the PA12/CF preform for OVB. Processing of two forms of semi-preg materials was first evaluated and optimised. The material selection for part manufacturing was mainly defined by the materials' processability. The spar section was manufactured in two OVB stages and was then mechanically tested. The mechanical test showed a linear strain response of the prototype up to the maximum load and validated the optimised layup configuration of the composite structure.

The extended fibers of the alar part of the nasalis connect to the levator labii superioris, zygomaticus minor, and skin adjacent to the nasal ala.

PURPOSE: This study aimed to elucidate the positions of the extended fibers of the alar part of the nasalis (Na), and their connections to the levator labii superioris (LLS), zygomaticus minor (Zmi), and adjacent skin near the nasal ala. METHODS: The extended fibers of the Na were investigated in 54 specimens obtained from 27 embalmed adult South Korean cadavers. RESULTS: In 51.9% of the specimens, some fibers of the Na extended over the alar facial crease, intermingling or blending with the LLS or Zmi, and attached to the skin lateral to the nasal ala. The quantity and distribution of these extended fibers varied: some fibers of the Na extended and intermingled or blended with the LLS in 25.9%, while another 25.9% exhibited the Na extending in a distinctive fan shape with longer fibers, and combining with both the LLS and Zmi. However, the Na had no extended fibers that reached the LLS, Zmi, or skin near the nasal ala in 48.1%. CONCLUSION: Contraction of the Na and its extended fibers can influence the nasal ala and also the laterally located skin and muscles, directing them inferomedially toward the incisive fossa of the maxilla, which is the origin of the nasalis. These insights offer a deeper understanding of the role and actions of facial muscles in facial expression. They will be instrumental in the comprehension and analysis of nose and mouth movements, and in conducting electromyographic analyses in this region.

Folklore use of medicinal plants for the treatment of gynecological diseases in Pakistan-a review.

Background: Gynecological issues and sexually transmitted infections (STIs) pose significant challenges to women's health, particularly in developing nations. These challenges are exacerbated by limited access to modern reproductive healthcare facilities, economic constraints, and entrenched cultural norms. Consequently, most of the Pakistani population relies on traditional ethno-medicinal healthcare systems. This preference stems from the ease of access, affordability, widespread availability, and inherent trust placed in these alternative healthcare methods. Aim/objective: The inquiry aimed to report details on the application of conventional uses of plants in the health field in rural areas that could contribute to advancing the natural discovery of drugs. The objective of this analysis is to provide researchers with information on conventional and empirical knowledge of plant species concerning women's diseases. Methodology: Information on the common use of medicinal plants in treating women's diseases was gathered from electronic databases. As a keyword for the quest, ethnobotany, and ethnopharmacology were used together with gynecological complications. Result: The work of the current analysis has revealed that 217 plant species belonging to 89 families have been used in Pakistan's rural communities. The majority of plant species belong to the Apiaceae family, followed by the Asteraceae, Fabaceae, Solanaceae, Rosaceae, Lamiaceae, and other families. The biological interpretation of plants used in rural communities of Pakistan revealed that herbs and trees are the dominant forms with 58 % and 23 % respectively while shrubs and sub-shrubs with a low percentage of 17 % and 2 %. In natural preparation, leaves 29 %, flowers 22 %, seeds 14 %, fruits 14 %, roots 13 %, bark 7 %, and stems 5 % were the most used parts respectively and aerial parts, dried pericarp, bulb, bud, berry, latex, wood, rhizome, husk, fruit coat, oil, resins, twigs, and shoot were also used in minimum percentage. A multitude of plant species have found extensive application in the management of diverse women's health issues. These encompass concerns such as fatigue, mood fluctuations attributed to menstrual problems, gonorrhea, complications related to pregnancy, cravings for specific foods, throbbing breast pain, abdominal and pelvic cramps, excessive vaginal discharge, mastitis, irritability, abortion-related matters, headaches, uterine hemorrhage, Menorrhagia, Amenorrhea, Menopause, Vomiting Abortion, infertility and lactation challenges, as well as the regulation of lochia flow. Conclusion: This review provides remarkable information about the use of medicinal plants against women's diseases in the rural communities of Pakistan. It opens the gateway for the discovery of natural drug development.

Efficacy and safety of nivolumab monotherapy in patients with unresectable clear cell sarcoma and alveolar soft part sarcoma (OSCAR Trial/NCCH1510).

BACKGROUND: Clear cell sarcoma (CCS) and alveolar soft part sarcoma (ASPS) are rare, and standard systemic therapy is not established except for sunitinib in ASPS. It is known that CCS and ASPS have a common biological feature of melanoma and Xp11.2/TFE3 translocation renal cell carcinoma, and immune-checkpoint inhibitors (ICIs) are effective in these tumors. The authors conducted a phase 2 trial to evaluate the efficacy and safety of nivolumab for CCS and ASPS. METHODS: The number of patients expected to be enrolled was 15-25 and was determined based on the Bayesian design. The primary end point was the confirmed objective response rate (ORR) according to the central review and the secondary end points included ORR, progression-free survival (PFS), overall survival (OS), and safety. RESULTS: A total of 26 patients (CCS, 12; ASPS, 14) were enrolled. Efficacy and safety were analyzed on 25 and 26 patients, respectively. The minimum number of responses required for a positive conclusion regarding the efficacy was four. However, only one patient (4.0%) with ASPS had a partial response. Complete response, stable disease, progression disease, and not evaluable were 0%, 60%, 32%, and 4.0%, respectively. Adverse events of grade 3 or 4 occurred in 57.7% (15 of 26). The median PFS was 4.9 months (95% confidence interval [CI], 3.7-8.6 months) and the median OS was 15.8 months (95% CI, 8.2-not reached). CONCLUSIONS: The primary end point of the ORR was not met for CCS and ASPS on the central review. Further studies are needed to evaluate ICIs in patients with ASPS.

Cycle-Based Control of Injection Moulding Process in Presence of Material Dual Sourcing Using Mass Feedback.

The low cost and precise tolerances of plastic injection moulded products are a major reason for the popularity of the manufacturing method. The tolerances are greatly influenced by the equipment, raw material and moulding process. One challenge is the raw material variation. This paper presents a production process using cycle based feedback of cycle mass, for control of part properties in the presence of material variation from dual sourcing. The part properties considered are part mass and outer dimensions. The process uses direct cycle mass feedback without additional process measurements in the proposed controller structure. The designed controller structure is tested in a multi-cavity mould while using raw materials from multiple vendors, encompassing five different grades. The results show a total decrease in part mass variance of approximately 50% and a decrease of length and width variance of approximately 40% compared to a moulding process with fixed settings.

Examining the relationship between flexible working arrangements and employee performance: a mini review.

This research aims to determine the relationship between flexible working arrangements (FWAs) and employee performance (EP). The research was conducted by reviewing studies in Web of Science (WoS), EBSCO and Google Scholar databases between 2010 and 2024. The research was screened in the databases in line with the inclusion criteria, which were determined as studies written in English, where data were collected by survey technique, data were analyzed by correlation, and those that met the criteria were included in the research. As a result of the preliminary screening, second screening, and screening in line with the inclusion criteria, the remaining 21 studies constituted the data set of this study. The correlation between FWAs and EP was [r (20) = 0.596, p < 0.05]. This value can be interpreted a significant and high-level relationship between them. According to the random model, Fisher's Z and 95% CI (LL = 0.52 and UL = 0.84), Z = 8.45, measured an effect size of 0.35 p = 0.000. This value shows a moderate effect size according to Cohen's d. FWAs have a positive effect on EP, productivity, job satisfaction, job stress, work-family harmony, and organizational commitment. It is recommended that organizations, managers, organizational psychology, and human resources professionals (HRP) should include FWAs in job analysis, job design, and planning.

Optimization Strategy for Process Design in Rubber Injection Molding: A Simulation-Based Approach Allowing for the Prediction of Mechanical Properties of Vulcanizates.

Selecting the optimal settings for the production of rubber goods can be a very time-consuming and resource-intensive process. A promising method for optimizing rubber processing in a short period of time is the use of simulation routines. However, process simulations have only recently enabled meaningful predictions of not only the part's state of cure but also its mechanical characteristics. As a first approach, second-order polynomials were considered suitable for describing the properties of compression-molded parts. However, more precision is required for injection molding due to the narrower distribution of mechanical characteristics of parts produced at different vulcanization temperatures. This became evident when the approximation of mechanical data with second order models partly revealed significant failures of part behavior prediction. To tackle this issue, a combined approach for approximation is proposed in this contribution by means of logistic growth function in addition to second order polynomials. To feed the model, an experimental plan was designed for producing injection-molded parts from an SBR compound at various temperatures and to different degrees of cure. The parts obtained were then characterized mechanically, and the results were opposed to varying degrees of cure and extents of reaction to calculate the model coefficients. Once available, a simulation-based calculation of the mechanical part quality is possible. The comparison of test results from the simulation and the real process has shown a reliable prediction, as simulation results were found within the natural deviation of the real measurements.

Alveolar Soft Tissue Sarcoma: Correlation of MRI Features With Histological Grading and Patient Prognosis.

BACKGROUND: Alveolar Soft Part Sarcoma (ASPS) is a rare, aggressive cancer whose diagnosis and treatment depend on histological grading. However, tumor variability can lead to underestimation, affecting treatment, and patient survival. OBJECTIVE: To evaluate MRI features associated with Grade III ASPS and to determine the relationship between MRI features and patient prognosis. STUDY TYPE: Retrospective analysis. SUBJECTS: Sixty-seven patients with ASPS were included with 37 males and 30 females (M/F = 1.23) follow-up and survival analysis on 50 patients. FIELD STRENGTH/SEQUENCE: A 3.0 T, T1WI-FSE, T2WI-FSE, DWI-EPI, DCE-MRI (gradient echo). ASSESSMENT: MRI features (margin, peritumoral oedema, peritumoral enhancement, necrosis, vascular flow void signal, heterogeneous signal intensity [SI] at T1WI and T2WI, ADCmean, time-intensity curve [TIC] type, distant metastasis, and bone invasion) and histological grading were independently evaluated by three radiologists and two pathologists, with Grade III considered high-grade. STATISTICAL TESTS: The chi-square or Fisher's exact test was used to assess the correlation between MRI features and histological grading. Multivariable binary logistic regression identified independent factors associated with high-grade tumors. The Kaplan-Meier method and Cox proportional hazards model were used to calculate hazard ratios for MRI features. RESULTS: Tumor necrosis, heterogeneous SI at T2WI ≥50%, and ADCmean were associated with high-grade ASPS. Tumor necrosis was an independent factors associated with local relapse-free survival (odds ratio [OR], 3.88). TIC type was associated with 5-year survival rate (OR, 2.80) and local relapse-free survival (OR, 2.69). Heterogeneous SI at T2WI ≥50% was associated with 5-year survival (OR, 4.00), local relapse-free survival (OR, 5.58), and local relapse-free survival (OR, 4.84). DATA CONCLUSION: MRI features including tumor necrosis, heterogeneity of SI at T2WI, ADCmean, and TIC type aid in assessing ASPS grading and prognosis. EVIDENCE LEVEL: 4 TECHNICAL EFFICACY: Stage 5.

Differential symptom cluster responses and predictors to repetitive transcranial magnetic stimulation treatment in Parkinson's disease: A retrospective study.

Background: Repetitive transcranial magnetic stimulation (rTMS) is an effective noninvasive neuromodulation technique for Parkinson's disease (PD). However, the efficacy of rTMS varies widely between individuals. This study aimed to investigate the factors related to the response to rTMS in PD patients. Methods: We retrospectively analyzed the response of 70 idiopathic PD patients who underwent rTMS for 14 consecutive days targeting the supplementary motor area (SMA) in either an open-label trail (n = 31) or a randomized, double-blind, placebo-controlled trial (RCT) (n = 39). The motor symptoms of PD patients were assessed by the United Parkinson's Disease Rating Scale Part III (UPDRSIII). Based on previous studies, the UPDRSIII were divided into six symptom clusters: axial dysfunction, resting tremor, rigidity, bradykinesia affecting right and left extremities, and postural tremor. Subsequently, the efficacy of rTMS to different motor symptom clusters and clinical predictors were analyzed in these two trails. Results: After 14 days of treatment, only the total UPDRSIII scores and rigidity scores improved in both the open-label trial and the RCT. The results of multiple linear regression analysis indicated that baseline rigidity scores (ß = 0.37, p = 0.047) and RMT (ß = 0.30, P = 0.02) positively predicted the improvement of UPDRSIII. The baseline rigidity score (ß = 0.55, P < 0.0001) was identified as an independent factor to predict the improvement of rigidity. Conclusion: This study demonstrated significant improvements in total UPDRSIII scores and rigidity after 14-day treatment, with baseline rigidity scores and RMT identified as predictors of treatment response, underscoring the need for individualized therapy.

Complete Response to Pembrolizumab in Stage IV Alveolar Soft Part Sarcoma After Failure of Four Lines of Treatment: A Case Report and Literature Review.

Alveolar soft part sarcoma (ASPS) is a rare malignant tumor that manifests as a slow-growing soft tissue mass and frequently presents with distant metastasis. The prognosis is variable, and complete remission of metastatic disease has rarely been reported. Our patient was diagnosed with metastatic ASPS at the age of 17, with a primary forearm lesion and metastasis to the lungs. She underwent surgical resection of her forearm mass, followed by adjuvant chemotherapy and radiation to target the lung metastasis. Over the next decade, she had a complicated course of treatment. Her disease continued to slowly progress despite treatment with sunitinib, pazopanib, and a combination of docetaxel and gemcitabine. We eventually treated her with immune checkpoint inhibitors (ICIs). Pembrolizumab, initially in combination with bevacizumab and later as monotherapy, resulted in significant tumor shrinkage, especially in the pulmonary lesions, within the first three months. Subsequent imaging reported complete remission within 15 months and no disease recurrence at her three-year follow-up. Our case highlights one of the very few reported cases of complete remission achieved in metastatic ASPS after treatment with ICIs. ICIs could offer hope for disease remission in advanced ASPS, a rare malignancy that has proven difficult to treat successfully in the past. More studies need to be conducted to further evaluate the efficacy and any associated predictors of successful treatment.

Getting the right tail right: Modeling tails of health expenditure distributions.

Health expenditure data almost always include extreme values, implying that the underlying distribution has heavy tails. This may result in infinite variances as well as higher-order moments and bias the commonly used least squares methods. To accommodate extreme values, we propose an estimation method that recovers the right tail of health expenditure distributions. It extends the popular two-part model to develop a novel three-part model. We apply the proposed method to claims data from one of the biggest German private health insurers. Our findings show that the estimated age gradient in health care spending differs substantially from the standard least squares method.

The heterogeneity of holistic processing profiles in developmental prosopagnosia: holistic processing is impaired but not absent.

Although it is generally assumed that face recognition relies on holistic processing, whether face recognition deficits observed in Developmental Prosopagnosics (DPs) can be explained by impaired holistic processing is currently under debate. The mixed findings from past studies could be the consequence of DP's heterogeneous deficit nature and the use of different measures of holistic processing-the inversion, part-whole, and composite tasks-which showed a poor association among each other. The present study aimed to gain further insight into the role of holistic processing in DPs. Groups of DPs and neurotypicals completed three tests measuring holistic face processing and non-face objects (i.e., Navon task). At a group level, DPs showed (1) diminished, but not absent, inversion and part-whole effects, (2) comparable magnitudes of the composite face effect and (3) global precedence effect in the Navon task. However, single-case analyses showed that these holistic processing deficits in DPs are heterogeneous.

Preoperative computed tomography semantic features in predicting lymph node metastasis of part-solid nodules in non-small cell lung cancer: a multicenter retrospective study.

Background: Lymph node metastasis (LNM) is the most common route of metastasis for lung cancer, and it is an independent risk factor for long-term survival and recurrence in patients with non-small cell lung cancer (NSCLC). The purpose of this study was to explore the value of preoperative computed tomography (CT) semantic features in the differential diagnosis of LNM in part-solid nodules (PSNs) of NSCLC. Methods: A total of 955 patients with NSCLC confirmed by postoperative pathology were retrospectively enrolled from January 2019 to March 2023. The clinical, pathological data and preoperative CT images of these patients were investigated and statistically analyzed in order to identify the risk factors for LNM. Multivariate logistic regression was used to select independent risk factors and establish different prediction models. Ten-fold cross-validation was used for model training and validation. The area under the curve (AUC) of the receiver operating characteristic (ROC) curve was calculated, and the Delong test was used to compare the predictive performance between the models. Results: LNM occurred in 68 of 955 patients. After univariate analysis and adjustment for confounding factors, smoking history, pulmonary disease, solid component proportion, pleural contact type, and mean diameter were identified as the independent risk factors for LNM. The image predictors model established by the four independent factors of CT semantic features, except smoking history, showed a good diagnostic efficacy for LNM. The AUC in the validation group was 0.857, and the sensitivity, specificity, and accuracy of the model were all 77.6%. Conclusions: Preoperative CT semantic features have good diagnostic value for the LNM of NSCLC. The image predictors model based on pulmonary disease, solid component proportion, pleural contact type, and mean diameter demonstrated excellent diagnostic efficacy and can provide non-invasive evaluation in clinical practice.

PART1 facilitates tumorigenesis and inhibits ferroptosis by regulating the miR-490-3p/SLC7A11 axis in hepatocellular carcinoma.

BACKGROUND: Ferroptosis is associated with cancer progression and has a promising application for treating hepatocellular carcinoma (HCC). Long non-coding RNA (lncRNA) participates widely in the regulation of ferroptosis, but the key lncRNA regulators implicated in ferroptosis and their molecular mechanisms remain to be identified. METHODS: Bioinformatic analysis was performed in R based on The Cancer Genome Atlas Program (TCGA) public database. The relative expression of genes was detected by real-time quantitative PCR. Cell viability was assessed by the CCK8 assay. The cell cycle and apoptosis were detected by flow cytometry. Migration and invasion of HCC cells were detected by Transwell assay and wound healing assay. Expression of relevant proteins was detected by Western blotting. A dual-luciferase reporter assay was used to detect interactions between PART1 (or SLC7A11) and miR-490-3p. RESULTS: The PART1/miR-490-3p/SLC7A11 axis was identified as a potential regulatory pathway of ferroptosis in HCC. PART1 silencing reduced HCC cell proliferation, migration, and metastasis and promoted apoptosis and erastin-reduced ferroptosis. Further investigation revealed that PART1 acted as a competitive endogenous RNA (ceRNA) for miR-490-3p to enhance SLC7A11 expression. Overexpression of miR-490-3p downregulated the expression of SLC7A11, inhibiting the proliferation, invasion, and metastasis of HCC cells while promoting apoptosis and erastin-induced ferroptosis. Knockdown of PART1 in HCC cells significantly improved the sensitivity of HCC cells to sorafenib. CONCLUSION: Our results revealed that the PART1/miR-490-3p/SLC7A11 axis enhances HCC cell malignancy and suppresses ferroptosis, which provides a new perspective for understanding of the function of long chain non-coding RNAs in HCC. The PART1/miR-490-3p/SLC7A11 axis may be target for improving sorafenib sensitivity in HCC.

Chloroplast Cell-Free Systems from Different Plant Species as a Rapid Prototyping Platform.

Climate change poses a significant threat to global agriculture, necessitating innovative solutions. Plant synthetic biology, particularly chloroplast engineering, holds promise as a viable approach to this challenge. Chloroplasts present a variety of advantageous traits for genetic engineering, but the development of genetic tools and genetic part characterization in these organelles is hindered by the lengthy time scales required to generate transplastomic organisms. To address these challenges, we have established a versatile protocol for generating highly active chloroplast-based cell-free gene expression (CFE) systems derived from a diverse range of plant species, including wheat (monocot), spinach, and poplar trees (dicots). We show that these systems work with conventionally used T7 RNA polymerase as well as the endogenous chloroplast polymerases, allowing for detailed characterization and prototyping of regulatory sequences at both transcription and translation levels. To demonstrate the platform for characterization of promoters and 5' and 3' untranslated regions (UTRs) in higher plant chloroplast gene expression, we analyze a collection of 23 5'UTRs, 10 3'UTRs, and 6 chloroplast promoters, assessed their expression in spinach and wheat extracts, and found consistency in expression patterns, suggesting cross-species compatibility. Looking forward, our chloroplast CFE systems open new avenues for plant synthetic biology, offering prototyping tools for both understanding gene expression and developing engineered plants, which could help meet the demands of a changing global climate.

CP-Net: Instance-aware part segmentation network for biological cell parsing.

Instance segmentation of biological cells is important in medical image analysis for identifying and segmenting individual cells, and quantitative measurement of subcellular structures requires further cell-level subcellular part segmentation. Subcellular structure measurements are critical for cell phenotyping and quality analysis. For these purposes, instance-aware part segmentation network is first introduced to distinguish individual cells and segment subcellular structures for each detected cell. This approach is demonstrated on human sperm cells since the World Health Organization has established quantitative standards for sperm quality assessment. Specifically, a novel Cell Parsing Net (CP-Net) is proposed for accurate instance-level cell parsing. An attention-based feature fusion module is designed to alleviate contour misalignments for cells with an irregular shape by using instance masks as spatial cues instead of as strict constraints to differentiate various instances. A coarse-to-fine segmentation module is developed to effectively segment tiny subcellular structures within a cell through hierarchical segmentation from whole to part instead of directly segmenting each cell part. Moreover, a sperm parsing dataset is built including 320 annotated sperm images with five semantic subcellular part labels. Extensive experiments on the collected dataset demonstrate that the proposed CP-Net outperforms state-of-the-art instance-aware part segmentation networks.