Analysis of Treatment Response With Proteins Related to Tumor Immunity in Postoperative Irradiated Cervical Cancer Patients.

BACKGROUND/AIM: This study evaluated the association between programmed cell death-ligand 1 (PD-L1) and prognosis in patients with cervical cancer treated with postoperative radiation and the impact of neoadjuvant chemotherapy (NAC) on this association. PATIENTS AND METHODS: Immunohistochemical analysis was performed on biopsy specimens from 42 patients who did not receive NAC and from paired samples before (biopsies) and after (resected tissues) chemotherapy from 46 patients who received NAC to determine the association of PD-L1 with radiotherapy outcomes. RESULTS: In the non-NAC group, patients with ≥10% PD-L1-expressing tumor cells prior to treatment had better recurrence-free survival (RFS) than those with <10% PD-L1-expressing tumor cells (p=0.001). In the NAC group, RFS was significantly lower (p=0.005) in the group with a ≥5% reduction of PD-L1 expression in tumor cells after chemotherapy than in those with <5% reduction. In multivariate analysis, only PD-L1 expression (non-NAC group) and the change in PD-L1 expression (NAC group) were associated with RFS. CONCLUSION: Low PD-L1 expression in a cervical tumor prior to treatment was identified as a risk factor for a poor outcome after postoperative radiotherapy. Furthermore, NAC induces an immunological shift that reduces PD-L1 levels in tumor cells, thereby negatively impacting treatment outcomes.

Prediction of Mismatch Repair Status in Endometrial Cancer from Histological Slide Images Using Various Deep Learning-Based Algorithms.

The application of deep learning algorithms to predict the molecular profiles of various cancers from digital images of hematoxylin and eosin (H&E)-stained slides has been reported in recent years, mainly for gastric and colon cancers. In this study, we investigated the potential use of H&E-stained endometrial cancer slide images to predict the associated mismatch repair (MMR) status. H&E-stained slide images were collected from 127 cases of the primary lesion of endometrial cancer. After digitization using a Nanozoomer virtual slide scanner (Hamamatsu Photonics), we segmented the scanned images into 5397 tiles of 512 × 512 pixels. The MMR proteins (PMS2, MSH6) were immunohistochemically stained, classified into MMR proficient/deficient, and annotated for each case and tile. We trained several neural networks, including convolutional and attention-based networks, using tiles annotated with the MMR status. Among the tested networks, ResNet50 exhibited the highest area under the receiver operating characteristic curve (AUROC) of 0.91 for predicting the MMR status. The constructed prediction algorithm may be applicable to other molecular profiles and useful for pre-screening before implementing other, more costly genetic profiling tests.

Mailing human papillomavirus self-sampling kits to women under-screened for cervical cancer improved detection in cervical cancer screening in a general population study in Japan.

BACKGROUND: One cause of the increase in cervical cancer rates in Japan is the long-term stagnation in the cervical cancer screening consultation rate. Therefore, improving the screening consultation rate is of urgent concern to reduce cervical cancer incidence. Self-collected human papilloma virus (HPV) tests have been successfully adopted in several countries, such as the Netherlands and Australia, as a measure of individuals who have not undergone cervical cancer screening in national programs. This study aimed to verify whether self-collected HPV tests presented an effective countermeasure for individuals who had not undergone the recommended cervical cancer screenings. METHODS: This study was conducted from December 2020 to September 2022 in Muroran City, Japan. The primary evaluated endpoint was the percentage of citizens who underwent cervical cancer screening at a hospital with positive self-collected HPV test results. The secondary endpoint was the percentage of included participants who were diagnosed with cervical intraepithelial neoplasia (CIN) or higher among those who visited a hospital and underwent cervical cancer screening. RESULTS: The included study participants were 7,653 individuals aged 20-50 years with no record of previous cervical cancer examination in the past 5 years. We mailed these participants information on self-administered HPV tests as an alternative screening procedure and sent the kit to 1,674 women who requested the test. Among them, 953 returned the kit. Among the 89 HPV-positive individuals (positive rate, 9.3%), 71 (79.8%) visited the designated hospital for an examination. A closer examination revealed that 13 women (18.3% of hospital visits) had a CIN finding of CIN2 or higher, among whom one each had cervical cancer and vulvar cancer, eight presented with CIN3, and three presented with CIN2; two cases of invasive gynecologic cancer were also identified. CONCLUSIONS: We conclude that the self-collected HPV tests showed a certain efficacy as a measure of individuals who had not undergone the recommended cervical cancer screening. We devised ways to have the unexamined patients undergo HPV testing and ensure that HPV-positive individuals visited the hospital. Despite a few limitations, our findings suggest the effectiveness of this public health intervention.

Predictive value of an exosomal microRNA-based signature for tumor immunity in cervical cancer patients treated with chemoradiotherapy.

Resistance of cervical cancer to radiotherapy with concurrent chemotherapy (CCRT) results in a poor prognosis. To identify new biomarkers for predicting the treatment response and prognosis, we explored exosomal microRNA (miRNA) expression signatures associated with the outcome of cervical cancer patients treated with CCRT. Exosomes were isolated from the plasma of 45 patients prior to CCRT during 2014-2020, and miRNA analysis was performed by next-generation sequencing. At a median follow-up of 38 months, 26 patients were recurrence free, 15 patients had died of the disease, and 4 patients received salvage chemotherapy due to distant metastasis. Of the 2522 miRNAs detected, 9 (miR-148a-5p, 1915-3p, 3960, 183-5p, 196b-5p, 200c-3p, 182-5p, 374a-5p, and 431-5p) showed differential expression between the recurrence-free and recurrence groups. Patients were divided into high- and low-risk groups according to the cutoff of the miRNAs-based risk score calculated from respective expression levels. The high-risk group had significantly worse disease-specific survival than the low-risk group (p < 0.001). In addition, miR-374a-5p and miR-431-5p expression showed a weak inverse correlation with tumor-infiltrating CD8+ and FOXP3+ T cells, suggesting a potential inhibitory effect on CCRT by suppressing tumor immunity. This miRNA signature could improve non-invasive monitoring and personalized treatment for cervical cancer.

Pathological classification of desmoplastic reaction is prognostic factor in cervical adenocarcinoma.

Desmoplastic reaction (DR) and inflammation are significant pathological manifestations of tumorigenesis in several cancers. However, the correlation between these stromal reactions and cervical adenocarcinoma has been poorly documented. This investigation elucidated whether DR is a prognostic indicator in early cervical adenocarcinoma patients. Fifty-nine patients with early stage cervical adenocarcinoma (stages I/II) were included in the study. DR was divided into three groups, mature, intermediate, and immature, based on the presence of myxoid stroma and hyalinized keloid-like collagen. Inflammatory cell responses were classified as mild, moderate, and severe. Those stromal reactions were separately evaluated in the invasion front stroma and intratumoral stroma. In both the intratumor and invasion front stroma, intermediate/immature DR was correlated with tumor size, T stage, N stage, lymphovascular invasion, and parametrial infiltration (p < 0.001 to p < 0.05). In addition, in the intratumoral stroma, intermediate/immature DR led to short relapse-free survival and overall survival (p < 0.001). In the invasion front stroma, inflammatory cell responses were associated with DR immaturity and FIGO stage (p < 0.01). These results suggest that the classification of DR maturity is a potential prognostic biomarker in early stage cervical adenocarcinoma patients. DR can be evaluated by routine H&E staining without immunohistochemistry, making it convenient and economical in clinical practice.

Radiotherapy for HPV-related cancers: prediction of therapeutic effects based on the mechanism of tumor immunity and the application of immunoradiotherapy.

Human papillomavirus (HPV)-related cancer is one of the diseases entities for which the applications of radiotherapy have been increasing. Recently, the process of carcinogenesis from HPV infection and the mechanism of tumor immunity that develops during disease progression have been elucidated. In this review, we will describe the mechanism of tumor immunity and how chemoradiotherapy may overcome and improve the efficacy of tumor immunity. We will also discuss the usefulness of proteins involved with tumor immunity as a predictive marker of radiotherapy response, and present an overview of ongoing clinical trials of combinations of immune checkpoint inhibitors and radiotherapy to demonstrate the promising combination therapy that has been currently emerging.

Prediction of treatment response from the microenvironment of tumor immunity in cervical cancer patients treated with chemoradiotherapy.

To supplement clinical decision-making in the management of cervical cancer, various prognostic factors, including tumor immune microenvironments, were examined in patients with cervical cancer treated with definitive chemoradiotherapy. We retrospectively analyzed the expression of CD8, FoxP3, HLA-1, PD-L1, and XRCC4 in 100 cases of cervical cancer. The observed tumor immune microenvironments were also classified into three types: inflamed, excluded, and cold type. Less FoxP3+ T cells and cold-type tumor were found to be poor prognostic factors in addition to non-SCC, large pre-treatment tumor volume, and three or less cycles of concurrent chemotherapy based on multivariate analysis. Cold-type tumors had significantly worse prognoses than the other two types, whereas inflamed- and excluded-type tumors showed similar 5-year disease-specific survival (P < 0.001; 0% vs. 60.3% vs. 72.3%). Radiotherapy could overcome the inhibitory immune microenvironment that occurs in excluded type. Individualized combination therapy adapted to pre-treatment tumor immunity may be necessary to improve radiotherapy outcomes in cervical cancer.

Anaerobic co-digestion of dairy manure and Japanese knotweed (Fallopia japonica) under thermophilic condition: Optimal ratio for biochemical methane production.

Anaerobic co-digestion of animal manure and lignocellulosic biomass is a potent approach for sustainable biomethane production. Co-digestion of dairy manure (DM) and Japanese knotweed (JK), which was collected from a riverbank, was investigated at five different DM-to-JK mixing ratios (100:0, 90:10, 80:20, 60:40, and 0:100; wet weight basis) under thermophilic condition. The results showed that the methane yields obtain from the co-digestion of DM and JK were much higher than that obtained from JK alone (104 ml/gVS), which indicates the synergistic effect and the benefits of co-digesting JK with DM. The highest methane yield (232 ml/gVS) was obtained from the DM-to-JK ratio of 90:10, which was 14.9% and 123.1% higher than that from DM and JK alone, respectively. It also showed the highest synergistic effect (61 ml/gVS). However, further increase in JK ratios led to the decrease in methane yield and synergistic effect. Therefore, applying the co-digestion of DM and JK at a ratio of 90:10 is recommended for biomethane production.

Radical Vaginal Trachelectomy.

Recently, radical vaginal hysterectomy (RVH) has developed into laparoscopically assisted radical vaginal hysterectomy (LARVH), which is associated with the laparoscopical procedure, and it is applied as radical vaginal trachelectomy and semi-radical vaginal hysterectomy. LARVH is indicated for patients with stage IB1 and IIA1 cervical carcinoma, especially those with a tumor size of less than 2 cm, because the cardinal ligaments cannot be resected widely. Although RVH that is associated with laparoscopic pelvic lymphadenectomy is the most used surgical procedure, radical trachelectomy may be performed either abdominally or vaginally (laparoscopic or robotic). One report found that the pregnancy rate was higher in patients who underwent minimally invasive or radical vaginal trachelectomy than in those who underwent radical abdominal trachelectomy.

ERα increases endometrial cancer cell resistance to cisplatin via upregulation of BAG3.

Endometrial cancer is a leading cause of cancer-associated mortality in women and has a poor prognosis in advanced stages. Our previous study revealed that BCL-2-associated athanogene 3 (BAG3) may contribute to enhancing cell viability through downregulation of microRNA (miR)-29b in endometrial cancer cell lines. In addition, a relationship between estrogen receptor α (ERα) and BAG3 was recently reported in several cancer cell types. The present study investigated the relationship between ERα and BAG3 in endometrial cancer cell lines. The results demonstrated that exogenous ERα overexpression enhanced BAG3 expression in the EMTOKA endometrial cancer cell line, which does not endogenously express ERα, but had no effect on BAG3 expression levels in the Ishikawa cell line, which does endogenously express ERα. In addition, ERα overexpression suppressed miR-29b expression and enhanced the expression of Mcl-1, a mediator situated downstream of BAG3, in EMTOKA cells, but not Ishikawa cells. ERα overexpression also enhanced EMTOKA, but not Ishikawa, endometrial cancer cell viability in the presence of cisplatin. These findings suggested that ERα may contribute to enhancing endometrial cancer cell resistance to anticancer agents through BAG3 overexpression.

Association between cancer immunity and treatment results in uterine cervical cancer patients treated with radiotherapy.

OBJECTIVE: To evaluate proteins related to tumor immune response and treatment outcome from radiotherapy for uterine cervical cancer patients. METHODS: We performed a retrospective immunohistochemical staining of 81 patients with uterine cervical cancer who underwent definitive radiotherapy. We examined the expression of programmed death ligand 1, human leukocyte antigen class I, tumor-infiltrating CD8+, and forkhead box P3+ (FoxP3+) T cells in tumor tissues. RESULTS: In biopsy specimen, patients with a higher number of CD8+ T cells and FoxP3+ T cells had a better disease-specific survival than patients with a lower number of CD8+ T cells and FoxP3+ cells (P = 0.018 and P = 0.009). Multivariate analysis showed that equivalent dose in 2 Gy fractions (EQD2) of the minimum dose to 90% of the high-risk clinical target volume, FoxP3+ T cells and expression of human leukocyte antigen class I were significant prognostic factors. When the EQD2 is 70 Gy or more, a higher local control rate is obtained regardless of the number of CD8- or FoxP3-positive cells. When EQD2 is <70 Gy, the number of CD8-positive cells has a significant impact on treatment outcome: the recurrence rate (local recurrence rate + distant metastasis rate) was 46.2% in the group with a CD8 value of 230 or higher, whereas the recurrence rate was 75.7% in the group with a CD8 value of less than 230. CONCLUSION: The combination of CD8 or FoxP3 with EQD2 can be potentially useful to predict the treatment results of radiotherapy for cervical cancer, leading to individualized optimal selection of treatment for cervical cancer.

Effect of solid-liquid separation on anaerobic digestion of dairy manure in semi-continuous stirred tank reactors: Process performance and digestate characteristics.

The objective of this study was to investigate the effect of solid-liquid separation on anaerobic digestion of dairy manure in semi-continuous stirred tank reactors. Reactors fed with liquid fraction of dairy manure (screened liquid manure) were kept in water baths at mesophilic (Run 1) and thermophilic (Run 3) temperatures, respectively, while reactors fed with water diluted manure and kept at mesophilic (Run 2) and thermophilic (Run 4) temperatures as control reactors. The performances of reactors were compared in terms of biogas and methane production, and digestate characteristics. The methane yields obtained from screened manure were higher than those from diluted manure at both mesophilic and thermophilic temperatures, while the highest methane yield was 185 L/kg VSadded under thermophilic temperature. Solid-liquid separation also had improved the effect on digestate fertilizer characteristics. Among four digestates from reactors, the highest contents of nutrients, N (4.12 g/kg) and P (2.36 g/kg) were found in Run 3, while the highest content of K (3.42 g/kg) was found in Run 1. These results showed the benefits of solid-liquid separation of dairy manure on process performance and digestate characteristics.

Potential of biogas production from manure of dairy cattle fed on natural soil supplement rich in iron under batch and semi-continuous anaerobic digestion.

This study provides a novel method for improving the anaerobic digestion (AD) of Holstein dairy manure (HDM) by the direct addition of Mineraso (MnS), a natural soil-derived supplement, to the feed of Holstein dairy cattle (HDC). MnS is chiefly composed of approximately 69.08% Fe3O4 and was supplemented at rates of 0 (F1), 25 (F2), and 50 (F3) g/head of HDC/d for two months. The HDM was then examined for non-absorbed iron prior to the batch and semi-continuous bench AD experiments. The results revealed that MnS enhanced CH4 generation in F2 and F3 by 25% and 42%, respectively, in the batch experiments compared to that of F1. Additionally, the gas yield improved in F2 and F3 by 45% and 66%, respectively, over the control after 7 d in the bench experiments. Therefore, supplementing animals with MnS represents a sustainable and economic approach to enhancing CH4 yields.

Are all wines made from various grape varieties beneficial in the prevention of myocardial infarction and stroke?

AIM: Epidemiologic studies support the assumption (French paradox hypothesis) that drinking red wine is beneficial in the prevention of cardiovascular diseases. Our recent works however cast doubt on such claim. Earlier we have shown that the antithrombotic activity of various fruits and vegetables mainly depends on their varieties. For this reason, several varieties of red and white grapes were tested for antithrombotic effect in animal experiments. RESULTS: Antithrombotic effect of 45 red and white grape varieties were assessed in the present study. Out of the 45, one red grape variety showed antithrombotic effect, while the majority of red and white grape varieties enhanced thrombosis. CONCLUSION: Most red and white grape varieties enhanced thrombotic activity of blood.

Prospects for biogas production and H2S control from the anaerobic digestion of cattle manure: The influence of microscale waste iron powder and iron oxide nanoparticles.

Improving the quality and quantity of biogas usually requires pre-treatment to maximize methane yields and/or post-treatment to remove H2S, which involves considerable energy consumption and higher costs. Therefore, this study proposes a cost-effective method for the enhanced anaerobic digestion (AD) of dairy manure (DM) without pre/post-treatment by directly adding waste iron powder (WIP) and iron oxide nanoparticles (INPs) to batch digesters. The results showed that the addition of iron in the form of microscale WIP (generated from the laser cutting of iron and steel) at concentrations of 100 mg/L, 500 mg/L, and 1000 mg/L improved methane yields by 36.99%, 39.36%, and 56.89%, respectively. In comparison, the equivalent dosages of INPs improved yields by 19.74%, 18.14%, and 21.11%, respectively. Additionally, the highest WIP dose (1000 mg/L) achieved the maximum improvement in the rate of hydrolysis (k), which was 1.25 times higher than in control reactions, and a maximum biomethane production rate (Rmax) of 0.045 L/gVS/d according to kinetic analysis models (i.e., first-order and the Gompertz kinetic models). The rate of H2S production was also significantly reduced (by 45.20%, 58.16%, and 77.24%) using the three WIP concentrations in comparison with INPs (which achieved reductions of 33.59%, 46.30%, and 53.52%, respectively). Therefore, the direct mixing of WIP with cattle manure is proposed as a practical and economical means of addressing complex and high-cost pre- and post-treatments that are otherwise required in the digestion process.

Impacts of iron oxide and titanium dioxide nanoparticles on biogas production: Hydrogen sulfide mitigation, process stability, and prospective challenges.

Anaerobic digestion for biogas production is one of the most used technology for bioenergy. However, the adoption of nanoparticles still needs further studies. Therefore, this study was designed to examine the effect of metal oxide nanoparticles (MONPs) at four different concentrations in two different combinations, 20 (R1) and 100 (R2) mg/L for Fe2O3, 100 (R3) and 500 (R4) mg/L for TiO2, and a mixture of Fe2O3 and TiO2 at rates of 20, 500 (R5) and 100, and 500 (R6), on hydrogen sulfide (H2S) mitigation, biogas, and methane (CH4) yield during the anaerobic digestion of cattle manure (CM) using an anaerobic batch system. The results showed that H2S production was 2.13, 2.38, 2.37, 2.51, 2.64, and 2.17 times lower than that of the control (R0), respectively, when the CM was treated by the aforementioned MONPs. Additionally, biogas and CH4 production were 1.09 and 1.105, 1.15 and 1.191, 1.07 and 1.097, 1.17 and 1.213, 1.10 and 1.133, and 1.13 and 1.15 times higher than those of R0 when R1, R2, R3, R4, R5, and R6 were supplemented with MONPs, respectively. The highest specific production of biogas and CH4 was 336.25 and 192.31 mL/gVS, respectively, which was achieved by R4 supplemented with 500 mg/L TiO2 NPs, while the corresponding values in the case of R0 were 286.38 and 158.55 mL/gVS.

The survival of pathogenic bacteria and plant growth promoting bacteria during mesophilic anaerobic digestion in full-scale biogas plants.

The introduction of biogas plants is a promising way to recycle organic wastes with renewable energy production and reducing greenhouse gas. Application of anaerobic digestate as a fertilizer reduces the consumption of chemical fertilizers. In this study, the survival of pathogenic bacteria and plant growth promoting bacteria (PGPB) in two full-scale biogas plants operated at mesophilic condition were investigated. Feedstock and anaerobic digestate samples were collected from biogas plants and bacteria load in samples were detected using standard dilution plate method. Pathogenic bacteria were reduced to not detected level through mesophilic digestion tank except for Campylobacter. However, it could be reduced by 98.7% through a sterilization tank. Bacillus was detected at 8.00 and 7.81 log10  CFU/g dry matter in anaerobic digestates, and it was also resistant to sterilization tank. Bacillus spp. is considered to be the safe bacteria that hold remarkable abilities for promoting plant growth. The results showed that treatment at biogas plants is effective to reduce pathogenic bacteria in dairy manure, and sterilization could further reduce the sanitary risks of pathogenic bacteria relating to anaerobic digestate application. Anaerobic digestates could also be utilized as bio-fertilizer as the high load of plant growth promoting bacteria.

Efficacy of liquid-based genetic diagnosis of endometrial cancer.

Although liquid-based cytology (LBC) has increased the sensitivity of cytological diagnosis of endometrial cancer (EC) compared with conventional smear cytology, the sensitivity of LBC for the detection of EC is between 70% and 96% and remains unsatisfactory. In the present study, we compared the efficacy of LBC with liquid-based genetic diagnosis (LBGDx) by amplicon sequencing of five genes including PTEN, PIK3CA, CTNNB1, KRAS, and TP53 in 48 LBC subjects who underwent endometrial screening. Consequently, LBC classified 15 samples as "positive or suspicious for malignancy" and the 15 were later confirmed as EC. However, LBC failed to identify five cases who were diagnosed as EC by additional transvaginal ultrasound and endometrial curettage, indicating that the sensitivity of cytology alone was 75% (15/20). LBGDx identified 11 pathogenic PTEN variants in 10 subjects, six PIK3CA variants in nine, three CTNNB1 variants in five, two KRAS variants in four, and three TP53 variants in three. Collectively, at least one pathogenic variant was identified in 19 subjects, which included 17 EC (15 endometrioid carcinoma and 2 endometrial carcinosarcomas), and one cervical adenocarcinoma. However, LBGDx did not identify any pathogenic mutations in three of the 20 EC, indicating that the sensitivity of LBGDx alone was 85% (17/20). Although five EC were negative for malignancy by LBC and three were negative for pathogenic mutations by LBGDx, the combination of LBC and LBGDx would successfully diagnose all 20 EC. These data suggested that LBGDx is a useful strategy to improve the sensitivity of screening of EC by LBC.

Potential of anaerobic digestate of dairy manure in suppressing soil-borne plant disease.

Frequent use of pesticides to control soil-borne plant disease leads to environmental pollution and the development of pesticide resistance in phytopathogens. Soil amendment is considered to have the potential of suppressing plant disease because of its biological properties. However, information on anaerobic digestate is limited. In this study, potential of antagonistic activities of anaerobic digestate against phytopathogens were investigated by detecting the amounts of antagonistic bacteria (Bacillus and Pseudomonas) in anaerobic digestates of dairy manure. The results showed that anaerobic digestion increased the total amounts of Bacillus and Pseudomonas in digestate. Bacillus suppressed growth of phytopathogens, while Pseudomonas did not show any antagonistic activities. These results indicated that Bacillus was an effective antagonistic bacterium in digestate against phytopathogens. Furthermore, two selected isolates, B11 (Bacillus subtilis) and B59 (Bacillus licheniformis), were applied in field experiments and showed significant reduction in percent infection of potato late blight (Phytophthora infestans). These results demonstrate the benefits of digestate in suppressing soil-borne plant diseases caused by antagonistic bacteria.