The emerging predictive and prognostic role of HER2 in HER2-negative early breast cancer: a retrospective study.

PURPOSE: Many patients with early breast cancer (eBC) undergoing neoadjuvant chemotherapy do not achieve pathological complete response (pCR), which is a prognostic factor. We examined the role of HER2-low expression in predicting pCR and prognosis in HER2-negative eBC. METHODS: We evaluated patients with stage I-III HER2-negative BC, treated between 2013 and 2023 at The Royal Marsden NHS Foundation Trust, London. Tumors were classified based on estrogen receptor (ER) status and into HER2-low and HER2-zero subgroups. We analyzed pCR rates, relapse-free survival (RFS) and overall survival (OS). RESULTS: 754 patients were included in the analysis. pCR rate was 8.9% in the ER+ /HER2-low, 16.5% in the ER+ /HER2-zero, 38.9% in the ER- ER-/HER2-low and 35.9% in the ER-/HER2-zero eBC (p < 0.001). Multivariable analysis showed a significantly lower pCR rate in HER2-low compared to HER2-zero BC in the ER+ subgroup. At a median follow-up of 63.8 months (59.9-67.4), we observed longer OS in HER2-low compared to HER2-zero patients in the overall and in the ER+ population. There was no predictive or prognostic impact of HER2-low status in the ER- population. CONCLUSION: This study supports the interpretation of HER2 status as a possible prognostic and predictive biomarker for HER2-negative eBC, especially among patients with ER+ disease.

Expanding the reach of commercial cell therapies requires changes at medical centers.

The clinical application of cell therapies is becoming increasingly important for the treatment of cancer, congenital immune deficiencies, and hemoglobinopathies. These therapies have been primarily manufactured and used at academic medical centers. However, cell therapies are now increasingly being produced in centralized manufacturing facilities and shipped to medical centers for administration. Typically, these cell therapies are produced from a patient's own cells, which are the critical starting material. For these therapies to achieve their full potential, more medical centers must develop the infrastructure to collect, label, cryopreserve, test, and ship these cells to the centralized laboratories where these cell therapies are manufactured. Medical centers must also develop systems to receive, store, and infuse the finished cell therapy products. Since most cell therapies are cryopreserved for shipment and storage, medical centers using these therapies will require access to liquid nitrogen product storage tanks and develop procedures to thaw cell therapies. These services could be provided by the hospital pharmacy or transfusion service, but the latter is likely most appropriate. Another barrier to implementing these services is the variability among providers of these cell therapies in the processes related to handling cell therapies. The provision of these services by medical centers would be facilitated by establishing a national coordinating center and a network of apheresis centers to collect and cryopreserve the cells needed to begin the manufacturing process and cell therapy laboratories to store and issue the cells. In addition to organizing cell collections, the coordinating center could establish uniform practices for collecting, labeling, shipping, receiving, thawing, and infusing the cell therapy.

Healthcare center-based cell therapy laboratories supporting off-site manufactured cell therapies: The experiences of a single academic cell therapy laboratory.

BACKGROUND: Healthcare center-based cell therapy laboratories (HC CTLs) evolved from solely processing hematopoietic stem cells for transplantation to manufacturing various advanced cellular therapies. With increasing interest in cellular therapy applications, off-site manufactured products are becoming more common. HC CTLs play a critical role in supporting these products by shipping out cellular starting material (CSM) for further manufacturing and/or receiving, storing, and distributing final products. The experiences and challenges encountered by a single academic HC CTL in supporting these products are presented. METHODS: All off-site manufacturing protocols supported before 2023 were reviewed. Collected data included protocol characteristics (treatment indication, product type), process logistics (shipping, receiving, storage, thawing, distribution, documentation), and product handling volumes (CSM shipping and final product infusions). RESULTS: Between 2012 and 2022, 15 off-site manufactured cellular therapy early-phase, single- and multicenter clinical trials were supported. Trials were sponsored by academic/research and commercial entities. The number of protocols supported annually increased each year, with few ending. Products included cancer immunotherapies and gene therapies. Autologous CSM was collected and shipped, while autologous and allogeneic final products were received, stored, thawed, and distributed. Process differences among protocols included CSM shipping conditions, laboratory analyses, final product thaw conditions and procedures, number of treatments, and documentation. DISCUSSION: HC CTLs must contend with several challenges in supporting off-site manufacturing protocols. As demand for cellular therapies increases, stakeholders should collaborate from the early phases of clinical trials to streamline processes and standardize procedures to increase value, improve safety, and reduce the burden on HC CTLs.

Optimization Conditions of Ultrasound-Assisted Extraction for Phenolic Compounds and Antioxidant Activity from Rubus alceifolius Poir Leaves.

Rubus alceifolius Poir (R.A. Poir) leaves are rich in phenolic compounds, offering many health benefits due to their incredible antioxidant potential. In this study, conditions for the ultrasound-assisted extraction (UAE) of phenolic compounds and antioxidant activity from R.A. Poir leaves were optimized using response surface methodology (RSM). This methodology assessed the effects of ultrasound power (X1: 100-500 W), extraction temperature (X2: 30-60°C), and extraction time (X3: 5-55 min). The optimized UAE conditions were then compared with conventional extraction methods (Soxhlet extraction: SE and maceration extraction: ME) for extracting total phenolics. A phenolic profile using GC-MS and antioxidant activity (ABTS) was also compared. According to the RSM, the best conditions for UAE to extract the highest total polyphenol content and ABTS radical scavenging activity were 320 W ultrasound power, 40°C extraction temperature, and 35.5 min sonication duration. Under these optimal conditions, the TPC and antioxidant activity reached 16.68 mg GAE/g dm and 21.9 mg TE/g, respectively, closely aligning with the predicted values. The UAE extraction technique proved to be more efficient in extracting phenolics and antioxidant capacity (ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)) radical scavenging activity, and enzyme inhibition) compared to the conventional extraction methods (SE and ME). A GC-MS analysis identified 12 components, including 5 phenolics and 3 flavonoids, which likely contribute to the antioxidant activity. Consequently, the UAE method improved extraction efficiency within a shorter time frame, suggesting that UAE is a promising, efficient, and ecofriendly technology for extracting bioactive compounds from R.A. Poir leaves.

An RNA Damage Response Network Mediates the Lethality of 5-FU in Clinically Relevant Tumor Types.

5-fluorouracil (5-FU) is a successful and broadly used anti-cancer therapeutic. A major mechanism of action of 5-FU is thought to be through thymidylate synthase (TYMS) inhibition resulting in dTTP depletion and activation of the DNA damage response. This suggests that 5-FU should synergize with other DNA damaging agents. However, we found that combinations of 5-FU and oxaliplatin or irinotecan failed to display any evidence of synergy in clinical trials, and resulted in sub-additive killing in a panel of colorectal cancer (CRC) cell lines. In seeking to understand this antagonism, we unexpectedly found that an RNA damage response during ribosome biogenesis dominates the drug's efficacy in tumor types for which 5-FU shows clinical benefit. 5-FU has an inherent bias for RNA incorporation, and blocking this greatly reduced drug-induced lethality, indicating that accumulation of damaged RNA is more deleterious than the lack of new RNA synthesis. Using 5-FU metabolites that specifically incorporate into either RNA or DNA revealed that CRC cell lines and patient-derived colorectal cancer organoids are inherently more sensitive to RNA damage. This difference held true in cell lines from other tissues in which 5-FU has shown clinical utility, whereas cell lines from tumor tissues that lack clinical 5-FU responsiveness typically showed greater sensitivity to the drug's DNA damage effects. Analysis of changes in the phosphoproteome and ubiquitinome shows RNA damage triggers the selective ubiquitination of multiple ribosomal proteins leading to autophagy-dependent rRNA catabolism and proteasome-dependent degradation of ubiquitinated ribosome proteins. Further, RNA damage response to 5-FU is selectively enhanced by compounds that promote ribosome biogenesis, such as KDM2A inhibitors. These results demonstrate the presence of a strong RNA damage response linked to apoptotic cell death, with clear utility of combinatorially targeting this response in cancer therapy.

Effects of extended transport on cryopreserved allogeneic hematopoietic progenitor cell (HPC) product quality and optimal methods to assess HPC stability.

BACKGROUND: Since the beginning of the COVID-19 pandemic, cryopreservation of hematopoietic progenitor cell (HPC) products has been increasingly used to ensure allogeneic donor graft availability prior to recipient conditioning for transplantation. However, in addition to variables such as graft transport duration and storage conditions, the cryopreservation process itself may adversely affect graft quality. Furthermore, the optimal methods to assess graft quality have not yet been determined. STUDY DESIGN AND METHODS: A retrospective review was performed on all cryopreserved HPCs processed and thawed at our facility from 2007 to 2020, including both those collected onsite and by the National Marrow Donor Program (NMDP). HPC viability studies were also performed on fresh products, retention vials, and corresponding final thawed products by staining for 7-AAD (flow cytometry), AO/PI (Cellometer), and trypan blue (manual microscopy). Comparisons were made using the Mann-Whitney test. RESULTS: For HPC products collected by apheresis (HPC(A)), pre-cryopreservation and post-thaw viabilities, as well as total nucleated cell recoveries were lower for products collected by the NMDP compared to those collected onsite. However, there were no differences seen in CD34+ cell recoveries. Greater variation in viability testing was observed using image-based assays compared to flow-based assays, and on cryo-thawed versus fresh samples. No significant differences were observed between viability measurements obtained on retention vials versus corresponding final thawed product bags. DISCUSSION: Our studies suggest extended transport may contribute to lower post-thaw viabilities, but without affecting CD34+ cell recoveries. To assess HPC viability prior to thaw, testing of retention vials offers predictive utility, particularly when automated analyzers are used.

Reference gene selection for clinical chimeric antigen receptor T-cell product vector copy number assays.

BACKGROUND AIMS: Reference genes are an essential part of clinical assays such as droplet digital polymerase chain reaction (ddPCR), which measure the number of copies of vector integrated into genetically engineered cells and the loss of plasmids in reprogrammed cells used in clinical cell therapies. Care should be taken to select reference genes, because it has been discovered that there may be thousands of variations in copy number from genomic segments among different individuals. In addition, within the same person in the context of cancer and other proliferative disorders, substantial parts of the genome also can differ in copy number between cells from diseased and healthy people. The purpose of this study was to identify reference genes that could be used for copy number variation analysis of transduced chimeric antigen receptor T cells and for plasmid loss analysis in induced pluripotent stem cells using ddPCR. METHODS: We used The Cancer Genome Atlas (TCGA) to evaluate candidate reference genes. If TCGA found a candidate gene to have low copy number variance in cancer, ddPCR was used to measure the copy numbers of the potential reference gene in cells from healthy subjects, cancer cell lines and patients with acute lymphocytic leukemia, lymphoma, multiple myeloma and human papillomavirus-associated cancers. RESULTS: In addition to the rPP30 gene, which we have has been using in our copy number assays, three other candidate reference genes were evaluated using TCGA, and this analysis found that none of the four gene regions (AGO1, AP3B1, MKL2 and rPP30) were amplified or deleted in all of the cancer cell types that are currently being treated with cellular therapies by our facility. The number of copies of the genes AP3B1, AGO1, rPP30 and MKL2 measured by ddPCR was similar among cells from healthy subjects. We found that AGO1 had copy number alteration in some of the clinical samples, and the number of copies of the genes AP3B1, MKL2 and rPP30 measured by ddPCR was similar among cells from patients with the cancer cell types that are currently being treated with genetically engineered T-cell therapies by our facility. CONCLUSIONS: Based on our current results, the three genes, AP3B1, MKL2 and rPP30, are suitable for use as reference genes for assays measuring vector copy number in chimeric antigen receptor T cells produced from patients with acute leukemia, lymphoma, multiple myeloma and human papillomavirus-associated cancers. We will continue to evaluate AGO1 on our future samples.

Longitudinal transcriptional analysis of peripheral blood leukocytes in COVID-19 convalescent donors.

BACKGROUND: SARS-CoV2 can induce a strong host immune response. Many studies have evaluated antibody response following SARS-CoV2 infections. This study investigated the immune response and T cell receptor diversity in people who had recovered from SARS-CoV2 infection (COVID-19). METHODS: Using the nCounter platform, we compared transcriptomic profiles of 162 COVID-19 convalescent donors (CCD) and 40 healthy donors (HD). 69 of the 162 CCDs had two or more time points sampled. RESULTS: After eliminating the effects of demographic factors, we found extensive differential gene expression up to 241 days into the convalescent period. The differentially expressed genes were involved in several pathways, including virus-host interaction, interleukin and JAK-STAT signaling, T-cell co-stimulation, and immune exhaustion. A subset of 21 CCD samples was found to be highly "perturbed," characterized by overexpression of PLAU, IL1B, NFKB1, PLEK, LCP2, IRF3, MTOR, IL18BP, RACK1, TGFB1, and others. In addition, one of the clusters, P1 (n = 8) CCD samples, showed enhanced TCR diversity in 7 VJ pairs (TRAV9.1_TCRVA_014.1, TRBV6.8_TCRVB_016.1, TRAV7_TCRVA_008.1, TRGV9_ENST00000444775.1, TRAV18_TCRVA_026.1, TRGV4_ENST00000390345.1, TRAV11_TCRVA_017.1). Multiplexed cytokine analysis revealed anomalies in SCF, SCGF-b, and MCP-1 expression in this subset. CONCLUSIONS: Persistent alterations in inflammatory pathways and T-cell activation/exhaustion markers for months after active infection may help shed light on the pathophysiology of a prolonged post-viral syndrome observed following recovery from COVID-19 infection. Future studies may inform the ability to identify druggable targets involving these pathways to mitigate the long-term effects of COVID-19 infection. TRIAL REGISTRATION: https://clinicaltrials.gov/ct2/show/NCT04360278 Registered April 24, 2020.

Growth and Distribution of Boron in Oilseed Rape (Brassica napus L.) as Affected by Boron Supply.

Oilseed rape (Brassica napus L.) is one of the most important oilseed crops. It has relatively high boron (B) requirements for growth. In this study, a hydroponic experiment was performed to determine the critical B requirement and B distribution in B. napus. The plants were grown for four weeks at a range of B levels (from 0.25 to 1000 µM) supplied in a nutrient solution. The results showed significant differences in the root and shoot dry matter and B accumulation in these tissues among the supplied B levels. Severe visible symptoms of B deficiency were observed on the leaves at levels lower than 1 µM B and toxicity at 1000 µM B in the nutrient solution. The maximum shoot and root dry matter were recorded at 25 µM B in the nutrient solution. The plants supplied with the lowest and the highest B levels produced 35% and 37% less shoot dry matter than those supplied with 25 µM B, while the corresponding decreases in the root dry matter were 48% and 36%, respectively. The critical concentration of B, which is the lowest concentration at which plants produce 90% of the maximum shoot dry matter, was proven to be 1 µM B for oilseed rape. At this level of external B supply, the B concentration in the shoot was 26.9 mg kg-1 DM. It was found that with the increase in B levels in the nutrient solution, the relative distribution of B between the roots and the shoots shifted in favor of the shoots.

The need for uniform and coordinated practices involving centrally manufactured cell therapies.

Cellular therapies have become an important part of clinical care. The treatment of patients with cell therapies often involves the collection of autologous cells at the medical center treating the patient, the shipment of these cells to a centralized manufacturing site, and the return of the cryopreserved clinical cell therapy to the medical center treating the patient for storage until infusion. As this activity grows, cell processing laboratories at many academic medical centers are involved with many different autologous products manufactured by several different centralized laboratories. The handling of these products by medical center-based cell therapy laboratories is complicated and resource-intensive since each centralized manufacturing laboratory has unique methods for labeling, storing, shipping, receiving, thawing, and infusing the cells. The field would benefit from the development of more uniform practices. The development of a coordinating center similar to those established to facilitate the collection, shipping, and transplantation of hematopoietic stem cells from unrelated donors would also be beneficial. In summary, the wide range of practices involved with labeling, shipping, freezing, thawing, and infusing centrally manufactured autologous cellular therapies lack efficiency and consistency and puts patients at risk. More uniform practices are needed.

Fetal ECG Extraction From Maternal ECG Using Attention-Based CycleGAN.

A non-invasive fetal electrocardiogram (FECG) is used to monitor the electrical pulse of the fetal heart. Decomposing the FECG signal from the maternal ECG (MECG) is a blind source separation problem, which is hard due to the low amplitude of the FECG, the overlap of R waves, and the potential exposure to noise from different sources. Traditional decomposition techniques, such as adaptive filters, require tuning, alignment, or pre-configuration, such as modeling the noise or desired signal to map the MECG to the FECG. The high correlation between maternal and fetal ECG fragments decreases the performance of convolution layers. Therefore, the masking region of interest based on the attention mechanism was performed to improve the signal generators' precision. The sine activation function was also used to retain more details when converting two signal domains. Three available datasets from the Physionet, including the A&D FECG, NI-FECG, and NI-FECG challenge, and one synthetic dataset using FECGSYN toolbox, were used to evaluate the performance. The proposed method could map an abdominal MECG to a scalp FECG with an average of 98% R-Square [CI 95%: 97%, 99%] as the goodness of fit on the A&D FECG dataset. Moreover, it achieved 99.7% F1-score [CI 95%: 97.8-99.9], 99.6% F1-score [CI 95%: 98.2%, 99.9%] and 99.3% F1-score [CI 95%: 95.3%, 99.9%] for fetal QRS detection on the A&D FECG, NI-FECG and NI-FECG challenge datasets, respectively. Also, the distortion was in the "very good" and "good" ranges. These results are comparable to the state-of-the-art results; thus, the proposed algorithm has the potential to be used for high-performance signal-to-signal conversion.

Giant-Cell Ependymoma of the Cervical Spinal Cord With Syringomyelia and Pathological Presentation: A Case Report and Review of the Literature.

Ependymomas are unusual neuroepithelial tumors of the central nervous system that arise from clusters of ependymal cells. In adults, ependymomas are the most common primary spinal cord tumors. Nevertheless, only a few cases of large-cell ependymoma have been documented; these cases often involve the brain. Here, we report the case of a 43-year-old man who had a cervical spinal cord ependymoma with syringomyelia. The giant-cell ependymoma (GCE) in the spinal cord discussed in this case emphasizes the characteristics of GCE and the discrepancy between the pathological appearance, the surgical results, and the clinically good prognosis.

One-Time Foliar Application and Continuous Resupply via Roots Equally Improved the Growth and Physiological Response of B-Deficient Oilseed Rape.

Oilseed rape (Brassica napus L.) is a high-boron (B)-demanding crop, and initially, normal growing plants might show B deficiency at advanced growth stages on soils with marginal B availability. Hence, we compared the effects of B resupply via roots and leaves on growth and physiological response, and relative expression of B transporters in B-deficient oilseed rape plants. Four-week-old plants initially grown with inadequate B (1 µM B for the first two weeks and 0.25 µM B for the next two weeks) were later grown either as such with 0.25 µM B, with 25 µM B in nutrient solution or foliar sprayed with 7 mL of 30, 60 and 150 mM B solution plant-1 as boric acid. Plants grown with 25 µM B in the nutrient solution from the beginning were included as adequate B treatment. Results showed that B resupply to B-deficient plants via roots and leaves (60 mM B) equally improved root and shoot dry matter, but not to the level of plants grown with adequate B supply. Foliar-applied 150 mM B proved toxic, causing leaf burn but not affecting dry matter. Resupply of B via roots increased B concentration in roots and leaves, while leaf-applied B did so only in leaves. Net carbon assimilation had a positive relationship with dry matter accumulation. Except for the highest foliar B level, B resupply via roots and leaves increased the accumulation of glucose, fructose and sucrose in leaves. Boron-deficient plants showed significant upregulation of BnaNIP5;1 in leaves and roots and of BnaBOR1;2 in roots. Boron resupply via roots reversed the B-deficiency-induced upregulation of BnaNIP5;1 in roots, whereas the expression of BnaBOR1;2 was reversed by both root and foliar B resupply. In leaves, B resupply by both methods reversed the expression of BnaNIP5;1 to the level of B-adequate plants. It is concluded that B resupply to B-deficient plants via roots and leaves equally but partially corrected B deficiency in B. napus grown in hydroponics.

Next-generation sequencing technologies: An overview.

Since the days of Sanger sequencing, next-generation sequencing technologies have significantly evolved to provide increased data output, efficiencies, and applications. These next generations of technologies can be categorized based on read length. This review provides an overview of these technologies as two paradigms: short-read, or "second-generation," technologies, and long-read, or "third-generation," technologies. Herein, short-read sequencing approaches are represented by the most prevalent technologies, Illumina and Ion Torrent, and long-read sequencing approaches are represented by Pacific Biosciences and Oxford Nanopore technologies. All technologies are reviewed along with reported advantages and disadvantages. Until recently, short-read sequencing was thought to provide high accuracy limited by read-length, while long-read technologies afforded much longer read-lengths at the expense of accuracy. Emerging developments for third-generation technologies hold promise for the next wave of sequencing evolution, with the co-existence of longer read lengths and high accuracy.

Boron uptake and distribution by oilseed rape (Brassica napus L.) as affected by different nitrogen forms under low and high boron supply.

Ammonium (NH4+) and nitrate (NO3-) conversely alter pH of the rooting medium, and thus differentially affect the equilibrium between boric acid and borate in soil solution. This can alter boron (B) uptake by plants, which is passive under high, but facilitated (boric acid) or active (borate) under low B supply. Therefore, the effect of NH4+ and NO3- forms was investigated on the growth, 10B uptake rate and accumulation, and expression of B transporters in Brassica napus grown with low (1 µM) or high (100 µM) 10B for five days in the nutrient solution. At the low 10B level, NO3--fed plants had the same specific 10B uptake rate, 10B accumulation and xylem 10B concentration as NH4NO3-fed plants but these attributes were reduced at the high 10B level. BnaBOR1;2 and BnaNIP5;1 were upregulated in roots of NO3-fed plants at low 10B supply. NH4+-fed plants had substantially lower dry matters; due to nutrient solution acidification (2.0 units)-induced deficiency of nitrogen, potassium, magnesium, and iron in plant shoots. Reduced transpiration rates resulted in lower 10B uptake rate and accumulation in the roots and shoots of NH4+-fed plants. BnaNIP5;1 in roots, while both BnaBOR1;2 and BnaNIP5;1 in shoots were upregulated in NH4+-fed plants at low 10B level. Collectively, NH4+-induced acidity and consequent lowering of 10B uptake induced the upregulation of B transport mechanisms, even at marginal 10B concentrations, while NO3--induced alkalinization resulted in altered B distribution between roots and shoots due to restricted B transport, especially at higher 10B supply.

Complex Linkage Disequilibrium Effects in HLA-DPB1 Expression and Molecular Mismatch Analyses of Transplantation Outcomes.

BACKGROUND: HLA molecular mismatch (MM) is a risk factor for de novo donor-specific antibody (dnDSA) development in solid organ transplantation. HLA expression differences have also been associated with adverse outcomes in hematopoietic cell transplantation. We sought to study both MM and expression in assessing dnDSA risk. METHODS: One hundred three HLA-DP-mismatched solid organ transplantation pairs were retrospectively analyzed. MM was computed using amino acids (aa), eplets, and, supplementarily, Grantham/Epstein scores. DPB1 alleles were classified as rs9277534-A (low-expression) or rs9277534-G (high-expression) linked. To determine the associations between risk factors and dnDSA, logistic regression, linkage disequilibrium (LD), and population-based analyses were performed. RESULTS: A high-risk AA:GX (recipient:donor) expression combination (X = A or G) demonstrated strong association with HLA-DP dnDSA (P = 0.001). MM was also associated with HLA-DP dnDSA when evaluated by itself (eplet P = 0.007, aa P = 0.003, Grantham P = 0.005, Epstein P = 0.004). When attempting to determine the relative individual effects of the risk factors in multivariable analysis, only AA:GX expression status retained a strong association (relative risk = 18.6, P = 0.007 with eplet; relative risk = 15.8, P = 0.02 with aa), while MM was no longer significant (eplet P = 0.56, aa P = 0.51). Importantly, these risk factors are correlated, due to LD between the expression-tagging single-nucleotide polymorphism and polymorphisms along HLA-DPB1. CONCLUSIONS: The MM and expression risk factors each appear to be strong predictors of HLA-DP dnDSA and to possess clinical utility; however, these two risk factors are closely correlated. These metrics may represent distinct ways of characterizing a common overlapping dnDSA risk profile, but they are not independent. Further, we demonstrate the importance and detailed implications of LD effects in dnDSA risk assessment and possibly transplantation overall.

Enhancing chemotherapy response through augmented synthetic lethality by co-targeting nucleotide excision repair and cell-cycle checkpoints.

In response to DNA damage, a synthetic lethal relationship exists between the cell cycle checkpoint kinase MK2 and the tumor suppressor p53. Here, we describe the concept of augmented synthetic lethality (ASL): depletion of a third gene product enhances a pre-existing synthetic lethal combination. We show that loss of the DNA repair protein XPA markedly augments the synthetic lethality between MK2 and p53, enhancing anti-tumor responses alone and in combination with cisplatin chemotherapy. Delivery of siRNA-peptide nanoplexes co-targeting MK2 and XPA to pre-existing p53-deficient tumors in a highly aggressive, immunocompetent mouse model of lung adenocarcinoma improves long-term survival and cisplatin response beyond those of the synthetic lethal p53 mutant/MK2 combination alone. These findings establish a mechanism for co-targeting DNA damage-induced cell cycle checkpoints in combination with repair of cisplatin-DNA lesions in vivo using RNAi nanocarriers, and motivate further exploration of ASL as a generalized strategy to improve cancer treatment.

Utilizing nanopore sequencing technology for the rapid and comprehensive characterization of eleven HLA loci; addressing the need for deceased donor expedited HLA typing.

The comprehensive characterization of human leukocyte antigen (HLA) genomic sequences remains a challenging problem. Despite the significant advantages of next-generation sequencing (NGS) in the field of Immunogenetics, there has yet to be a single solution for unambiguous, accurate, simple, cost-effective, and timely genotyping necessary for all clinical applications. This report demonstrates the benefits of nanopore sequencing introduced by Oxford Nanopore Technologies (ONT) for HLA genotyping. Samples (n = 120) previously characterized at high-resolution three-field (HR-3F) for 11 loci were assessed using ONT sequencing paired to a single-plex PCR protocol (Holotype) and to two multiplex protocols OmniType (Omixon) and NGSgo®-MX6-1 (GenDx). The results demonstrate the potential of nanopore sequencing for delivering accurate HR-3F typing with a simple, rapid, and cost-effective protocol. The protocol is applicable to time-sensitive applications, such as deceased donor typings, enabling better assessments of compatibility and epitope analysis. The technology also allows significantly shorter turnaround time for multiple samples at a lower cost. Overall, the nanopore technology appears to offer a significant advancement over current next-generation sequencing platforms as a single solution for all HLA genotyping needs.

Sustained effects of a multimodal campaign aiming at hand hygiene improvement on compliance and healthcare-associated infections in a large gynaecology/obstetrics tertiary-care centre in Vietnam.

BACKGROUND: Hung Vuong Hospital (HVH) is a 900-bed maternity hospital in Ho-Chi-Minh-City, Vietnam. Due to low compliance, a quasi-experimental, observational study was conducted with the aim to improve hand hygiene. METHODS: A multimodal promotion strategy was established in 2010 and further developed towards ongoing, repetitive and inventive campaigns including patient participation. Hand hygiene compliance was monitored by direct observation and healthcare-associated infections (HAIs) by applying standard definitions. RESULTS: Between 2010 and 2018, a total of 43,711 hand hygiene opportunities were observed. Compliance improved from 21.5% (95%CI: 20.2-22.8%) in 2010 to 75.1% (73.9-76.2%) in 2018 (incidence rate ratio, IRR , 1.10; 95%CI, 1.10-1.11). This was achieved through increasing recourse to alcohol-based hand rubbing. A total of 554,720 women were admitted to HVH during the study period for 353,919 deliveries (198,679 vaginal; 155,240 by C-section) and 257,127 surgical procedures. The HAI-incidence decreased significantly from 1.10 episodes per 1000 patient-days in 2010 to 0.45 per 1000 patient-days in 2018 (IRR 0.85; 95%CI, 0.79-0.90). Significant improvement was observed also for surgical site infections after gynaecological surgery (IRR 0.95; 95%CI, 0.92-0.99) and endometritis after abortion (IRR 0.80; 95%CI, 0.68-0.93). CONCLUSIONS: A multimodal strategy aiming at behaviour change significantly improved and sustained hand hygiene, which contributed to the reduction of healthcare-associated infections.

A Cost-Effective and Portable Optical Sensor System to Estimate Leaf Nitrogen and Water Contents in Crops.

Non-invasive determination of leaf nitrogen (N) and water contents is essential for ensuring the healthy growth of the plants. However, most of the existing methods to measure them are expensive. In this paper, a low-cost, portable multispectral sensor system is proposed to determine N and water contents in the leaves, non-invasively. Four different species of plants-canola, corn, soybean, and wheat-are used as test plants to investigate the utility of the proposed device. The sensor system comprises two multispectral sensors, visible (VIS) and near-infrared (NIR), detecting reflectance at 12 wavelengths (six from each sensor). Two separate experiments were performed in a controlled greenhouse environment, including N and water experiments. Spectral data were collected from 307 leaves (121 for N and 186 for water experiment), and the rational quadratic Gaussian process regression (GPR) algorithm was applied to correlate the reflectance data with actual N and water content. By performing five-fold cross-validation, the N estimation showed a coefficient of determination () of 63.91% for canola, 80.05% for corn, 82.29% for soybean, and 63.21% for wheat. For water content estimation, canola showed an of 18.02%, corn showed an of 68.41%, soybean showed an of 46.38%, and wheat showed an of 64.58%. The result reveals that the proposed low-cost sensor with an appropriate regression model can be used to determine N content. However, further investigation is needed to improve the water estimation results using the proposed device.