Search

SARS-CoV-2 is the causative agent of COVID-19. The main viral protease (Mpro) is an attractive target for antivirals. The clinically approved drug nirmatrelvir and the clinical candidate ensitrelvir have so far showed great potential for treatment of viral infection. However, the broad use of antivirals is often associated with resistance generation. Herein, we enzymatically characterized 14 naturally occurring Mpro polymorphisms that are close to the binding site of these antivirals. Nirmatrelvir retained its potency against most polymorphisms tested, while mutants G143S and Q189K were associated with diminished inhibition constants. For ensitrelvir, diminished inhibition constants were observed for polymorphisms M49I, G143S, and R188S, but not for Q189K, suggesting a distinct resistance profile between inhibitors. In addition, the crystal structures of selected polymorphisms revealed interactions that were critical for loss of potency. In conclusion, our data will assist the monitoring of potential resistant strains, support the design of combined therapy, as well as assist the development of the next generation of Mpro inhibitors. [ABSTRACT FROM AUTHOR]

Different immunohistochemical programmed death-ligand 1 (PD-L1) assays and scorings have been reported to yield variable results in triple-negative breast cancer (TNBC). We compared the analytical concordance and reproducibility of four clinically relevant PD-L1 assays assessing immune cell (IC) score, tumor proportion score (TPS), and combined positive score (CPS) in TNBC. Primary TNBC resection specimens (n = 104) were stained for PD-L1 using VENTANA SP142, VENTANA SP263, DAKO 22C3, and DAKO 28–8. PD-L1 expression was scored according to guidelines on virtual whole slide images by four trained readers. The mean PD-L1 positivity at IC-score ≥1% and CPS ≥1 ranged between 53% and 75% with the highest positivity for SP263 and comparable levels for 22C3, 28–8, and SP142. Inter-assay agreement was good between 28–8 and 22C3 across all scores and cut-offs (kappa 0.68–0.74) and for both assays with SP142 at IC-score ≥1% and CPS ≥1 (kappa 0.61–0.67). The agreement between SP263 and all other assays was substantially lower for all scores. Inter-reader agreement for each assay was good to excellent for IC-score ≥1% (kappa 0.73–0.78) and CPS ≥1 (kappa 0.68–0.74), fair to good for CPS ≥10 (kappa 0.52–0.67) and TPS ≥1% (kappa 0.53–0.72). The percentage of overlapping cases in the positive/negative category was >90% between IC-score ≥1% and CPS ≥1 but below when comparing IC-score ≥1% with CPS ≥10. We demonstrate an overall good inter-reader agreement for all PD-L1 assays in TNBC along with assay specific differences in positivity and concordances, which may aid to select the right test strategy in routine diagnostics. • Different PD-L1 IHC assays and scorings may show variable results in TNBC. • Overall good assay concordance between SP142, 22C3, and 28–8 at IC-score 1%. • Overall good assay concordance between SP142, 22C3, and 28–8 at CPS 1. • SP142 is less optimal for CPS assessment at higher cut-offs. • SP263 assay is not interchangeable with the other three PD-L1 assays. [ABSTRACT FROM AUTHOR]

Data-based methods are capable to monitor machine components. Approaches for semi-supervised anomaly detection are trained using sensor data that describe the normal state of machine components. Thus, such approaches are interesting for industrial practice, since sensor data do not have to be labeled in a time-consuming and costly way. In this work, an ensemble approach for semi-supervised anomaly detection is used to detect anomalies. It is shown that the ensemble approach is suitable for condition monitoring of ball screws. For the evaluation of the approach, a data set of a regular test cycle of a ball screw from automotive industry is used. [ABSTRACT FROM AUTHOR]

Adjuvant chemotherapy decision in patients with hormone receptor positive, HER2 negative breast cancer (BC) is challenging. Ki-67 is widely used for adjuvant therapy decision in BC. The multigene assay EndoPredict (EP) has shown to provide valid and additional information about the risk of recurrence compared to traditional pathological factors. In this study, we compared Ki-67 with EP assay generated risk groups. We analyzed the results from prospective EP testing (n = 373) and tumor proliferation assessed by Ki-67 staining in luminal breast cancer. We statistically investigated the association of both parameters and probed for equivalence in risk stratification. Evaluation of Ki-67 was feasible in 307 (82%) BC specimens with known EP test results. The EPscore (now called 12-gene molecular score) delineated 140 low and 167 high scores. After combining the EPscore with pathological tumor stage and nodal status, we received 203 EPclin low-risk and 104 EPclin high-risk classifications. EPscore and EPclin were significantly associated with Ki-67 indices and tumor grade (p < 0.001). Overall, we observed a moderate correlation between Ki-67 and the EPscore (r = 0.63) as well as the EPclin score (r = 0.59). Ki-67 values above 25% partly overlap with EP test results and therefore indicate a high-risk profile. In these cases, the additional prognostic information from EP testing might be rather low. However, low and intermediate Ki-67 values (less than 25%) alone were not reliable in predicting a low risk EP profile, indicating that EP testing is useful in this subgroup. • Comparison of prospective EndoPredict tests with Ki-67 in a large breast cancer cohort. • EPscore and EPclin were significantly associated with Ki-67 indices. • Ki-67 values above 25% partly overlap with high-risk EP test results. • Ki-67 less than 25% was not reliable in predicting a low-risk EP profile. • EP testing is useful, particularly in breast cancer with low and intermediate Ki-67 levels. [ABSTRACT FROM AUTHOR]

Immune cell infiltration in breast cancer is important for the patient's prognosis and response to systemic therapies including immunotherapy. We sought to investigate the prevalence of tumour-infiltrating lymphocytes (TILs) and their association with immune checkpoints such as programmed cell death 1 (PD-1) and programmed cell death ligand 1 (PD-L1) in high-risk, node-positive breast cancer of the adjuvant German Adjuvant Intergroup Node–positive (GAIN-1) trial. We evaluated TILs by haematoxylin and eosin staining and PD-1 and PD-L1 (SP263 assay) expression by immunohistochemistry in 1318 formalin-fixed, paraffin-embedded breast carcinomas. The association of TILs with PD-1, PD-L1, molecular intrinsic subtypes, outcome and therapy regimens (dose-dense [dd] epirubicin, paclitaxel and cyclophosphamide [EPC] and dd epirubicin, cyclophosphamide, paclitaxel and capecitabine [EC-PwX]) was statistically tested. Overall TILs density was significantly associated with the expression of PD-1 and PD-L1 in immune cells (each p < 0.0001) and PD-L1 in tumour cells (p = 0.0051). TILs were more common in triple-negative breast cancer (TNBC) and human epidermal growth factor receptor 2 (HER2)-positive tumours (each p < 0.0001). On multivariate Cox regression analyses, patients with breast cancer without TILs had an unfavourable disease-free survival (DFS) in the EPC arm compared with the EC-PwX arm (hazard ratio [HR] = 0.69 [0.44–1.06], p = 0.0915); but no differences were seen in tumours with TILs (HR = 1.24 [0.92–1.67], p = 0.1566, interaction p = 0.0336). PD-1–positive immune cells in TNBC were associated with a significantly better DFS (HR = 0.50 [0.25–0.99], p = 0.0457). PD-L1 expression had no impact on patient outcome. TILs predict the benefit of intensified ddEPC compared with ddEC-PwX therapy in node-positive, high-risk breast cancer. TILs, PD-1 and PD-L1 are linked to each other indicating tumour immunogenicity. Moreover, PD-1–positive immune cells have a positive prognostic impact in TNBC. NCT00196872. • Tumour infiltrating lymphocyts (TILs) are predictive for dose-dense EPC therapy as compared to dose dense EC-PwX in node-positive breast cancer. • TILs correlate with PD-1 and PD-L1 expression. • TILs, PD-1 and PD-L1 show the highest levels in triple-negative breast cancer (TNBC). • TILs and PD-1–positive immune cells have prognostic impact in TNBC. • PD-L1 expression is more common in immune than in tumor cells but has no prognostic value. [ABSTRACT FROM AUTHOR]

Background: Cocoa flavanol intake, especially that of (2)-epicatechin, has been linked to beneficial effects on human cardiovascular function. However, cocoa also contains the methylxanthines theobromine and caffeine, which may also affect vascular function. Objective: We sought to determine whether an interaction between cocoa flavanols and methylxanthines exists that influences cocoa flavanol-dependent vascular effects. Design: Test drinks that contained various amounts of cocoa flavanols (0-820 mg) and methylxanthines (0-220 mg), either together or individually, were consumed by healthy volunteers (n = 47) in 4 different clinical studies--3 with a randomized, double-masked crossover design and 1 with 4 parallel crossover studies. Vascular status was assessed by measuring flow-mediated vasodilation (FMD), brachial pulse wave velocity (bPWV), circulating angiogenic cells (CACs), and blood pressure before and 2 h after the ingestion of test drinks. Results: Although cocoa flavanol intake increased FMD 2 h after intake, the consumption of cocoa flavanols with methylxanthines resulted in a greater enhancement of FMD. Methylxanthine intake alone did not result in statistically significant changes in FMD. Cocoa flavanol ingestion alone decreased bPWVand diastolic blood pressure and increased CACs. Each of these changes was more pronounced when cocoa flavanols and methylxanthines were ingested together. It is important to note that the area under the curve of the plasma concentration of (2)-epicatechin metabolites over time was higher after the co-ingestion of cocoa flavanols and methylxanthines than after the intake of cocoa flavanols alone. Similar results were obtained when pure (2)-epicatechin and the methylxanthines theobromine and caffeine were consumed together. Conclusion: A substantial interaction between cocoa flavanols and methylxanthines exists at the level of absorption, in which the methylxanthines mediate an increased plasma concentration of (2)-epicatechin metabolites that coincides with enhanced vascular effects commonly ascribed to cocoa flavanol intake. This trial was registered at clinicaltrials.gov as NCT02149238. [ABSTRACT FROM AUTHOR]

In models of triple-negative breast cancer (TNBC), it has recently been shown that the epidermal growth factor receptor (EGFR) pathway is up-regulated in the aldehyde dehydrogenase 1 (ALDH1)--positive cancer stem cell fraction. Because high-grade serous ovarian carcinoma (HGSC) reveals strong molecular similarities to TNBC, we aimed to investigate the potential link between ALDH1 and EGFR in this entity. Expression of ALDH1 was investigated in 131 primary HGSCs using immunohistochemistry. Expression data were correlated with EGFR expression as well as with clinicopathologic parameters and survival. Forty-two carcinomas (32.1%) were positive for ALDH1 protein expression. Data on EGFR expression were available for 112 tumors. In these cases ALDH1 was significantly linked to EGFR expression (P < .0001). ALDH1 positivity was a significant negative prognostic factor for overall survival both in univariate (P = .010) and in multivariate survival analyses (P = .041). Tumors that were positive for both ALDH1 and EGFR had an exceptionally bad prognosis as compared with carcinomas with 1 or both markers negative in univariate analysis (P < .0001) and in the multivariate setting (P = .004). Our study suggests that similar to TNBC, there is a link between ALDH1 and EGFR expression in HGSC. Double positivity for both markers identifies a subgroup of highly aggressive, poor-prognosis cancers for which alternative treatment options--potentially EGFR-targeting drugs--should be evaluated. [ABSTRACT FROM AUTHOR]

Abstract: Recombinant adenoviruses are widely used in basic virology research, therapeutic applications, vaccination studies or simply as a tool for genetic manipulation of eukaryotic cells. Dependent on the application, transient or stable maintenance of the adenoviral genome and transgene expression are required. The newest generation of recombinant adenoviral vectors is represented by high-capacity adenoviral vectors (HC-AdVs) which lack all viral coding sequences. HC-AdVs were shown to result in long-term persistence of transgene expression and phenotypic correction in small and large animal models with negligible toxicity. Although there is evidence that adenoviral vectors predominantly persist as episomal DNA molecules with a low integration frequency into the host genome, detailed information about the nuclear fate and the molecular status of the HC-AdV genome once inside the nucleus is lacking. In recent years we have focused on analyzing and modifying the nuclear fate of HC-AdVs after infection of mammalian cells. We have focused on investigating the molecular DNA forms of HC-AdV genomes and we have designed strategies to excise and stably integrate a transgene from an episomal adenovirus vector genome into the host chromosomes by recombinases. This review article provides a state-of-the art overview of the current knowledge of episomal HC-AdV persistence and it discusses strategies for changing the nuclear fate of a transgene inserted into the HC-AdV genome by somatic integration into host chromosomes. [Copyright &y& Elsevier]

Abstract: Salvage harvesting may represent an important additional impact on sediment and nutrient transfer in forest environments affected by wildfire. However, few studies have examined the effect of this management practice on stream water quality and catchment nutrient exports. In this paper, we investigate nutrient losses following post-wildfire salvage harvesting of a radiata pine plantation catchment compared to an adjacent natural eucalypt forest catchment that was also burnt but not harvested. The study catchments form part of the long-term Cropper Creek Hydrology Project (established in 1975) that is situated in south-eastern Australia. Post-fire monitoring (2007–2009) involved collection of both weekly and flow proportional water samples that were compared with previously reported data from samples collected prior to the fire (1997–2003). It was found that median values of total suspended solids (TSS) and turbidity returned to pre-fire levels within 3years in both catchments, whereas maximum levels during storm events in the harvested pine catchment continued to exceed the eucalypt catchment. This reflected a previously reported large increase in post-fire sediment exports from the harvested pine catchment that was a minimum 180 times the eucalypt catchment over the study period. In contrast, the impact of harvesting on solute concentrations (nitrate-N, P, S, Cl, Na, K, Ca, Mg) was minor and most solutes returned to pre-fire levels within 2–3years in both catchments. Nutrient exports from the pine catchment exceeded the eucalypt catchment by 102 times for particulate P associated with suspended material compared to 1.9–4 times for solutes. The post-fire changes in solute concentrations were generally similar for both catchments and the increase in solute exports was largely a result of greater discharge after the fire and harvesting compared to the burnt eucalypt catchment. The post-fire loss of particulate P in suspended sediment and bedload from the pine catchment for the study period was estimated at a minimum of 11kgha−1 and together with the estimated loss of P from burning and the removal of the pines represented approximately 6% of the total P in surface soil and fertilizer applied to the plantation. [Copyright &y& Elsevier]

Abstract: Erosion studies at multiple scales have shown selectivity in particle transport and delivery can be inferred from nutrient enrichment. Most studies reveal the enrichment ratio (ER) of phosphorus on sediment is high, demonstrating the deposition of coarser particles over the transport pathway. Experimental work on burnt landscapes often takes place at the hillslope plot scale (1–10m2) by necessity, with inferences made on system sediment and nutrient responses from small areas. Scale effects on generation and delivery need to be considered in such cases. In a study in wet eucalypt forests burnt by wildfire, phosphorus concentration on mineral sediment and organic material at the point (0.01m2), plot (1–10m2) and catchment (106 m2) was used to estimate ERs over two scales. The data revealed ERs of 1.5 from point to plot and 2 from plot to catchment. These ratios are relatively low compared with other studies. We suggest the principal reason is short transport pathways which act to decrease deposition, due to the spatial heterogeneity of post-burn soil hydraulic properties. The association of total phosphorus with mineral material was slightly higher than with organic matter. The study suggests that using plot scale hillslope experiments to infer sediment and nutrient loss after wildfire would have overestimated losses by around 100%. [Copyright &y& Elsevier]

Summary: Antitumor inflammatory response is known to inhibit tumor growth in colorectal carcinoma. The density and functionality of tumor-infiltrating lymphocytes (TIL) is regulated by the antigen processing machinery through regulator proteins such as transporters associated with antigen processing (TAP) and major histocompatibility complex (MHC) class I antigen. We aimed to investigate the in vivo association of those factors and their impact on prognosis in colorectal cancer. TAP1, TAP2 and MHC class I antigen expression, inflammatory infiltrate and TIL (CD4+, CD8+, and CD20+) were assessed by immunohistochemistry in 336 sporadic colorectal carcinomas. The factors were correlated with each other and with clinic-pathological parameters and patient outcome. We found TAP1 and TAP2 expression to be significantly associated with MHC class I antigen expression (TAP1: r = 0.363, P < .001; TAP2: r = 0.393, P < .001). Increased density of CD8+ TIL was predominantly found in TAP1, TAP2 and MHC class I antigen–positive cases. Increased density of CD4+ TIL was linked with TAP1 and TAP2, but not with MHC class I antigen. High CD4+ and CD8+ cell count but not TAP1, TAP2 and MHC class I antigen expression had favorable prognostic impact in colorectal cancer (P = .003 and P = .003, respectively). In conclusion, our data show that the expression of key components of the antigen processing machinery is tightly linked to the density of TIL, which are positive prognostic factors in colorectal cancer in vivo. This implies that modulation of these factors may help to enhance antitumor inflammatory response which in turn may improve patient prognosis. [Copyright &y& Elsevier]

Abstract: Isothermal titration microcalorimetry (ITC) is shown here to be a sensitive and accurate method for assaying the steady-state enzyme activity of the Na+,K+-ATPase. Single ATP injection experiments yield an apparent enthalpy change for the ATP hydrolysis reaction catalyzed by the enzyme of −51 (±1) kJ mol− 1. This value is independent of the amount of ADP accumulated in the sample cell, which indicates that under the experimental conditions studied here (saturating Na+ and K+ concentrations) ADP does not inhibit enzyme activity by reversal of the phosphorylation reaction and resynthesizing ATP. Multiple ATP injection titration experiments in which varying concentrations of ADP were initially included in the sample cell could be adequately explained by a Michaelis–Menten kinetic model incorporating noncompetitive inhibition. This suggests that ADP inhibits the enzyme by binding to more than one enzyme intermediate and inhibiting forward reactions of the enzyme. Values of K m and K I obtained for the fits agree with literature values obtained by other methods. Because ITC is a direct method of continually monitoring enzyme activity, it is a valuable supplement to less direct or noncontinuous methods such as colorimetric, enzyme-coupled or radioactivity-based assays. [Copyright &y& Elsevier]

Summary: The biological behavior and the optimal management of benign breast lesions with uncertain malignant potential, the so-called B3 lesions, found in breast needle core biopsies is still under debate. We addressed this study to compare histologic findings in B3 needle core biopsies with final excision specimens to determine associated rates of malignancy. Consecutive needle core biopsies were performed in a 3-year period (January 1, 2006-December 31, 2008). Biopsies were image-guided (31 by ultrasound, 85 stereotactic vacuum-assisted, 6 unknown) for evaluation of breast abnormalities. We reviewed 122 needle core biopsies with B3 lesions of 91 symptomatic patients and 31 screen-detected women and compared the B3 histologic subtypes with the final excision histology. A total of 1845 needle core biopsies were performed and B3 lesions comprised 6.6% of all B categories. The most common histologic subtype in biopsies was flat epithelia atypia in 35.2%, followed by papillary lesions in 21% and atypical ductal hyperplasia in 20%. Reports on excision specimens were available in 66% (81 patients). Final excision histology was benign in 73 (90.2%) and malignant in 8 (9.8%) patients (2 invasive cancer, 6 ductal carcinoma in situ). Of all B3 subtypes, atypical ductal hyperplasia and flat epithelial atypia were associated with malignancy, whereas only atypical ductal hyperplasia was accompanied by invasive cancer. Of all lesions, flat epithelial atypia was most frequently found in excision specimens (18%). In our study, flat epithelial atypia and atypical ductal hyperplasia are common lesions of the B3 category in needle core biopsies of the breast. Both lesions are associated with malignancy, whereas only atypical ductal hyperplasia was related to invasive cancer. We conclude that an excision biopsy after diagnosis of flat epithelial atypia is recommended depending on clinical and radiologic findings. [Copyright &y& Elsevier]

Abstract: The measurement of soil erosion rates under natural rainfall conditions is costly and time consuming. Data provided by rainfall simulation and static site measurements can be used to predict erosion rates under natural conditions, however the accuracy of this method is largely untested. This is especially true for erosion rates from unsealed forest roads. In this study, the values for a range of erodibility indices calculated from rainfall simulation experiments are compared to observed erodibility index values from 1 year of detailed in-situ erosion monitoring of seven different forest road types. The prediction of phosphorus generation rates from rainfall simulation and/or soil sampling was also evaluated. The results showed that a series of commonly used erodibility indices such as sediment per unit rainfall, sediment per unit runoff, sediment per unit EI 30, and sediment per unit rainfall energy were poorly predicted from the rainfall simulation experiments. Five of the six indices tested substantially overpredicted the observed erodibility at one site, a gravel road subjected to minimal traffic. For the other six road sites predictions were poor and highly variable, the coefficient of efficiency ranging from −13.32 to 0.17 for these erodibility indices. A modified index, the ratio of sediment per unit rainfall energy to the mean rate of rainfall energy input, was able to predict annual erosion rates from six different road surfaces using rainfall simulation data with a coefficient of efficiency of 0.9. The results indicate that existing erodibility indices are not suitable for predicting observed erosion rates on forest roads using rainfall simulation data as collected in this study. It is argued that the modified index is more suited to sites (such as compacted roads) where interrill processes dominate, and erosion rates are less sensitive to peak flows. With respect to nutrient generation rates, rainfall simulation was able to accurately predict the observed proportion by mass of total phosphorus (TP) in runoff with a coefficient of efficiency of 0.96. Direct soil sampling of the road surface could also be used to predict the proportion by mass of TP in runoff. Concentrations of total nitrogen in forest road materials were found to be at the lower detection limit of the laboratory instruments. [Copyright &y& Elsevier]

Abstract: The protein kinase AKT is involved in several signaling pathways that are important for tumor development and progression, suggesting that AKT might be an interesting target for a molecular tumor therapy. In this study, we investigated the AKT expression in ovarian carcinomas and the role of the AKT isoforms to ovarian cancer cell proliferation. We observed an increased AKT expression in 58% of the primary ovarian carcinomas as compared to normal ovaries by immunohistochemistry. AKT expression was significantly associated with positive lymph node status (P=0.002) and advanced FIGO stage (P=0.009). In western blot analysis, total AKT was expressed in all ovarian cancer cell lines and HOSE cells, while phosphorylated AKT was only observed in OVCAR-3 and SKOV-3 cells. The isoforms AKT1 and AKT2 were expressed at the mRNA level in all cell lines, while no relevant AKT3 mRNA levels were detected by conventional and quantitative RT-PCR. To determine the effects on cell proliferation, we used the unselective PI3K-inhibitor LY294002 as well as RNA interference to selectively inhibit the AKT isoforms. Treatment with LY294002 and the AKT2 siRNA reduced proliferation of OVCAR-3 cells. Our results show that AKT is expressed in a subpopulation of advanced ovarian carcinomas suggesting a role for this protein in the progression of this entity. Deactivation of AKT, especially AKT2 can result in reduction of cell growth. Accordingly, AKT is an interesting target for therapeutic intervention in ovarian cancer. [Copyright &y& Elsevier]

Abstract: Background: Using cerebral microdialysis, baseline values for energy-related chemical markers have been reported in awake patients. Radionuclide studies have demonstrated a locally decreased metabolism, thought to be the result of brain retraction. These baseline values, however, may not be applicable to patients after surgical aneurysm repair following a subarachnoid hemorrhage (SAH). We assessed metabolic chemical marker levels in World Federation of Neurological Surgeons Committee (WFNS) grade I SAH patients after aneurysm surgery and compared them with previously reported baseline values. Methods: In 5 WFNS grade I SAH patients, energy-related chemical marker levels were obtained using microdialysis in the area of brain retraction after aneurysm surgery. In addition, an [18F]2-deoxy-d-glucose positron emission tomography (FDG-PET) was performed. Results: The FDG-PET showed a decrease of glucose metabolism in the frontotemporal area. Comparing the mean values for chemical markers of this study with reported baseline values, the most striking difference was a mild decrease of pyruvate and an increase of the lactate/pyruvate ratio. In individual patients, some markers indicated possible ischemia. A consistent pattern or ischemic profile for all markers, however, was not found. Conclusion: FDG-PET scanning confirmed postoperative metabolic changes found in previous studies. Mean interstitial chemical marker levels ranged from normal to mildly deviant compared with reference chemical marker levels for awake patients and are likely to be applicable in SAH patients after aneurysm repair. [Copyright &y& Elsevier]

We sought to evaluate serum concentrations of cathepsin K in peripheral blood and to determine whether they correlated with bone-mineral density (BMD) and the incidence of nontraumatic fractures. We took blood samples from 162 patients (101 with osteoporosis, 48 with osteopenia) and 13 healthy controls, then conducted quantitative measurements of cathepsin K using a commercially available enzyme-linked immunosorbent assay. Cathepsin K concentrations were correlated with the incidence of nontraumatic fracture, BMD, markers of bone turnover (alkaline phosphatase, bone-specific alkaline phosphatase, osteocalcin, parathyroid hormone, and C-telopeptide). The correlations between cathepsin K concentrations in subjects without fractures and in those with multiple nontraumatic fractures were statistically significant (t = ?2.1, degrees of freedom = 107, P = .036). The cathepsin K levels of controls and patients with osteoporosis were significantly different (t = ?3.7, degrees of freedom = 58.9, p>0.0001) These results suggest that the serum level of cathepsin K could serve as a marker for fracture prediction and BMD. [Copyright &y& Elsevier]

Abstract: The ubiquitously expressed actin-binding protein, gelsolin, is known to play a role in the modulation of the actin network and in the regulation of cell growth and cell motility. In the present study, we analysed the expression of gelsolin in 241 matched cDNA pairs from human normal and tumour tissues using a Cancer Profiling Array. We found a decreased expression of gelsolin in cancer tissue from female reproductive organs, including the ovary. On a protein level, we examined the expression of gelsolin in human ovarian cancer cell lines and in a set of 110 cases of human benign and malignant ovarian tumours. Low levels of gelsolin protein were observed in four of six ovarian carcinoma cell lines, in contrast to its expression in normal ovarian surface epithelial cells. In addition, we found a reduced expression of gelsolin in borderline tumours and ovarian carcinomas compared with the epithelium of normal ovaries and benign adenomas. Decreased gelsolin expression was associated with poorly differentiated carcinomas (p=0.014). No significant association between gelsolin expression and other clinicopathological markers or patient survival could be established. In addition, we investigated the growth regulatory function of gelsolin in human ovarian cancer cell lines using cDNA transfections. Re-expression of gelsolin in OAW42 and ES-2 cells resulted in a suppression of tumour cell survival in vitro. To explore the mechanism responsible for the downregulation of gelsolin expression in ovarian carcinoma cells, we treated cells with inhibitors of DNA methylation and histone deacetylation. We observed an upregulation of gelsolin in ovarian cancer cells after treatment with both types of inhibitor. Our results suggest that gelsolin might be involved in the growth regulation of human ovarian cancer. [Copyright &y& Elsevier]

[Display omitted] • MSCs offer substantial therapeutic potential, but are notoriously hard to transfect. • Various tyrosine-modified linear/branched PEI/PPI-G4 were tested for siRNA delivery. • Some candidates show high knockdown efficacy and biocompatibility. • PPI-G4-Y dendrimers are found most efficient for siRNA transfection into MSCs. • High gene knockdown is achieved in the absence of cytotoxic and genotoxic effects. Mesenchymal stromal cells (MSCs) are multipotent cells derived from different sources and able to differentiate into distinct cell lineages. For their possible biomedical application, the "tuning" of MSCs also involves the specific knockdown of defined target genes. A major limitation, however, is the notoriously low transfection efficacy especially of primary MSCs. In this paper, we systemically analyze a large set of tyrosine-modified linear or branched low molecular weight polyethylenimines (PEIs) of different sizes, as well as the tyrosine-modified polypropylenimine dendrimer PPI-G4, for their capacity of non-viral siRNA transfection into umbilical cord-derived MSCs from two different donors. Knockdown efficacies are determined on the molecular level and confirmed in functional assays. Beyond the determination of cell viabilities, acute cytotoxicity, induction of apoptosis/necrosis and mitochondrial membrane alterations are also studied. On the molecular level, caspase activation, ROS induction and genotoxic effects are analyzed. Major differences are observed between the various tyrosine-modified PEIs, with some candidates showing high knockdown efficacy and biocompatibility. PPI-G4-Y dendrimers, however, are identified as most efficient for siRNA transfection into MSCs. PPI-G4-Y/siRNA nanoparticles lead to particularly high gene knockdown, without cytotoxic and genotoxic effects on the cellular and molecular level, and are thus particularly well-suited for the tuning of MSCs. [ABSTRACT FROM AUTHOR]

Therapeutic gene silencing by RNA interference relies on the safe and efficient in vivo delivery of small interfering RNAs (siRNAs). Polyethylenimines are among the most studied cationic polymers for gene delivery. For several reasons including superior tolerability, small linear PEIs would be preferable over branched PEIs, but they show poor siRNA complexation. Their chemical modification for siRNA formulation has not been extensively explored so far. We generated a set of small linear PEIs bearing tyrosine modifications (LP x Y), leading to substantially enhanced siRNA delivery and knockdown efficacy in vitro in various cell lines, including hard-to-transfect cells. The tyrosine-modified linear 10 kDa PEI (LP10Y) is particularly powerful, associated with favorable physicochemical properties and very high biocompatibility. Systemically administered LP10Y/siRNA complexes reveal antitumor effects in mouse xenograft and patient-derived xenograft (PDX) models, and their direct application into the brain achieves therapeutic inhibition of orthotopic glioma xenografts. LP10Y is particularly interesting for therapeutic siRNA delivery. For therapeutic siRNA delivery, we generated and analyzed a set of small linear polyethylenimines bearing tyrosine modifications (LP x Y). Substantially enhanced siRNA delivery and knockdown efficacy, favorable physicochemical nanoparticle properties, very high biocompatibility and therapeutic efficacy were observed in vitro and in different tumor models in vivo. [Display omitted] • Polyethylenimines (PEIs) are attractive systems for siRNA delivery in vitro/in vivo. • Here, a set of small linear PEIs bearing tyrosine modifications (LP x Y) is explored. • LP x Y show markedly enhanced transfection efficacy also in hard-to-transfect cells. • LP x Y/siRNA complexes offer very favorable physical properties and biocompatibility. • Nanoparticle efficacies are seen in vitro , ex vivo and in three tumor models in vivo. [ABSTRACT FROM AUTHOR]

The protozoan Giardia lamblia (GL) and the bacterium Helicobacter pylori (HP) are common causes of gastrointestinal disease. Coinfection is common and has been reported in studies from Africa, Europe, North America and Asia, but data for Switzerland are scarce. To investigate GL and HP prevalence and coinfection rate in gastrointestinal biopsies from the Zurich area of Switzerland. Cases were retrieved from the laboratory information system (Medica Institute of Clinical Pathology, Zurich, Switzerland). Histological slides of cases with GL were reviewed, as were the concurrent gastric biopsies, where available. Between January 1, 2013 and December 31, 2020, GL was found in 88 (0.14%) of 62,402 patients with a small intestine biopsy and HP in 10,668 (15.5%) of 68,961 patients with a gastric biopsy. 74/88 (84.1%) of patients with GL had unremarkable small intestine biopsies, 13/88 (14.8%) had increased intraepithelial lymphocytes, 5/88 (5.7%) showed villous atrophy and 2/88 (2.3%) acute inflammation. 71/88 patients (80.7%) with GL had an available gastric biopsy, of which 12/71 (16.9%) were unremarkable, 28/71 (39.4%) had HP-associated gastritis, 11/71 (15.5%) showed reactive gastropathy and 1/71 (1.4%) had autoimmune gastritis. Coinfection with HP is common in patients with GL in gastrointestinal biopsies from the Zurich area of Switzerland. Therefore, gastroenterologists should consider sampling the stomach when GL is suspected for evaluation of possible concurrent HP-associated gastritis. Likewise, pathologists should scrutinize any small intestine biopsy for the presence of GL when HP-associated gastritis is seen, and vice versa. • For Switzerland, data on G. lamblia and H. pylori coinfection are scarce. • In our series, G. lamblia was found in ≈1 of 700 small intestine biopsies. • Likewise, ≈1 of 7 gastric biopsies harbored H. pylori -associated gastritis. • Coinfection was common and was seen in ≈4 of 10 G. lamblia positive cases. • Gastroenterologists and pathologists should be aware of this in daily practice. [ABSTRACT FROM AUTHOR]

Atezolizumab (an anti–PD-L1 antibody) has shown clinical activity alone or in combination with nab-paclitaxel in patients (pts) with first-line metastatic TNBC who have PD-L1 expression on their tumour-infiltrating immune cells (IC). We analysed the performance of 4 PD-L1 IHC assays for PD-L1 IC expression in TNBC. Thirty archival TNBC tissue specimens were selected from a set of 107 based on PD-L1 IC expression per VENTANA SP142 (<1%: 15 cases; 1–5%: 7 cases; >5%: 8 cases), to represent the distribution of PD-L1 IC–positivity in the pivotal atezolizumab studies (Emens et al., SABCS 2018; JAMA Oncol 2019). Serial histologic sections were stained with VENTANA SP142 and SP263, and DAKO 22C3 and 28-8, per manufacturer protocols. Slides were blinded for both assay and sample information and scored by trained readers at 7 sites for PD-L1 IC expression (% per tumour area), as well as tumour cell expression (≥1% vs < 1%), by online virtual microscopy. Adjusted means of PD-L1 IC staining ranged from 3.7% to 7.8% (Table); SP263 stained more IC than the other assays. Pairwise comparison of adjusted means showed small, non-significant differences (–1.2% to 0.6%) between SP142, 22C3 and 28-8, but a significant increase in PD-L1 staining for SP263 vs the other assays (3.0% to 4.2%). Intra-class correlations (ICC) for the assays showed moderate (0.460) to excellent (0.805) reader concordance (Table). Pre-specified allocation to a 1% binary IC cut-off revealed good-to-high inter-reader agreement (Kappa 0.589 to 0.789). Table 359P Table Assay PD-L1 on IC, % (95% CI) * Reader ICC (95% CI) † SP263 7.8 (7.1–8.6) 0.616 (0.477–0.758) SP142 4.3 (3.5–5.0) 0.805 (0.710–0.887) 22C3 3. 7 (2.9–4.4) 0.605 (0.474–0.755) 28-8 4.9 (4.1–5.6) 0.460 (0.319–0.636) * Sample and reader adjusted means; † Between 7 readers The results of this first multicentre PD-L1 assay comparison study in TNBC indicate good-to-high reproducibility and concordance of PD-L1 IC expression between the SP142, 22C3 and 28-8 assays, while higher PD-L1 IC expression levels were detected with SP263. Hence, SP142, 22C3 and 28-8 may be considered analytically interchangeable for PD-L1 IC testing. Roche study ID SL41336. Support for third-party writing assistance for this abstract, furnished by Katie Wilson, PhD, of Health Interactions, was provided by Roche Pharma AG, Grenzach-Wyhlen, Germany. F. Hoffmann-La Roche Ltd. Roche Pharma AG, Grenzach-Wyhlen, Germany. A. Noske: Travel / Accommodation / Expenses: Roche; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. J. Ammann: Full / Part-time employment: Roche Pharma AG, Germany; Shareholder / Stockholder / Stock options: Roche; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. D. Wagner: Research grant / Funding (institution), Travel / Accommodation / Expenses: Roche; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. C. Denkert: Honoraria (institution): Teva; Honoraria (self): Roche; Honoraria (self): Celgene; Honoraria (self): Amgen; Advisory / Consultancy: Daiichi; Advisory / Consultancy: MSD; Shareholder / Stockholder / Stock options: Sividon Diagnostics; Shareholder / Stockholder / Stock options: Myriad; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG. A. Lebeau: Advisory / Consultancy: Roche Pharma AG, Germany; Advisory / Consultancy: Novartis; Research grant / Funding (self): Roche Pharma AG, Germany; Research grant / Funding (self): Sysmex Europe; Research grant / Funding (self): BioNTech Diagnostics; Travel / Accommodation / Expenses: Roche Pharma AG, Germany; Travel / Accommodation / Expenses: Novartis; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. P. Sinn: Honoraria (self), Research grant / Funding (self), Travel / Accommodation / Expenses: Roche; Honoraria (institution), Travel / Accommodation / Expenses: Nanostring; Honoraria (self), Travel / Accommodation / Expenses: Genomic Health; Research grant / Funding (self): Dietmar-Hopp-Stiftung; Full / Part-time employment: Univ. Heidelberg; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. H. Kreipe: Advisory / Consultancy: Roche Pharma AG, Germany; Advisory / Consultancy: Novartis; Advisory / Consultancy: AstraZeneca; Advisory / Consultancy: Genomic Health; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. G. Baretton: Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution), Travel / Accommodation / Expenses: Roche; Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution), Travel / Accommodation / Expenses: BMS; Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution): MSD; Honoraria (self), Advisory / Consultancy: AstraZeneca; Honoraria (self), Advisory / Consultancy: Pfizer; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. K. Steiger: Research grant / Funding (institution): Roche; Research grant / Funding (institution): BMS; Research grant / Funding (institution): MSD; Research grant / Funding (institution): Bruker Daltonics; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. M. Kiechle: Honoraria (self), Travel / Accommodation / Expenses: AstraZeneca; Honoraria (self), Travel / Accommodation / Expenses: Myriad; Research grant / Funding (self): German Cancer Aid; Research grant / Funding (self): DFG; Shareholder / Stockholder / Stock options: Therawis Diagnostics GmBH; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. S. Hieke-Schulz: Full / Part-time employment: Roche Pharma AG, Germany; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. W. Roth: Honoraria (self): Roche; Honoraria (self): AstraZeneca; Honoraria (self): MSD; Honoraria (self): Merck; Honoraria (self): Bayer; Honoraria (self): Novartis; Honoraria (self): Chugai; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. W. Weichert: Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution): MSD; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution): BMS; Advisory / Consultancy, Speaker Bureau / Expert testimony: AstraZeneca; Advisory / Consultancy: Pfizer; Advisory / Consultancy: Merck; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution): Roche; Advisory / Consultancy, Speaker Bureau / Expert testimony: Novartis; Advisory / Consultancy: Celgene; Speaker Bureau / Expert testimony: Boehringer; Speaker Bureau / Expert testimony: Lilly; Speaker Bureau / Expert testimony: Takeda; Speaker Bureau / Expert testimony: Amgen; Research grant / Funding (institution): Bruker Daltonics; Non-remunerated activity/ies, Support for third-party medical writing assistance: Roche Pharma AG, Germany. [ABSTRACT FROM AUTHOR]