A clinical-molecular update on azanucleoside-based therapy for the treatment of hematologic cancers

The azanucleosides azacitidine and decitabine are currently used for the treatment of acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) in patients not only eligible for intensive chemotherapy but are also being explored in other hematologic and solid cancers. Based on their capacity to interfere with the DNA methylation machinery, these drugs are also referred to as hypomethylating agents (HMAs). As DNA methylation contributes to epigenetic regulation, azanucleosides are further considered to be among the first true "epigenetic drugs" that have reached clinical application. However, intriguing new evidence suggests that DNA hypomethylation is not the only mechanism of action for these drugs. This review summarizes the experience from more than 10 years of clinical practice with azanucleosides and discusses their molecular actions, including several not related to DNA methylation. A particular focus is placed on possible causes of primary and acquired resistances to azanucleoside treatment. We highlight current limitations for the success and durability of azanucleoside-based therapy and illustrate that a better understanding of the molecular determinants of drug response holds great potential to overcome resistance

Prognostic value of indoleamine 2,3 dioxygenase in patients with higher‐risk myelodysplastic syndromes treated with azacytidine

Hypomethylating agents (HMAs) are widely used in patients with higher‐risk myelodysplastic syndromes (MDS) not eligible for stem cell transplantation; however, the response rate is <50%. Reliable predictors of response are still missing, and it is a major challenge to develop new treatment strategies. One current approach is the combination of azacytidine (AZA) with checkpoint inhibitors; however, the potential benefit of targeting the immunomodulator indoleamine‐2,3‐dioxygenase (IDO‐1) has not yet been evaluated. We observed moderate to strong IDO‐1 expression in 37% of patients with high‐risk MDS. IDO‐1 positivity was predictive of treatment failure and shorter overall survival. Moreover, IDO‐1 positivity correlated inversely with the number of infiltrating CD8+ T cells, and IDO‐1+ patients failed to show an increase in CD8+ T cells under AZA treatment. In vitro experiments confirmed tryptophan catabolism and depletion of CD8+ T cells in IDO‐1+ MDS, suggesting that IDO‐1 expression induces an immunosuppressive microenvironment in MDS, thereby leading to treatment failure under AZA treatment. In conclusion, IDO‐1 is expressed in more than one‐third of patients with higher‐risk MDS, and is predictive of treatment failure and shorter overall survival. Therefore, IDO‐1 is emerging as a promising predictor and therapeutic target, especially for combination therapies with HMAs or checkpoint inhibitors

Numerical Investigations of Mixed Convection of Incompressible Viscous Fluid in LNG Storage with a Various Locations of Input and Output Mass

The article shows the results of mathematical simulation of mixed convection in the low-temperature storage of liquefied natural gas with a regenerative cooling. The regimes of mixed convection in a closed area with the different arrangement of the input and output sections of the masses are investigated. Two-dimensional nonstationary problem in the model of the Navier-Stokes in dimensionless variables "vorticity - stream function - temperature" was examined. Are obtained distributions of the hydrodynamic parameters and temperatures, characteristic basic laws governing the processes being investigated. Detailed circulating currents and carried out analysis of the mechanism of vortices formation and the temperature distribution in the solution for mixed convection mode with low Reynolds and Grashof numbers (Gr=10{6}, 100<Re<1000). Is established the significant influence of the geometrical arrangement of the input and output mass sections and input stream velocity on the structure of liquid flow and temperature in the low temperature LNG storage tanks

Transfer of plantar pressure from the medial to the central forefoot in patients with hallux valgus

Background: The aim of the study was to evaluate changes in plantar pressure distribution in feet affected by hallux valgus compared with their contralateral non-affected feet and with the feet of healthy control subjects. Methods: Thirty-six patients with unilateral hallux valgus who were indicated for surgery and 30 healthy subjects were assessed on a pedobarographic instrumented treadmill for step length and width, mean stance phase, and plantar foot pressure distribution. Plantar pressure distribution was divided into eight regions. Results: Significantly higher plantar pressures were observed in hallux valgus feet under the second and third metatarsal heads (p = .033) and the fourth and fifth toes (p &lt; .001) than in the healthy control feet. Although decreased pressures were measured under the hallux in affected feet (197 [82–467] kPa) in contrast to the contralateral side (221 [89–514] kPa), this difference failed to reach statistical significance (p = .055). The gait parameters step width, step length, and single-limb support did not show any differences between hallux valgus and control feet. Conclusion: Although the literature on changes in plantar pressures in hallux valgus remains divided, our findings on transferring load from the painful medial to the central and lateral forefoot region are consistent with the development of transfer metatarsalgia in patients with hallux valgus

Correlation of cytomorphology and histopathology in the diagnostic process of myeloid malignancies

Bone marrow cytomorphology and histopathology are the cornerstones for the initial diagnosis of myelodysplastic syndromes (MDS) and other related myeloid disorders. They provide a rapid first insight into diagnostic categories and thus help in clinical decision making. However, difficulties in the morphologic assessment of MDS exist due to inter- and intra-observer variability. In this study, we directly compared the results of cytomorphology and histopathology obtained in a real-world diagnostic scenario in 90 patients with myeloid malignancies aiming to evaluate their validity for diagnosing and classifying various myeloid malignancies. While both techniques placed 80% of our bone marrow samples into the same diagnostic category and thus showed a good correlation, our study also demonstrates the limitations in correlating marrow cytomorphology and histopathology, even following stringent and repetitive diagnostic assessments. This was particularly true for CMML, where not only additional diagnostic tools such as molecular genetics or clinical evaluation but also the analysis of the peripheral blood smears aided in finding the correct diagnosis. Overall, our data emphasize the need for a comprehensive diagnostic review in a patient-for-patient setting when a myeloid malignancy is suspected or confirmed. We propose that the combination of cytomorphologic and histopathologic assessment with clinical, laboratory, and genetic parameters is essential in achieving high diagnostic accuracy in an interdisciplinary setting

Bone marrow mesenchymal stromal cell-derived extracellular matrix displays altered glycosaminoglycan structure and impaired functionality in Myelodysplastic Syndromes

Myelodysplastic syndromes (MDS) comprise a heterogeneous group of hematologic malignancies characterized by clonal hematopoiesis, one or more cytopenias such as anemia, neutropenia, or thrombocytopenia, abnormal cellular maturation, and a high risk of progression to acute myeloid leukemia. The bone marrow microenvironment (BMME) in general and mesenchymal stromal cells (MSCs) in particular contribute to both the initiation and progression of MDS. However, little is known about the role of MSC-derived extracellular matrix (ECM) in this context. Therefore, we performed a comparative analysis of in vitro deposited MSC-derived ECM of different MDS subtypes and healthy controls. Atomic force microscopy analyses demonstrated that MDS ECM was significantly thicker and more compliant than those from healthy MSCs. Scanning electron microscopy showed a dense meshwork of fibrillar bundles connected by numerous smaller structures that span the distance between fibers in MDS ECM. Glycosaminoglycan (GAG) structures were detectable at high abundance in MDS ECM as white, sponge-like arrays on top of the fibrillar network. Quantification by Blyscan assay confirmed these observations, with higher concentrations of sulfated GAGs in MDS ECM. Fluorescent lectin staining with wheat germ agglutinin and peanut agglutinin demonstrated increased deposition of N-acetyl-glucosamine GAGs (hyaluronan (HA) and heparan sulfate) in low risk (LR) MDS ECM. Differential expression of N-acetyl-galactosamine GAGs (chondroitin sulfate, dermatan sulfate) was observed between LR- and high risk (HR)-MDS. Moreover, increased amounts of HA in the matrix of MSCs from LR-MDS patients were found to correlate with enhanced HA synthase 1 mRNA expression in these cells. Stimulation of mononuclear cells from healthy donors with low molecular weight HA resulted in an increased expression of various pro-inflammatory cytokines suggesting a contribution of the ECM to the inflammatory BMME typical of LR-MDS. CD34+ hematopoietic stem and progenitor cells (HSPCs) displayed an impaired differentiation potential after cultivation on MDS ECM and modified morphology accompanied by decreased integrin expression which mediate cell-matrix interaction. In summary, we provide evidence for structural alterations of the MSC-derived ECM in both LR- and HR-MDS. GAGs may play an important role in this remodeling processes during the malignant transformation which leads to the observed disturbance in the support of normal hematopoiesis

Isolation of human MHC class II-restricted T cell receptors from the autologous T-cell repertoire with potent anti-leukaemic reactivity

Summary Adoptive transfer of T cells genetically modified with tumour-specific T-cell receptors (TCR) is a promising novel approach in the treatment of cancer. We have previously isolated an allorestricted MHC class I-restricted TCR with specificity for Formin-like protein 1 (FMNL1) with potent activity against chronic lymphocytic leukaemia cells. CD4 + T cells have been described to be highly important for tumour elimination although TCR derived from CD4 + T cells with anti-tumour reactivity have been only rarely described. In this study we aimed to isolate MHC class-II-restricted CD4 + T cells and TCR with specificity for leukaemia antigens. We used professional antigen-presenting cells pulsed with the leukaemiaassociated and tumour-associated antigen FMNL1 for stimulation of autologous T cells in vitro. We isolated two CD4 + HLA-DR-restricted T-cell clones and T-cell-derived TCR with so far unknown specificity but high reactivity against lymphoma cells and native malignant cells derived from HLA-matched patients with diverse leukaemias. Moreover, characterization of the TCR after TCR gene transfer revealed that specific characteristics of isolated TCR as reactivity in response to Toll-like receptors were transferable on effector cells. Our results have a major impact on the development of novel immunotherapies. They demonstrate that TCR with potent HLA-DR-restricted anti-leukaemic reactivity against so far undefined self-restricted antigens can be isolated from the healthy autorestricted CD4 + T-cell repertoire and these TCR are highly interesting candidate tools for novel immunotherapies

Selinexor in Advanced, Metastatic Dedifferentiated Liposarcoma: A Multinational, Randomized, Double-Blind, Placebo-Controlled Trial

PURPOSE Antitumor activity in preclinical models and a phase I study of patients with dedifferentiated liposarcoma (DD-LPS) was observed with selinexor. We evaluated the clinical benefit of selinexor in patients with previously treated DD-LPS whose sarcoma progressed on approved agents. METHODS SEAL was a phase II-III, multicenter, randomized, double-blind, placebo-controlled study. Patients age 12 years or older with advanced DD-LPS who had received two-five lines of therapy were randomly assigned (2:1) to selinexor (60 mg) or placebo twice weekly in 6-week cycles (crossover permitted). The primary end point was progression-free survival (PFS). Patients who received at least one dose of study treatment were included for safety analysis (ClinicalTrials.gov identifier: ). RESULTS Two hundred eighty-five patients were enrolled (selinexor, n = 188; placebo, n = 97). PFS was significantly longer with selinexor versus placebo: hazard ratio (HR) 0.70 (95% CI, 0.52 to 0.95; one-sided P = .011; medians 2.8 v 2.1 months), as was time to next treatment: HR 0.50 (95% CI, 0.37 to 0.66; one-sided P < .0001; medians 5.8 v 3.2 months). With crossover, no difference was observed in overall survival. The most common treatment-emergent adverse events of any grade versus grade 3 or 4 with selinexor were nausea (151 [80.7%] v 11 [5.9]), decreased appetite (113 [60.4%] v 14 [7.5%]), and fatigue (96 [51.3%] v 12 [6.4%]). Four (2.1%) and three (3.1%) patients died in the selinexor and placebo arms, respectively. Exploratory RNA sequencing analysis identified that the absence of CALB1 expression was associated with longer PFS with selinexor compared with placebo (median 6.9 v 2.2 months; HR, 0.19; P = .001). CONCLUSION Patients with advanced, refractory DD-LPS showed improved PFS and time to next treatment with selinexor compared with placebo. Supportive care and dose reductions mitigated side effects of selinexor. Prospective validation of CALB1 expression as a predictive biomarker for selinexor in DD-LPS is warranted. (C) 2022 by American Society of Clinical Oncolog

The IMiD target CRBN determines HSP90 activity toward transmembrane proteins essential in multiple myeloma

The complex architecture of transmembrane proteins requires quality control (QC) of folding, membrane positioning, and trafficking as prerequisites for cellular homeostasis and intercellular communication. However, it has remained unclear whether transmembrane protein-specific QC hubs exist. Here we identify cereblon (CRBN), the target of immunomodulatory drugs (IMiDs), as a co-chaperone that specifically determines chaperone activity of HSP90 toward transmembrane proteins by means of counteracting AHA1. This function is abrogated by IMiDs, which disrupt the interaction of CRBN with HSP90. Among the multiple transmembrane protein clients of CRBN-AHA1-HSP90 revealed by cell surface proteomics, we identify the amino acid transporter LAT1/CD98hc as a determinant of IMiD activity in multiple myeloma (MM) and present an Anticalin-based CD98hc radiopharmaceutical for MM radio-theranostics. These data establish the CRBN-AHA1-HSP90 axis in the biogenesis of transmembrane proteins, link IMiD activity to tumor metabolism, and nominate CD98hc and LAT1 as attractive diagnostic and therapeutic targets in MM

Studies on the suitability of rumen fill scoring as an antepartal indicator of health, fertility and milk yield in Holstein-Friesian cows

In den ersten Wochen nach der Kalbung liegt eine hohe Erkrankungsrate der Milchkühe von 35 bis 66 % vor. Ein wichtiges Ziel in der modernen Milchviehhaltung muss daher das frühzeitige Erkennen potentieller Risikotiere sein, noch bevor sie erkranken. Bereits die Futteraufnahme vor der Kalbung steht mit Erkrankungen und Abgängen nach der Kalbung in Beziehung. Die genaue Futteraufnahme jedes Einzeltieres a.p. könnte demzufolge von den Landwirten als Indikator für Erkrankungen a.p. und p.p. genutzt werden. Derzeit wird diskutiert, ob die Bewertung der äußerlich sichtbaren Pansenfüllung über ein Notensystem als Messinstrument genutzt werden kann, um die Futteraufnahme der individuellen Kuh zu schätzen. Ziel dieser Arbeit war es, den Verlauf der Pansenfüllung innerhalb von 24 Stunden sowie bei täglich einmaliger Benotung zu analysieren. Es sollte untersucht werden, welche Ursachen einem abweichenden Pansenfüllungsverlauf zugrunde liegen, welche Einflussfaktoren auf die Pansenfüllung wirken und ob die Pansenfüllung ein geeigneter Parameter zur Schätzung der Futteraufnahme sowie zur Vorhersage von Futteraufnahme, Leistung und Tiergesundheit in der Folgelaktation ist. Dazu fand in einem Milchviehbetrieb in Sachsen-Anhalt über einen Zeitraum von Mai 2015 bis April 2016 bei 109 pluriparen Kühen der Rasse Holstein-Friesian die Datenerhebung statt. Grundlage zur Benotung der Pansenfüllung war das Schema von Zaaijer und Noordhuizen (2003). Alle Kühe wurden täglich zur selben Uhrzeit während der Transitperiode bonitiert. Zusätzlich wurden auch Tagesprofile von 21 Kühen a.p. und 10 Kühen p.p. erstellt. Die Futteraufnahme konnte tierindividuell über elektronische Wiegetröge erfasst werden. Weiterhin wurde regelmäßig die Rückenfettdicke und Lebendmasse gemessen, der BCS bestimmt und die Kühe nach einem festen Schema klinisch allgemein untersucht. Es wurden zu festgelegten Zeitpunkten Blut- und Harnproben genommen und analysiert. Tägliche Milchmengen, wöchentliche Milchinhaltsstoffe, Krankheits- und Fruchtbarkeitsdaten wurden erhoben. A.p. besteht eine geringe Variabilität der Pansenfüllungsnoten, wobei über 97 Prozent aller Noten zwischen 3,5 und 4,0 liegen. P.p. ist die tierindividuelle Variabilität der Noten sehr viel ausgeprägter. Es wirken sowohl a.p. als auch p.p. mehrere Einflussfaktoren auf die Pansenfüllung. Dabei handelt es sich um die Futteraufnahme, den BCS, die 2. Laktation und p.p. zusätzlich den Lahmheitsgrad. Ein gestörter Pansenfüllungsverlauf a.p. steht im Zusammenhang mit den Blutwerten Harnstoff, Phosphat und Magnesium sowie mit den Harnwerten Kalzium und Kalium. P.p. besteht eine Beziehung zwischen einem gestörten Pansenfüllungsverlauf und den Blutparametern Harnstoff, Bilirubin, Natrium und den Harnwerten Kreatinin und Chlorid. Es liegen sehr geringe bis geringe positive signifikante Korrelationen zwischen der Pansenfüllung und der Frischmasseaufnahme a.p. und p.p. vor. Der Laktationszeitpunkt hat einen signifikanten Einfluss auf den Zusammenhang zwischen Futteraufnahme und Pansenfüllung. In den ersten 9 Tagen p.p. reagiert die Pansenfüllung schon bei 7 kg zusätzlicher Frischmasseaufnahme mit einer halben Note. Eine Woche a.p. korreliert die Pansenfüllung mit der Futteraufnahme eine und zwei Wochen p.p. Eine Beziehung besteht auch zu verschiedenen Erkrankungen a.p. und p.p. Kein Zusammenhang besteht zwischen der Pansenfüllung a.p. und der Milchleistung oder der Fruchtbarkeit der Folgelaktation. Die Pansenfüllung ist bei hochtragenden Kühen ein stabiler Parameter, da eine Notenvergabe im Bereich kleinerer Noten aufgrund der schlechten Beurteilbarkeit der Hungergrube schwierig ist. P.p. ist das Vergeben kleinerer Noten einfacher. Der Einfluss der Faktoren Futteraufnahme, BCS und 2. Laktation auf die Pansenfüllung ist a.p. nur gering und p.p. von größerer Bedeutung. Abweichende Pansenfüllungsverläufe stehen in Beziehung mit einer verminderten Futteraufnahme. Bei auffälligen Kühen sollte bewusst auf die Futteraufnahme geachtet und Heu und Propylenglykol verabreicht werden. In den ersten 9 Tagen p.p. reagiert die Pansenfüllung sehr sensibel auf die Futteraufnahme. Daher wird das tägliche Benoten aller Kühe in der ersten Woche p.p. zur selben Uhrzeit empfohlen. Ab Tag 10 p.p. ist die Pansenfüllung nur noch mäßig zur Futteraufnahmeschätzung geeignet. Das einmalige Benoten aller Kühe an Tag 28 oder 60 p.p. und ein Vergleich der Noten auf Herdenbasis kann angewendet werden. A.p. ist die Pansenfüllung nur geeignet, um Kühe mit deutlich reduzierter Futteraufnahme zu erkennen. Daher genügt das einmalige Benoten der Kühe an Tag 14 (± 3) a.p. Die Pansenfüllung kann eine Woche a.p. als Indikator für die Futteraufnahme in den ersten beiden Wochen p.p. herangezogen werden. A.p. und p.p. ist die Pansenfüllung als Indikator für Erkrankungen geeignet, sollte aber vorsichtig interpretiert werden. Innerhalb von 24 Stunden schwanken die Pansenfüllungsnoten a.p. wenig. P.p. liegt eine größere Variabilität der Noten vor. A.p. und p.p. haben die Faktoren Futteraufnahme, BCS, 2. Laktation und p.p. zusätzlich der Lahmheitsgrad signifikanten Einfluss auf die Pansenfüllung. Weicht die Pansenfüllung vom durchschnittlichen Verlauf aller Kühe ab, weist dies auf eine verminderte Futteraufnahme hin. Zwischen der Pansenfüllung und der Futteraufnahme liegt a.p. und p.p eine Beziehung vor, wobei der Laktationszeitpunkt einen großen Einfluss auf diesen Zusammenhang hat. Unmittelbar p.p. reagiert die Pansenfüllung sehr sensibel auf die Futteraufnahme. Das tägliche Benoten der Hungergrube während der ersten Woche p.p. sowie das einmalige Benoten aller Kühe a.p. werden empfohlen.During the first weeks after calving, there are high disease rates of 35 – 66 % in dairy cows. An important aim of a modern dairy livestock must be the early identification of risk animals before they become ill. Antepartal feed intake is already related to diseases and culling rates after calving. Therefore, the exact feed intake a.p. of each single animal could be used by farmers as an indicator of disease a.p. and p.p. It is currently being discussed whether the evaluation of the externally visible rumen fill can be used as a measuring device via a scoring system to estimate the feed intake of the individual cow. The objective of this study is to analyze the course of rumen fill within 24 hours as well as on a daily basis. The aim is to investigate the causes of a different course of rumen fill, the factors influencing rumen fill, and whether rumen fill is a suitable parameter for estimating the feed intake as well as for predicting feed intake, performance and animal health in the subsequent lactation. In a dairy farm in Sachsen- Anhalt, data collection took place on 109 pluriparous cows of the Holstein- Friesian breed over a period from May 2015 to April 2016. The basis for scoring the rumen fill was the scoring-system of Zaaijer and Noordhuizen (2003). All cows were scored daily at the same time during the transition period. In addition, 24-hours-profiles of 21 cows a.p. and 10 cows p.p. were recorded. Feed intake was recorded individually by electronic weighing troughs. In addition, the back fat thickness and live mass were measured regularly, the BCS was determined and the cows were clinically examined according to a fixed plan. Blood and urine samples were taken and analyzed at fixed times, and daily milk amounts, weekly milk contents, disease and fertility data were collected. There is little variability in rumen fill scores a.p., with over 97% of all grades being between 3.5 and 4.0. The individual variability of the animals’ notes p.p. is much more distinctive. Several influencing factors have an effect on the rumen fill a.p. as well as p.p. The influencing factors are feed intake, BCS, second lactation and, in addition, the degree of lameness for the measuring periode p.p. A disturbed rumen filling course a.p. is associated with the blood parameters urea, phosphate and magnesium as well as the urine parameters calcium and potassium. There is a relationship between a disturbed rumen filling course p.p. and the blood parameters urea, bilirubin, sodium and the urinary values creatinine and chloride. There are very low to low positive, significant correlations between the rumen fill scores and as-fed feed intake a.p. and p.p. The number of days in milk has a significant influence on the relationship between feed intake and rumen fill. During the first nine days p.p. the rumen fill already reacts with half a note every aditional seven kilograms of as-fed feed intake. There is a correlation between the rumen fill score one week a.p. and the feed intake one and two weeks p.p. A relationship also exists to various disorders a.p. as well as p.p. There is no connection between the rumen fill score a.p. and milk yield or fertility during the following lactation. The rumen fill score is a solid parameter in high-yielding cows. Since it is difficult to assess the fossa paralumbalis in high-yielding cows, grading of small notes is challenging. It is easier to grade smaller scores p.p. The influence of the factors feed intake, BCS and second lactation on the rumen fill is a.p. low and p.p. of greater importance for daily herd management. Divergent rumen fill courses are related to reduced feed intake. In conspicuous cows, attention should be paid to the feed intake and hay or propylene glycol should be fed. During the first nine days p.p., the rumen fill reacts very sensitively to feed intake. Therefore, daily grading of all cows during the first week p.p. at the same time is recommended. From day 10 p.p. the rumen fill is only of limited suitability for estimating the food intake. A single rating of all cows on day 28 or 60 p.p. and comparison of the notes on a herd basis can be applied. The rumen fill a.p. is only suitable to recognize cows whose feed intake is highly reduced. Therefore, it is sufficient to score all cows once on day 14 (± 3) a.p. The rumen fill in the first week a.p can be used as an indicator of feed intake in the first two weeks p.p. The rumen fill a.p. and p.p. is suitable as an indicator for diseases, but should be interpreted carefully. The rumen fill notes a.p. vary only slightly within 24 hours. P.p. there is greater variability of the notes. A.p. and p.p., the factors feed intake, BCS, second lactation and, in addition, the lameness degree p.p. have a significant influence on the rumen fill. If the rumen fill course differs from the average course of all cows, this indicates a reduced feed intake. There is a relationship between rumen fill and feed intake, both a.p. as well as p.p. The number of days in milk has a great influence on this connection. Immediately p.p., rumen fill reacts very sensitively to feed intake. It is recommended to score the fossa paralumbalis of all cows daily during the first week p.p. and once a.p