Strategies to reduce relapse risk in patients undergoing allogeneic stem cell transplantation for acute myeloid leukaemia.

Allogeneic stem cell transplantation is a centrally important curative strategy in adults with acute myeloid leukaemia; however, relapse occurs in a significant proportion of patients and remains the leading cause of treatment failure. The prognosis for patients who relapse post-transplant remains poor, and the development of new strategies with the ability to reduce disease recurrence without increasing transplant toxicity remains a priority. In this review, within the context of our understanding of disease biology and the graft-versus-leukaemia (GVL) effect, we will discuss established, evolving and novel approaches for increasing remission rates, decreasing measurable residual disease pretransplant, future methods to augment the GVL effect and the opportunities for post-transplant maintenance. Future progress depends upon the development of innovative trials and networks, which will ensure the rapid assessment of emerging therapies in prospective clinical trials.

Adenylate kinase 9 is essential for sperm function and male fertility in mammals.

Despite passing routine laboratory tests for semen quality, bulls used in artificial insemination exhibit significant variation in fertility. Routine analysis of fertility data identified a dairy bull with extreme subfertility (10% pregnancy rate). To characterize the subfertility phenotype, a range of in vitro, in vivo, and molecular assays were carried out. Sperm from the subfertile bull exhibited reduced motility and severely reduced caffeine-induced hyperactivation compared to controls. Ability to penetrate the zona pellucida, cleavage rate, cleavage kinetics, and blastocyst yield after IVF or AI were significantly lower than in control bulls. Whole-genome sequencing from semen and RNA sequencing of testis tissue revealed a critical mutation in adenylate kinase 9 (AK9) that impaired splicing, leading to a premature termination codon and a severely truncated protein. Mice deficient in AK9 were generated to further investigate the function of the gene; knockout males were phenotypically indistinguishable from their wild-type littermates but produced immotile sperm that were incapable of normal fertilization. These sperm exhibited numerous abnormalities, including a low ATP concentration and reduced motility. RNA-seq analysis of their testis revealed differential gene expression of components of the axoneme and sperm flagellum as well as steroid metabolic processes. Sperm ultrastructural analysis showed a high percentage of sperm with abnormal flagella. Combined bovine and murine data indicate the essential metabolic role of AK9 in sperm motility and/or hyperactivation, which in turn affects sperm binding and penetration of the zona pellucida. Thus, AK9 has been found to be directly implicated in impaired male fertility in mammals.

Loss of the importin Kpna2 causes infertility in male mice by disrupting the translocation of testis-specific transcription factors.

Karyopherins mediate the movement between the nucleus and cytoplasm of specific proteins in diverse cellular processes. Through a loss-of-function approach, we here examine the role of Karyopherin Subunit Alpha 2 (Kpna2) in spermatogenesis. Knockout male mice exhibited reduced body size and sperm motility, increased sperm abnormalities, and led to the dysregulation of testis gene expression and ultimately to infertility. Impaired mRNA expression mainly affected clusters of genes expressed in spermatids and spermatocytes. Downregulated genes included a set of genes that participate in cell adhesion and extracellular matrix (ECM) organization. We detected both the enrichment of some transcription factors that bind to regions around transcription start sites of downregulated genes and the impaired transport of specific factors to the nucleus of spermatid cells. We propose that Kpna2 is essential in the seminiferous tubules for promoting the translocation of testis-specific transcription factors that control the expression of genes related to ECM organization.

Effect of fermented soy beverage in aged female mice model.

Soy beverage is a rich source of phytoestrogens isoflavones, with potential benefits on health. The effect of those compounds depends greatly on their bacterial metabolization into their aglycone forms. This study evaluated the health effects of two soy beverages, non-fermented (SB) and fermented with Bifidobacterium pseudocatenulatum INIA P815 (FSB), in acyclic and cyclic C57BL/6J aged female mice as a model of menopause and premenopause, respectively. SB and FSB treatments were administrated for 36 days and, subsequently, body weight, lipid and inflammatory profile and fertility were analyzed and compared. In addition, hepatic gene expression and faecal microbiota composition were also assessed. After fermentation, FSB presented a high content in the aglycones daidzein and genistein and a higher antioxidant activity. FSB treated cyclic mice showed a significant increase in the number of retrieved oocytes and zigotes. Differences in serum lipids were observed in triglycerides, which were lower in FSB than in SB groups. None of the treatments influenced the inflammatory profile or caused a dramatic change in the intestinal microbiota profile or hepatic gene expression in any of the groups. Our data showed that FSB provided greater health benefits than SB in lipid profile and fertility in cyclic mice. These beneficial effects could be attributed to the fermentation process, which produces more bioavailable and bioactive compounds, achieving a greater impact on health.

The technology of high-power ultrasound and its effect on the color and aroma of rosé wines.

BACKGROUND: The application of high-power ultrasound (US) has been studied extensively to obtain value-added red wines using short maceration times. It is a technique of wide interest for the extraction of aromatic compounds and compounds responsible for color, so it could also be an important tool to use in the elaboration process of rosé wines. Thus, this study focused on the effect of the application of US on the phenolic, aromatic, and sensorial profile of rosé wines. For this, three different types of rosé wine were produced: a control with direct pressing after crushing, another wine obtained from a 4 h macerated must, and a wine whose crushed and destemmed grape was sonicated and subsequently pressed. RESULTS: The results showed a higher color intensity and a higher total polyphenol and anthocyanin content in the wine obtained from sonicated grape compared to both control wine and that obtained from grape macerated for 4 h. Ultrasound treatment enhanced the extraction of varietal volatile compounds in must, especially the free faction of terpenes and norisoprenoids. It also gave rise to wines with a higher concentration of C6 alcohols and other compounds such as guaiacol and 4-vinylguaiacol, but led to fewer fatty acids, especially C6, C4 and C8 acids. Sensorially these wines showed intense aromas of red fruits and flowers, while no defects in aroma or astringency were found. CONCLUSIONS: The color and sensory profile of the resulting wines indicate that the US may be an interesting tool for obtaining quality rosé wines, replacing the maceration stage and reducing potential oxidation problems. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

An ancient testis-specific IQ motif-containing H gene regulates specific transcript isoform expression during spermatogenesis.

Spermatogenic cells express more alternatively spliced RNAs than most whole tissues; however, the regulation of these events remains unclear. Here, we have characterized the function of a testis-specific IQ motif-containing H gene (Iqch) using a mutant mouse model. We found that Iqch is essential for the specific expression of RNA isoforms during spermatogenesis. Using immunohistochemistry of the testis, we noted that Iqch was expressed mainly in the nucleus of spermatocyte and spermatid, where IQCH appeared juxtaposed with SRRM2 and ERSP1 in the nuclear speckles, suggesting that interactions among these proteins regulate alternative splicing (AS). Using RNA-seq, we found that mutant Iqch produces alterations in gene expression, including the clear downregulation of testis-specific lncRNAs and protein-coding genes at the spermatid stage, and AS modifications - principally increased intron retention - resulting in complete male infertility. Interestingly, we identified previously unreported spliced transcripts in the wild-type testis, while mutant Iqch modified the expression and use of hundreds of RNA isoforms, favouring the expression of the canonical form. This suggests that Iqch is part of a splicing control mechanism, which is essential in germ cell biology.

Bull spermatozoa selected by thermotaxis exhibit high DNA integrity, specific head morphometry, and improve ICSI outcome.

BACKGROUND: Sperm migration by thermotaxis is a guidance mechanism that operates along the oviduct and it has proved to be a valid method for selecting spermatozoa with low DNA fragmentation (SDF) in mice, humans, and stallions. This study aimed to analyse if bull spermatozoa could be selected by thermotaxis and to assess their quality in terms of SDF as well as determine the presence of a specific sperm subpopulation based on sperm morphometry and assess their fertilizing capacity by ICSI. METHODS: We used frozen-thawed sperm from 6 bulls and sperm selection by thermotaxis was performed with TALP medium supplemented with 25 mmol/L of HEPES and 5 mmol/L of caffeine. In these conditions, sperm selection was achieved, obtaining a net thermotaxis of 3.6%. Subsequently, we analysed the SDF of the migrated and not-migrated spermatozoa using the neutral COMET assay, and we evaluated the size of the sperm head using Hemacolor® staining with Motic Images Plus 3 software. Additionally, migrated and not-migrated spermatozoa by thermotaxis were used to fertilize bovine in vitro matured (IVM) oocytes by ICSI, a very inefficient procedure in cattle that is only successful when the oocyte is artificially activated. RESULTS: The results showed lower SDF (χ², P < 0.001, 13.3% reduction, n = 8) and lower head size parameters (length and width, P < 0.01; and perimeter and area, P < 0.001; n = 4) in those spermatozoa migrated in comparison to those not-migrated. The distribution of sperm subpopulations structure varied between groups, highlighting cluster 2, characterized by spermatozoa with small head size, and high ellipticity and elongated heads, as the most abundant in the thermotaxis migrated group. When performed ICSI (without oocyte artificial activation) with the thermotactic sperm, the blastocyst rate was 32.2% ± 9.3% in the group microinjected with the thermotactic spermatozoa vs. 8.3% ± 7.8% in the group of not-migrated sperm (χ², P < 0.05). CONCLUSION: Our results showed that bull sperm selection by thermotaxis has a much higher DNA integrity, small and elongated head size parameters, and different sperm subpopulation structure than the not-selected spermatozoa. Additionally, we evidenced that thermotactic spermatozoa improve ICSI success rates.

The Added Value of Somatosensory Potential N70 in Neurological Prognosis After Coma by Acute Brain Structural Injury: A Retrospective Study.

Background: Coma after acute brain structural injury (ABI) are associated with high mortality and disability. Somatosensory evoked potentials (SSEP) N20 and N70 are used to predict prognosis. Purpose: We assessed the utility of SSEP (N20-N70) as an early indicator of long-term functional prognosis in these patients. Methods: We conducted a retrospective cohort study of patients admitted to the intensive care unit (ICU) with a diagnosis of coma after ABI (n=60). An SSEP study including N20 and N70 was performed 24-72 hours after coma onset. Functional recovery was evaluated 6 to 12 months later using the Modified Glasgow Scale (mGS). The study was approved by our local research ethics committee. Results: The absence of N20 (41% specificity=100%) or N70 (78%) was a strong indicator of a poor outcome. In contrast, the presence of N70 was an indicator of a good outcome (specificity=64.2% sensitivity=91.3%). Conclusion: SSEP N20 and N70 are useful early prognostic markers with high specificity (N20) and sensitivity (N70). N70 has potential additional value for improving the prediction of good functional outcomes in the long term.

Rapid and Non-Destructive Analysis of Corky Off-Flavors in Natural Cork Stoppers by a Wireless and Portable Electronic Nose.

This article discusses the use of a handheld electronic nose to obtain information on the presence of some aromatic defects in natural cork stoppers, such as haloanisoles, alkylmethoxypyrazines, and ketones. Typical concentrations of these compounds (from 5 to 120 ng in the cork samples) have been measured. Two electronic nose prototypes have been developed as an instrumentation system comprise of eight commercial gas sensors to perform two sets of experiments. In the first experiment, a quantitative approach was used whist in the second experiment a qualitative one was used. Machine learning algorithms such as k-nearest neighbors and artificial neural networks have been used in order to test the performance of the system to detect cork defects. The use of this system tries to improve the current aromatic defect detection process in the cork stopper industry, which is done by gas chromatography or human test panels. We found this electronic nose to have near 100 % accuracy in the detection of these defects.

Use of Microwave Maceration in Red Winemaking: Effect on Fermentation and Chemical Composition of Red Wines.

The objective of this study was to evaluate the effect of microwave treatment of crushed grapes on the yeast population of the must and on the development of alcoholic fermentation, as well as on the extraction of different compounds from the grapes such as polysaccharides and amino acids that can affect the organoleptic quality and stability of the wine. This study demonstrated for the first time the effect of the microwave treatment of grapes on native yeast species and their diversity, producing an increase in fermentation kinetics and a decrease in the lag phase. The microwave treatment produced a positive effect on the extraction of amino acids and polysaccharides from the grapes, resulting in significantly higher amounts of the main amino acids of the must and some major volatile compounds in the treated samples. The polysaccharides most affected by the microwave treatment were the PRAGs, the main polysaccharides liberated from grapes during the maceration.

Effect of Microwave Maceration and SO2 Free Vinification on Volatile Composition of Red Wines.

This study evaluates the effect of microwave treatment in grape maceration at laboratory scale on the content of free and glycosidically bound varietal compounds of must and wines and on the overall aroma of wines produced with and without SO2. The volatile compounds were extracted by solid phase extraction and analyzed by gas chromatography-mass spectrometry, carrying out a sensory evaluation of wines by quantitative descriptive analysis. Microwave treatment significantly increased the free and bound fraction of most varietal compounds in the must. Wines from microwave maceration showed faster fermentation kinetics and shorter lag phase, resulting in an increase in some volatile compounds of sensory relevance. The absence of SO2 caused a decrease in concentration of some volatile compounds, mainly fatty acids and esters. The sensory assessment of wines from microwave treatment was higher than the control wine, especially in wines without SO2, which had higher scores in the "red berry" and "floral" odor attributes and a more intense aroma. This indicates that the pre-fermentative treatment of grapes with microwaves could be used to increase the wine aroma and to reduce the occurrence of SO2.

Effect of Power Ultrasound Treatment on Free and Glycosidically-Bound Volatile Compounds and the Sensorial Profile of Red Wines.

This study presents the effect of the application of high-power ultrasound to crushed grapes, at a winery-scale, on the content of varietal volatile compounds (free and glycosidically-bound) in musts and on the overall aroma of wines. Two different frequencies (20 kHz and 28 kHz) were tested and the combination of grape sonication and different maceration times on wine aroma was also evaluated. The volatile compounds were isolated by solid phase extraction and analyzed by gas chromatography-mass spectrometry, carrying out a sensory evaluation of wines by quantitative descriptive analysis. Sonication produced an increase in the concentration of free varietal compounds such as C6 alcohols, terpenes and norisoprenoids in musts and also in wines made by 48 h of skin maceration, being less efficient in the extraction of the bound fraction. Fermentation compounds were also positively affected by ultrasound treatment, although this effect was variable depending on the frequency used, the maceration time and the type of compound. All the wines made from sonicated grapes had better scores in the evaluated olfactory attributes with respect to the control wines. Our results indicate that sonication could produce an increase in the content of some volatile compounds of sensory relevance, obtaining wines with an aroma quality similar or higher than those elaborated with longer maceration times.

Evaluation of the Storage Conditions and Type of Cork Stopper on the Quality of Bottled White Wines.

The effects of different storage conditions, light exposure, temperature and different commercially available cork stoppers on the phenolic, volatile and sensorial profile of Verdejo wines were studied. Two natural corks of different visual quality and a microgranulated cork stopper were investigated over one year at two different storage conditions. One simulating light exposure and temperature in retail outlets and the other simulating optimal cellar conditions (darkness and 12 °C). The wines stored under commercial conditions showed greater losses of total and free SO2 and higher levels of brown-yellowish tones, related to the oxidation of flavan-3-ols. Although these wines underwent a decrease in the total content of stilbenes, a significant increase in trans-piceid was observed. In addition, these wines suffered important changes in their volatile and sensory profile. Volatile compounds with fruity and floral aromas decreased significantly, while volatile compounds related to aged-type characters, as linalool oxides, vitispirane, TDN or furan derivatives increased. Wines stored in darkness at 12 °C underwent minor changes and their sensory profiles were similar to wine before bottling. The high-quality natural corks and microgranulated corks better preserved the quality of the white wines from a sensory point of view. These results showed that temperature and light exposure conditions (diffuse white LEDs and 24 ± 2 °C) in retail outlets considerably decrease the quality of bottled white wines and, consequently, their shelf life, due to the premature development of aged-type characters.

Effects of the pre-fermentative addition of chitosan on the nitrogenous fraction and the secondary fermentation products of SO2 -free red wines.

BACKGROUND: Different red winemaking were carried out to evaluate the effects of the prefermentative addition of chitosan, as an alternative to the use of SO2 , on the secondary products of alcoholic fermentation, yeast available nitrogen (YAN), biogenic amines and ethyl carbamate. RESULTS: The wines made with chitosan presented higher total acidity and higher content of tartaric and succinic acids than those made only with SO2 . The use of chitosan in winemaking resulted in wines with higher glycerol and diacetyl content without increasing the concentration of ethanol, acetic acid, acetaldehyde or butanediol. YAN was lower in wines made with chitosan, which may mean an advantage for the microbial stability of the wines. Furthermore, the use of chitosan at the beginning of alcoholic fermentation did not increase the concentration of biogenic amines or the formation of ethyl carbamate in SO2 -free red wines. CONCLUSION: The total or partial substitution of SO2 for chitosan at the beginning of the alcoholic fermentation gives rise to quality red wines without negatively affecting their nitrogen fraction or their very important secondary fermentation products such as acetic acid or acetaldehyde. © 2020 Society of Chemical Industry.

Effect of Wine Lees as Alternative Antioxidants on Physicochemical and Sensorial Composition of Deer Burgers Stored during Chilled Storage.

Wine lees from two grape varieties (Vitis vinifera L. Cv. "verdejo" and "palomino") were studied as natural preservatives in deer burgers compared with the traditional additive sodium ascorbate. Burgers packed in modified atmosphere packaging and stored in refrigeration were analyzed at 0, 4, 8, and 12 days. The addition of lees (2.5% and 5%) produced a reduction of pH and variations in color (L* and a*), higher antioxidant capacity and phenolic content, lower lipid and protein oxidation, and the inhibition of psychotrophic aerobic bacteria and enterobacteria during the storage time. Likewise, burgers with lees kept the aldehydes concentration (volatile compounds indicators of lipid oxidation) over storage time, while esters, acids, and other compounds, previously present in lees, increased. These changes provided new odor and taste attributes like wine, bakery, and raisin notes. Therefore, the addition of wine lees had an antioxidant and antimicrobial effect and produced new sensory attributes in deer burgers.

Mango by-products as a natural source of valuable odor-active compounds.

BACKGROUND: The aromatic potential of mango by-products was evaluated to seek natural and cheap sources of odor-active compounds. Volatile compounds in mango peel and seed were chemically characterized and compared with those in mango pulp using headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry (HS-SPME/GC-MS). RESULTS: More than 60 volatile compounds were detected in mango by-products, whose aromatic activity was estimated using odorant activity values (OAVs). The results indicated that mango peel was a valuable matrix of odor-active compounds, which were found in even larger quantities than in edible mango fractions. 3-Carene was the predominant compound, although other compounds such as decanal, 1-octen-3-one, nonanal, limonene, ß-damascenone, and 2-nonenal were the most odor-active compounds in mango peel. The greatest aromatic impact was obtained from mango peel, with sensorial features described as fresh / herbaceous, fruity, floral and resinous. CONCLUSION: The exceptional flavoring potential of mango peel by-product opens a door for its use and revalorization as a natural flavoring ingredient in the food and cosmetic industries. © 2020 Society of Chemical Industry.

Impact of oenological antioxidant substances on the formation of 1-hydroxyethyl radical and phenolic composition in SO2 free red wines.

BACKGROUND: Different natural substances, chitosan, inactive dry yeasts and freeze-dried aqueous extracts from two wine industry by-products (stems and shoots) were used in red winemaking as possible alternatives to SO2 . The resistance to oxidation of wines was evaluated by electron paramagnetic resonance. The phenolic composition of wines was analyzed by high-performance liquid chromatography-diode array detection/electrospray ionization mass spectrometry, antioxidant activity was determined by DPPH (1,1-diphenyl-2-picrylhydrazyl radical) and ABTS [2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation] assays and spectrophotometric measurements of color were compared. RESULTS: The wines elaborated with chitosan and inactive dry yeast presented greater capacity to inhibit the formation of the 1-hydroxyethyl radical compared to the wines elaborated with stem or shoot extracts. The total content of anthocyanins was higher in the wines elaborated with SO2 ; however, the concentration of flavan-3-ols was higher in the wines with chitosan. In addition, the wines with chitosan and inactive dry yeast presented the highest % polymerization. Wines elaborated with stem extract had a lower concentration of flavonols and stilbenes. CONCLUSION: Chitosan and inactive dry yeast, which are used as an alternative to SO2 in winemaking, allow the control of the formation of 1-HER in red wines. Wines with stem and shoot extracts showed a lower resistance to oxidation. © 2020 Society of Chemical Industry.

Molecular Basis of the Mechanisms Controlling MASTL.

The human MASTL (Microtubule-associated serine/threonine kinase-like) gene encodes an essential protein in the cell cycle. MASTL is a key factor preventing early dephosphorylation of M-phase targets of Cdk1/CycB. Little is known about the mechanism of MASTL activation and regulation. MASTL contains a non-conserved insertion of 550 residues within its activation loop, splitting the kinase domain, and making it unique. Here, we show that this non-conserved middle region (NCMR) of the protein is crucial for target specificity and activity. We performed a phosphoproteomic assay with different MASTL constructs identifying key phosphorylation sites for its activation and determining whether they arise from autophosphorylation or exogenous kinases, thus generating an activation model. Hydrogen/deuterium exchange data complements this analysis revealing that the C-lobe in full-length MASTL forms a stable structure, whereas the N-lobe is dynamic and the NCMR and C-tail contain few localized regions with higher-order structure. Our results indicate that truncated versions of MASTL conserving a cryptic C-Lobe in the NCMR, display catalytic activity and different targets, thus establishing a possible link with truncated mutations observed in cancer-related databases.

Natural extracts from fresh and oven-dried winemaking by-products as valuable source of antioxidant compounds.

Winemaking by-products are a natural source of antioxidant components; however, due to their highly perishable and seasonal nature, they may require a prior conservation step before being processed. Natural extracts from fresh and oven-dried red grape agro-industrial by-products were obtained by ultrasound assisted extraction (UAE), using a hydroalcoholic solution as extracting solvent. Extracts were analyzed by HPLC-DAD-ESI-MS, to know the feasibility of winemaking by-products as natural sources of phenolic compounds, as well as the effect of the oven-drying treatment on the phenolic composition. Oven-drying at 45°C caused a significant decrease on the total phenolic content, which implied a reduction of the antioxidant capacity of the extracts. Also, it produced a decrease in total and individual flavan-3-ols, stilbenes, and flavonols, being greater in those extracts from stems. Respect to anthocyanins, which were only identified in grape pomace extracts, oven-drying caused an important decrease, being the peonidin-3-O-glucoside the more thermosensitive compound. Natural extracts from fresh or oven-dried winemaking by-products could be used in other food industries as a valuable source of phenolic compounds with antioxidant properties. However, further studies on other drying methods are required for addressing the preservation of phenolic compounds from winery by-products successfully.

Thrombocytopenia-associated mutations in Ser/Thr kinase MASTL deregulate actin cytoskeletal dynamics in platelets.

MASTL, a Ser/Thr kinase that inhibits PP2A-B55 complexes during mitosis, is mutated in autosomal dominant thrombocytopenia. However, the connections between the cell-cycle machinery and this human disease remain unexplored. We report here that, whereas Mastl ablation in megakaryocytes prevented proper maturation of these cells, mice carrying the thrombocytopenia-associated mutation developed thrombocytopenia as a consequence of aberrant activation and survival of platelets. Activation of mutant platelets was characterized by hyperstabilized pseudopods mimicking the effect of PP2A inhibition and actin polymerization defects. These aberrations were accompanied by abnormal hyperphosphorylation of multiple components of the actin cytoskeleton and were rescued both in vitro and in vivo by inhibiting upstream kinases such as PKA, PKC, or AMPK. These data reveal an unexpected role of Mastl in actin cytoskeletal dynamics in postmitotic cells and suggest that the thrombocytopenia-associated mutation in MASTL is a pathogenic dominant mutation that mimics decreased PP2A activity resulting in altered phosphorylation of cytoskeletal regulatory pathways.