Dehydrogenase Gene Research Articles

Clinical Implementation of Rare and Novel DPYD Variants for Personalizing Fluoropyrimidine Treatment: Challenges and Opportunities.

Fluoropyrimidines (FLs) [5-Fluorouracil, Capecitabine] are used in the treatment of several solid tumors. Dihydropyrimidine dehydrogenase (DPD) is the rate-limiting enzyme for FL detoxification, and its deficiency could lead to severe, life-threatening or fatal toxicity after FL administration. Testing with a pharmacogenetic panel of four deleterious variants in the dihydropyrimidine dehydrogenase gene (DPYD) (DPYD*2A, DPYD*13, c.2846A > T, c.1129-5923C > G) prior to FL treatment, is recommended by scientific consortia (e.g., CPIC, DPWG) and drug regulatory agencies (e.g., EMA). However, this panel identifies < 20% of patients at risk of severe FL-related toxicity. Cumulative recent evidence highlights the potential clinical value of rare (minor allele frequency < 1%) and novel DPYD genetic variants for identifying an additional fraction of DPD-deficient patients at increased risk of severe FL-related toxicity. In this review, we aimed to comprehensively describe the available evidence regarding the potential clinical predictive role of novel and rare DPYD variants as toxicity markers in FL-treated patients, and to discuss the challenges and opportunities in tailoring FL treatment based upon clinical application of such markers. Although we must overcome existing barriers to the clinical implementation, the available data support that comprehensive assessment of the DPYD sequence, including rare and novel genetic variants, may significantly enhance the pre-emptive identification of at-risk patients, compared to the current targeted approach.

Effect of expansion of Shine-Dalgarno sequence for expression of malate- and aldehyde-dehydrogenase genes from Deinococcus geothermalis in Escherichia coli

The thermostability of thermophilic enzymes makes them ideal for use in biotechnology and medical science research. The overexpression of heterogeneous genes from thermophiles, such as the mesophilic Deinococcus geothermalis, in Escherichia coli is a genetic engineering technique for producing stable and useful proteins. In our previous study, 1- or 2-base expansion of the Shine-Dalgarno sequence, which is a ribosome-binding site in mRNA, was effective for the overexpression of the nicotinamide adenine dinucleotide oxidase gene from Deinococcus. geothermalis in Escherichia coli. In the present study, we examined the effect of expanding the Shine-Dalgarno sequence of the malate dehydrogenase gene and three genes from the aldehyde dehydrogenase family. Our results revealed that a 1- or 2-base expansion of the Shine-Dalgarno sequence was effective for the overexpression of all genes from Deinococcus geothermalis in Escherichia coli. However, the effects of the expansion of Shine-Dalgarno sequence from 3 to 5 bases were different between these genes.

Multiple Acyl-CoA Dehydrogenase Deficiency: Phenotypic and Genetic Features of a Malaysian Cohort.

Multiple acyl-CoA dehydrogenase deficiency (MADD) is an inherited disorder of fatty acid oxidation that causes lipid storage myopathy (LSM). This is the first report on MADD that describes the phenotypic and genetic features of a Malaysian cohort. Among the >2,500 patients in a local muscle biopsy database, patients with LSM were identified and their genomic DNA were extracted from muscle samples and peripheral blood. All 13 exons of the electron-transfer flavoprotein dehydrogenase gene (ETFDH) were subsequently sequenced. Fifty controls were included to determine the prevalence of identified mutations in the normal population. Fourteen (82%) of the 17 LSM patients had MADD with ETFDH mutations. Twelve (86%) were Chinese and two were Malay sisters. Other unrelated patients reported that they had no relevant family history. Nine (64%) were females. The median age at onset was 18.5 years (interquartile range=16-37 years). All 14 demonstrated proximal limb weakness, elevated serum creatine kinase levels, and myopathic changes in electromyography. Three patients experienced a metabolic crisis at their presentation. Sanger sequencing of ETFDH revealed nine different variants/mutations, one of which was novel: c.998A>G (p.Y333C) in exon 9. Notably, 12 (86%) patients, including the 2 Malay sisters, carried a common c.250G>A (p.A84T) variant, consistent with the hotspot mutation reported in southern China. All of the patients responded well to riboflavin therapy. Most of our Malaysian cohort with LSM had late-onset, riboflavin-responsive MADD with ETFDH mutations, and they demonstrated phenotypic and genetic features similar to those of cases reported in southern China. Furthermore, we report a novel ETFDH mutation and possibly the first ever MADD patients of Malay descent.

Cloning and functional analysis of 3β-hydroxysteroid dehydrogenase gene involved in biosynthesis of digitoxin

Digitoxin, an important secondary metabolite of Digitalis purpurea, is a commonly used cardiotonic in clinical practice. 3β-Hydroxysteroid dehydrogenase(3βHSD) is a key enzyme involved in the biosynthesis of digitoxin. It belongs to the short-chain dehydrogenase/reductase(SDR) family, playing a role in the biosynthesis of cardiac glycosides by oxidizing and isomerizing the precursor sterol. In this study, two 3βHSD genes were cloned from D. purpurea. The results showed that the open reading frame(ORF) of Dp3βHSD1 was 780 bp, encoding 259 amino acid residues. The ORF of Dp3βHSD2 was 774 bp and encoded 257 residues. Dp3βHSD1/2 had the cofactor binding site TGxxxA/GxG and the catalytic site YxxxK. In vitro experiments confirmed that Dp3βHSD1/2 catalyzed the generation of progesterone from pregnenolone, and Dp3βHSD1 had stronger catalytic capacity than Dp3βHSD2. The expression level of Dp3βHSD1 was much higher than that of Dp3βHSD2 in leaves, and digitoxin was only accumulated in leaves. The results implied that Dp3βHSD1 played a role in the dehydrogenation of pregnenolone to produce progesterone in the biosynthesis of digitoxin. This study provides a reference for further exploring the biosynthetic pathway of cardiac glycosides in D. purpurea.

Rice NADP-dependent malate dehydrogenase gene OsMDH8.2 is involved in heat tolerance

Convergent and divergent evolution lead to plants with stronger adaptability to higher temperatures, thus averting crop biomass yield reductions. NADP-dependent malate dehydrogenase (NADP-MDH) is a redox-regulated enzyme that catalyzes the reversible reduction of oxaloacetate to malate. The sole NADP-MDH gene in rice, OsMDH8.2, is expressed in mesophyll cells of photosynthetic organs and various sink tissues. However, it is unknown whether NADP-MDH functions in heat stress in rice. We characterized a transgenic OsMDH8.2 overexpression line under thermal stress treatment (40 °C). The transgenic line exhibited better adaptability to heat stress than the wild type; it better maintained biomass and a lower surface temperature through stomatal closure, and photosynthetic activity was less affected. OsMDH8.2 was found to affect peroxidase activity by reducing the hydrogen peroxide content in flag leaves after 3 and 5 days of thermal stress. Analysis results of OsMDH8.2 knockout lines confirmed that OsMDH8.2 contributes to heat tolerance. Transcriptome and metabolome analyses demonstrated that OsMDH8.2 plays a key role in energy homeostasis by reducing tricarboxylic acid cycle activity while inducing the glyoxylate cycle to produce more energy, and by regulating amino acid metabolism to rescue heat-stress damage. Collectively, these results suggest that OsMDH8.2 enhances glyoxylate cycle efficiency, resulting in lower carbon dioxide release into the environment through stomatal closure, providing an excellent strategy for improving plant heat tolerance through genetic engineering.

Genome-wide characterization of soybean malate dehydrogenase genes reveals a positive role of GmMDH2 in salt stress response

Malate dehydrogenase (MDH) is a widely expressed enzyme that plays a key role in plant growth, development, and the stress response. However, information on MDH genes in the soybean genome is limited. Seventeen members of the soybean MDH family were identified by genome-wide analysis, and the presence of conserved protein motifs was analyzed. The genes were divided into five clusters according to their phylogenetic relationships. The intracellular localizations of six GmMDHs were determined by confocal microscopy on Arabidopsis mesophyll protoplasts. Transcripts of GmMDHs were significantly increased by abiotic stress (drought, salt, and alkalinity) and hormone treatments, as shown by analysis of cis-regulatory elements and quantitative real-time polymerase chain reaction (qRT-PCR). GmMDHs displayed unique expression patterns in diverse soybean tissues. It is noteworthy that under salt stress, the expression levels of a chloroplast isoform (GmMDH2) were unusually high, presumably indicating a critical role in soybean responses to salinity. Expression of GmMDH2 in Escherichia coli showed that the recombinant enzyme had NADP-dependent MDH activity. The redox states of the nicotinamide adenine dinucleotide phosphate (NADPH) pool and antioxidant activities were shown to be modulated by GmMDH2 gene overexpression, which in turn reduced reactive oxygen species (ROS) formation in transgenic soybean, significantly enhancing the salt stress resistance. Gene-based association analysis showed that variations in GmMDH2 were strongly linked to seedling salt tolerance. A polymorphism possibly associated with salt tolerance was discovered in the promoter region of GmMDH2. These findings not only improve our understanding of the stress response mechanism by identifying and characterizing the MDH gene family throughout the soybean genome but it also identified a potential candidate gene for the future enchancement of salt tolerance in the soybean.

СПОСОБЫ ПОЛУЧЕНИЯ МОЛОЧНОЙ КИСЛОТЫ С ПОМОЩЬЮ ДРОЖЖЕВЫХ ШТАММОВ, СКОНСТРУИРОВАННЫХ СОВРЕМЕННЫМИ МЕТОДАМИ ГЕНЕТИЧЕСКОЙ ИНЖЕНЕРИИ

Methods for producing lactic acid using yeast strains constructed by cutting-edge methods of genetic engineering are discussed in the review. Some of the latest studies and patents on increasing the productivity of yeast strains producing lactic acid and improving their characteristics using genome editing methods, as well as imparting the ability to synthesize lactic acid to new yeast strains that have not previously been used for this purpose are presented. New yeast strains producing lactic acid are obtained by transforming the original strains with plasmid vectors carrying a foreign lactate dehydrogenase gene or an existing enhanced one. To introduce a foreign gene into a plasmid, homologous recombination is mainly used, which underlies the modern method of genome editing by CRISPR-Cas9. To enhance an existing gene, plasmids in which genes encoding enzymes of competing metabolic pathways are knocked out, or plasmids with increased copies of the LDH gene and an enhanced corresponding promoter are developed. In addition, by editing the genome of yeast cells, it is possible to increase their acid tolerance, reduce the synthesis of by-products, and impart the ability to use new substrates for the synthesis of lactic acid.

HYPERTROPHIC OSTEOARTHROPATHY IN A PATIENT WITH HETEROZYGOUS MUTATION IN THE SLCO2A1 GENE: A CASE REPORT

Hypertrophic osteoarthropathy (HOA) is a condition characterized by aberrant skin and osseous tissue proliferation in the distal extremities. Mutations in the 15-hydroxyprostaglandin dehydrogenase gene (HPGD) and the soluble carrier organic anion carrier family member 2A1 gene (SLCO2A1) were associated with primary HOA. Secondary HOA, which is also called as 'hypertrophic pulmonary osteoarthropathy' is responsible for 95-97% of cases. Herein, we present a 19-year-old female patient with primary HOA and heterozygous mutation in the SLCO2A1 gene.

The chromatin remodeler Ino80 regulates yeast stress tolerance and cell metabolism through modulating nitrogen catabolite repression

Chromatin remodelers are important in maintaining the dynamic chromatin state in eukaryotic cells, which is essential for epigenetic regulation. Among the remodelers, the multi-subunits complex INO80 plays crucial roles in transcriptional regulation. However, current knowledge of chromatin regulation of the core subunit Ino80 on stress adaptation remains mysterious. Here we revealed that overexpressing the chromatin remodeler Ino80 elevated tolerance to multiple stresses in budding yeast Saccharomyces cerevisiae. Analyses of differential chromatin accessibility and global transcription levels revealed an enrichment of genes involved in NCR (nitrogen catabolite repression) under acetic acid stress. We demonstrated that Ino80 overexpression reduced the histone H3 occupancy in the promoter region of the glutamate dehydrogenase gene GDH2 and the allantoinase gene DAL1. Consistently, the decreased occupancy of nucleosome was revealed in the Ino80-inactivation mutant. Further analyses showed that Ino80 was recruited to the specific DNA locus in the promoter region of GDH2. Consistently, Ino80 overexpression facilitated the utilization of non-preferred nitrogen source to enhance ethanol yield under prolonged acetic acid stress. These results demonstrate that Ino80 plays a crucial role in coordinating carbon and nitrogen metabolism during stress adaptation.

MOLECULAR IDENTIFICATION OF A LOCAL LACTIC ACID BACTERIA (LAB) ISOLATE (B21) AND PRIMER CONFIRMATION FOR D-Lactate Dehydrogenase (D-LDH) GENE ISOLATION

This study aims to identify a lactic acid bacteria (BAL) isolated from native chicken (Gallus gallus domesticus) intestinum (B21) based on 16S rRNA gene sequence and also to confirm primers for PCR-based D-Lactate dehydrogenase (D-LDH) gene isolation. The genomic DNA of B21 was isolated then amplified using 16S rRNA. The PCR product then sequenced and aligned. The sequencing was done in 1st Base Pte. Malaysia. The sequence then aligned using the Basic Local Alignment Search Tool (BLAST) in NCBI to know the similar genome species. Isolation D-LDH gene was done through PCR using 5 primers (idb 0101, idb 1147, idb 0813, idb 1010, and idb 2021) of B21 and also J15 isolate genome. The PCR product were then sequenced and aligned with the D-LDH gene sequences in NCBI.&#x0D; The results of molecular identification based on 16S rRNA markers showed that B21 has 99.45% similarity to Bacillus proteolyticus. Amplification of the LDH gene with idb 1010 primer on J15 isolate yielded 4 fragments (i.e. 650, 1400, 1500, and 1700bp). The results of phylogenetic tests showed that the 1700bp fragment has a high similarity to the D-LDH gene in Myobacterium tuberculosis species with a bootstrap value of 84%. While other size fragments cannot be trusted to have similarity to LDH sequence because they have not reached the minimum bootstrap value that meets the qualifications.

First report of Colletotrichum gloeosporioides causing severe anthracnose on Litsea deccanensis seedlings in India

Litsea deccanensis, commonly known as Jiggat, finds commercial use in India for the production of agarbatti or incense sticks, with its stem bark serving as a crucial raw material. The plant seedlings are important for conservation efforts, biodiversity, and potential uses in medicine, horticulture, and research. Within the campus nursery of the Institute, distinct signs of anthracnose were observed in the seedlings of L. deccanensis. The incidence of this disease was documented at 50 % among the hundred assessed seedlings maintained in the KFRI nursery. Through rigorous isolation, purification, and analysis, the responsible pathogen was identified as Colletotrichum sp. Phylogenetic analysis of concatenated sequence alignment of Internal Transcribed Spacer (ITS) region, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and beta-tubulin (TUB2) genes supported by cultural morphological characteristics revealed Colletotrichum gloeosporioides as causal agent. To affirm the pathogenic nature of the fungus, pathogenicity tests were conducted, both in controlled environments and within the living plant, supported by subsequent re-isolation and identification of the pathogen, in accordance with Koch's postulates. To the best of our knowledge, this is the first report of anthracnose caused by C. gloeosporioides on Litsea deccanensis in India.

Effect of the overexpression of external alternative NADH dehydrogenase gene in Arabidopsis on the resistance of transformed tobacco plants to negative temperatures

The study aims to confirm the functional activity and localization of the At_NDB2 transgenic protein of Arabidopsis in tobacco cells and to evaluate the effect produced by the permanent expression of external alternative NADH dehydrogenase on the resistance of a heat-loving plant to low temperatures. Proteins and mitochondria were isolated from the leaves of tobacco plants grown at 25 °С (day/night) at the 7-leaf stage. At_NDB2 protein localization in mitochondria was determined via electrophoresis and immunoblotting. The functional activity of At_NDB2 was confirmed through the polarography of isolated mitochondria and the specific mitochondrial complex I inhibitor (rotenone). It was also found that the cyanide-resistant respiration rate and the activity of an alternative oxidase enzyme were significantly higher in transgenic plants than in wild-type plants. In order to determine the resistance to low temperatures, the parent and transgenic tobacco plants were grown in soil until the 2–3 and 6–7 leaf stages, after which they were kept at 3 to -3 °С for a day in the dark and left to regrow at 25 °С for seven days. Although it was previously shown that oxidative stress is reduced in transgenic plants at low temperatures as compared to wild-type plants, it was established that the tolerance of transgenic and nontransformed plants does not differ. Thus, alternative NADH dehydrogenase activity was found to reduce oxidative stress and increase alternative oxidase activity, without enhancing the resistance of Nicotiana tabacum to negative temperatures.

Deteksi dan Identifikasi Molekuler Kapang Berpotensi sebagai Penghasil Patulin pada Buah Tropis

Production and export of tropical fruits in Indonesia have an increasing trend every year. The important factor in the production and export of fruits is food safety. One of the contaminants that may be found in fruits is mycotoxin, especially patulin. Patulin mainly found in fruits such as apple, orange, grape, and pear. This study aimed to detect molds from tropical fruits and to identify potentially patulin-producing molds. Detection of potentially patulin-producing molds obtained from tropical fruits was carried out using the isoepoxydon dehydrogenase (idh) gene. Species identification was carried out using internal transcribed spacer (ITS) region. The mold isolate sequences of ITS rDNA region were analyzed for their homology using both BLAST search and phylogenetic tree. A total of 26 molds were isolated from four types of fruit (malang apple, ambon banana, medan orange, and avocado) obtained from five different places in Bogor including fruit stalls, traditional markets, and supermarkets. The positive results of idh gene were mold isolates that have an amplicon at 620 bp. The result showed that 5 of 26 (19,2%) isolates were positive for idh gene, namely A11, A34, A43, A51 and B23. The positive isolates for idh gene were identified as Aspergillus aculeatus, Aspergillus niger, Cladosporium anthropophilum, Cladosporium tenuissimum, and Talaromyces verruculosus.

Increases in anterograde axoplasmic transport in neurons of the hyper-glutamatergic, glutamate dehydrogenase 1 (Glud1) transgenic mouse: Effects of glutamate receptors on transport.

The excitatory neurotransmitter glutamate has a role in neuronal migration and process elongation in the central nervous system (CNS). The effects of chronic glutamate hyperactivity on vesicular and protein transport within CNS neurons, that is, processes necessary for neurite growth, have not been examined previously. In this study, we measured the effects of lifelong hyperactivity of glutamate neurotransmission on axoplasmic transport in CNS neurons. We compared wild-type (wt) to transgenic (Tg) mice over-expressing the glutamate dehydrogenase gene Glud1 in CNS neurons and exhibiting increases in glutamate transmitter formation, release, and synaptic activation in brain throughout the lifespan. We found that Glud1 Tg as compared with wt mice exhibited increases in the rate of anterograde axoplasmic transport in neurons of the hippocampus measured in brain slices exvivo, and in olfactory neurons measured in vivo. We also showed that the in vitro pharmacologic activation of glutamate synapses in wt mice led to moderate increases in axoplasmic transport, while exposure to selective inhibitors of ion channel forming glutamate receptors very significantly suppressed anterograde transport, suggesting a link between synaptic glutamate receptor activation and axoplasmic transport. Finally, axoplasmic transport in olfactory neurons of Tg mice in vivo was partially inhibited following 14-day intake of ethanol, a known suppressor of axoplasmic transport and of glutamate neurotransmission. The same was true for transport in hippocampal neurons in slices from Glud1 Tg mice exposed to ethanol for 2 h exvivo. In conclusion, endogenous activity at glutamate synapses regulates and glutamate synaptic hyperactivity increases intraneuronal transport rates in CNS neurons.

Using In silico Tools to Analyze the 5ʹ Untranslated Regions of the Alcohol Dehydrogenase Gene from Arabidopsis thaliana and Omega Sequence

Using In silico Tools to Analyze the 5ʹ Untranslated Regions of the Alcohol Dehydrogenase Gene from Arabidopsis thaliana and Omega Sequence

Genetic characterization and description of Leishmania (Leishmania) ellisi sp. nov.: a new human-infecting species from the USA

In a 2018 report, an unusual case of cutaneous leishmaniasis was described in a 72-year-old female patient residing in Arizona, United States of America (USA). Preliminary analysis of the 18S rDNA and glyceraldehyde-3-phosphate dehydrogenase genes supported the conclusion that the Leishmania strain (strain 218-L139) isolated from this case was a novel species, though a complete taxonomic description was not provided. Identification of Leishmania at the species level is critical for clinical management and epidemiologic investigations so it is important that novel human-infecting species are characterized taxonomically and assigned a unique scientific name compliant with the ICZN code. Therefore, we sought to provide a complete taxonomic description of Leishmania strain 218-L139. Phylogenetic analysis of several nuclear loci and partial maxicircle genome sequences supported its position within the subgenus Leishmania and further clarified the distinctness of this new species. Morphological characterization of cultured promastigotes and amastigotes from the original case material is also provided. Thus, we conclude that Leishmania (Leishmania) ellisi is a new cause of autochthonous cutaneous leishmaniasis in the USA.

Compilation of Genotype and Phenotype Data in GCDH-LOVD for Variant Classification and Further Application.

Glutaric aciduria type 1 (GA-1) is a rare but treatable autosomal-recessive neurometabolic disorder of lysin metabolism caused by biallelic pathogenic variants in glutaryl-CoA dehydrogenase gene (GCDH) that lead to deficiency of GCDH protein. Without treatment, this enzyme defect causes a neurological phenotype characterized by movement disorder and cognitive impairment. Based on a comprehensive literature search, we established a large dataset of GCDH variants using the Leiden Open Variation Database (LOVD) to summarize the known genotypes and the clinical and biochemical phenotypes associated with GA-1. With these data, we developed a GCDH-specific variation classification framework based on American College of Medical Genetics and Genomics and the Association for Molecular Pathology guidelines. We used this framework to reclassify published variants and to describe their geographic distribution, both of which have practical implications for the molecular genetic diagnosis of GA-1. The freely available GCDH-specific LOVD dataset provides a basis for diagnostic laboratories and researchers to further optimize their knowledge and molecular diagnosis of this rare disease.

Identification of IDH and TERTp mutations using dynamic susceptibility contrast MRI with deep learning in 162 gliomas

PurposeIsocitrate dehydrogenase (IDH) and telomerase reverse transcriptase gene promoter (TERTp) mutations play crucial roles in glioma biology. Such genetic information is typically obtained invasively from excised tumor tissue; however, these mutations need to be identified preoperatively for better treatment planning. The relative cerebral blood volume (rCBV) information derived from dynamic susceptibility contrast MRI (DSC-MRI) has been demonstrated to correlate with tumor vascularity, functionality, and biology, and might provide some information about the genetic alterations in gliomas before surgery. Therefore, this study aims to predict IDH and TERTp mutational subgroups in gliomas using deep learning applied to rCBV images. MethodAfter the generation of rCBV images from DSC-MRI data, classical machine learning algorithms were applied to the features obtained from the segmented tumor volumes to classify IDH and TERTp mutation subgroups. Furthermore, pre-trained convolutional neural networks (CNNs) and CNNs enhanced with attention gates were trained using rCBV images or a combination of rCBV and anatomical images to classify the mutational subgroups. ResultsThe best accuracies obtained with classical machine learning algorithms were 83 %, 68 %, and 76 % for the identification of IDH mutational, TERTp mutational, and TERTp-only subgroups, respectively. On the other hand, the best-performing CNN model achieved 88 % accuracy (86 % sensitivity, 91 % specificity) for the IDH-mutational subgroups, 70 % accuracy (73 % sensitivity and 67 % specificity) for the TERTp-mutational subgroups, and 84 % accuracy (86 % sensitivity, 81 % specificity) for the TERTp-only subgroup using attention gates. ConclusionsDSC-MRI can be utilized to noninvasively classify IDH- and TERTp-based molecular subgroups of gliomas, facilitating preoperative identification of these genetic alterations.

Sex-Chromosome-Related Dimorphism in Steroidogenic Enzymes and Androgen Receptor in Response to Testosterone Treatment: An In Vitro Study on Human Primary Skeletal Muscle Cells.

Gender-related methodology in biomedical sciences receives considerable attention, with numerous studies highlighting biological differences between cisgender males and females. These differences influence the clinical symptoms of various diseases and impact therapeutic approaches. In this in vitro study, we investigate the potential role of sex-chromosome-related dimorphism on steroidogenic enzymes, androgen receptor (AR) expression, and cellular translocation in primary human skeletal muscle cells before and after exposure to testosterone. We analyzed 46XY and 46XX cells for 17β-hydroxysteroid dehydrogenase (17β-HSD), 5α-reductase (5α-R2), aromatase (Cyp-19), and AR gene expression. We also compared AR expression and intracellular translocation after increasing exposure to testosterone. At baseline, we observed higher mRNA expression for 5α-R2 and AR in 46XY cells and higher Cyp-19 mRNA expression in 46XX cells. Following testosterone exposure, we observed an increase in AR expression and translocation in 46XX cells, even at the lowest dose of 0.5 nM, while significant changes in 46XY cells were observed only from 10 nM. Our in vitro results demonstrate that the diverse sex chromosome assets reflect important differences in muscle steroidogenesis. They support the concept that chromosomal disparities between males and females, even in vitro, lead to pivotal variations in cellular physiology and response. This understanding represents a crucial starting point in gender medicine, ensuring a precise approach in clinical practice, sports, and exercise settings and facilitating the translation of in vitro data to in vivo applicability.

Comparative characterization and expression analysis revealed cinnamyl alcohol dehydrogenase genes differentially responding to Colletotrichum fructicola in woodland and cultivated strawberries

BACKGROUND: Strawberry (Fragaria×ananassa) production is hampered by anthracnose caused by Colletotrichum spp. Cinnamyl alcohol dehydrogenase (CAD) catalyzes lignin biosynthesis to fortify plant cell wall upon various stresses, but their involvement in strawberry resistance is elusive. OBJECTIVE: Current work aims to uncover the involvement of CAD genes in both woodland and cultivated strawberry responses to Colletotrichum spp. METHODS: Genome-wide identification of CAD genes was updated in cultivated strawberry. Comparative molecular characterization integrated with pathogenesis phenotyping after C. fructicola inoculation, were followed by RT-qPCR analysis and RNAseq data retrieval. RESULTS: Cultivated strawberry comprises 32 CAD genes of three classes. Thirteen syntenic relationships were observed between cultivated and woodland strawberry, with high homology in protein coding regions while promoters being greatly diversified. F. vesca ‘Hawaii4’ was least susceptible, followed by octoploid cultivar ‘ShenQi’, and ‘Benihoppe’ the most susceptible to C. fructicola. The class I CAD4 and CAD5, preferentially expressed in leaves and crowns, respectively, were up-regulated in susceptible varieties at 72 h post inoculation (hpi). Of class II, CAD8 was up-regulated in three genotypes at distinct timepoints, while CAD12 was quickly down-regulated in ‘ShenQi’ upon inoculation. CAD1 and CAD2 of class III were down-regulated in ‘ShenQi’ and ‘Hawaii4’ at distinct stages. The strong involvement of CAD genes in strawberry response to Colletotrichum spp. sets a basis for future functional study. CONCLUSION: F. vesca is less susceptible to Colletotrichum spp. than cultivated strawberries but the expression of CAD family in F. vesca and F.×ananassa were differentially provoked in timing and magnitude hinting no correlation with strawberry resistance to this pathogen.