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Recently, the siRNAs pathways, and especially reproductive phasiRNAs, have attracted attention in eudicots since their biological roles are still unknown and their biogenesis took different evolutionary pathways compared to monocots. In this work, we used Coffea arabica L., a recent allotetraploid formed from the hybridization of Coffea canephora and C. eugenioides unreduced gametes, to explore microsporogenesis and small RNAs-related pathways in a eudicot crop. First, we identified the microsporogenesis stages during anther development revealing that pre-meiosis occurs in anthers of 1.5 mm inside floral buds (FBs), whereas meiosis between 1.5 and 4.2 mm FBs, and post-meiosis in FBs larger than 4.2 mm. These stages coincide with the Brazilian winter, a period of FBs reduced growth which suggests temperature sensitivity. Next, we identified and quantified the expression of reproductive 21- and 24-nt phasiRNAs during coffee anther development together with their canonical and novel miRNA triggers, and characterized the DCL and AGO families. Our results showed that the pattern of reproductive phasiRNA abundance in C. arabica is unique among described eudicots and the canonical trigger car-miR2275 is involved in the processing of both 21- and 24-nt phasiRNAs. Fourteen DCL genes were identified, but DCL5, related to phasiRNA biosynthesis in monocots, was not, according to its specificity for monocots. Thus, our work explored the knowledge gap about microsporogenesis and related siRNAs pathways in coffee, contributing to the control of reproductive development and the improvement of fertility in eudicots.

The use of transcriptomic data to make inferences about plant metabolomes is a useful tool to help the discovery of important compounds in the available biodiversity. To unveil previously undiscovered metabolites of Coffea, of phytotherapeutic and economic value, we employed 24 RNAseq libraries. These libraries were sequenced from leaves exposed to a diverse range of environmental conditions. Subsequently, the data were meticulously processed to create models of putative metabolic networks, which shed light on the production of potential natural compounds of significant interest. Then, we selected one of the predicted compounds, the L-3,4-dihydroxyphenylalanine (L-DOPA), to be analyzed by LC-MS/MS using three biological replicates of flowers, leaves, and fruits from Coffea arabica and Coffea canephora. We were able to identify metabolic pathways responsible for producing several compounds of economic importance. One of the identified pathways involved in isoquinoline alkaloid biosynthesis was found to be active and producing L-DOPA, which is a common product of POLYPHENOL OXIDASES (PPOs, EC 1.14.18.1 and EC 1.10.3.1). We show that coffee plants are a natural source of L-DOPA, a widely used medicine for treatment of the human neurodegenerative condition called Parkinsons disease. In addition, dozens of other compounds with medicinal significance were predicted as potential natural coffee products. By further refining analytical chemistry techniques, it will be possible to enhance the characterization of coffee metabolites, enabling a deeper understanding of their properties and potential applications in medicine.

MADS-box transcription factors (TFs) are involved in multiple plant development processes and are most known during the reproductive transition and floral organ development. Very few genes have been characterized in the genome of Humulus lupulus L. (Cannabaceae), an important crop for the pharmaceutical and beverage industries. The MADS-box family has not been studied in this species yet. We identified 65 MADS-box genes in the hop genome, of which 29 encode type-II TFs (27 of subgroup MIKCC and 2 MIKC*) and 36 type-I proteins (26 α, 9 β, and 1 γ). Type-II MADS-box genes evolved more complex architectures than type-I genes. Interestingly, we did not find FLOWERING LOCUS C (FLC) homologs, a transcription factor that acts as a floral repressor and is negatively regulated by cold. This result provides a molecular explanation for a previous work showing that vernalization is not a requirement for hop flowering, which has implications for its cultivation in the tropics. Analysis of gene ontology and expression profiling revealed genes potentially involved in the development of male and female floral structures based on the differential expression of ABC homeotic genes in each whorl of the flower. We identified a gene exclusively expressed in lupulin glands, suggesting a role in specialized metabolism in these structures. In toto, this work contributes to understanding the evolutionary history of MADS-box genes in hop, and provides perspectives on functional genetic studies, biotechnology, and crop breeding.

Orbital exenteration is a severe and disabling surgical procedure that involves the removal of all orbital contents. Effective reconstruction is crucial to managing the resulting defects. This study aims to propose a reconstructive algorithm utilizing free flaps derived from the lateral circumflex femoral artery (LCFA) system for orbital exenteration defects, based on our clinical experience. Patients who underwent orbital exenteration and subsequent reconstruction using LCFA-based free flaps between January 2013 and January 2022 were included in this study. A total of 44 patients underwent orbital exenteration, followed by LCFA free flap reconstruction. The choice of reconstructive technique depended on defect type and dura mater exposure, adhering to the proposed algorithm. The procedures included 20 standard ALT (Anterolateral thigh) flaps, 13 chimeric ALT flaps, 1 SALT chimeric flap with skin grafting, 7 SALT flaps with skin grafting, and 3 buried SALT flaps. Complications were recorded in 10 cases at the recipient site and 2 cases at the donor site. Orbital exenteration presents significant challenges both surgically and reconstructively, with a high risk of local recurrence and postoperative mortality. The reconstruction algorithm using LCFA-based free flaps, as proposed in this study, may guide surgeons in selecting the most appropriate technique, tailored to the specific defect and considering postoperative treatments such as adjuvant radiotherapy. [ABSTRACT FROM AUTHOR]

Hypoalbuminemia, a biomarker of malnutrition, has been associated with adverse surgical outcomes;, however, its impact on breast reduction surgery is not yet well-documented. We queried the American college of surgeons national surgical quality improvement program database to identify patients who underwent breast reduction surgery between 2008 and 2022. Patients were grouped by preoperative normal albumin levels (≥3.5 g/dL) and hypoalbuminemia (<3.5 g/dL). Preoperative, intraoperative, and 30-day postoperative outcomes, including complications and readmissions, were compared using the univariate tests and multivariable logistic regression. We included a total of 7277 cases, among whom 96% (n = 6964) had normal albumin values and 4% (n = 298) had hypoalbuminemia (n = 298). Patients with hypoalbuminemia showed a significantly higher body mass index (37.1 ± 8.1 vs. 33.3 ± 6.3 kg/m², p < 0.001) and were more likely to be Black or African American (49.0 vs. 27.8%, p < 0.001). Comorbidities such as diabetes (14.7 vs. 7.4%, p < 0.001), chronic obstructive pulmonary disease (4.0 vs. 1.0%, p < 0.001), and hypertension (35.2 vs. 26.3%, p = 0.002) were significantly more prevalent in the hypoalbuminemia group. Hypoalbuminemia was associated with a significantly increased risk of complications (13.8 vs. 6.1%, p < 0.001), with higher rates of superficial incisional infections (7.0 vs. 2.6%, p = 0.001) and unplanned readmissions (3.4 vs. 1.4%, p = 0.05). Multivariable analysis confirmed hypoalbuminemia as an independent predictor of postoperative complications (OR 1.96, p = 0.001), medical complications (OR 2.62, p = 0.02), and surgical complications (OR 1.91, p = 0.02). Hypoalbuminemia significantly raises the risk of 30-day postoperative complications in breast reduction surgery. Preoperative nutritional assessment and optimization are crucial in improving surgical outcomes, particularly in patients with high body mass index and comorbidities. [ABSTRACT FROM AUTHOR]

microRNAs (miRNAs) are derived from self-complementary hairpin structures, while small-interfering RNAs (siRNAs) are derived from double-stranded RNA (dsRNA) or hairpin precursors. The core mechanism of sRNA production involves DICER-like (DCL) in processing the smallRNAs (sRNAs) and ARGONAUTE (AGO) as effectors of silencing, and siRNA biogenesis also involves action of RNA-Dependent RNA Polymerase (RDR), Pol IV and Pol V in biogenesis. Several other proteins interact with the core proteins to guide sRNA biogenesis, action, and turnover. We aimed to unravel the components and functions of the RNA-guided silencing pathway in a non-model plant species of worldwide economic relevance. The sRNA-guided silencing complex members have been identified in the Coffea canephora genome, and they have been characterized at the structural, functional, and evolutionary levels by computational analyses. Eleven AGO proteins, nine DCL proteins (which include a DCL1-like protein that was not previously annotated), and eight RDR proteins were identified. Another 48 proteins implicated in smallRNA (sRNA) pathways were also identified. Furthermore, we identified 235 miRNA precursors and 317 mature miRNAs from 113 MIR families, and we characterized ccp-MIR156, ccp-MIR172, and ccp-MIR390. Target prediction and gene ontology analyses of 2239 putative targets showed that significant pathways in coffee are targeted by miRNAs. We provide evidence of the expansion of the loci related to sRNA pathways, insights into the activities of these proteins by domain and catalytic site analyses, and gene expression analysis. The number of MIR loci and their targeted pathways highlight the importance of miRNAs in coffee. We identified several roles of sRNAs in C. canephora, which offers substantial insight into better understanding the transcriptional and post-transcriptional regulation of this major crop.

Neoumbilicoplasty aims to reconstruct an aesthetically pleasing new umbilicus following agenesis, malignancy, anatomical distortion, or umbilicus loss. Despite the wide variety of surgical techniques described, literature is scarce when it comes to standardized categorization of these as well as the clear definition of patients' selections, specific indications, final outcomes, and possible complications. According to available literature, this work aims to evaluate different surgical approaches, and correlate them to specific surgical needs, to simplify the surgical choice and patient management. A systematic review was performed in December 2020 in PubMed, Web of Science, and MedLine Ovid databases according to the PRISMA guidelines. A total of 41 studies and 588 patients were finally included. On the basis of the evidence of the literature collected, we divided the studies into four groups according to the neoumbilicoplasty techniques: single suture or purse-string suture, single flap, multiple flap, and skin graft. Patients' surgical comorbidities, neoumbilicoplasty indications, and aesthetic and surgical outcomes were investigated. Direct suture and single and multiple flap techniques assured overall, satisfactory cosmetic outcomes with a low rate of surgical complications. Whereas suture-only techniques were chosen mostly by general surgeons/urologists in laparoscopic surgery, the single flap was the preferred method to reconstruct the umbilicus in open abdominal surgery or combined abdominoplasty with herniorrhaphy. Multiple flap and skin grafts were adopted in abdominoplasty-related umbilicus reconstruction, although the latter option showed impactful aesthetic and surgical complications. Umbilicoplasty can assure generally pleasant aesthetic outcomes with relatively low complication rates. Indications for specific techniques correspond to different patient populations and surgical scenarios. [ABSTRACT FROM AUTHOR]