Sustainability‐Oriented Businesses and Earnings Management. What Is the True Relationship? Evidence From Italian Benefit Corporations

This study examines how benefit corporations (BCs)—a type of sustainability-­ oriented hybrid business dedicated to corporate social responsibility (CSR) goals—are associated with earnings management (EM). Two theoretical perspectives—long-­ term strategy and managerial opportunism—shape the hypotheses on the association between BC adoption and EM practices. An empirical analysis is conducted on a panel dataset of 2449 Italian firms (354 BCs and 2095 non-­ BCs) from 2014 to 2021. Using Stubben's (2010) conditional earnings model and propensity score matching, the findings suggest that adopting the BC model is linked to a higher likelihood of managerial engagement in EM. Although the BC model emphasizes transparency and accountability, it might unintentionally enable opportunistic behavior, calling into question the assumption that CSR-­ oriented business models inherently limit financial manipulation. These findings shed light on the impact of legal frameworks for hybrid organizations like BCs on corporate behavior, offering valuable insights for policymakers and practitioners working to foster ethical business practices

Tackling redox pathways in eukaryotic parasites by a structural biology approach

All aerobic organisms are exposed to reactive oxygen species (ROS) generated as byproducts of their metabolism during their lifetime. To prevent and repair ROS-derived damage and maintain cell homeostasis, both prokaryotic and eukaryotic organisms have evolved enzymatic and non-enzymatic antioxidant systems. Among them, thioredoxin reductase (TrxR) and glutathione reductase (GR), belonging to the pyridine nucleotide-disulfide oxidoreductase family, play key roles in the regulation of redox pathways across all organisms. In particular, parasites are repeatedly exposed to the host defense machinery in their invasive stages and have to cope with oxidative stress generated both by their metabolic reactions and by the host immune system. On the global scale of public health impact, infectious diseases still deserve attention, and new therapies are urgently needed to combat devastating parasitic infections worldwide. Given their involvement in parasite survival, redox pathways represent an outstanding source of druggable target for novel antiparasitic drug discovery. The present thesis work undertakes the investigation by a structural biology approach of key thiol-dependent enzymes from two human eucaryotic parasites: Thioredoxin Glutathione Reductase from Schistosoma mansoni (SmTGR) and Thioredoxin Reductase from Cryptosporidium parvum (CpTrxR). The two studies are at different stages of research but are both grounded in a thorough characterization of the structure-function relationship of the identified drug targets, with the goal of exploiting them in a structure-based drug design approach. SmTGR is a validated and well-characterized drug target against Schistosomiasis, a neglected tropical disease affecting 280 million people and resulting in 200 000 deaths annually. To date, only praziquantel is available for the treatment of schistosomiasis and, due to massive drug administration, less sensitive parasite strains are emerging, making the identification of new therapies urgent. However, selective drug development for this class of enzyme is challenging, mainly due to the reliance on irreversible and/or covalent inhibition strategies, which are associated with unacceptable off-targets effects. This challenge was further exacerbated, until a few years ago, by the lack of structural data. Recently, a breakthrough by means of an X-ray crystallography fragment screening allowed us to identify the so-called "doorstop pocket", a novel regulatory and druggable site of TGR. The initial molecular fragments identified were ligated and partially optimized, but further attempts to determine the binding mode of these inhibitors by X-ray crystallography were unsuccessful. Thus, we settle on an integrative structural biology approach by switching to the cutting-edge cryo-EM technique that enables us to solve the first structure of TGR-inhibitor complex by this method, validating the doorstop pocket as a druggable site. Since this allosteric site is present in the whole protein family, this breakthrough offers opportunities for selectively inhibiting other pyridine nucleotide-disulfide oxidoreductases essential for various pathogens and holds implications for combating cancer. The experimental work carried out on this project has been supported by grants of the National Institute of Allergy and Infectious diseases (NIH/NIAID). On the other hand, TrxR from the apicomplexan Cryptosporidium parvum has attracted increasing attention as a promising drug target against cryptosporidiosis, the leading cause of diarrheal disease worldwide. Despite the established association with pediatric morbidity and mortality in low- and middle-income countries and with a chronic and life-threatening enteric disease in HIV-AIDS patients, there are no vaccines and treatments options are severely limited. Drug development is further hindered by the lack of many conventional target exploited for other parasitic diseases and limited ex vivo Cryptosporidium models. Compared to the related and most known malaria parasite Plasmodium falciparum, too little is known about antioxidant defense systems in Cryptosporidium. While P. falciparum relies on both thioredoxin (Trx) and glutathione (GSH) systems, no GR is encoded in the genome of C. parvum, and TrxR is the cornerstone of the antioxidant defense, supplying electrons to both the Trx and the GSH pathways. Given its role in parasite's redox homeostasis, we focus on functional and structural characterization of CpTrxR, illustrating the unique C-terminal -CGGGKCG motif, exclusive to apicomplexan parasites. Our study reports crystal structures of the enzyme with the C-terminal tail captured in different competent catalytic position, unveiling new aspects of the apicomplexan TrxR's mechanism of action. Moreover, has been shown that Auranofin (AF), a gold-containing compound and FDA-orphan drug, kills parasites in culture and here we validate the inhibition mechanism by enzymatic assays and providing the crystal structure of CpTrxR in complex with AF. These results offer crucial insights for the design of selective inhibitors, being the apicomplexan C-terminal motif notably distinct from the -GCU/CG one found in mammalian and insect TrxRs and emphasize CpTrxR as a critical target for anti-parasitic drug development

The analgesic evaluation of an opioid-free anaesthesia protocol in cats undergoing ovariectomy

Opioid-free protocols are increasingly used in veterinary medicine although there are few studies concerning the feline species. In 2022, Rufiange et al. (1) compared the analgesic power of an opioid-free protocol with that of a standard protocol (using opioids) in ovariohysterectomized female cats, evaluating the degree of analgesia and sedation using the Feline Grimace Scale (FGS), obtaining as a result an equivalence between the opioid-free and standard protocols in the management of intraoperative pain and the degree of anesthesia achieved. Among the most commonly used drugs in opioid-free protocols is dexmedetomidine. Gupert A. et al. (2), have shown that this causes no side effects to the respiratory system, as well as having an analgesic, sedative and anxiolytic effect. The use in continuous intraoperative infusion of dexmedetomidine in cats was studied in 2018 by authors who demonstrated its usefulness also in reducing the volatile anaesthetic agent used to maintain general anaesthesia (isofluorane), thus decreasing its undesirable effects systemically (3). The objective of the study is to evaluate the analgesic and anesthesiologic capacity of an opioid-free protocol using dexmedetomidine, alfaxalone, and lidocaine intra-operatively in female cats undergoing ovariectomy, comparing it with a standard protocol. The study is based on 30 subjects randomly assigned to experimental group 1 and standard group 2. Group 1 received administration of dexmedetomidine at 15 μg/kg IM and alfaxalone at 2 mg/kg IM for induction of anesthesia, followed by maintenance of anaesthesia in isofluorane and administration by continuous infusion (CRI) of dexmedetomidine at 1 μg/kg/h IV. Physiological parameters were measured for each patient during nine well-defined stages of surgery. Patients in standard group 2 received an additional dose of methadone at 0.1 mg/kg IM during anaesthesia induction and did not receive intraoperative dexmedetomidine CRI. Both groups received a dose of intra-ligamentous lidocaine during surgery before ovariectomy. The parameters recorded for both groups and the FGS resulted in the good outcome from both anesthesiological and analgesic point of view of the experimental group compared with the control group, demonstrating in fact the analgesic power of intraoperative dexmedetomidine and of the exsperimental protocol

Case Report: Reproductive evaluation of a Murgese stallion with obstructive azoospermia, accumulation of hyaline material in the ampullae ducts, and corpora amylacea in vesicular glands

This report aims to present a case of obstructive azoospermia in a stud stallion diagnosed with an alkaline phosphatase (SPAP) assessment. A 20-year-old Murgese stallion is referred for acquired azoospermia. History is negative for reproductive disorders, and clinical examinations and ultrasonography of internal and external genitalia do not reveal significant alterations. Semen collection highlights the absence of spermatozoa in the ejaculate and the urinalysis is negative for spermatozoa. SPAP assay is performed on seminal plasma, with a value of 30 IU/L, compatible with obstructive azoospermia. A biopsy is performed, detecting the presence of complete germ lines in both testes. A resolution is attempted endoscopically, gently insufflating ampullae, with negative results, so the stallion is excluded from breeding. Time afterwards, the stallion dies of natural causes, and necropsy and histopathological analyses are performed. Corpora amylacea are highlighted in both seminal vesicles; the right and left ampullae show ectasic lumen, with the diffuse presence of hyaline material. Ampullae obstruction is an uncommon pathology, which can affect stallions and jacks, generally caused by the accumulation of spermatozoa, but, unfortunately, this case was unresponsive to attempted treatments. Interestingly, to the authors' knowledge, this is the first report of corpora amylacea in equine stallion sexual glands

The Crucial Dynamics of Lipid Droplets in Early Ovine Embryonic Development

Lipid droplets (LDs) are highly abundant in early mammalian embryos, yet their functional relevance to preimplantation development and cell fate decisions remains largely unexplored. Given their prevalence, we hypothesized that LDs contribute to developmental competence and cell fate. This thesis investigates the role of LDs in sheep embryos, combining physical manipulation, molecular interference, and phenotypic analysis. Using hyperosmotic centrifugation and micromanipulation, we removed LDs from zygotes, which led to significantly reduced blastocyst formation (20% vs. 38.7%, P < 0.05), despite normal cleavage and restored LD content at the blastocyst stage. These blastocysts lacked the typical LD enrichment in the inner cell mass (ICM), exhibited altered SOX2-positive cell distribution, a higher ICM-to-trophectoderm ratio, and increased histone H3K9 acetylation and lysine crotonylation—pointing to a link between lipid availability, cell identity, and epigenetic regulation. To further dissect the contribution of lipid biosynthesis, we targeted ATP citrate lyase (ACLY) and/or acetyl-CoA synthetase short family member 2 (ACSS2), key enzymes in cytosolic/nuclear acetyl-CoA production. ACLY knockdown impaired blastocyst development and arrested embryos at early stages, whereas chemical inhibition of ACLY did not affect blastocyst formation or lipid biosynthesis but altered the pattern of histone acylation. Importantly, inhibition of ACLY and ACSS2 following LD removal further compromised lipid recovery and exacerbated disruptions in histone acylation, underscoring a synergistic effect between lipid availability and acetyl-CoA metabolism in supporting chromatin regulation and embryonic development. Together, these findings demonstrate that LDs and lipid metabolism influence not only early embryo viability but also cell lineage allocation and chromatin remodelling. This work positions lipid dynamics as a critical, previously underappreciated layer of developmental regulation