Experimental Medicine and Biology Institute

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive cancers, primarily due to its complex tumor microenvironment (TME), which drives both disease progression and therapy resistance. Understanding the molecular mechanisms governing TME dynamics is essential for developing new treatment strategies for this devastating disease. In this study, we uncover an oncogenic role for Galectin-1 (Gal1), a glycan-binding protein abundantly expressed by activated pancreatic stellate cells (PSCs), a key component of the PDAC TME that orchestrates tumor progression. Our findings reveal that Gal1 expression is elevated in the nucleus of human PSCs in both tissue samples and cultured cell lines. Using chromatin immunoprecipitation followed by sequencing analysis (ChIP-seq), we identify Gal1 occupancy at the promoters of several cancer-associated genes, including KRAS , a pivotal oncogene involved in PDAC pathogenesis. We demonstrate that Gal1 binds to the KRAS promoter, sustaining KRAS expression in PSCs, which, in turn, maintains PSC activation and promotes the secretion of protumorigenic cytokines. Mechanistically, Gal1 is required to preserve histone H3 lysine 4 monomethylation levels and to recruit the histone methyltransferase MLL1 to target promoters. Collectively, our findings define a nuclear function of Gal1 in modulating the transcriptional landscape of cancer-associated genes in PSCs within the PDAC TME, mediated through an epigenetic mechanism. These insights enhance our understanding of PDAC pathology and open potential avenues for therapeutic interventions targeting intracellular Gal1.

Las alteraciones del color en el marsupial Didelphis albiventris, han sido escasamente documentadas. Los entornos urbanos ofrecen buenas oportunidades para estudiar animales nocturnos sinantrópicos como las zarigüeyas. En un barrio de Mendoza, Argentina, un residente informó de la presencia de una hembra de D. albiventris. El animal fue capturado utilizando una trampa Tomahawk y anestesiado para un examen clínico detallado, que reveló leucismo y mala condición corporal. Se estudiaron medidas morfométricas, parámetros reproductivos y observaciones clínicas y ecológicas para investigar posibles vínculos con la anomalía de color. Las deficiencias nutricionales en entornos urbanos podrían estar asociadas a la coloración inusual.

CRISP2 is enriched in the male reproductive system of mammals and plays roles in spermatogenesis, sperm motility, and fertilization. Although extensively investigated in rodents and boars, human CRISP2 (hCRISP2) remains poorly studied, particularly concerning its localization in testicular and epididymal tissues and its molecular features. In this study, we used immunofluorescence to determine the localization of hCRISP2 in testis, epididymis, and ejaculated sperm. While no expression was observed in the epididymal epithelium, hCRISP2 was detected at different stages during spermatogenesis. Specifically, hCRISP2 was found in the nucleus of primary spermatocytes and of both round and early elongated spermatids. In elongated spermatids, it was additionally observed in the cytoplasm, the flagellum, and the equatorial segment of the acrosome (EqS). The presence of aggregated material with hCRISP2 immunoreactivity in the apical pole of Sertoli cells suggests that most of the hCRISP2 involved in spermatogenesis is phagocytized by these cells during spermiation. In ejaculated sperm, hCRISP2 was found in the cytoplasmic droplet, flagellum, and EqS, consistent with its described roles in sperm motility and gamete fusion. Native and SDS-PAGE combined with western blot analyses depicted the ability of hCRISP2 to form stable high molecular weight complexes and mass spectrometry revealed that these complexes likely consist exclusively of hCRISP2. Furthermore, we showed that hCRISP2 undergoes only limited post-translational modifications. These findings shed light into the dynamic localization of hCRISP2 throughout spermatogenesis and in ejaculated sperm, as well as its molecular features, enhancing our understanding of its pivotal functional roles and relevance for male fertility.

Introduction Maternal lifestyle impacts reproductive performance. Previously, we demonstrated that maternal environmental enrichment promotes pregnancy success in BALB/c mice. As progesterone regulates gestation, we decided to study the effect of maternal environmental enrichment on ovarian physiology during early gestation. Methods For this, six-week-old female mice were housed in enriched or control cages for six weeks and then mated with control fertile males. Females with a mucus plug were returned to their respective control or enriched cages. Pregnant mice were euthanized on day 7 of pregnancy, and ovaries and progesterone levels were investigated. Results Hematoxylin and eosin slices showed no differences in the area (μm²) of the ovaries between control and enriched females. Also, the number of primordial, primary, preantral, antral, and atretic follicles was similar for both treatments. However, the number and area (μm²) of corpora lutea were increased in the ovaries from the enriched group. Moreover, enriched females presented higher progesterone serum levels and increased 3β-HSD expression. Discussion Therefore, maternal environmental enrichment regulates ovarian physiology, and this could promote the benefits previously reported.

Breast cancer is the leading cause of cancer deaths in women worldwide, with about 20,000 cases annually in Argentina. While age, diet, and genetics are known risk factors, most breast cancer cases have unknown causes, necessitating the discovery of new risk factors. The aim of this study was the analysis of the prognostic relevance of the oncobiome in Argentinean breast cancer patients. Sequencing of the V4 region 16S rRNA gene was performed on 34 primary breast tumor samples, using bioinformatic and statistical analyses to identify bacteria and hypothetical pathways. Each sample presented a unique microbial profile, with Proteobacteria being the most abundant phylum. Tumors >2 cm showed greater alpha diversity with increased nucleotide biosynthesis. Moreover, progesterone-receptor tumors showed differences in beta diversity, being progesterone receptor-positive tumors that had the highest expression of Acinetobacter and Moraxella . In disease progression, the phylum Chloroflexi was prevalent in tumors of surviving patients. Acinetobacter and Cloacibacterium genera were significantly higher in patients without events and those without metastasis. We found that nucleotide and cell-structure biosynthesis, and lipid metabolism pathways were enriched in tumors with poor progression, whereas amino-acid degradation was increased in tumors of surviving patients. This finding is an indication that tumor cells are taking advantage of this effect of the microbiome during tumor progression. We conclude that oncobiome is dysbiotic in these patients, with distinct patterns in those with poor progression. Suggesting a link between the oncobiome and cancer progression, paving the way for new therapies to improve patient quality of life and survival. IMPORTANCE This is the first study to investigate the relevance of the oncobiome in the evolution of breast cancer in a cohort of Argentine patients. It also highlights the need for further research in this area to improve our understanding of the role of the microbiome in this disease and potentially identify new therapeutic targets or prognostic indicators. Understanding the complex interaction between the microbiome, the tumor microenvironment, and the pathogenesis of breast cancer holds the promise of more personalized and effective treatment approaches in the future.

Obesity, a global epidemic, is linked to adverse reproductive outcomes, including infertility and ovulation dysfunction. The cafeteria diet (CAF) serves as an animal model mirroring Western diet habit. Coenzyme Q10 (CoQ10), known for enhancing reproductive outcomes in various pathologies, is not fully understood for its effects on obesity treatment. Here, obesity was modeled using CAF-fed rats to assess CoQ10's impact on metabolic and ovarian disruptions caused by obesity. Wistar rats were divided into control (standard diet) and obese (CAF diet) groups. After 75 days, half of each group received oral CoQ10 (5 mg/kg) for 13 days, while the rest received a vehicle. Animals were euthanized during the estrus phase, and blood and ovaries were collected for analysis. CAF caused increased body weight gain (p < 0.01) associated with hyperglycemia, hypertriglyceridemia, and hypercholesterolemia (p < 0.05). Moreover, it caused a reduction in the number of AMH + follicles (p < 0.001), increasing follicular atresia (p < 0.05) and serum estradiol levels (p < 0.05). Obesity also altered the estrous cycle and reduced the ovulation rate (p < 0.05). CoQ10 administration showed beneficial effects on all ovarian disruptions but had no effect on the metabolic alterations induced by obesity. In summary, CoQ10 could be an additional treatment for obesity-related infertility in patients with normal metabolic profiles. While CoQ10 does not affect metabolic parameters influenced by obesity, crucial for reproductive issues and offspring health, it is recommended as part of a treatment plan that includes a balanced diet and increased physical activity for obese individuals with metabolic alterations seeking pregnancy.

Background/Objectives: Endometriosis has a marked impact on fertility, although the mechanisms behind this relationship remain poorly understood, particularly in cases without significant anatomical distortions or in the context of ovarian endometriomas. This study aimed to investigate the effect of peritoneal endometriosis on ovarian function by assessing ovarian reserve and apoptosis. Methods: Peritoneal endometriosis was surgically induced in Sprague Dawley rats through the autotransplantation of uterine fragments onto the bowel mesothelium. One month post-surgery, ovarian structures were counted, follicle and corpora lutea apoptosis was evaluated by TUNEL, and apoptotic-related protein expression in ovaries was assessed by Western blot. Additionally, a co-culture system using 12Z endometriotic and KGN granulosa cell lines was utilized to evaluate gene expression by RT-qPCR. Results: Rats with peritoneal endometriosis exhibited a significant reduction in ovarian structures characterized by a low number of total follicles, particularly primordial, primary, preantral, and late-antral follicles. Consistently, AMH protein expression was decreased in ovaries in the presence of endometriosis. In addition, this disease led to a significant increase in late-antral follicles that were TUNEL-positive and in the number of apoptotic cells in corpora lutea, indicating higher apoptosis in endometriosis ovaries. Concomitantly, the altered expression of apoptosis-related proteins was observed, with increased procaspase 3 and decreased BCL-2 expression. In addition, KGN granulosa cells co-cultured with 12Z endometriotic cells displayed reduced KITLG mRNA expression and increased AMHR2 mRNA expression. Conclusions: Peritoneal endometriosis significantly impairs ovarian health by disrupting folliculogenesis, reducing ovarian reserve, and increasing apoptosis, potentially accelerating ovarian aging and contributing to infertility. These results underscore the need for further research to identify the molecular pathways involved and to develop targeted therapeutic strategies.

In brief The cyclic adenosine monophosphate (cAMP) pathway is essential for maintaining sperm physiology. This study examines a cAMP analog and a phosphodiesterase inhibitor that effectively enhance human sperm motility, thereby improving the efficiency of in vitro sperm selection. Abstract cAMP plays a central role in sperm physiology. Various cAMP upregulators, both cAMP analogs and phosphodiesterase (PDE) inhibitors, have been used in handling human sperm in vitro , although conflicting results and variable responses among patients have been reported. This study aims to evaluate the ability of two compounds – Sp-5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole 3′-5′-cyclic monophosphothioate (cBiMPs), a cAMP analog, and TAK-063, a specific PDE10A inhibitor – to enhance human sperm motility parameters and, consequently, improve sperm preparation procedures. Our results showed that both cBiMPs and TAK-063 significantly enhanced human sperm motility and hyperactivation compared to the control (dimethyl sulphoxide). They also increased protein phosphorylation levels without inducing premature acrosomal exocytosis or DNA fragmentation. The enhancement of sperm motility persisted for 4 h after their removal, surpassing the effects of known cAMP analogs (8-bromo-adenosine-3′, 5′-cAMP (8-Br-cAMP) or dibutyryl cAMP (db-cAMP)) or PDE inhibitors (3-isobutyl-1-methylxanthine (IBMX) or pentoxifylline (PTX)). Furthermore, the presence of cBiMPs or TAK-063 during sperm selection resulted in higher recovery rates in comparison to the control, and these compounds effectively improved sperm motion in both fresh and cryopreserved samples with impaired motility. In conclusion, cBiMPs and TAK-063 exhibit potent and sustained effects on human sperm motility, enhancing the efficiency of sperm preparation techniques. The ability to improve sperm motility holds significant implications for male infertility treatment, facilitating the use of low complexity techniques such as intrauterine insemination or in vitro fertilization, and may also aid in selecting viable testicular sperm for intracytoplasmic sperm injection.

The assessment of research performance is widely seen as a vital tool in upholding the highest standards of quality, with selection and competition believed to drive progress. Academic institutions need to take critical decisions on hiring and promotion, while facing external pressure by also being subject to research assessment1, 2, 3–4. Here we present an outlook on research assessment for career progression with specific focus on promotion to full professorship, based on 314 policies from 190 academic institutions and 218 policies from 58 government agencies, covering 32 countries in the Global North and 89 countries in the Global South. We investigated how frequently various promotion criteria are mentioned and carried out a statistical analysis to infer commonalities and differences across policies. Although quantitative methods of assessment remain popular, in agreement with what is found in more geographically restricted studies5, 6, 7, 8–9, they are not omnipresent. We find differences between the Global North and the Global South as well as between institutional and national policies, but less so between disciplines. A preference for bibliometric indicators is more marked in upper-middle-income countries. Although we see some variation, many promotion policies are based on the assumption of specific career paths that become normative rather than embracing diversity. In turn, this restricts opportunities for researchers. These results challenge current practice and have strategic implications for researchers, research managers and national governments.

In brief Artificial oocyte activation (AOA) with Ca ²⁺ ionophores is a valuable tool in assisted reproduction but may affect cellular events critical for embryo development. This study shows the impact of different AOA protocols on embryonic development efficiency and meiotic progression. Abstract AOA with Ca ²⁺ ionophores is an experimental procedure that benefits patients who fail to obtain fertilized eggs. However, the impact of non-physiological Ca ²⁺ increases on cellular events involved in the egg–embryo transition and early development remains poorly understood. Using the mouse model, this study compares common Ca ²⁺ ionophore protocols applied in clinical practice – one or two exposures to A23187 or a single exposure to ionomycin – focusing on embryonic development and cellular events associated with egg activation. All groups of ionophore-activated eggs exhibit lower levels of first mitotic division compared to those activated by spermatozoa or SrCl 2 , attributable to variations in Ca ²⁺ dynamics during activation. At the cellular level, these eggs presented defects in spindle morphology and chromosome segregation during meiosis progression, associated with lower levels of cytoplasmic ATP, without changes in reactive oxygen species. These findings highlight the importance of optimizing Ca ²⁺ management in AOA protocols.

Aging, mitochondria, and neurodegenerative diseases Aging is often viewed as the buildup of changes that lead to the gradual transformations associated with getting older, along with a rising likelihood of disease and mortality. Although organism-wide deterioration is observed during aging, organs with high metabolic demand, such as the brain, are more vulnerable. As a consequence, most neurodegenerative diseases occur in the aged population. Even in the healthy brain, the normal aging process is associated with several features of the neurodegenerative process, including neuroinflammation, and brain shrinkage, with a progressive decline in physiological functions (Lee and Kim, 2022). The neurodegenerative process, therefore, is a complex and multifactorial process driven by age-related changes involving a combination of genetic, environmental, and lifestyle factors that collectively contribute to the progressive loss of structure and function of neurons. Among the mechanisms involved in neurodegeneration, the excessive formation of reactive oxygen species (ROS) in mitochondria plays a key role. During aging, the efficiency of antioxidant defense mechanisms declines, and mitochondria become more prone to ROS-induced damage, characterized by a decrease in mitochondrial DNA (mtDNA) integrity and reduced energy production. ROS can damage all mitochondrial components, creating a "vicious cycle" which activates the mitochondrial stress response (mtSR), a process by which cells respond to stressors that alter mitochondrial physiology (Mottis, Herzig and Auwerx, 2019). The mtSR plays a role in inter-tissue communication, with mitochondria releasing molecules and cell non-autonomous factors into the cytosol or the extracellular milieu, including mtROS, mtDNA, ATP, cardiolipin, and Ca2+. These molecules are identified as damage-associated molecular

In the present contribution, a complete revision of the Moruloidea species found in the Southwest Atlantic is provided, including two new species: M. urocerata n. sp. and M. pentacantha n. sp. from the Marine-Protected Area Namuncurá-Burdwood Bank, Argentina and adjacent waters. Both species can be distinguished from the remaining congeners by their dorsal ornamentation. Moruloidea urocerata n. sp. presents two rounded lobes on the head and pereonite 1 each, and one large, curved process on the pleotelson in dorsal view, whereas M. pentacantha n. sp. presents two spines on the head and pereonite 1 each, and one spine on the pleotelson in dorsal view. Cassidinopsis tuberculata Schultz, 1978 is transferred to the genus Moruloidea, new combination M. tubercuata (Schultz 1978). The original description is augmented with fully described males. The geographic and bathymetric ranges of M. tuberculata (Schultz 1978) n. comb. are expanded. The intraspecific morphological variation of M. darwinii (Cunningham 1871) is reported. The geographical distribution of all the Moruloidea species is discussed and an identification key is provided.

Introduction Gastropod hemocyanins are potent immunostimulants in mammals, a trait associated with their large molecular size and unusual glycosylation patterns. While the hemocyanin from the marine snail keyhole limpet (KLH), has been widely studied and successfully employed as a carrier/adjuvant in several immunological applications, as well as a non-specific immunostimulant for bladder cancer treatment, few other gastropod hemocyanins have been biochemically and immunologically characterized. In this work, we investigated the immunogenic properties of the hemocyanin from Pomacea canaliculata (PcH), an invasive south American freshwater snail. This species, known for its high reproductive rate and easy rearing, represents a promising source of potential biomedical compounds, including hemocyanin. Methods Employing flow cytometry, fluorescence microscopy, immunoassays, and quantitative PCR, we analysed the effects of PcH on THP-1 monocytes and their derived macrophages, as well as its ability to induce humoral response on C57BL/6 mice. Additionally, we evaluated the structural stability of PcH across a wide range of temperature and pH values. Results and discussion Our findings demonstrate that PcH is a structurally stable protein that not only triggers a pro-inflammatory effect on THP-1 derived-macrophages by increasing IL1-β and TNF-α levels, but also promotes phenotypic changes associated with the monocyte-to-macrophage differentiation. Moreover, the humoral response induced by PcH in mice was indistinguishable from that of KLH, highlighting the promising immunostimulatory properties of this freshwater snail hemocyanin.