Margarida Fardilha

University of Aveiro | UA · Signal Transduction Laboratory; Department of Medical Sciences; Institute of Biomedicine - iBiMED

Nek2 is a cell cycle-regulated serine/threonine protein kinase that is up-regulated in human cancers. Functionally, it is implicated in control of centrosome separation and bipolar spindle formation in mitotic cells and chromatin condensation in meiotic cells. Two major splice variants have been described in vertebrates, Nek2A and Nek2B, that diffe...

It is now widely accepted that abnormal processing of the Alzheimer's amyloid precursor protein (APP) can contribute significantly to Alzheimer's disease (AD). APP can be processed proteolytically to give rise to several fragments, including toxic beta-amyloid (Abeta) fragments that are subsequently deposited as amyloid plaques in brains of AD pati...

Background: Dental caries are one of the most prevalent chronic diseases worldwide affecting all age groups. Current therapeutic strategies are technically demanding and invasive procedures that only address the restoration of dental pulp. Upon damage, mineralized tissues and dental pulp are affected, activating signalling cascades that promote the...

Protein Phosphatase 1 (PP1) is a major serine/threonine phosphatase in eukaryotes, participating in several cellular processes and metabolic pathways. Due to their low substrate specificity, PP1's catalytic subunits do not exist as free entities but instead bind to Regulatory Interactors of Protein Phosphatase One (RIPPO), which regulate PP1's sub-...

Studying proteins associated with sex chromosomes can provide insights into sex-specific proteins. Membrane proteins accessible through the cell surface may serve as excellent targets for diagnostic, therapeutic, or even technological purposes, such as sperm sexing technologies. In this context, proteins encoded by sex chromosomes have the potentia...

The sex of the animals is of paramount importance in many animal production systems. This is particularly evident in the production of milk or in breeding programs focused on the production of female animals. In some cases, slaughter or euthanasia of animals of the unwanted sex becomes the only solution, highlighting ethical and economic concerns....

The trend to delay parenthood is increasing, impacting fertility and reproductive outcomes. Advanced paternal age (APA), defined as men’s age above 40 years at conception, has been linked with testicular impairment, abnormal semen parameters, and poor reproductive and birth outcomes. Recently, the significance of sperm microRNA for fertilization an...

Simple Summary Rabbit farming plays a significant role in the global production of animal-derived protein, with hundreds of millions of rabbits being produced every year. However, research on feto-maternal monitoring in rabbits remains limited. This review aims to explore intricate factors affecting maternal and fetal health, providing valuable ins...

Male infertility is a prevalent concern affecting couples worldwide. While genetic factors, hormonal imbalances, and reproductive system defects play significant roles, emerging evidence suggests that lifestyle choices also profoundly impact male fertility. This study aimed to explore the effects of several lifestyle factors, including tobacco and...

Simple Summary The rabbit production industry would greatly benefit from a technology that allows producers to obtain a more predominantly male or female offspring, accordingly to the goals of each production farm. Our review highlights the current and future developments in rabbit sperm sexing technologies, as well as the potential impact of imple...

Djeya1 (RKLAFRYRRIKELYNSYR) is a very effective cell penetrating peptide (CPP) that mimics the α5 helix of the highly conserved Eya domain (ED) of eyes absent (Eya) proteins. The objective of this study was to bioengineer analogues of Djeya1 that, following effective translocation into planarian tissues, would reduce the ability of neoblasts (totip...

Djeya1 (RKLAFRYRRIKELYNSYR), is a very effective cell penetrating peptide (CPP) which mimics the α5 helix of the highly conserved Eya domain (ED) of eyes absent (Eya) proteins. The objective of this study was to bioengineer analogues of Djeya1 that, following effective translocation into planarian tissues, would reduce the ability of neoblasts (tot...

Galectin-3 (Gal-3) is a carbohydrate-binding protein associated with the development and progress of heart failure. Here, we report the first colorimetric and low-cost approach for detecting and quantifying Gal-3 using gold nanoparticles (AuNPs) bioconjugated with Gal-3 antibody. The interaction of Gal-3 with the resulting nanoprobes led to a linea...

This work aimed to understand how lifelong exercise training promotes the remodelling of the immune system and prostate signalome in a rat model of PCa. Fifty-five male Wistar rats were divided into four groups: control sedentary, control exercised, induced PCa sedentary and induced PCa exercised. Exercised animals were trained in a treadmill for 5...

Background: The high rates of unintended pregnancy and the ever-growing world population impose health, economic, social, and environmental threats to countries. Expanding contraceptive options, including male methods, are urgently needed to tackle these global challenges. Male contraception is limited to condoms and vasectomy, which are unsuitabl...

Purpose: The advent of proteomics provides new opportunities to investigate the molecular mechanisms underlying male infertility. The selection of relevant targets based on a single analysis is not always feasible, due to the growing number of proteomic studies with conflicting results. Thus, this study aimed to systematically review investigations...

Despite advances in understanding the molecular mechanisms underlying prostate cancer progression, the development of effective therapeutic approaches remains a major challenge. In this context, the protein phosphatase 1 (PP1) and its complexes have been recognized as potential drug targets. Herein, we designed and synthetized a peptide sequence ba...

Gal-3 plays an important role in cell survival, mRNA splicing, and cell–cell and cell–matrix interactions. Depending on its cellular localization and cancer type, Gal-3 may have tumour-suppressive or tumour-promoting activities. Given the promising diagnostic role of Gal-3 in the urine of PCa patients found in our previous study, its concordant gen...

Galectin-3 (Gal-3) belongs to galectin protein family, a type of β-galactose-binding lectin having more than one evolutionarily conserved domain of carbohydrate recognition. Gal-3 is mainly located in the cytoplasm, but it also enters the nucleus and is secreted into the extracellular environment and biological fluids such as urine, saliva, and ser...

Male fertility relies on the ability of spermatozoa to fertilize the egg in the female reproductive tract (FRT). Spermatozoa acquire activated motility during epididymal maturation; however, to be capable of fertilization, they must achieve hyperactivated motility in the FRT. Extensive research found that three protein phosphatases (PPs) are crucia...

Most studies on the effects of physical exercise have focused on its influence on muscle tissue, forgetting its interference in liver function. Ageing leads to the progressive impairment of hepatic functions. Several biochemical and bioenergetics parameters were determined to test the impact of a lifelong aerobic training program in the hepatic age...

Bottom-up proteomics is a popular approach in molecular biomarker research. However, protein analysts have realized the limitations of protein-based approaches for identifying and quantifying proteins in complex samples, such as the identification of peptides sequences shared by multiple proteins and the difficulty in identifying modified peptides....

Organoids are units of function of a given organ able to reproduce, in culture, a biological structure similar in architecture and function to its counterpart in vivo. Today, it is possible to develop an organoid from a fragment of tissue, a stem cell located in an adult organ, an embryonic stem cell, or an induced pluripotent stem cell. In the pas...

G protein-coupled receptors (GPCRs) are involved in several physiological processes, and they represent the largest family of drug targets to date. However, the presence and function of these receptors are poorly described in human spermatozoa. Here, we aimed to identify and characterize the GPCRs present in human spermatozoa and perform an in sili...

Aging is associated with testicular morphological and functional alterations, but the underlying molecular mechanisms and the impact of physical exercise are poorly understood. In this study, we examined the effects of age and lifelong moderate-intensity exercise on rat testis. Mature adults (35 weeks) and middle-aged (61 weeks) Wistar Unilever mal...

In Laboratories in Biomedicine 5, a 3rd year class of the Biomedical Sciences bachelor (University of Aveiro), the cellular, molecular, and biochemical basis of male infertility diagnosis is addressed (Module 3; M3), usually involving 4 practical classes in the laboratory. In October 2020, due to the COVID-19 constrains, the practical classes neede...

Prostate cancer (PCa) is one of the most lethal diseases in men, which justifies the search for new diagnostic tools. The aim of the present study was to gain new insights into the progression of prostate carcinogenesis by analyzing the urine proteome. To this end, urine from healthy animals and animals with prostate adenocarcinoma was analyzed at...

The Ser/Thr-protein phosphatase PP1 (PP1) is a positive regulator of the androgen receptor (AR), which suggests major roles for PP1 in prostate carcinogenesis. However, studies dedicated to the characterization of PP1 in PCa are currently scarce. Here we analyzed the expression and localization of the PP1 catalytic (PP1c) isoforms in formalin-fixed...

Study question Are there ageing-related alterations in human sperm protein and small RNA content that can be responsible for the age-associated decline in male fertility? Summary answer Our results revealed a set of proteins and small RNAs, particularly miRNAs, that are altered in older men. What is known already Male infertility is a common heal...

Prostate cancer (PCa) is one of the most common cancers among men, and its incidence has been rising through the years. Several risk factors have been associated with this disease and unhealthy lifestyles and inflammation were appointed as major contributors for PCa development, progression, and severity. Despite the advantages associated with the...

Simple Summary Prostate cancer (PCa) is one of the most common cancers. Due to the limited and invasive approaches for PCa diagnosis, it is crucial to identify more accurate and non-invasive biomarkers for its detection. The aim of our study was to non-invasively uncover new protein targets for detecting PCa using a proteomics and proteogenomics ap...

Background and Aim: Male hypogonadism results from failure to produce physiological levels of testosterone. Testosterone in men is essential in masculine development, sperm production, and adult man's health. Osteoporosis is one of the consequences of hypogonadism. Regular physical exercise and exogenous testosterone administration are frequently u...

Despite the increasing life expectancy, an individual’s later years tends to be accompanied by a decrease in the quality of life. Though biological changes that occur through the natural process of aging cannot be controlled, the risk factors associated with lifestyle can. Thus, the main goal of this systematic review was to evaluate how nutrition...

Currently, two conventional freezing techniques are used in sperm cryopreservation: slow freezing (SF) and rapid freezing (RF). Despite the protocolar improvements, cryopreservation still induces significant alterations in spermatozoon that are poorly understood. Here, available proteomic data from human cryopreserved sperm was analyzed through bio...

The mature spermatozoon, a highly differentiated cell equipped for the sole purpose of fertilization, lacks the protein machinery required for conventional endocytotic mechanisms. Perhaps contrary to expectation, cell-penetrating peptides (CPPs) rapidly translocate across the unique sperm plasma membrane to accrete within distinct intracellular com...

Protein phosphatase 1 (PP1) regulates several cellular events via interaction with multiple regulatory subunits. The human prostate proteome includes various PP1-interacting proteins; however, a very limited number of interactions is yet characterized and their role in prostate tumorigenesis remains poorly understood. Tctex1 domain-containing prote...

Background: Spermatogenesis generates a small and highly specialised type of cell that is apparently incapable of transcription and translation. For many years, this dogma was supported by the assumption that (i) the compact sperm nucleus, resulting from the substitution of histones by protamine during spermatogenesis, renders the genome inaccessi...

Cancer is the second leading cause of death worldwide. Despite the availability of numerous therapeutic options, tumor heterogeneity and chemoresistance have limited the success of these treatments, and the development of effective anticancer therapies remains a major focus in oncology research. The serine/threonine-protein phosphatase 1 (PP1) and...

Prostate cancer (PCa) is the second most frequent cancer diagnosed in men worldwide. Among the common treatment options, photodynamic therapy (PDT) is being considered a promising local therapy to treat this cancer. Although PDT is an established treatment modality approved for several types of cancer, the low solubility, the reduced tumor selectiv...

WebRePRO - I Reproductive Sciences & Fertility Virtual Summit

The protein quality control network, including autophagy, the proteasome and the unfolded protein response (UPR), is triggered by stress and is overactive in acquired antiestrogen therapy resistance. We show for the first time that the aggresome load correlates with apoptosis and is increased in antiestrogen-sensitive cells compared to endocrine-re...

Objective: To design and implement a laboratory teaching/learning strategy applied to male infertility diagnostic during COVID-19 pandemic. Methods: Seventy-five students attending Module 3 (M3): Male Infertility Diagnostic Laboratories, of curricular unit Laboratories in Biomedicine 5 were divided into 8 groups to reduce students’ number in class....

Several strands of evidence indicate the presence of marked similarities between human brain and testis. Understanding these similarities and their implications has become a topic of interest among the scientific community. Indeed, an association of intelligence with some semen quality parameters has been reported and a relation between dysfunction...

The use of sexed semen in dairy and beef farms ensures the production of animals of the desired sex, resulting in a reduction of costs and an improvement of environmental sustainability. Several methods have been developed over the years, but most of them were abandoned due to their limited efficacy. Currently, the only commercially available metho...

Bisphenol A (BPA), a well-known endocrine disruptor present in epoxy resins and polycarbonate plastics, negatively disturbs the male reproductive system affecting male fertility. In vivo studies showed that BPA exposure has deleterious effects on spermatogenesis by disturbing the hypothalamic–pituitary–gonadal axis and inducing oxidative stress in...

Prostate cancer (PCa) is one of the most common cancer types among men. The quantification of prostate-specific antigen used for PCa detection has revealed limited applicability. Thus, it is crucial to identify new minimally invasive biomarkers for PCa. It is believed that the integration of proteomics data from different studies is vital for ident...

Purpose: Prostate cancer is a major cause of cancer-related death in males worldwide and, in addition to impairing prostate function, also causes testicular adaptations. In this study, we aim to investigate the preventive effect of exercise training on PCa-induced testicular dysfunction. Methods: As a model, we used fifty Wistar Unilever male rats,...

Objective: To design protein phosphatase 1 (PP1)-disrupting peptides covalently coupled to inert cell-penetrating peptides (CPPs) as sychnologically organized bioportide constructs as a strategy to modulate sperm motility. Design: Experimental study. Setting: Academic research laboratory. Patient(s)/animal(s): Normozoospermic men providing s...

Cancer cells take advantage of signaling cascades to meet their requirements for sustained growth and survival. Cell signaling is tightly controlled by reversible protein phosphorylation mechanisms, which require the counterbalanced action of protein kinases and protein phosphatases. Imbalances on this system are associated with cancer development...

Prostate cancer (PCa) is one of the most common male-specific cancers worldwide, with high morbidity and mortality rates associated with advanced disease stages. The current treatment options of PCa are prostatectomy, hormonal therapy, chemotherapy or radiotherapy, the selection of which is usually dependent upon the stage of the disease. The devel...

Aim: This work intended to improve the knowledge of the rat model of prostate cancer (PCa) by ultrasonographic monitoring. Materials and methods: Male Wistar rats were divided into control (n=8) and PCa (n=14) groups. PCa development was induced in the PCa group through the sequential administration of the anti-androgenic drug flutamide, testost...

Prostate cancer (PCa) treatment remains challenging, especially in advanced stages, where the lack of sensitivity and specificity of available biomarkers makes it difficult to establish an accurate prognosis. Therefore, it is imperative to study PCa biology to identify key molecules that can improve PCa management. In this study, eight prostate tum...

The sperm cell is unique in its function. It is the only human cell that must leave the body where it is produced and fulfills its goal in a different organism being, thus, a highly specialized cell. Sperm cells are produced in the testis, acquire motility during the epididymis journey and fertilize the oocyte in the female reproductive system. Mor...

The prostate is the major exocrine gland of the male reproductive system. The prostatic epithelium secretes an alkaline fluid, the prostatic fluid, that constitutes about 20–30% volume of the seminal fluid. It provides proteins and ions essential to control the ejaculation process and to regulate proteins involved in sperm maturation (e.g. human ka...

Signal transduction comprises the intracellular biochemical signals which induce the appropriate cell response to an external stimulus. The players in signal transduction are diverse, from small molecules as first messengers, to proteins, receptors, transcription factors, among many others. The different signaling pathways and the crosstalk between...

The unfolded protein response (UPR) is a conserved and essential cellular pathway involved in protein quality control which is activated in response to several cellular stressors such as diseases states, ageing, infection and toxins. The cytosol, endoplasmic reticulum (ER) and mitochondria are continuously exposed to new proteins and in situations...

The unfolded protein response (UPR) is involved in protein quality control and is activated in response to several stressors. Although in testis the UPR mechanisms are well described, their presence in spermatozoa is contentious. We aimed to investigate the presence of UPR-related proteins in human sperm and the impact of oxidative stress induction...

An efficient access to porphyrin derivatives bearing uracil-alditol moieties at the β-pyrrolic position was developed. The synthetic strategy involved a hetero Diels-Alder reaction between uracil-alditol based orthoquinodimethanes using 2-vinyl-5,10,15,20-tetraphenylporphyrin as the scaffold. The preliminary evaluation of their photodynamic effecti...

Understanding how age affects fertility becomes increasingly relevant as couples delay childbearing toward later stages of their lives. While the influence of maternal age on fertility is well established, the impact of paternal age is poorly characterized. Thus, this study aimed to understand the molecular mechanisms responsible for age-dependent...

Sulfate-based lipids (SL) have been proposed as players in inflammation, immunity and infection. In spite of the many biochemical processes linked to SL, analysis on this class of lipids has only focused on specific SL sub-classes in individual fluids or cells leaving a range of additional SL in other biological samples unaccounted for. This study...

The rat has been frequently used as a model to study several human diseases, including cancer. In many research protocols using cancer models, researchers find it difficult to perform several of the most commonly used techniques and to compare their results. Although the protocols for the study of carcinogenesis are based on the macroscopic and mic...

Teaching methodologies used in biochemistry classes at the University level are traditionally dependent on theorical classes. The assessment is usually based on written tests performed at the end of the semester. However, most students who learn metabolism by this traditional method consider the study of metabolic biochemistry a terrifying and unfo...

The unfolded protein response (UPR) is involved in protein quality control and activated in response to cellular stressors. Although in testis the UPR mechanisms are well described, the presence of these mechanisms in spermatozoa is contentious. We aimed to investigate the presence of UPR-related proteins in human sperm and the impact of oxidative...

In mouse and bovine sperm, GSK3 activity is inversely proportional to motility. Targeted disruption of the GSK3A gene in testis results in normal spermatogenesis, but mature sperm present a reduced motility, rendering male mice infertile. On the other hand, GSK3B testis-specific KO is fertile. Yet in human sperm, an isoform-specific correlation bet...

Purpose The impact of exercise training on testicular function is relatively ill-defined. To gain new insights into this important topic, published data, deriving from both humans and animal studies, were critically analyzed. Results and conclusions The effects of exercise on the hypothalamus–pituitary–gonadal axis, influenced by the type, intensi...

Resistant Breast and Prostate cancer remain a major clinical problem and new therapeutic approaches and better predictors of therapeutic response are clearly needed. Due to the involvement of the Unfolded Protein Response (UPR) in cell proliferation and apoptosis evasion, an increasing number of publications support the hypothesis that impairments...

Introduction: Prostate Cancer (PCa) is the second most common neoplasia in men. Because it is often diagnosed at a late stage, mortality rates remain high. Studying cancer metabolome, which reflects early changes that occur in cells, has gained relevance and may contribute to the identification of early diagnostic biomarkers and understanding tumor...

Germ cell proliferation is epigenetically controlled, mainly through DNA methylation and histone modifications. However, the pivotal epigenetic regulators of germ cell self-renewal and differentiation in postnatal testis are still poorly defined. The histone methyltransferase enhancer of zeste homolog 2 (EZH2) is the catalytic subunit of Polycomb r...

Gac Sanit 2018;32 Supl C:7-140 349. EFFECT OF MERCURY EXPOSURE IN HUMAN FERTILITY: A SYSTEMATIC REVIEW

In this study we unraveled human spermatozoa signaling pathways and correlated the activity of sperm signaling proteins with clinical data. Thirty seven semen samples were used and basic semen parameters, sperm DNA fragmentation and the expression levels of 75 protein kinases and the phosphorylation/cleavage patterns of 18 signaling proteins were e...

Treadmill training as a preventive measure for Prostate cancer-induced testicular dysfunction

Globally, cancer is one of the leading causes of death, so the development of strategies for an early diagnosis of cancer is of great importance. Biochemical alterations precede morphological changes in cells and tissues, so studying cancer metabolome seems like a reasonable approach for early diagnosis, prognosis and to follow treatment progressio...

The trend in parenthood at an older age is increasing for both men and women in developed countries, raising concerns about the reproductive ability and the consequences for the offspring’s health. While reproductive activity in women stops with menopause, a complete cessation of the reproductive potential does not occur in men. Although several st...

Most techniques to study protein-protein interactions, gene expression or signal transduction, among others, in Sertoli cells, depend on obtaining a protein extract of such cells. This is accomplished by lysing the Sertoli cells and solubilizing the intracellular proteins. Depending on the purpose of your study, the technique used to lyse and conse...