Fernando Dominguez

• University of Santiago de Compostela

Despite extensive efforts from drug developers, physicians, and patients, the prognosis for glioblastoma (GBM) remains one of the poorest in adult oncology. Current oncology drug development often revisits established technologies, with few truly novel treatments emerging to tackle challenging diseases like GBM. ARJUNA Therapeutics is breaking new...

Background: Primary bilateral uveal melanoma (BUM) is an exceptionally rare form of uveal melanoma (UM). This study aimed to explore the potential existence of a genetic predisposition towards the development of BUM. Methods: We employed an exome sequencing approach on germline DNA from four unrelated patients diagnosed with BUM, seeking pathogenic...

Cancers are driven by multiple genetic mutations but evolve to evade treatments targeting specific mutations. Nonetheless, cancers cannot evade a treatment that targets mitochondria, which are essential for tumor progression. Iridium complexes have shown anticancer properties, but they lack specificity for their intracellular targets, leading to un...

Antitumoral Agents In article number 2113028, M. Arturo López‐Quintela, Fernando Domínguez, and co‐workers report that Ag5 clusters show unique catalytic activities to oxidize thiols. They catalyze the complete oxidation of thiols present in cysteine, glutathione, and thioredoxin. This oxidation is tunable by relevant biological oxidants species (h...

Low atomicity clusters present properties dependent on the size, due to the quantum confinement, with well‐defined electronic structures and high stability. Here it is shown that Ag5 clusters catalyze the complete oxidation of sulfur to S⁺⁶. Ag5 catalytic activity increases with different oxidant species in the order O2 ≪ H2O2 < OH•. Selective oxid...

Synthesis and purification of metal clusters without strong binding agents by wet chemical methods are very attractive for their potential applications in many research areas. However, especially challenging is the separation of uncharged clusters with only a few number of atoms, which renders the usual techniques very difficult to apply. Herein, w...

p>Bioenergetics of cancer results in an increased oxidation state and toxic by-products such as reactive oxygen species (ROS). ROS-mediated thiol oxidation of specific sensory proteins triggers the antioxidant response in order to decrease ROS levels and to trigger apoptosis when the oxidative stress cannot be overcome. This metabolic reprogramming...

Background Uveal melanoma (UM) has a high tendency to cause liver metastases. Metastatic disease is fatal, with a low survival rate. There are two large groups of UMs that, according to their risk of metastatic disease, can be divided into risk subgroups based on histopathological, cytogenetic and molecular characteristics. The presence of somatic...

[Background on medical need in hematology]Even though significant progress has been made in a number of hematological malignancies in recent years, an unmet medical need still remains in indications such as multiple myeloma (MM), chronic lymphocytic leukemia (CLL) or acute myeloid leukemia (AML) due to refractory disease, severe adverse events or t...

Half-sandwich ruthenium(II) complexes [(g 6-p-cymene)Ru(C^N)-(X)] 0/+ (X = Cl, py or 4-NMe 2-py) containing a cyclometalated 2-ppy or 1-ppz with a non-coordinated CHO group as a handle for further functionalization have been synthesized to achieve selective cytotoxicity to cancer cells, the more potent compounds acting as proteosynthesis inhibitors...

A new family of neutral ruthenium(II) arene complexes of the type [Ru(η6-arene)X(κ2- O, N-L)] (η6-arene = p-cym, bz; X = Cl-, SCN-; HL1 = 2-(2'-hydroxyphenyl)benzimidazole, HL2 = 2-(2'-hydroxyphenyl)benzothiazole) has been synthesized and characterized. The cytotoxic activity of the Ru(II) complexes was evaluated in several tumor cell lines (A549,...

Evidence of the significance of targeting chromatin compaction to increase the therapeutic index of chemotherapy is provided by Arturo López‐Quintela, Fernando Domínguez, and co‐workers in article number 1801317. Co‐administration of silver quantum clusters of three atoms, which increases chromatin accessibility, raises the efficacy of DNA‐binding...

Nanomaterials with very low atomicity deserve consideration as potential pharmacological agents owing to their very small size and to their properties that can be precisely tuned with minor modifications to their size. Here, it is shown that silver clusters of three atoms (Ag3‐AQCs)—developed by an ad hoc method—augment chromatin accessibility. Thi...

Essential processes for living cells such as transcription and replication depend on the formation of specific protein-DNA recognition complexes. Proper formation of such complexes requires suitable fitting between the protein surface and the DNA surface. Adopting doxorubicin (DOX) as a model probe, we report here that Ag3 clusters (Ag-AQCs) inhibi...

Subnanometric samples, containing exclusively Ag2 and Ag3 clusters, were synthesized for the first time by kinetic control using an electrochemical technique without the use of surfactants or capping agents. By combination of thermodynamic and kinetic measurements and theoretical calculations, we show herein that Ag3 clusters interact with DNA thro...

Subnanometric samples, containing exclusively Ag2 and Ag3 clusters, were synthesized for the first time by kinetic control using an electrochemical technique without the use of surfactants or capping agents. By combination of thermodynamic and kinetic measurements and theoretical calculations, we show herein that Ag3 clusters interact with DNA thro...

The complex [(η6-p-cymene)Ru(OH2)(κ2-N,N-HL1)](OTf)2 (HL1 = 2-pyridin-2-yl-1H-benzimidazole), [2c](OTf)2, undergoes easy deprotonation of the N–H group at physiological pH, producing [(η6-p-cymene)Ru(OH2)(κ2-N,N-L1)]+, [2c′]+, prone to dimerization both in solution and in the solid state. X-ray measurements on [2c′]BF4 have shown that dimer units a...

Biodegradable electrospun poly(ε-caprolactone) (PCL) scaffolds were coated with platelet-rich plasma (PRP) to improve cell adhesion and proliferation. PRP was obtained from human buffy coat, and tested on human adipose-derived mesenchymal stem cells (MSCs) to confirm cell proliferation and cytocompatibility. Then, PRP was adsorbed on the PCL scaffo...

The development and evaluation of PEGylated chitosan (CS) nanocapsules (NCs) conjugated to a monoclonal antibody anti-TMEFF-2 (CS-PEG-anti-TMEFF-2 mAb NCs) for targeted delivery of docetaxel (DCX) is presented. CS-PEG-Biotin NCs, displaying biotin tags at their surface, were obtained and efficiently functionalized with an anti-TMEFF-2 mAb through a...

Nanoparticles (NPs) have emerged as a potential tool to improve cancer treatment. Among the proposed uses in imaging and therapy, their use as a drug delivery scaffold has been extensively highlighted. However, there are still some controversial points which need a deeper understanding before clinical application can occur. Here the use of gold nan...

Different cellular internalization of conjugate and free drug (not drawn to scale). AuNPs-cisplatin are internalized via an endocytic pathway, hence cisplatin is only released at the acidic pH of the endosomes. Moreover AuNPs protect the drug from being deactivated by plasma proteins. Free cisplatin mainly enters via diffusion through the cell memb...

Therapeutic efficiency and body weight change of short treatment. (a) Differences in tumor volumes measured by caliper after two consecutive injections (day 0 and 3) of 3 and 1.5 mg cisplatin (kg mouse)–1 each of free cisplatin and AuNP–cisplatin, respectively. The antitumor activity of cisplatin was maintained after the drug was conjugated to the...

Experimental Details. (DOCX)

Confocal microscopy of A549 treated with AuNP-cisplatin. Cells were incubated with AuNP-cisplatin for (a) 30 min, (b) 1 h, (c) 3 h, and (d) 24 h. Afterwards, they were processed by staining nuclear DNA with DAPI, and α-tubulin microtubules with monoclonal mouse anti α-tubulin antibody and goat anti mouse antibody conjugated with Alexa 488. Nanopart...

Analysis of relevant biochemical markers. Aspartate transaminase (AST) and Alanine transaminase (ALT) levels indicate that there is no evidence of liver dysfunction. The renal function is usually determined by the levels of Blood Urea Nitrogen (BUN) and creatinine. Although the BUN seems to be higher than levels reported in other works [100], it sh...

Background: Tmeff2 is a protein with unclear function. It has been described that Tmeff2 promoter is silenced by hypermethylation in certain primary tumors. To date, no other mechanism regulating Tmeff2 expression has been described in tumor cells. In addition to other intracellular signaling pathways, the TGF-b pathway plays an important role in g...

An improved synthetic route to 1α,25-dihydroxyvitamin D(3) des-side chain analogues 2 a and 2 b with substituents at C18 is reported, along with their biological activity. These analogues display significant antiproliferative effects toward MCF-7 breast cancer cells and prodifferentiation activity toward SW480-ADH colon cancer cells; they are also...

To investigate the prevalence of pituitary stalk dysgenesis (PSD) in adult hypopituitary patients by describing the chronology of hormone deficiencies and their potential correlation with traumatic delivery, mutations in genes required for pituitary development and function and pituitary stalk visibility on MRI. Retrospective and prospective study...

Hepcidin acts as the main regulator of iron homeostasis through regulation of intestinal absorption and macrophage release. Hepcidin deficiency causes iron overload whereas its overproduction is associated with anaemia of chronic diseases. The aims of the study were: to identify genetic variants in the hepcidin gene (HAMP) promoter, to asses the as...

Context: Steroid 11β-hydroxylase (CYP11B1) deficiency (11OHD) is the second most common form of congenital adrenal hyperplasia (CAH). Cases of nonclassic 11OHD are rare compared with the incidence of nonclassic 21-hydroxylase deficiency. Objective: The aim of the study was to analyze the functional consequences of seven novel CYP11B1 mutations (p.M...

Congenital adrenal hyperplasia (CAH) due to steroid 21-hydroxylase (CYP21A2) deficiency is the commonest inborn error in steroid hormone biosynthesis. Functional in vitro assessment of mutant activity generally correlates well with clinical phenotype and therefore has contributed greatly to phenotype prediction in this CAH variant. Three CYP21A2 se...

Telomere fluorescence in situ hybridization (T-FISH) of metaphase spreads. Merged images of DAPI (blue) and telomeric probe (red). The selected regions of metaphase spreads from mouse astrocytes infected with HRASV12 are magnified to show centromere fragments (CF) and Robertsonian centromeric fusion (Rob), indicated by arrows. (3.00 MB TIF)

Acute deletion of Rb in astrocytes and analysis of pRb and p107 levels. Immunoblot analysis of proliferating cRbloxP/loxP astrocytes after infection at day 1 and 6 post-selection with puromycin. pRb was no longer detectable at day 1 after selection. The pRb family member p107 was significantly affected by loss of pRb. Compensation of pRb loss by p1...

Wip1 inhibition decreases the proliferation rate in cRb−/−/RasV12 astrocytes. All the experimental groups were treated with the Wip1 chemical inhibitors CCT007093 and Arsenic Trioxide (ATO). The relative cell number of early-passage cRbloxP/loxP conditional astrocytes co-infected with PIG/pBABE (vector), PIG/pBABE-HRasV12, PIG-Cre/pBABE and PIG-Cre...

Acute deletion of Rb in astrocytes and the effect on Wip1 and pp38 levels. Immunoblot analysis of proliferating cRb loxP/loxP astrocytes after infection at day 1 and 6 post-selection with puromycin. The lower level of pp38 is shown after 7 days but not at day 1 after selection. (3.00 MB TIF)

Asthma is a chronic inflammatory disease of the airways that involves many cell types, amongst which mast cells are known to be important. Adenosine, a potent bronchoconstricting agent, exerts its ability to modulate adenosine receptors of mast cells thereby potentiating derived mediator release, histamine being one of the first mediators to be rel...

Chitosan (CS) colloidal carriers, which consist of an oily core and a CS coating, were developed to facilitate a controlled intracellular delivery of docetaxel. The systems presented a particle size of <200 nm and a positive surface charge. As shown by the flow cytometry analysis, fluorescent CS carriers were rapidly internalized by human tumor cel...

Hypochondroplasia (HCH) is a skeletal dysplasia inherited in an autosomal dominant manner due, in most cases, to mutations in the fibroblast growth factor receptor 3 (FGFR3). Acanthosis nigricans (AN) is a velvety and papillomatous pigmented hyperkeratosis of the skin, which has been recognized in some genetic disorders more severe than HCH involvi...

Senescence is one of the main barriers against tumor progression. Oncogenic signals in primary cells result in oncogene-induced senescence (OIS), crucial for protection against cancer development. It has been described in premalignant lesions that OIS requires DNA damage response (DDR) activation, safeguard of the integrity of the genome. Here we d...

The systematic study of the human genome indicates that the inter-individual variability is greater than expected and it is not only related to sequence polymorphisms but also to gene copy number variants (CNVs). Congenital Adrenal Hyperplasia due to 21-hydroxylase deficiency (21OHD) is the most common autosomal recessive disorder with a carrier fr...

Correct diagnosis of 21-hydroxylase deficiency (21OHD) requires the identification of CYP21A2 gene deletions and CYP21A1P/CYP21A2 chimeric genes, which are disease-causing alleles, and gene duplications, which can lead to false-positive 21OHD allele results. Because lack of suitable CYP21A2 dosage assessment methods hampers correct 21OHD diagnosis,...

Recent evidences indicate new roles for the glycolytic protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in essential mammalian cell processes, such as apoptosis and proliferation. To clarify the involvement of this protein in growth and programmed cell death in the liver, cell models of hepatocytes in culture were used to study GAPDH expres...

Prothymosin alpha is an acidic protein, reported to be involved in cell proliferation and apoptosis, although its precise function in both processes are still unknown. Due to the importance of these processes in the pathogenesis of hepatic diseases and the need to understand the molecular mechanisms underlying these diseases we aimed to investigate...

To detect common as well as rare and novel CYP21A mutations in 21-hydroxylase deficiency patients. To estimate the distribution of mutations and compare them with other European studies. To construct haplotypes linked to a recurrent novel mutation. Genetic analysis by sequencing the entire CYP21A2 gene plus Southern blot. A total of 138 unrelated S...

Kallmann's syndrome (KS) refers to the association of hypogonadic hypogonadism and anosmia or hyposmia. The X-linked form of the disease is due to mutations in the KAL1 gene that encodes for the protein anosmin-1. We studied the KAL1 gene in a patient with KS and his family by PCR amplification and direct sequencing. A novel missense mutation (V263...

The main objective of the present work was the development of new nanoparticulate carrier systems for the delivery of plasmid DNA. These new carriers consist of a blend matrix formed by a poly(lactic-co-glycolic acid) (PLGA) copolymer and polyoxyethylene derivatives. More specifically, we have prepared nanostructures with different PLGA:poloxamer a...

Uveal malignant melanoma is the most frequent primary intraocular tumor in adult humans. The cellular events leading to neoplasic transformation of normal uveal melanocytes are not well known when compared to other cancers. In this study, we investigated the role of G1 and G1/S regulatory proteins of the cell cycle in human uveal melanoma (UM) prim...

Millions of people worldwide suffer goiter, a proliferative disease of the follicular cells of the thyroid that may become neoplastic. Thyroid neoplasms have low proliferative index, low apoptotic index and a high incidence of metastasis. TGF-beta is overexpressed in thyroid follicular tumor cells. To investigate the role of TGF-beta in thyroid tum...

The aim of this work was to culture human retinal pigment epithelium (hRPE) cells over human amniotic membrane (hAM). Human AM was studied for its viability as an adequate support for transplantation of an hRPE cell monolayer with preserved cell polarity to the subretinal space. Human AM was obtained from pregnant women during caesarean section. Th...

We show in this work that the inhibition of Cdk4 (6) in Rb(-/-) 3T3 cells enhances the accumulation of the p27(kip1) cyclin-dependent kinase inhibitor when these cells are induced into quiescence. Two different forms of inhibition of Cdk4 (6), namely overexpression of the Cdk4 (6) inhibitor p16 and overexpression of a dominant negative mutant of Cd...

Laron syndrome (LS) or growth hormone (GH) insensitivity syndrome (GHIS) is an autosomal recessive disease due to molecular defects in the GH receptor gene (GHR). Most of the identified mutations are located on the extracelular domain of the receptor. We studied the GHR gene in a patient with LS and found a homozygous missense mutation in exon 2. T...

Background: Isoprenoid biosynthesis is known to be essential for diverse cellular functions, including cell proliferation. The aim of this work was to study the effects caused by the addition of different inhibitors of isoprenylation (lovastatin, manumycin A, farnesyltransferase inhibitor III and N-acetyl-S-farnesyl-L-cysteine) to human retinal pig...

In a previous report we suggested that the estimation of prothymosin alpha (PTA) levels in primary breast tumours might be used to identify breast cancer patients at high risk for distant metastasis (Dominguez F et al (1993) Eur J Cancer 29A: 893-897). Here the role of tumour PTA levels as predictor was investigated with respect to both disease-fre...

The enormous expansion in the application of the technologies of molecular and cell biology with the objective of resolving medical problems is discussed, as well as the problems arising due to this revolution. Progress in methodology is also reported paying special attention to chip technology. The different types of chips, strategies for readout,...

Prothymosin alpha (PTA) was detected by immunocytological and biochemical methods in oocytes at different stages of oogenesis, and in early embryos of the amphibian Bufo anenarum. In all cases PTA was detected in the nucleus and was absent from the cytoplasm. This indicates that this protein could act at the level of regulating transcription. Weste...

Prothymosin alpha is a polypeptide whose function is still a matter of debate. The present paper represents the first morphological description of the prothymosin alpha distribution in the human and rat digestive tract tissues using immunohistochemical techniques. In these epithelia prothymosin alpha immunostaining was localized in the nuclei of th...

Pituitary-specific expression of the GH gene is dependent on a pituitary-specific transcription factor GH factor-1 (GHF-1), a homeodomain protein also known as pituitary-specific transcription factor-1 (Pit-1). The aim of this study was to investigate the regulation of GHF-1 messenger RNA (mRNA) levels in primary monolayer cultures of rat anterior...

Pituitary-specific expression of the GH gene is dependent on a pituitary-specific transcription factor GH factor-1 (GHF-1), a homeodomain protein also known as pituitary-specific transcription factor-1 (Pit-1). The aim of this study was to investigate the regulation of GHF-1 messenger RNA (mRNA) levels in primary monolayer cultures of rat anterior...

Prothymosin alpha (ProT) is a polypeptide widely distributed in the organism and expressed by cell types with a high proliferative capacity. The aim of the current work was to investigate if ProT was localized in the progenitor compartment of the adrenal cortex which, following the cell migration theory, corresponds to the zona glomerulosa. We stud...

Using flow cytometry we observed the effects that different hormonal treatments had on the progression of rat thyroid (FRTL-5) cells through the cell cycle. The absence of hormones or the addition of TSH (6 mU/ml) did not induce DNA synthesis; however, the addition of IGF-I (30 ng/ml) promoted cell proliferation. The number of cells recruited by IG...

In 71 patients with classic invasive ductal carcinomas, levels of prothymosin alpha (PT alpha), as assayed by a radioimmunoassay that detects thymosin alpha 1 (the NH2-terminal fragment of PT alpha), were significantly greater in tumour samples than in normal breast tissue. PT alpha levels were correlated with (a) the number of positive axillary ly...

The cellular distribution of prothymosin alpha (ProT) was studied in ovarian follicles of adult cycling rats. We found positive granulosa and theca cells throughout follicular maturation. When both ProT and proliferating cell nuclear antigen (PCNA) immunoreactivity was studied, we observed that both proteins were expressed in the same granulosa and...

Prothymosin alpha (PTA) mRNA and histone H4 (H4) mRNA levels were studied in various experimental conditions that affected GH1 pituitary tumor cell proliferation. Cell proliferation and progression through the cell cycle was assessed by counting cells, 3H-thymidine incorporation and flow cytometry. PTA mRNA levels were decreased in a time-dependent...

Parathymosin (ParaT) is an almost ubiquitious peptide first isolated from rat thymus, the highest concentration being in the liver. Its subcellular localization is controversial and both a cytoplasmic and a nuclear distribution have been reported. The aim of the present work was to study the intracellular localization of ParaT in rat and human live...

The aim of this study was to investigate the effect of in vivo estrogen administration on hypothalamic growth hormone releasing hormone (GHRH) and somatostatin (SS) gene expression. We found that estrogen administration (estradiol valerate 250 micrograms/every 3 days, subcutaneously) to male rats induced a decrease in both hypothalamic GHRH mRNA le...

Prothymosin alpha (PT-alpha) mRNA levels were evaluated at different stages during the cell cycle. NIH 3T3 cells were synchronized: (a) by serum deprivation, (b) by mitotic shake off after nocodazole arrest, and (c) by double thymidine block. Cell synchronism was estimated by flow cytometry. In cells grown in serum-free medium, PT-alpha mRNA levels...

Prothymosin-alpha (PT-alpha) mRNA levels were evaluated at different stages during the cell cycle. NIH 3T3 cells were synchronized: (a) by serum deprivation, (b) by mitotic shake off after nocodazole arrest, and (c) by double thymidine block. Cell synchronism was estimated by flow cytometry. In cells grown in serum-free medium, PT-alpha-mRNA levels...

The effects of free fatty acids (FFAs) on growth hormone releasing hormone (GHRH) secretion by fetal rat cortical and hypothalamic neurons in monolayer culture have been investigated. Treatment with caprylic or oleic acids for 90 min, induced a dose-dependent increase in GHRH secretion. The stimulatory effect of caprylic acid was time-dependent, be...

By means of immunohistochemical methods, we have investigated the cellular distribution of prothymosin alpha and parathymosin in rat thymus and spleen, using specific antibodies raised against thymosin alpha-1 and against parathymosin. We observed prothymosin alpha immunoreactivity in lymphoid cells both in thymus and spleen. In the thymus, prothym...

Neuroendocrine regulation is complex. Each pituitary hormone is controlled at different levels and by a large array of messengers, in order to achieve carefully modulated release with respect to both quantity and frequency. The fine tuning of growth hormone (GH) control is exerted at pituitary, hypothalamic and suprahypothalamic levels. At each lev...

A cDNA library was constructed from human spleen mRNA and screened for clones containing cDNAs coding for prothymosin alpha. A clone containing a 503-base-pair insert including the entire coding sequence for the translated portion of the mRNA was isolated. The deduced amino acid sequence confirms and completes the partial sequence of human prothymo...

Young normal control rats were compared with a group made diabetic by treatment with streptozotocin and with other normal rats maintained on a restricted diet to obtain a daily body weight change similar to that of the diabetics. In diabetic rats plasma levels of Val, Ile and Leu rose and those of Asp, Thr, Ser, Gln, Gly, Tyr, Orn, Lys, His and Arg...

The concentration of most amino acids was higher in the brains of 19- and 21-day rat fetuses than in their respective mothers. After an intraperitoneal load of tryptophan to the mother, the intracerebral concentration of several amino acids (including leucine) decreased not only in the mothers, but also in their fetuses. The in vitro incorporation...