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PALB2 variants have been scarcely described in Argentinian and Latin-American reports. In this study, we describe molecular and clinical characteristics of PALB2 mutations found in multi-gene panels (MP) from breast-ovarian cancer (BOC) families in different institutions from Argentina.We retrospectively identified PALB2 pathogenic (PV) and likely pathogenic (LPV) variants from a cohort of 1905 MP results, provided by one local lab (Heritas) and SITHER (Hereditary Tumor Information System) public database. All patients met hereditary BOC clinical criteria for testing, according to current guidelines.The frequency of PALB2 mutations is 2.78% (53/1905). Forty-eight (90.5%) are PV and five (9.5%) are LPV. Most of the 18 different mutations (89%) are nonsense and frameshift types and 2 variants are novel. One high-rate recurrent PV (Y551*) is present in 43% (23/53) of the unrelated index cases. From the 53 affected carriers, 94% have BC diagnosis with 14% of bilateral cases. BC phenotype is mainly invasive ductal (78%) with 62% of hormone-receptor positive and 22% of triple negative tumors. Self-reported ethnic background of the cohort is West European (66%) and native Latin-American (20%) which is representative of Buenos Aires and other big urban areas of the country.This is the first report describing molecular and clinical characteristics of PALB2 carriers in Argentina. Frequency of PALB2 PV in Argentinian HBOC families is higher than in other reported populations. Y551* is a recurrent mutation that seems to be responsible for almost 50% of PALB2 cases.

Cystic fibrosis (CF) is the main genetic cause of death among the Caucasian population. The disease is characterized by abnormal fluid and electrolyte mobility across secretory epithelia. The first manifestations occur within hours of birth (meconium ileus), later extending to other organs, generally affecting the respiratory tract. It is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR encodes a cyclic adenosine monophosphate (cAMP)-dependent, phosphorylation-regulated chloride channel required for transport of chloride and other ions through cell membranes. There are more than 2,000 mutations described in the CFTR gene, but one of them, phenylalanine residue at amino acid position 508 (p.F508del), a recessive allele, is responsible for the vast majority of CF cases worldwide. Here, we present the results of the application of genome-editing techniques to the restoration of CFTR activity in p.F508del patient-derived induced pluripotent stem cells (iPSCs). Gene-edited iPSCs were subsequently used to produce intestinal organoids on which the physiological activity of the restored gene was tested in forskolin-induced swelling tests. The seamless restoration of the p.F508del mutation resulted in normal expression of the mature CFTR glycoprotein, full recovery of CFTR activity, and a normal response of the repaired organoids to treatment with two approved CF therapies: VX-770 and VX-809., Graphical Abstract, Cystic Fibrosis (CF) is the main genetic cause of death among the Caucasian population. Here, we present the results of the application of genome-editing techniques to repair the CF defect in patient-derived cells and organoids. This protocol could be the foundation for future treatments based on cell repair and transplantation.

Background In countries where comparative genomic hybridization arrays (aCGH) and next generation sequencing are not widely available due to accessibility and economic constraints, conventional 400–500-band karyotyping is the first-line choice for the etiological diagnosis of patients with congenital malformations and intellectual disability. Conventional karyotype analysis can rule out chromosomal alterations greater than 10 Mb. However, some large structural abnormalities, such as derivative chromosomes, may go undetected when the analysis is performed at less than a 550-band resolution and the size and banding pattern of the interchanged segments are similar. Derivatives frequently originate from inter-chromosomal exchanges and sometimes are inherited from a parent who carries a reciprocal translocation. Case presentation We present two cases with derivative chromosomes involving a 9.1 Mb 5p deletion/14.8 Mb 10p duplication in the first patient and a 19.9 Mb 5p deletion/ 18.5 Mb 9p duplication in the second patient. These long chromosomal imbalances were ascertained by aCGH but not by conventional cytogenetics. Both patients presented with a deletion of the Cri du chat syndrome region and a duplication of another genomic region. Each patient had a unique clinical picture, and although they presented some features of Cri du chat syndrome, the phenotype did not conclusively point towards this diagnosis, although a chromosomopathy was suspected. Conclusions These cases highlight the fundamental role of the clinical suspicion in guiding the approach for the etiological diagnosis of patients. Molecular cytogenetics techniques, such as aCGH, should be considered when the clinician suspects the presence of a chromosomal imbalance in spite of a normal karyotype.

El artículo se inscribe en el Proyecto “Institucionalidades en construcción, sujetos y experiencias en contexto. Tensiones y aperturas en tiempos de demandas de igualdad” (Facultad de Filosofía y Humanidades, Secretaría de Ciencia y Técnica, Universidad Nacional de Córdoba, 2016-2017), y avanza en lecturas de material de campo desde una perspectiva de derechos. Con base en recorridos investigativos anteriores, el proyecto mencionado se propone como una etapa centrada en el estudio de las instituciones educativas, sus procesos y transformaciones recientes, en tiempos que hemos denominado de “demandas de igualdad”, y recupera para ello registros analizados en sus contextos locales. La metodología se encuadra en la perspectiva cualitativa, de orientación socio-antropológica; se apela al trabajo de campo para recuperar situaciones, relatos y reflexiones de los actores sociales y educativos que dan cuenta del modo en que la ampliación de derechos en diversos planos atravesó la vida institucional, generó nuevos posicionamientos y permitió el reconocimiento de niños y jóvenes como “sujetos de derechos”; estos procesos han dado lugar a tensiones que expresan la conflictividad de las transformaciones y las controversias latentes en el conjunto social. Se analizan tres registros: el primero se titula “Jóvenes y Memoria”, vinculado a las luchas por la Memoria, la Verdad y la Justicia, el segundo “Niñez y Escuela”, relativo a constitución de las niñas, niños y adolescentes como sujetos de derecho, y el tercero a “Vivencias de género” que incluye relatos de un adolescente y sus reflexiones luego de la sanción de la Ley de Matrimonio Igualitario. Nos parece oportuno, en el contexto político regresivo que Argentina afronta en la actualidad, realizar un aporte construido desde la investigación educativa en el espacio universitario.

El presente trabajo se inscribe en el proyecto “Institucionalidades en construcción, sujetos y experiencias en contexto. Tensiones y aperturas en tiempos de demandas de igualdad” (FFyH; Secyt-UNC). Con base en recorridos anteriores, dicho proyecto se centra en el estudio de “las instituciones, sus procesos y transformaciones recientes, recuperando el papel de los sujetos y las experiencias en contextos locales”. Focaliza la mirada en la producción de nuevas institucionalidades y en el lugar de los sujetos, contemplando el papel instituyente de las prácticas y de la pluralidad de experiencias y trayectorias. Más puntualmente, se ubica en una de las líneas del proyecto que apunta al estudio de la relación escuela-familia y comunidad; se recuperan registros producidos en el marco de una propuesta de formación solicitada por el sindicato, titulada “Escuelas, familias y contextos. Hacia la construcción de estrategias institucionales de trabajo en torno a una relación compleja”. A través de esas notas de campo, se recorren algunas situaciones relatadas por los docentes, para luego reflexionar en torno a diversas aristas de esta relación; planteamos algunas conceptualizaciones fértiles para ampliar el horizonte analítico y discutir estrategias institucionales de trabajo en el marco de las orientaciones previstas en la Ley Nacional de Educación. This paper is part of the project “Institutions under construction, subjects and experiences in context. Tensions and openings in times of demands of equality “ (FFyH; Secyt-UNC). Based on previous tours, this project focuses on the study of “institutions, their processes and recent transformations, recovering the role of subjects and experiences in local contexts.” It focuses on the production of new institutions and the place of the subjects, contemplating the institutional role of practices and the plurality of experiences and trajectories. More punctually, is located in one of the lines of the project that aims at the study of the relationship school-family and community; records are retrieved within the framework of a training proposal requested by the union, entitled “Schools, families and contexts. Towards the construction of institutional strategies of work around a complex relationship”. Through these field notes, we review some situations reported by teachers, and then reflect on several aspects of this relationship; we propose some fertile conceptualizations to expand the analytical horizon and discuss institutional work strategies within the framework of the guidelines provided for in the National Education Law. Fil: Avila, Olga Silvia. Universidad Nacional de Córdoba; Argentina.

Background Hand-foot-genital syndrome (HFGS) is a rare condition characterized by congenital malformations in the limbs and genitourinary tract. Generally, this syndrome occurs due to point mutations that cause loss of function of the HOXA13 gene, which is located on 7p15; however, there are some patients with HFGS caused by interstitial deletions in this region. Case presentation We describe a pediatric Mexican patient who came to the Medical Genetics Department at the National Institute of Pediatrics because he presented with genital, hand and feet anomalies, facial dysmorphisms, and learning difficulties. Array CGH reported a 12.7 Mb deletion that includes HOXA13. Conclusions We compared our patient with cases of HFGS reported in the literature caused by a microdeletion; we found a minimum shared region in 7p15.2. By analyzing the phenotype in these patients, we suggest that microdeletions in this region should be investigated in all patients with clinical characteristics of HFGS who also present with dysplastic ears, mainly low-set implantation with a prominent antihelix, as well as a low nasal bridge and long philtrum.

Recently, 2q24.2 deletion syndrome has emerged as a cause ofintellectual disability. Takatsuki et al. [2010] provided the firstreportwithadeletionin2q24.2q24.3,confirmedusingcomparativegenomic hybridization microarray (aCGH) analysis, who had alow-birth weight, hypotonia, myoclonic seizures, dysmorphic fea-tures, and pulmonary emphysema. Subsequently, two other caseswere reported showing severe hypotonia, joint laxity, and facialdysmorphism;oneofthemhadadenovointerstitialdeletionof5-Mb at 2q24.2q24.3 [Magri et al., 2011], and the other patient (ID4547) who was included in the European Cytogeneticists Associa-tion Register of Unbalanced Chromosome Aberrations database,carrying a deletion in 2q24.2. The emerging syndrome was firstsuggestedbyPalumboetal.[2012],whodescribedacasewitha7.5-Mb de novo deletion in 2q24.1q24.2 encompassing 59 genes, asdetectedusingSNParray.Theclinicalfeaturesincludedgeneralizedhypotonia, marked bruxism, and bilaterally hip dislocation.In addition, two patients with mosaicism have been reported;the first (ID 1576) in Database of Chromosomal Imbalance andPhenotype in Humans using Ensembl Resources, is carrier of adouble2q24.2 deletion, one segmentof0.63-Mb (meanratio 1)and other of 0.17-Mb (mean ratio 0.41) inside the same region,this can be interpreted as a mosaic of monosomy and nullisomywhose molecular and clinical description is incomplete, thereforeit is not possible to include her in the present study. The secondmosaic,reportedbyBurrageetal.[2013]isamildlyaffectedpatientwithshortstatureandmosaicismwith61%ofcellscarryinga0.4-Mbdeletionon2q24.2,whichencompassesonlythreegenes.Theysuggested that TBR1was responsible for the intellectual disabilityobserved and that the short stature was associated with thePSMD14 gene.Wepresentapatient,whowasrecruitedinaprospectiveresearchprojecttodetectgenomicabnormalitiesthroughaCGHinpatientswithintellectualdisability;theresearchwasreviewedandapprovedby the ethics committee of the Instituto Nacional de Pediatri´a.The patient, a 7-year-old boy is the first child of healthy andnonconsanguineous parents. He was born after an uneventful

Angiogenesis, the sprouting of new blood vessels from pre-existing vessels, is a remarkable feature of tumours growth and metastasis. The in vivo inhibition of this receptor by cyclic peptides containing RGD sequence may be used to selectively suppress these disease1. Our research group has developed a new methodology2 for the evaluation of new antiangiogenic compounds, based in the genetic expression analysis using CGH array system. The RGD mimetics activity can be modified by the presence of ester-bond3, Therefore, we decided to prepare some depsipeptides analogous to the RGD-β-lactam compounds and evaluate their activity performing a genetic expression analysis. We also have evaluated the activity of open-chain compounds without RGD formal structure. The cyclic depsipeptides have demonstrated a very effective inhibitory activity. In the other hand the open-chain compounds have a surprising behavior, demonstrating a similar gene activation. This way, we call into question the essential need of RGD sequence to have an interaction between ligand and the receptor of the integrin4, 5. da Ressurreicao, A. S. M.; Vidu, A.; Civera M.; Belvisi L.; Potenza, D.; Manzoni, L.; Ongeri, S.; Gennari, C.; Piarulli, U.; Eur. 2009, 15, 12184. Aizpurua, J. M.; Ganboa, J. I.; Palomo, C.; Loinaz, I.; Oyarbide, J.; Fernandez, X.; Belentova, E.; Fratila, R. M.; Jimenez A.; Miranda, J. I.; Laso, A.; Avila, S.; Castrillo, J. L.; ChemBioChem, 2001, 11, 401. Cupido, T.; Spengler, J.; Ruiz-Rodriguez, J.; Adan, J.; Mitjans, F.; Puilats, J.; Albericio, F.; Chem. Int. Ed., 2010, 49, 2732. Xiong, J. P.; Stehle, T.; Zhang, R.; Joachimiak, A.; Frech, M.; Goodman, S. L.; Arnaout, M. A.; Science, 2002, 296, 151. Craig, D.; Gao, M.; Schulten, K.; Vogel, V.; Structure, 2004, 12, 2049.

In humans, a germline mutation (c.309C>G) in the TWIST1 oncogene may predispose to breast cancer and its expression has been associated with tumour progression and metastasis. In this study, the feline TWIST1 gene was screened for sequence variations in 37 neoplastic and eight hyperplastic mammary gland lesions from cats. In addition, mRNA levels were examined in 15 mammary tumours and three cases of mammary hyperplasia by quantitative real-time reverse-transcriptase PCR. Feline mammary carcinomas had significantly lower levels of expression of TWIST1 mRNA than benign mammary tumours. No variations were identified in the TWIST1 coding region in feline mammary tumours and the mutation described in humans was not detected. However, two germline variants in the TWIST1 gene intron were identified in four and three carcinomas, respectively: GQ167299:g.535delG and GQ167299:g.460C>T. There was no association between these sequence alterations and TWIST1 mRNA levels.

Angiogenesis is a fundamental step in the transition of solid tumors from a dormant state to a malign one. Many of the low-molecular-weight anti-angiogenic drug candidates mimic the short peptide epitope Arg-Gly-Asp (RGD), disrupting the extracellular matrix/integrin adhesion and, ultimately, leading to tumor cell apoptosis. In contrast to the detailed structural information available for the extracellular adhesion inhibition of endothelial cells through the recognition of integrins (typically aVb3) by RGD peptidomimetics, [2] most aspects of possible intracellular angiogenic gene regulation caused by peptidomimetics remain unexplored. Importantly, dysfunction of this signaling system is suspected to be behind the resistance phenomena developed in anti-angiogenic therapies. Inside the endothelial cell, dozens of proteins mediate or control the signaling pathways of angiogenesis after integrin activation, but only a couple of kinases (JNK, ERK) and transcription factors (NFkB, FoxO) are able to promote gene regulation (Figure 1). In addition, large environmental ligands, such as vascular endothelial growth factors (VEGFs) or peptide hormones, are required to elicit proangiogenic gene regulation. In this context, we set out to design alternative lowmolecular-weight RGD probes for interaction with aVb3 integrin and gene regulation in HUVECs. Ideally, these peptidomimetics should: 1) contain a minimal scaffold to prevent undesired scaffold/integrin interactions, 2) have a uniform and predictable scaffold conformation, 3) bear a maximum of recognition groups, including hydrophobic or hydrophilic ones, and 4) permit the deletion of selected residues from the RGD triad without global shape change. We selected five-membered and four-membered small cyclic peptides for development. The residual flexibility of these cyclopeptides can be further constrained by incorporating lactam bridges between neighboring amino acids to stabilize protein secondary structure motifs characterized by combinations of b-turns and/or g-turns. Several lactam pseudopeptides resulting from such an extension of Freidinger’s concept (Scheme 1; 1!2) have been explored with the goals of mimicking receptor-bound conformations of bioactive peptides and of providing pharmacophore information for nonpeptide drug design. However, despite its apparent simplicity, this design is not always reliable for the precise positioning of a maximum number of recognition groups around the pseudopeptide cyclic core. Because the interresidual lactam bridge created by modification of the side chain (R) shares recognition and constraint functions, the design of mimetic libraries becomes difficult and non-general, when synthetically achievable. An alternative way to constrain cyclic peptides is based on the incorporation of a d-amino acid and an N-alkyl-amino acid into the macrocycle, as illustrated by the remarkable aVb3 antagonist cilengitide (3, cyclo-[Arg-Gly-Asp-d-Phe-N(Me)Val]) developed by Kessler et al. Here we report an alternative, more versatile solution to the above design problem, by introducing a-amino-b-lactam scaffolds originating from aCH/NH proton mimicry (Scheme 2; 1! 4). This “b-lactam scaffold-assisted design” (b-LSAD) approach is based on the separation of recognition and constraint groups, and has previously proven efficient for promotion of unusually stable type-II (II’) b-turn peptidomimetics from extended open native peptides. Accordingly, the straight application of the b-LSAD principle to RGD cyclic mimetics led us to the b-lactam pseudopeptides 4a–d, which fulfilled several structural requirements to trigger quite different signaling interactions with aVb3 integrin. The hydrophobic benzyl group in the d-Phe residue of cilengitide (3), for instance, was replaced by the strongly hydrophilic 1,2-dihydroxyethyl moiety in the mimetic 4a or by the poorly hydrophobic methyl group in compound 4b. Conversely, the hydrophobic R–R substituents Figure 1. Signaling pathways initiated by integrins affecting the gene regulation of angiogenesis. ERK=extracellular-signal-regulated kinase, FAK= focal adhesion kinase, FoxO= forkhead box O, JNK=c-Jun N-terminal kinase, NFkB=nuclear factor k of B-cells.

Many chromosomal deletions encompassing the 2q23.1 region have been described ranging from small deletions of 38 kb up to >19 Mb. Most phenotypic features of the 2q23.1 deletion syndrome are due to a MBD5 gene loss independent of the size of the deletion. Here, we describe a male patient harboring a novel interstitial deletion encompassing the 2q22.3 q23.3 chromosomal region. Array-CGH revealed a 7.1 Mb deletion causing haploinsufficiency of several genes including MBD5, ACVR2, KIF5C, and EPC2. This patient presents with additional findings to those already described in individuals who have deletions of MBD5 including toes absence of halluces, pure red cell aplasia, and intestinal aganglionosis. Interestingly, in the deleted region there are previously identified regulatory sequences which are located upstream to ZEB2, which is associated with Hirschsprung disease (HSCR). Several genes have been associated with pure red cell aplasia, but to our knowledge, this is the first time that 2q deletion is associated with this phenotype. These additional findings should be added to the list of manifestations associated with 2q deletion, and provide support for the hypothesis that this individual has a true contiguous gene deletion syndrome. © 2015 Wiley Periodicals, Inc.

Expression of POU1F1 gene, a member of the POU homeodomain family of transcription factors, is necessary for normal differentiation, development and survival of three anterior pituitary cell types (thyrotrophs, somatotrophs and lactotrophs) and for the proper expression of growth hormone (GH), prolactin (PRL), thyroid-stimulating hormone (TSH) genes and POU1F1 gene itself. Alternative splicing forms of this gene have been reported in different species, with few functional studies. Apart from the POU1F1-Wild-type with the expected length, in this work we isolated three additional splicing variants: POU1F1-beta, with a 78 bp insert in the trans-activation domain; POU1F1-gamma that lacks exon 3 and POU1F1-delta that lacks exons 3, 4 and 5. Four different protein isoforms were also detected by Western blot in the sheep pituitary tissue. Functional assays were performed to study the trans-activation of GH and PRL promoters by the splicing variants. Regarding the PRL promoter, the beta variant presented only 12% of the Wild-type trans-activation capacity. Variants gamma and delta showed no capacity to trans-activate PRL promoter. Both gamma and delta variants acted as repressors of Wt, reducing significantly the trans-activation made by Wt alone (p

In the past, human endometrial receptivity has been investigated by chasing specific molecules throughout the menstrual cycle. Now the genomic approach allows us to investigate the hierarchical contribution of a high number of genes to a specific function. In this study, we analyzed differentially the gene expression pattern of 375 human cytokines, chemokines, and related factors, plus that of their receptors, in endometrial receptivity. To do this, we used a combined approach of human endometrium and cell lines. We have compared the gene expression pattern in receptive vs. prereceptive human endometria and contrasted the results with gene expression in the highly adhesive cell line (to JAR cells and mouse blastocysts) RL95-2 vs. HEC-1A, a cell line with markedly less adhesiveness. IGF-binding protein-related protein 1 (IGFBP-rP1), also known as IGFBP-7/mac 25, was the second most up-regulated gene in both of the investigated models. These results were corroborated by performing RT-PCR on the same RNA samples and validated by quantitative fluorescent RT-PCR and in situ hybridization in endometrium throughout the menstrual cycle. Interestingly, a 35-fold increase in expression during the receptive phase was compared with the prereceptive phase followed by a sharp increase in the late luteal. Further quantitative fluorescent RT-PCR experiments using the epithelial and stromal endometrial fraction throughout the menstrual cycle confirmed that IGFBP-rP1 expression was localized in the epithelial and stromal compartments and up-regulated mainly in the latter. In situ experiments confirmed the endometrial localization and regulation of IGFBP-rP1 mRNA. At the protein level, IGFBP-rP1 was localized by immunohistochemistry at the apical part of the luminal and glandular epithelium, stromal, and endothelial cells. In conclusion, using a genomic approach with a combined experimental design of receptivity in vivo and in vitro, we have discovered the implication of IGFBP-rP1 in endometrial physiology, which seems related to endometrial receptivity.

Endometrial receptivity is a self-limited period in which the endometrial epithelium (EE) acquires a functional and transient ovarian steroid-dependent status that allows blastocyst adhesion. Termed as "the window of implantation", this specific period opens 4-5 days after progesterone production or administration and closes after 9-10 days. Scientific knowledge on the endometrial receptivity process is fundamental for the understanding of human reproduction, but so far none of the proposed biochemical markers for endometrial receptivity has been proven to be clinically useful. In this work, we present strategies of cDNA analysis technologies that aim to clarify the fragmented information in this field. Specifically, the objective is the differential identification, cloning and sequencing of genes linked to endometrial receptivity in humans, combining differential display PCR and cDNA microarray analysis of endometrial epithelial-derived cell lines and endometrial samples obtained in the same patient 2 and 7 days after the luteinizing hormone (LH) surge (day LH+2) and (day LH+7), respectively.

A 4-month-old infant had two 3 cm × 4 cm hemangiomatous lesions on the scalp and back, present since birth, which contained peculiar white-yellowish small nodules. Histologically the lesions proved to have a hemangiomatous (capillary-type) component together with small keratin-containing epidermal cysts (milialike) which progressively extruded their contents. The lesions also contained embryonic-like hair structures. The hemangiomas resolved spontaneously. We were unable to find any reference in the literature describing this peculiar combination of features.