On the role of syllable structure in atypical phonological development: Evidence from the acquisition of /l/ by Portuguese children.

The alveolar lateral is phonetically and phonologically complex. Previous studies have shown that /l/ is one of the last segments to be acquired by typically developing Portuguese children. However, little is known about how Portuguese children with atypical development acquire /l/. In this paper, we investigate the acquisition of /l/ by Portuguese children with protracted phonological development (DLD; SSD). We explore the effect of syllable structure and segmental properties in the acquisition of /l/ and describe mismatches used for target /l/, thus contributing empirical evidence to the ongoing discussion on differential diagnoses for children with primary phonological disorders. Our results show that the lateral is more problematic in SSD than in DLD, with the manner of articulation being more problematic than its place. A syllable-segment interface effect was attested. Mismatches showed a preference for [w, ɾ, ø]. The results are discussed considering their implications for clinical practice and the role of target phonetic and phonological properties in the /l/ acquisition path.

Life beyond Loss: A Retrospective Analysis of the Impact of Meaning of Life Therapy on the Grieving Process of Cancer Patients' Family Caregivers.

Oncological disease in the palliative stage is a huge challenge for patients and their family caregivers (FCs) due to the fact that it confronts them with death, as well as physical, psychological, and existential suffering. Meaning of Life Therapy (MLT) is a brief structured psycho-existential intervention aiming to help patients in a meaning-making life review process, promoting end-of-life adaptation. The Life Letter (LL) resulting from MLT is an element that facilitates communication between the patient and their caregivers. The goal of this study was to understand the impact of MLT on the grieving processes of eight FCs and to study their perceptions of the role of the LL on grief through semi-structured interviews. The results of our qualitative analysis indicate that MLT was perceived by the FCs as a positive experience despite the conspiracy of silence being identified as a drawback. The LL was interpreted as a communicational element, promoting emotional closeness with the cancer patients and serving as a valuable tool in the FCs' adaptation to loss. Our research findings show that the needs of FCs, especially after experiencing the loss of their relative, are dynamic and specific. This is why it is urgent to develop interventions that consider the idiosyncrasies of end-of-life cancer patients and their FCs in order to avoid frustrated farewells, lonely deaths, and maladaptive grieving processes. This is the direction in which MLT should evolve.

Protracted phonological development of a Portuguese six-year-old from the perspective of nonlinear phonology.

As a contribution to a special issue with individual profiles in protracted phonological development (PPD), we present a European Portuguese-speaking six-year-old, "Vicente". By age six years, Portuguese-learning children have generally mastered most of the phonology. However, Vicente showed severe persisting PPD, which was negatively impacting his general socialisation. Although word length and stress often matched the adult targets as did many consonants and vowels, consonants were restricted in distribution and sequences. Consonant clusters showed a particularly high proportion of reduction. A constraints-based nonlinear phonological analysis led to a proposed intervention plan to address needs across the phonological hierarchy in the context of his greater need for enhanced socialisation: starting with the more attainable new word positions and sequences for consonants in his inventory, before addressing a major need for new word structure (clusters) and minor needs for segmental development (additional coronals).

Direct Costs of COVID-19 Inpatient Admissions in a Portuguese Tertiary Care University Centre.

Background: The COVID-19 pandemic has posed greater financial pressure on health systems and institutions that had to respond to the specific needs of COVID-19 patients while ensuring the safety of the diagnosis and treatment of all patients and healthcare professionals. To assess the financial impact of COVID-19 patients admitted to hospitals, we have characterized the cost of COVID-19 admissions, using inpatient data from a Portuguese Tertiary Care University Centre. Methods: We analysed inpatient data from adult patients diagnosed with COVID-19 who were admitted between March 1, 2020 and May 31, 2020. Admissions were eligible if the ICD-10-CM principal diagnosis was coded U07.1. We excluded admissions from patients under 18 years old, admissions with incomplete records, admissions from patients who had been transferred to or from other hospitals or those whose inpatient stay was under 24 h. Pregnancy, childbirth, and puerperium admissions were also excluded, as well as admissions from patients who had undergone surgery. Results: We identified 223 admissions of patients diagnosed with COVID-19. Most were men (64.1%) and aged 45-64 years (30.5%). Around 13.0% of patients were admitted to intensive care units and 9.9% died in hospital. The average length of hospital stay was 12.7 days (SD = 10.2) and the average estimated cost per admission was EUR 8,177 (SD = 11,534), which represents more than triple the inpatient base price (EUR 2,386). Human resources accounted for the highest proportion of the total costs per admission (50.8%). About 92.4% of the admissions were assigned to Diagnosis Related Group (DRG) 723, whose inpatient price is lower than COVID-19 inpatient costs for all degrees of severity. Conclusion: COVID-19 admissions represent a substantial financial burden for the Portuguese NHS. For each COVID-19 hospitalized patient it would have been possible to treat three other hospitalized patients. Also, the price set for DRG 723 is not adjusted to the cost of COVID-19 patients. These findings highlight the need for additional financial resources for the health system and, in particular, for hospitals that have treated high volumes of hospitalized patients diagnosed with COVID-19.

Introdução: A pandemia por COVID-19 colocou uma maior pressão financeira sobre os sistemas e instituições de saúde que tiveram de responder às necessidades específicas dos doentes com COVID-19, garantindo a segurança do diagnóstico e tratamento de todos os doentes e profissionais de saúde. Para avaliar o impacto financeiro dos doentes com COVID-19 internados em hospitais, caracterizámos o custo dos internamentos com COVID-19, utilizando dados de um Centro Hospitalar Universitário Português. Métodos: Analisaram-se os dados de internamento dos doentes adultos diagnosticados com COVID- 19 admitidos entre 1 de março de 2020 e 31 de maio de 2020. Foram considerados elegíveis os episódios de internamento com diagnóstico principal U07.1 (ICD-10- CM). Excluíram-se os internamentos de doentes com menos de 18 anos, internamentos com registos incompletos, internamentos de doentes que foram transferidos de/ou para outros hospitais ou aqueles com internamento inferior a 24 horas. Também foram excluídos os internamentos por gravidez, parto e puerpério, bem como internamentos de doentes operados. Resultados: Identificaramse 223 internamentos de doentes diagnosticados com COVID-19. A maioria era do sexo masculino (64,1%) e com idade entre 45 e 64 anos (30,5%). Cerca de 13,0% dos doentes foram internados em unidades de cuidados intensivos e 9,9% morreram no hospital. A média do tempo de internamento foi 12,7 dias (DP = 10,2) e a média do custo estimado por admissão foi 8.177€ (DP = 11.534), o que representa mais do triplo do preço base de internamento (2.386€). Os recursos humanos representaram a maior proporção do total dos custos por admissão (50,8%). Cerca de 92,4% dos internamentos foram atribuídos ao Grupo de Diagnóstico Homogéneo (GDH) 723, cujo preço de internamento é inferior aos custos de internamento com COVID-19 para todos os graus de severidade. Conclusão: Os internamentos por COVID-19 representam um encargo financeiro substancial para o SNS português. Por cada doente internado com COVID-19, seria possível tratar outros três doentes internados. Além disso, o preço definido para o GDH 723 não está ajustado ao custo dos doentes com COVID-19. Estes resultados destacam a necessidade de recursos financeiros adicionais para o sistema de saúde e, em particular, para os hospitais que trataram um elevado número de doentes internados e diagnosticados com COVID-19.

Disruption of protein phosphatase 1 complexes with the use of bioportides as a novel approach to target sperm motility.

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 samples for routine analysis and Holstein Frisian bulls. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Effect of the bioportides on the activity and interactions of PP1γ2-a PP1 isoform expressed exclusively in testicular germ cells and sperm-and on sperm vitality and motility. RESULT(S): PP1-disrupting peptides were designed based on the sequences from: 1) a sperm-specific PP1 interactor (A kinase anchor protein 4); and 2) a PP1 inhibitor (protein phosphatase inhibitor 2). Those sequences were covalently coupled to inert CPPs as bioportide constructs, which were successfully delivered to the flagellum of sperm cells to induce a marked impact on PP1γ2 activity and sperm motility. Molecular modeling studies further facilitated the identification of an optimized PP1-binding sequence and enabled the development of a modified stop-sperm bioportide with reduced size and increased potency of action. In addition, a bioportide mimetic of the unique 22-amino acid C-terminus of PP1γ2 accumulated within spermatozoa to significantly reduce sperm motility and further define the PP1γ2-specific interactome. CONCLUSION(S): These investigations demonstrate the utility of CPPs to deliver peptide sequences that target unique protein-protein interactions in spermatozoa to achieve a significant impact upon spermatozoa motility, a key prognostic indicator of male fertility.

Design, synthesis and biological evaluation of pyrazolo[3,4-d]pyrimidine-based protein kinase D inhibitors.

The multiple roles of protein kinase D (PKD) in various cancer hallmarks have been repeatedly reported. Therefore, the search for novel PKD inhibitors and their evaluation as antitumor agents has gained considerable attention. In this work, novel pyrazolo[3,4-d]pyrimidine based pan-PKD inhibitors with structural variety at position 1 were synthesized and evaluated for biological activity. Starting from 3-IN-PP1, a known PKD inhibitor with IC50 values in the range of 94-108 nM, compound 17m was identified with an improved biochemical inhibitory activity against PKD (IC50 = 17-35 nM). Subsequent cellular assays demonstrated that 3-IN-PP1 and 17m inhibited PKD-dependent cortactin phosphorylation. Furthermore, 3-IN-PP1 displayed potent anti-proliferative activity against PANC-1 cells. Finally, a screening against different cancer cell lines demonstrated that 3-IN-PP1 is a potent and versatile antitumoral agent.

Nurse managers' perceptions of nurse staffing and nursing care quality: A cross-sectional study.

AIM: To verify the association between the nurse staffing and the quality of nursing care, mediated by the care process, based on a hypothetical model, in Portuguese public hospitals. BACKGROUND: Nurse staffing influences health outcomes. Understaffing is associated with an increased risk for adverse events (AEs) and a reduction in the quality of care. METHOD: A cross-sectional study was conducted using a sample of 55 Portuguese nurse managers. A path model was developed to analyse potential causal mediation effects on care quality. RESULTS: Nurse staffing (number and competencies) and teamwork indirectly influence the quality of care. This process is mediated by the response capacity, the use of new techniques and work methods and patient's surveillance capacity. The AEs occurrence also has a mediating role, being negatively associated with the quality of care. CONCLUSIONS: Optimizing nursing care safety and quality requires an adequate nurse staffing level, both in terms of number and competencies, as well as teamwork. Process components seem to play a mediating role in these relations. IMPLICATIONS FOR NURSING MANAGEMENT: These results deserve the attention of nursing management for investment in the nursing staff and in the care process, to improve quality and create value in health care.

Phonological development in Portuguese deaf children with cochlear implants: preliminary study / Desarrollo fonológico en niños sordos portugueses con implantes cocleares: estudio preliminar

Introduction: children with severe to profound sensorineural hearing loss and prelingual onset of deafness exhibit less linguistic skills and poorer academic achievements when compared with their hearing peers. Recent studies show that cochlear implants may promote language development in this paediatric population, although phonological development does not always follow the patterns attested for typically developing children. Aim and method: the present study aims at describing the phonetic properties of segments and the phonological processes produced by 18 deaf Portuguese children with more than 2.0 years of implant use, divided into two groups according of their hearing age (A=2.10-4.04 and B=5.04-7.03) For this purpose, we used a formal articulation test validated for the Portuguese population. Results: results show that Group B, with a higher hearing age, exhibit a consonant repertoire more complete than Group A, whose hearing age was lower. Syllable and segmental difficulties tend to persist over a long period of time, even after 4 years of implant experience (i.e. hearing age). Discussion: the children's consonant repertoire of plosives and fricatives seems to take longer to establish in deaf children with cochlear implants than in their Portuguese hearing peer; other phonological patterns found in the data show an immature phonological development (e.g. syllable omission and onset deletion); these results match the ones reported for other languages in other studies on cochlear implanted children

Introducción: los niños con un nivel de hipoacusia neurosensitiva entre intensa y profunda, y en fases iniciales de sordera prelocutiva presentan conocimientos lingüísticos y resultados académicos inferiores en relación con otros niños de su misma edad con un cuadro auditivo normal. Estudios recientes muestran que los implantes cocleares pueden promover el desarrollo lingüístico de este segmento pediátrico, aun cuando el desarrollo fonológico no siempre siga la progresión estándar que se observa en niños con una evolución prototípica. Objetivo y metodología: este estudio tiene como objetivo describir las propiedades de los segmentos fonéticos y de las pautas fonológicas producidos por 18 niños sordos portugueses con más de 2 años de experiencia con el implante coclear, divididos en dos cohortes en función del tiempo que han utilizado este tipo de implantes (A = 2.10-4.04 y B = 5.04-7.03). Con este propósito nos hemos servido de una prueba de articulación formal validada para hablantes portugueses. Resultados: los resultados muestran que el grupo B, con mayor edad auditiva, presenta un repertorio de consonantes más completo que el grupo A. Las dificultades en la producción de sílabas y segmentos suelen persistir durante un largo período de tiempo, incluso después de los 4 años de experiencia con el implante, es decir, con 4 años de edad auditiva. Discusión: al parecer, el repertorio de consonantes obstruyentes (oclusivas y fricativas) tarda más tiempo en establecerse en niños sordos con implantes cocleares que en niños con audición normal. Los datos obtenidos revelan, además, pautas propias de un desarrollo fonológico inmaduro (p. ej., omisión total de sílabas y omisión de ataques silábicos); los resultados coinciden así con los que arrojan varios estudios sobre niños de otras lenguas que llevan también implante coclear

Use of bronchial blocker in emergent thoracotomy in presence of upper airway hemorrhage, and cervical spine fracture: a difficult decision / Uso de bloqueador brônquico em toracotomia de emergência na presença de hemorragia das vias aéreas superiores e fratura cervical: uma decisão difícil

Abstract Female, 85 y.o., weighting 60 kg, multiple trauma patient. After an initial laparotomy, an emergent thoracotomy was performed using a bronchial blocker for lung isolation (initial active suction was applied). During surgery, bronchial cuff was deflated, causing a self-limited tracheal blood flooding. A second lung isolation was attempted but it was not as effective as initially. Probably, a lung collapse with the same bronchial blocker was impaired in the second attempt because of the obstruction of bronchial blocker lumen by intraoperative endobronchial hemorrhage. Bronchial blocker active suction may contribute to obtain or accelerate lung collapse, particularly in patients that do not tolerate ventilator disconnection technique or lung surgical compression. The use of bronchial blockers technology was a valuable alternative to double lumen tubes in this case of emergent thoracotomy in the context of a patient having thoracic, abdominal trauma, severe laceration of tongue and apophysis odontoid fracture associated to massive hemorrhage, despite several pitfalls that could compromise its use. The authors intend to discuss the advantages and disadvantages of bronchial blockers comparing to double-lumen tubes for lung isolation, and the risks of our approach, in this complex multitrauma case.

Resumo Paciente do sexo feminino, 85 anos, 60 kg, com trauma múltiplo. Após uma laparotomia inicial, uma toracotomia de emergência foi feita com um bloqueador brônquico para isolamento pulmonar (sucção inicial ativa foi aplicada). Durante a cirurgia, o balonete brônquico foi desinflado, causou um derrame hemorrágico traqueal autolimitado. Reisolamento foi tentado, mas não foi tão eficaz como inicialmente. Provavelmente, o colapso do pulmão com o mesmo bloqueador brônquico foi prejudicado na segunda tentativa devido à obstrução do lúmen do bloqueador brônquico pela hemorragia endobrônquica intraoperatória. A sucção ativa do bloqueador brônquico pode contribuir para obter ou acelerar o colapso pulmonar, particularmente em pacientes que não toleram a técnica de desconexão do ventilador ou a compressão cirúrgica pulmonar. O uso da tecnologia de bloqueadores brônquicos foi uma opção valiosa para os tubos de duplo lúmen neste caso de toracotomia de emergência em paciente com trauma torácico e abdominal, laceração grave da língua e fratura da apófise odontoide associados a hemorragia maciça, apesar de vários riscos que poderiam comprometer seu uso. Os autores pretendem discutir as vantagens e desvantagens dos bloqueadores brônquicos em comparação com os tubos de duplo lúmen para isolamento pulmonar e quais foram os riscos de nossa abordagem neste complexo caso de múltiplo trauma.

Sertoli Cell Preparation for Co-immunoprecipitation.

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 consequently obtain protein extracts from Sertoli cells must be considered. In this chapter, we will focus on how to obtain a protein Sertoli cell extract suitable for co-immunoprecipitation technique.

Profiling Signaling Proteins in Sertoli Cells by Co-immunoprecipitation.

Sertoli cells are crucial for germ cell support, create a suitable environment for spermatogenesis, and integrate information from the nervous system and germ cell line. To fully understand the role of Sertoli cells, it is necessary to characterize the protein-protein interactions. Identifying the interactome of a given protein may provide leads about the role and molecular mechanism of such protein in Sertoli cells. One of the techniques to characterize protein interactomes consists of co-immunoprecipitation followed by mass spectrometry or Western blot. Co-immunoprecipitation enables the isolation of a protein target and interactome under physiological conditions. In this chapter, we described how to perform an interactomic study using the co-immunoprecipitation technique in Sertoli cells. Moreover, we will focus on how to analyze and interpret the results of a co-immunoprecipitation before mass spectrometry analysis.

[Use of bronchial blocker in emergent thoracotomy in presence of upper airway hemorrhage, and cervical spine fracture: a difficult decision]. / Uso de bloqueador brônquico em toracotomia de emergência na presença de hemorragia das vias aéreas superiores e fratura cervical: uma decisão difícil.

Female, 85 y.o., weighting 60kg, multiple trauma patient. After an initial laparotomy, an emergent thoracotomy was performed using a bronchial blocker for lung isolation (initial active suction was applied). During surgery, bronchial cuff was deflated, causing a self-limited tracheal blood flooding. A second lung isolation was attempted but it was not as effective as initially. Probably, a lung collapse with the same bronchial blocker was impaired in the second attempt because of the obstruction of bronchial blocker lumen by intraoperative endobronchial hemorrhage. Bronchial blocker active suction may contribute to obtain or accelerate lung collapse, particularly in patients that do not tolerate ventilator disconnection technique or lung surgical compression. The use of bronchial blockers technology was a valuable alternative to double lumen tubes in this case of emergent thoracotomy in the context of a patient having thoracic, abdominal trauma, severe laceration of tongue and apophysis odontoid fracture associated to massive hemorrhage, despite several pitfalls that could compromise its use. The authors intend to discuss the advantages and disadvantages of bronchial blockers comparing to double-lumen tubes for lung isolation, and the risks of our approach, in this complex multitrauma case.

Signaling mechanisms in mammalian sperm motility.

The goal of sperm is to fertilize the oocyte. To achieve that purpose, it must acquire motility in the epididymis and hyperactivated motility in the female reproductive tract. Motility is only achieved when the sperm presents a fully functional flagellum, is capable of producing energy to fuel the movement, and suffers epididymal maturation and capacitation. Since sperm is a transcriptionally silent cell, motility depends on the activation and/or inhibitions of key signaling pathways. This review describes and discusses the main signaling pathways involved in primary and hyperactivated motility, as well as the bioenergetic mechanisms necessary to produce energy to fuel sperm motility. Although the complete human sperm motility process is far from being fully known, we believe that in the upcoming decades extensive progress will be made. Understanding the signaling pathways behind sperm motility can help pinpoint the cause of male infertility and uncover targets for male contraception.

Study on the short-term effects of increased alcohol and cigarette consumption in healthy young men's seminal quality.

Many studies have reported a negative impact of lifestyle factors on testicular function, spermatozoa parameters and pituitary-gonadal axis. However, conclusions are difficult to draw, since studies in the general population are rare. In this study we intended to address the early and late short-term impact of acute lifestyle alterations on young men's reproductive function. Thirty-six healthy male students, who attended the Portuguese academic festivities, provided semen samples and answered questionnaires at three time-points. The consumption of alcohol and cigarette increased more than 8 and 2 times, respectively, during the academic festivities and resulted in deleterious effects on semen quality: one week after the festivities, a decrease on semen volume, spermatozoa motility and normal morphology was observed, in parallel with an increase on immotile spermatozoa, head and midpiece defects and spermatozoa oxidative stress. Additionally, three months after the academic festivities, besides the detrimental effect on volume, motility and morphology, a negative impact on spermatozoa concentration was observed, along with a decrease on epididymal, seminal vesicles and prostate function. This study contributed to understanding the pathophysiology underlying semen quality degradation induced by acute lifestyle alterations, suggesting that high alcohol and cigarette consumption are associated with decreased semen quality in healthy young men.

Unravelling the Power of Omics for the Infertile Aging Male.

The aging phenomenon is intrinsically responsible for diseases and lifestyle-associated conditions afflicting the aging male. In particular, male infertility seems to result from deleterious lifestyle choices. However, the aging effect at the individual gene/protein level is poorly discussed and valuable information is certainly missing. We have thus used an omics approach to identify differentially expressed proteins and genes from spermatozoa and seminal plasma samples across several conditions affecting adult male fertility. Our search resulted in 400 differentially expressed proteins in seminal plasma and 409 differentially expressed proteins in spermatozoa as well as, almost 6,000 differentially expressed spermatozoa mRNAs. We have functionally analyzed these proteins and genes to understand and discuss how biological processes and signaling pathways associated with aging might affect male fertility. Sperm and seminal fluid proteins from infertile males display significant alterations in i) processes previously implicated in the aging phenomena, such as mitochondrial dysfunction, DNA instability, oxidative stress, protein misfolding and intracellular mistrafficking, and ii) processes specifically involved in reproductive phenomena, such as sperm-egg recognition/acrosome reaction, embryo and morula development, blastocyst implantation and DNA methylation involved in embryo development. These proteins display a widespread distribution and seem to be significantly influenced by deleteriously lifestyle choices. Conventional assays to assess male fertility are inadequate to comprehensively reveal the broad-spectrum of alterations at the transcriptional and translational levels afflicting the infertile aging male. In turn, proteomics and transcriptomics are suitable options for addressing these key issues that may explain many poorly understood fertility-associated phenomena resulting from the aging process.

Non-stop lab week: A real laboratory experience for life sciences postgraduate courses.

At the Portuguese universities, practical classes of life sciences are usually professor-centered 2-hour classes. This approach results in students underprepared for a real work environment in a research/clinical laboratory. To provide students with a real-life laboratory environment, the Non-Stop Lab Week (NSLW) was created in the Molecular Biomedicine master program at the University of Aveiro, Portugal. The unique feature of the NSLW is its intensity: during a 1-week period, students perform a subcloning and a protein expression project in an environment that mimics a real laboratory. Students work autonomously, and the progression of work depends on achieving the daily goals. Throughout the three curricular years, most students considered the intensity of the NSLW a very good experience and fundamental for their future. Moreover, after some experience in a real laboratory, students state that both the techniques and the environment created in the NSLW were similar to what they experience in their current work situation. The NSLW fulfills a gap in postgraduate students' learning, particularly in practical skills and scientific thinking. Furthermore, the NSLW experience provides skills to the students that are crucial to their future research area. © 2016 by The International Union of Biochemistry and Molecular Biology, 44:297-303, 2016.

Physical Activity, Exercise, and Mammalian Testis Function: Emerging Preclinical Protein Biomarker and Integrative Biology Insights.

Exercise and physical activity have long been recognized for health promotion and to delay the onset of many pathological situations such as diabetes and cancers. Still, there appears to be an upper limit on the beneficial health effects regarding intensity and frequency of exercise training. In humans, the effect of exercise training in the male reproductive system has been studied mainly through the analysis of semen quality parameters, with inconsistent results. Less is known on molecular biomarkers of exercise-related changes in testis at the protein/proteome level. This review offers an in-depth analysis on the small scale protein studies available primarily from the preclinical studies and interprets their functional impact on the reproductive health with a view to humans. In all, exercise training in preclinical models seems to negatively modulate, in the course of health, critical functions that directly affect spermatogenesis, such as testosterone biosynthesis, energy supply, and antioxidant system components. Exercise training induces apoptosis, leading to the impairment of spermatogenesis and, consequently, to male infertility. In pathological conditions, an improvement in the testicular functions is observed by increases in steroidogenic enzymes and antioxidant defenses, and reductions in activity of inflammatory pathways. Importantly, the mechanisms by which exercise training modulates the reproductive function are far from being fully understood. The analyses of the testis proteome in varying exercise conditions would inform the molecular mechanisms involved and identify putative theranostics opportunities. Such future research is a cornerstone for health promotion in the pursuit of reproductive health informed by omics systems sciences.

Profiling signaling proteins in human spermatozoa: biomarker identification for sperm quality evaluation.

OBJECTIVE: To determine the correlation between semen basic parameters and the expression and activity of signaling proteins. DESIGN: In vitro studies with human spermatozoa. SETTING: Academic research institute. PATIENT(S): Thirty-seven men provided semen samples for routine analysis. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Basic semen parameters tracked included sperm DNA fragmentation (SDF), the expression levels of 75 protein kinases, and the phosphorylation/cleavage patterns of 18 signaling proteins in human spermatozoa. RESULT(S): The results indicated that the phosphorylated levels of several proteins (Bad, GSK-3ß, HSP27, JNK/SAPK, mTOR, p38 MAPK, and p53), as well as cleavage of PARP (at D214) and Caspase-3 (at D175), were significantly correlated with motility parameters. Additionally, the percentage of morphologically normal spermatozoa demonstrated a significant positive correlation with the phosphorylated levels of p70 S6 kinase and, in turn, head defects and the teratozoospermia index (TZI) showed a significant negative correlation with the phosphorylated levels of Stat3. There was a significant positive correlation between SDF and the teratozoospermia index, as well as the presence of head defects. In contrast, SDF negatively correlated with the percentage of morphologically normal spermatozoa and the phosphorylation of Akt and p70 S6 kinase. Subjects with varicocele demonstrated a significant negative correlation between head morphological defects and the phosphorylated levels of Akt, GSK3ß, p38 MAPK, and Stat1. Additionally, 34 protein kinases were identified as expressed in their total protein levels in normozoospermic samples. CONCLUSION(S): This study contributed toward establishing a biomarker "fingerprint" to assess sperm quality on the basis of molecular parameters.

The power of the yeast two-hybrid system in the identification of novel drug targets: building and modulating PPP1 interactomes.

Since the description of the yeast two-hybrid (Y2H) method, it has become more and more evident that it is the most commonly used method to identify protein-protein interactions (PPIs). The improvements in the original Y2H methodology in parallel with the idea that PPIs are promising drug targets, offer an excellent opportunity to apply the principles of this molecular biology technique to the pharmaceutical field. Additionally, the theoretical developments in the networks field make PPI networks very useful frameworks that facilitate many discoveries in biomedicine. This review highlights the relevance of Y2H in the determination of PPIs, specifically phosphoprotein phosphatase 1 interactions, and its possible outcomes in pharmaceutical research.

TGF-ß cascade regulation by PPP1 and its interactors -impact on prostate cancer development and therapy.

Protein phosphorylation is a key mechanism by which normal and cancer cells regulate their main transduction pathways. Protein kinases and phosphatases are precisely orchestrated to achieve the (de)phosphorylation of candidate proteins. Indeed, cellular health is dependent on the fine-tune of phosphorylation systems, which when deregulated lead to cancer. Transforming growth factor beta (TGF-ß) pathway involvement in the genesis of prostate cancer has long been established. Many of its members were shown to be hypo- or hyperphosphorylated during the process of malignancy. A major phosphatase that is responsible for the vast majority of the serine/threonine dephosphorylation is the phosphoprotein phosphatase 1 (PPP1). PPP1 has been associated with the dephosphorylation of several proteins involved in the TGF-ß cascade. This review will discuss the role of PPP1 in the regulation of several TGF-ß signalling members and how the subversion of this pathway is related to prostate cancer development. Furthermore, current challenges on the protein phosphatases field as new targets to cancer therapy will be addressed.