Characterisation of cis-regulatory modules controlling gene expression within sox17+ lineages during zebrafish embryonic development

Precise spatial and temporal regulation of gene expression is critical for embryonic development. This is achieved in part via transcription factor binding to cis-regulatory modules (CRMs). However, in contrast to other germ layers, the gene regulatory network involved in endoderm formation is relatively understudied in vertebrates. In this study we use genomic approaches to study the chromatin landscape that governs endoderm specification and early endoderm organ formation in zebrafish embryos. Since the endoderm is a minor cell population, we employed two distinct methods to find to endoderm-specific CRMs: 1) to capture CRMs that regulate early endoderm-specific genes during early specification, we performed overexpression of the master regulator of endoderm formation, sox32, 2) to capture CRMs that function during early organogenesis, we have generated a three-colour transgenic line to specifically FACS isolate endoderm cells and study endoderm-specific CRMs at 28 hours post-fertilisation. Our results show that sox32 overexpression not only induces ectopic endoderm formation but also ectopic formation of progenitors of the zebrafish left/right organiser known as dorsal forerunner cells (DFCs), a phenomenon not previously discussed in literature. We also identified 16, 000 regions that exhibit enhanced accessibility by ATAC seq in embryos injected with sox32 mRNA compared to control and that a large number of these regions harboured motifs of transcription factors that play important roles during endoderm specification. Our findings also suggest that sorted GFP+ cells from the triple transgenic line comprise both endoderm and transdifferentiated Kupffer’s vesicle cells, which are implicated in left/right asymmetry in zebrafish. We also show by ATAC-seq analysis that a large number of accessible regions are likely shared by GFP+ cells and cells that are devoid of fluorescence. Finally, we show that changes in the chromatin landscape correlate with changes in gene expression at both stages of endoderm development through comparison of data from ATAC-seq and RNA-seq, but also that the chromatin landscape changes between endoderm specification and early organogenesis. Our results provide inside into how the chromatin landscape changes in the endoderm during development and also reveals a number of potentially important CRMs that may help shed light into how endoderm is formed during embryogenesis

EFL Teachers’ Phonological Awareness Beliefs and Practices: Help or Prevent EFL Children Developing Reading

EFL teachers’ proficiency seems to contribute to the reading difficulties that early graders encounter. This paper investigates the knowledge, beliefs, practices and awareness in phonological awareness (PA) of two-hundred and ten ramdonly selected EFL in-service teachers and then examines the impact of teachers’ experiences, qualifications, and gender on shaping teachers’ instruction. The researchers used a four-section survey to collect teachers’ demographic information, perceived and actual knowledge of phonological awareness and classroom practices related to PA, phonics, and syllabication. The results reported teachers as moderate level in the beliefs, practice and awareness of PA. In terms of teachers’ knowledge in PA, however, results showed teachers lacking the basics in teaching reading. Implications for EFL teacher education and preparation are highlighted. Keywords: EFL, in-service teachers, deficits, reading, phonological awareness. DOI: 10.7176/JEP/10-12-09 Publication date: April 30th 201

Investigating the molecular guts of endoderm formation using zebrafish

The vertebrate endoderm makes major contributions to the respiratory and gastrointestinal tracts and all associated organs. Zebrafish and humans share a high degree of genetic homology and strikingly similar endodermal organ systems. Combined with a multitude of experimental advantages, zebrafish are an attractive model organism to study endoderm development and disease. Recent functional genomics studies have shed considerable light on the gene regulatory programs governing early zebrafish endoderm development, while advances in biological and technological approaches stand to further revolutionize our ability to investigate endoderm formation, function and disease. Here, we discuss the present understanding of endoderm specification in zebrafish compared to other vertebrates, how current and emerging methods will allow refined and enhanced analysis of endoderm formation, and how integration with human data will allow modeling of the link between non-coding sequence variants and human disease

Cyclo (Nα-dinicotinoyl)-bis-[(L-valinyl)-L-lysinyl acid hydrazide]: Assessment of its Role in Cancer and Kinase Activity Inhibition

292–296Current research aimed at evaluating the in vitro as well as in vivo anticancer activities of a newly synthesized peptide hydrazide; i.e. 4,14-diisopropyl-2,5,13,16-tetraoxo-3,6,12,15-tetraaza-1(3,5)-pyridinacyclohexadecaphane-7-carbohydrazide. The hydrazide was synthesized from methyl 4,14-diisopropyl-2,5,13,16-tetraoxo-3,6,12,15-tetraaza-1(3,5)-pyridinacyclohexadecaphane-7-carboxylate 2 by acting of hydrazine hydrate. It showed significant anticancer effects against different tested cell lines, where cervical, breast, liver, colon, prostate, brain, fibrosarcoma, leukemia and melanoma cell lines were the most affected cell types. The prepared derivative also inhibited VEGF-2 kinase enzyme significantly and exhibited an in vivo tumorigenic effects in mice model

Cyclo (Nα-dinicotinoyl)-bis-[(L-valinyl)-L-lysinyl acid hydrazide]: Assessment of its Role in Cancer and Kinase Activity Inhibition

Current research aimed at evaluating the in vitro as well as in vivo anticancer activities of a newly synthesized peptide hydrazide; i.e. 4,14-diisopropyl-2,5,13,16-tetraoxo-3,6,12,15-tetraaza-1(3,5)-pyridinacyclohexadecaphane-7-carbohydrazide. The hydrazide was synthesized from methyl 4,14-diisopropyl-2,5,13,16-tetraoxo-3,6,12,15-tetraaza-1(3,5)-pyridinacyclohexadecaphane-7-carboxylate 2 by acting of hydrazine hydrate. It showed significant anticancer effects against different tested cell lines, where cervical, breast, liver, colon, prostate, brain, fibrosarcoma, leukemia and melanoma cell lines were the most affected cell types. The prepared derivative also inhibited VEGF-2 kinase enzyme significantly and exhibited an in vivo tumorigenic effects in mice model

Impact of opioid-free analgesia on pain severity and patient satisfaction after discharge from surgery: multispecialty, prospective cohort study in 25 countries

Background: Balancing opioid stewardship and the need for adequate analgesia following discharge after surgery is challenging. This study aimed to compare the outcomes for patients discharged with opioid versus opioid-free analgesia after common surgical procedures.Methods: This international, multicentre, prospective cohort study collected data from patients undergoing common acute and elective general surgical, urological, gynaecological, and orthopaedic procedures. The primary outcomes were patient-reported time in severe pain measured on a numerical analogue scale from 0 to 100% and patient-reported satisfaction with pain relief during the first week following discharge. Data were collected by in-hospital chart review and patient telephone interview 1 week after discharge.Results: The study recruited 4273 patients from 144 centres in 25 countries; 1311 patients (30.7%) were prescribed opioid analgesia at discharge. Patients reported being in severe pain for 10 (i.q.r. 1-30)% of the first week after discharge and rated satisfaction with analgesia as 90 (i.q.r. 80-100) of 100. After adjustment for confounders, opioid analgesia on discharge was independently associated with increased pain severity (risk ratio 1.52, 95% c.i. 1.31 to 1.76; P < 0.001) and re-presentation to healthcare providers owing to side-effects of medication (OR 2.38, 95% c.i. 1.36 to 4.17; P = 0.004), but not with satisfaction with analgesia (beta coefficient 0.92, 95% c.i. -1.52 to 3.36; P = 0.468) compared with opioid-free analgesia. Although opioid prescribing varied greatly between high-income and low- and middle-income countries, patient-reported outcomes did not.Conclusion: Opioid analgesia prescription on surgical discharge is associated with a higher risk of re-presentation owing to side-effects of medication and increased patient-reported pain, but not with changes in patient-reported satisfaction. Opioid-free discharge analgesia should be adopted routinely

MR Arthrography versus conventional MRI in evaluation of labral and chondral lesions in different types of femoroacetabular impingement

Objective: To detect the value of MR Arthrography over MRI in evaluation of labral and chondral lesions in all types of FAI, and to correlate the findings with arthroscopy as a gold standard. Patients and methods: 50 patients including 33 males and 17 females, age ranges from 19 to 54 years old (mean age 39 ± 5.5), underwent MRI and MR Arthrography of the hip joint followed by arthroscopy correlation. Images were evaluated for labral abnormalities, cartilage and osseous abnormalities associated in FAI. α angle. Acetabular lateral edge angle and degree of focal retroversion were measured. Results: Cam type detected in 20, Mixed type in 28, pincer type in 2, MRI detected 28 labral injury, 51 cartilage affection in 33 cases, MR Arthrography detected 38 labral injury, fraying of the L/C zone in 23 cases, 53 cartilage affection in 33 cases, cam type ch.ch by large α angle, anterosuperior femoral cartilage lesion and osseous bump formation; mixed type include the previous cam findings with a deep acetabulum and posteroinferior cartilage lesions. Anterosuperior labral (AS) tears are more common than postersuperior (PS). Conclusion: Hip MR Arthrography is a faithful evaluation modality for diagnosing the acetabular labral tears, and cartilage abnormalities associated with different types of FAI

Rac1 controls cell turnover and reversibility of the involution process in postpartum mammary glands

Cell turnover in adult tissues is essential for maintaining tissue homeostasis over a life span and for inducing the morphological changes associated with the reproductive cycle. However, the underlying mechanisms that coordinate the balance of cell death and proliferation remain unsolved. Using the mammary gland, we have discovered that Rac1 acts as a nexus to control cell turnover. Postlactational tissue regression is characterised by the death of milk secreting alveoli, but the process is reversible within the first 48 h if feeding recommences. In mice lacking epithelial Rac1, alveolar regression was delayed. This defect did not result from failed cell death but rather increased cell turnover. Fitter progenitor cells inappropriately divided, regenerating the alveoli, but cell death also concomitantly accelerated. We discovered that progenitor cell hyperproliferation was linked to nonautonomous effects of Rac1 deletion on the macrophageal niche with heightened inflammation. Moreover, loss of Rac1 impaired cell death with autophagy but switched the cell death route to apoptosis. Finally, mammary gland reversibility failed in the absence of Rac1 as the alveoli failed to recommence lactation upon resuckling

Rac1 controls cell turnover and reversibility of the involution process in postpartum mammary glands.

Cell turnover in adult tissues is essential for maintaining tissue homeostasis over a life span and for inducing the morphological changes associated with the reproductive cycle. However, the underlying mechanisms that coordinate the balance of cell death and proliferation remain unsolved. Using the mammary gland, we have discovered that Rac1 acts as a nexus to control cell turnover. Postlactational tissue regression is characterised by the death of milk secreting alveoli, but the process is reversible within the first 48 h if feeding recommences. In mice lacking epithelial Rac1, alveolar regression was delayed. This defect did not result from failed cell death but rather increased cell turnover. Fitter progenitor cells inappropriately divided, regenerating the alveoli, but cell death also concomitantly accelerated. We discovered that progenitor cell hyperproliferation was linked to nonautonomous effects of Rac1 deletion on the macrophageal niche with heightened inflammation. Moreover, loss of Rac1 impaired cell death with autophagy but switched the cell death route to apoptosis. Finally, mammary gland reversibility failed in the absence of Rac1 as the alveoli failed to recommence lactation upon resuckling

RON, ROR1 and SUSD2 expression in tissues of endometrial carcinoma patients. Clinicopathological and prognostic implications

INTRODUCTION: Endometrial carcinoma is now considered a common female gynecologic cancer with increasing incidence, with 13–25% of patients being still liable to recurrence and metastasis, which needs further studies to detect novel targets and new therapies. The aim of the study was evaluate tissue expression of RON, ROR1 and SUSD2 in endometrial carcinoma and atypical endometrial hyperplasia using immunohistochemistry and correlate their expression with clinical, pathological and prognostic parameters of patients. MATERIAL AND METHODS: We included samples from 100 patients with endometrial carcinoma. Sections from paraffin blocks were stained with RON, ROR1 and SUSD2 using immunohistochemistry. Correlations between marker expression, clinicopathological features and prognostic samples were evaluated. RESULTS: Upregulation of RON and ROR1 and downregulation of SUSD2 expression were found in endometrial carcinoma more than atypical endometrial hyperplasia (p < 0.001). High RON and ROR1 expression levels were significantly associated with high grade (p < 0.001), presence of lymph node metastases (p = 0.003), distant metastases (p = 0.009), advanced International Federation of Gynecology and Obstetrics stage (p = 0.002), poor response to therapy (p = 0.046), and lower recurrence-free survival (RFS) rate (p = 0.002), progression-free survival (PFS) rate (p = 0.008), distant metastasis-free survival (DMFS) rate (p = 0.019) and overall survival rate (p < 0.001). Low SUSD2 expression was significantly associated with older patient age (p = 0.002), large tumor size (p = 0.003), high grade (p = 0.005), presence of adnexal invasion (p = 0.023), presence of lympho-vascular invasion (p = 0.021), extent of myometrial invasion (p = 0.002), lower RFS rate (p = 0.008), lower PFS rate (p = 0.023), and lower DMFS rate (p < 0.001). CONCLUSIONS: Upregulation of RON and ROR1 and downregulation of SUSD2 lead to promotion of endometrial cancer cell proliferation, migration, epithelial-mesenchymal transition, and invasion