Comprehensive analysis of DNA replication timing across 184 cell lines suggests a role for MCM10 in replication timing regulation.

Cellular proliferation depends on the accurate and timely replication of the genome. Several genetic diseases are caused by mutations in key DNA replication genes; however, it remains unclear whether these genes influence the normal program of DNA replication timing. Similarly, the factors that regulate DNA replication dynamics are poorly understood. To systematically identify trans-acting modulators of replication timing, we profiled replication in 184 cell lines from three cell types, encompassing 60 different gene knockouts or genetic diseases. Through a rigorous approach that considers the background variability of replication timing, we concluded that most samples displayed normal replication timing. However, mutations in two genes showed consistently abnormal replication timing. The first gene was RIF1, a known modulator of replication timing. The second was MCM10, a highly conserved member of the pre-replication complex. Cells from a single patient carrying MCM10 mutations demonstrated replication timing variability comprising 46% of the genome and at different locations than RIF1 knockouts. Replication timing alterations in the mutated MCM10 cells were predominantly comprised of replication delays and initiation site gains and losses. Taken together, this study demonstrates the remarkable robustness of the human replication timing program and reveals MCM10 as a novel candidate modulator of DNA replication timing.

Tumor start-up: mitochondrial fusion makes it happen.

Whether changes in cellular metabolism precede tumor formation and trigger malignant properties or simply serve as a bioenergetic adaptation of cancer during disease progression remains debated. Bonnay et al (2020) now show that a metabolic reprogramming toward increased oxidative phosphorylation is required for irreversible cell immortalization and subsequent tumor formation.

Therapeutic Polymer-Based Cannabidiol Formulation: Tackling Neuroinflammation Associated with Ischemic Events in the Brain.

Cannabidiol (CBD) is the most relevant nonpsychostimulant phytocompound found in Cannabis sativa. CBD has been extensively studied and has been proposed as a therapeutic candidate for neuroinflammation-related conditions. However, being a highly lipophilic drug, it has several drawbacks for pharmaceutical use, including low solubility and high permeability. Synthetic polymers can be used as drug delivery systems to improve CBD's stability, half-life, and biodistribution. Here, we propose using a synthetic polymer as a nanoparticulate vehicle for CBD (NPCBD) to overcome the pharmacological drawbacks of free drugs. We tested the NPCBD-engineered system in the context of ischemic events in a relevant oxygen and glucose deprivation (OGD) model in primary cortical cells (PCC). Moreover, we have characterized the inflammatory response of relevant cell types, such as THP-1 (human monocytes), HMC3 (human microglia), and PCC, to NPCBD and observed a shift in the inflammatory state of the treated cells after the ischemic event. In addition, NPCBD exhibited a promising ability to restore mitochondrial function after OGD insult in both HMC3 and PCC cells at low doses of 1 and 0.2 µM CBD. Taken together, these results suggest the potential for preclinical use.

dMyc-dependent upregulation of CD98 amino acid transporters is required for Drosophila brain tumor growth.

Tumor cells have an increased demand for nutrients to sustain their growth, but how these increased metabolic needs are ensured or how this influences tumor formation and progression remains unclear. To unravel tumor metabolic dependencies, particularly from extracellular metabolites, we have analyzed the role of plasma membrane metabolic transporters in Drosophila brain tumors. Using a well-established neural stem cell-derived tumor model, caused by brat knockdown, we have found that 13 plasma membrane metabolic transporters, including amino acid, carbohydrate and monocarboxylate transporters, are upregulated in tumors and are required for tumor growth. We identified CD98hc and several of the light chains with which it can form heterodimeric amino acid transporters, as crucial players in brat RNAi (brat IR) tumor progression. Knockdown of these components of CD98 heterodimers caused a dramatic reduction in tumor growth. Our data also reveal that the oncogene dMyc is required and sufficient for the upregulation of CD98 transporter subunits in these tumors. Furthermore, tumor-upregulated dmyc and CD98 transporters orchestrate the overactivation of the growth-promoting signaling pathway TOR, forming a core growth regulatory network to support brat IR tumor progression. Our findings highlight the important link between oncogenes, metabolism, and signaling pathways in the regulation of tumor growth and allow for a better understanding of the mechanisms necessary for tumor progression.

Investigating academic performance and perceptions of human physiology and exercise physiology courses among undergraduate students of physical education at a Brazilian public university.

PURPOSE: To compare the academic performance of undergraduate students in physical education who studied exercise physiology before and after studying human physiology and investigate students' perceptions of human physiology and exercise physiology courses. METHODS: This study included 311 undergraduate students pursuing a bachelor's degree in physical education. Participants were divided into two groups: those who had previously attended and completed the human physiology course (group 1, n = 212, 68.2%) and those who had not previously attended or had attended but failed the human physiology course (group 2, n = 99, 31.8%). The prevalence ratio (PR) and 95% confidence interval (95% CI) were calculated using a Poisson regression model with a robust variance estimator. The second aim comprised 67 students with bachelor's degrees in physical education who completed an electronic questionnaire about their perceptions of human physiology and exercise physiology curriculum. RESULTS: Compared with those who attended human physiology and passed, those who had not previously attended or had attended but failed the human physiology course have a higher PR of 2.37 (95% CI, 1.68-3.34) for failing exercise physiology. Regarding the students' perceptions of human physiology and exercise physiology courses, most students reported that they were challenging (58.2% and 64.2%, respectively), but they also recognized the importance of these courses for professional practice (59.7% and 85.1%, respectively). CONCLUSION: Human physiology should be considered a prerequisite for an undergraduate course leading to a bachelor's degree in physical education. Furthermore, students considered human physiology and exercise physiology courses important yet challenging. Therefore, continuous student assessment is vital for improving the teaching-learning process.

Esophagopleural fistula due to empyema necessitans.

Empyema necessitans is a rare entity that consists on the development of an abscess that begins in the pleural space and then extends to the adjacent tissues. This case shows a rare and very late complication of a total pneumonectomy, emphasizing the importance of the multidisciplinary approach and the potential of endoscopic therapy with over-the-scope clips.

Terminal ileitis - When all factors come together.

A 45-year-old male, with a recent surgery for odontoid fracture, presented to the emergency department with a 15-days history of abdominal pain associated with fever and weight loss. He reported a recent history of antibiotic therapy due to respiratory infection and a frequent use of anti-inflammatory drugs (NSAIDs) in the last three months.

Endoscopic ampullectomy of a giant pedunculated ampullary adenoma.

The authors present a case of a 72-year-old woman with a personal history of arterial hypertension, dyslipidemia and gallstones. For suspected choledocholithiasis, a MR-Cholangiopancreatography was performed, which revealed the presence of an endoluminal polypoid structure in the third duodenal portion.

A rare cause of obscure gastrointestinal bleeding.

A 62-year-old woman was referred to Gastroenterology appointment due to severe iron deficiency anemia (5.9 g/dL), complaining of asthenia and requiring blood transfusion. The patient denied blood loss. Initial blood test showed a severe iron deficiency, with ferritin of 5ng/mL and transferrin saturation of 2.7%. Folic acid and vitamin B12 were normal. Upper gastroscopy and colonoscopy didn't show any lesions. Abdominopelvic CT and capsule endoscopy were, also, normal.

An uncommon colonic polyp.

A 50-year-old male was referred to a Gastroenterology appointment after a screening colonoscopy with a 25mm exophytic lesion, with a depressed central area, on the transverse colon. Histologic examination of the biopsy specimen showed low-grade dysplasia. The patient was submitted to a new colonoscopy and what was seen was a flat lesion with central depression, with no lift-sign (Figure 1a and 1b) and, therefore, endoscopic resection was not performed. New endoscopic biopsies were taken and showed no dysplasia.

Distinct Glycosylation Responses to Spinal Cord Injury in Regenerative and Nonregenerative Models.

Traumatic spinal cord injury (SCI) results in disruption of tissue integrity and loss of function. We hypothesize that glycosylation has a role in determining the occurrence of regeneration and that biomaterial treatment can influence this glycosylation response. We investigated the glycosylation response to spinal cord transection in Xenopus laevis and rat. Transected rats received an aligned collagen hydrogel. The response compared regenerative success, regenerative failure, and treatment in an established nonregenerative mammalian system. In a healthy rat spinal cord, ultraperformance liquid chromatography (UPLC) N-glycoprofiling identified complex, hybrid, and oligomannose N-glycans. Following rat SCI, complex and outer-arm fucosylated glycans decreased while oligomannose and hybrid structures increased. Sialic acid was associated with microglia/macrophages following SCI. Treatment with aligned collagen hydrogel had a minimal effect on the glycosylation response. In Xenopus, lectin histochemistry revealed increased levels of N-acetyl-glucosamine (GlcNAc) in premetamorphic animals. The addition of GlcNAc is required for processing complex-type glycans and is a necessary foundation for additional branching. A large increase in sialic acid was observed in nonregenerative animals. This work suggests that glycosylation may influence regenerative success. In particular, loss of complex glycans in rat spinal cord may contribute to regeneration failure. Targeting the glycosylation response may be a promising strategy for future therapies.

Integrating animal development: How hormones and metabolism regulate developmental transitions and brain formation.

Our current knowledge on how individual tissues or organs are formed during animal development is considerable. However, the development of each organ does not occur in isolation and thus their formation needs to be done in a coordinated manner. This coordination is regulated by hormones, systemic signals that instruct the simultaneous development of all organs and direct tissue specific developmental programs. In addition, multi- and individual-organ development requires the integration of the nutritional state of the animal, since this affects nutrient availability necessary for the progression of development and growth. Variations in the nutritional state of the animal are normal during development, as the sources and access to nutrients greatly differ depending on the animal stage. Furthermore, adversities of the external environment also exert major alterations in extrinsic nutritional conditions. Thus, both in normal and malnutrition circumstances, the animal needs to trigger metabolic changes to maintain energy homeostasis and sustain growth and development. This metabolic flexibility is mediated by hormones, that drive both developmental encoded metabolic transitions throughout development and adaptation responses according to the nutritional state of the animal. This review aims to provide a comprehensive summary of the current knowledge of how endocrine regulation coordinates multi-organ development by orchestrating metabolic transitions and how it integrates metabolic adaptation responses to starvation. We also focus on the particular case of brain development, as it is extremely sensitive to hormonally induced metabolic changes. Finally, we discuss how brain development is prioritized over the development of other organs, as its growth can be spared from nutrient deprivation.

Germ cell tumor with duodenal involvement: a rare case of gastrointestinal bleeding.

A 19-year-old male presented to the emergency department with a 7-day history of melena, anorexia and asthenia. Blood tests revealed a hemoglobin of 5.8 g/dL. Upper endoscopy showed a large ulcerated and stenosing lesion in the duodenum. The histologic examination of the biopsy specimen showed a neoplasia with epithelioid cells, accentuated atypia and pleomorphism, expressing MNF 116 cytokeratin, CD30, glypican 3 and alpha-fetoprotein on immunohistochemistry, suggesting of a germ cell tumor metastasis.

Pyoderma gangrenosum in ulcerative colitis - An exuberant and painful complication.

A 41-years-old female, with ulcerative colitis, presented to the emergency department with 7-days history of abdominal pain, bloody stools (> 10/day). The patient referred the appearance of a cutaneous lesion, on her left thigh, with subsequent appearance of similar lesions on the lower limbs. No improvement after amoxicillin/clavulanic acid. On admission, she was febrile (38.2 ºC) and tachycardic. She had three cutaneous lesions, the largest one with 8cm in the left thigh - a deep and painful lesion, with extensive ulceration, necrosis, exudative edges and with marked pathergia, compatible with pyoderma gangrenosum.

A rare endoscopic pattern in a patient with gastroesophageal reflux disease.

A 21-year-old man, with personal history of asthma and no usual medication, was referred to gastroenterology appointment due to dysphagia for solids and previous episodes of food impaction in the last 5 months. He also reported nocturnal heartburn without any other warning signs. Upper gastroscopy revealed a peptic stenosis in the distal esophagus. Histopathologic examination showed hyperplasia and numerous intraepithelial eosinophils, without dysplasia or malignancy. Therapy with a double-dose proton pump inhibitor (PPI) was started.

The O-Glycome of Human Nigrostriatal Tissue and Its Alteration in Parkinson's Disease.

O-Glycosylation changes in misfolded proteins are of particular interest in understanding neurodegenerative conditions such as Parkinson's disease (PD) and incidental Lewy body disease (ILBD). This work outlines optimizations of a microwave-assisted nonreductive release to limit glycan degradation and employs this methodology to analyze O-glycosylation on the human striatum and substantia nigra tissue in PD, ILBD, and healthy controls, working alongside well-established reductive release approaches. A total of 70 O-glycans were identified, with ILBD presenting significantly decreased levels of mannose-core (p = 0.017) and glucuronylated structures (p = 0.039) in the striatum and PD presenting an increase in sialylation (p < 0.001) and a decrease in sulfation (p = 0.001). Significant increases in sialylation (p = 0.038) in PD were also observed in the substantia nigra. This is the first study to profile the whole nigrostriatal O-glycome in healthy, PD, and ILBD tissues, outlining disease biomarkers alongside benefits of employing orthogonal techniques for O-glycan analysis.

Complete spatial characterisation of N-glycosylation upon striatal neuroinflammation in the rodent brain.

BACKGROUND: Neuroinflammation is an underlying pathology of all neurological conditions, the understanding of which is still being comprehended. A specific molecular pathway that has been overlooked in neuroinflammation is glycosylation (i.e., post-translational addition of glycans to the protein structure). N-glycosylation is a specific type of glycosylation with a cardinal role in the central nervous system (CNS), which is highlighted by congenital glycosylation diseases that result in neuropathological symptoms such as epilepsy and mental retardation. Changes in N-glycosylation can ultimately affect glycoproteins' functions, which will have an impact on cell machinery. Therefore, characterisation of N-glycosylation alterations in a neuroinflammatory scenario can provide a potential target for future therapies. METHODS: With that aim, the unilateral intrastriatal injection of lipopolysaccharide (LPS) in the adult rat brain was used as a model of neuroinflammation. In vivo and post-mortem, quantitative and spatial characterisation of both neuroinflammation and N-glycome was performed at 1-week post-injection of LPS. These aspects were investigated through a multifaceted approach based on positron emission tomography (PET), quantitative histology, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), liquid chromatography and matrix-assisted laser desorption ionisation mass spectrometry imaging (MALDI-MSI). RESULTS: In the brain region showing LPS-induced neuroinflammation, a significant decrease in the abundance of sialylated and core fucosylated structures was seen (approximately 7.5% and 8.5%, respectively), whereas oligomannose N-glycans were significantly increased (13.5%). This was confirmed by MALDI-MSI, which provided a high-resolution spatial distribution of N-glycans, allowing precise comparison between normal and diseased brain hemispheres. CONCLUSIONS: Together, our data show for the first time the complete profiling of N-glycomic changes in a well-characterised animal model of neuroinflammation. These data represent a pioneering step to identify critical targets that may modulate neuroinflammation in neurodegenerative diseases.

Sleep Disturbance in Inflammatory Bowel Disease Is Associated with Disease Activity and Adverse Outcome.

BACKGROUND: There is growing evidence about the relationship between sleep quality (SQ) and disease activity in inflammatory bowel disease (IBD). This study aimed to identify the prevalence of sleep disturbance in IBD and its predictive factors and to assess its association with worse outcome. METHODS: IBD patients were prospectively enrolled. Clinical activity, inflammatory activity (high C-reactive protein or fecal calprotectin), and SQ (assessed using the Pittsburgh Sleep Quality Index) were evaluated, and logistic regression was used to identify predictors of poor SQ at baseline. The development of disability or disease progression at 6 months (surgery, hospitalization, development of stenosis, penetrating or perianal disease, steroid dependency, or start/change immunosuppression) was compared between patients with and without poor SQ. RESULTS: Two hundred and five patients were enrolled, with 44.9% (n = 92) reporting poor SQ. On multivariate analysis, current smoking (OR 2.80), extraintestinal manifestations (OR 2.68), clinical activity (OR 3.31), and inflammatory activity (OR 4.62) were predictive factors of poor SQ. Cox proportional hazards model showed that poor SQ was predictive of worse prognosis at 6 months (HR 2.470). CONCLUSION: There is a high prevalence of poor SQ in IBD patients, highlighting the importance of its inclusion in patient-reported outcomes. Sleep disturbance seems to have prognostic value in IBD.

Morphological characterization of blue-and-yellow macaw (Ara ararauna) seasonal testicular variations in captivity.

The reproduction of monogamous wild birds in captivity it`s difficult and the apparent low fertility in males requires more investigations. The objective of the study was to test the hypothesis that wild bird species in captivity would present low reproductive potential, through the analysis of the morphological characteristics of Ara ararauna testicles, maintained in captivity, correlating them with the climate variations in the Cerrado Biome. For that, testicles were captured in April (dry) and October (rainy). The right and left testicles showed mean weight, gonadosomatic index, longer axis, and volume similar between the dry and rainy season. Only the shorter axis demonstrated higher values during the rainy season. The morphometric variables of the seminiferous tubules have also higher values during the rainy season. By these histological and morphometric characteristics of the seminiferous epithelium we can conclude that, during the rainy season, the testicles were in gonadal recrudescence, which precedes the reproduction phase. During the dry season, the testicles were in the rest phase of the seminiferous epithelium. Therefore, we concluded that the species in captivity, under Cerrado environmental conditions, have kept their reproductive potential, presenting a complete spermatogenic cycle during the rainy season, which can guarantee the species perpetuation.

Suicidal behaviour after first-episode psychosis: results from a 1-year longitudinal study in Portugal.

BACKGROUND: Suicide is one of the main causes of excess of premature death in psychotic patients. Published studies found that suicide risk begins in ultra-high risk of psychosis and continues in early years of the disease. Previous studies identifying predictive and risk factors associated with suicidality in first-episode psychosis (FEP) are highly inconsistent. Also, there are relatively few longitudinal studies on suicidal behaviour in FEP. The aim of this study was to examine prevalence, evolution and predictors of suicidal behaviour at baseline and the 12-month follow-up in patients presenting with FEP. METHODS: One hundred and eighteen patients presenting with FEP were recruited from two early psychosis units in Portugal. A comprehensive assessment examining socio-demographic and clinical characteristics was administered at baseline and the 12-month follow-up. Odds ratio were calculated using logistic regression analyses. McNemar test was used to evaluate the evolution of suicidal behaviour and depression prevalence from baseline to 12 months of follow-up. RESULTS: Follow-up data were available for 60 participants from the 118 recruited. Approximately 25.4% of the patients had suicidal behaviour at the baseline evaluation, with a significant reduction during the follow-up period to 13.3% (p = 0.035). A multivariate binary logistic regression showed that a history of suicidal behaviour and depression at baseline independently predicted suicidal behaviour at baseline, and a history of suicidal behaviour and low levels of total cholesterol predicted suicidal behaviour at the 12-month follow-up. A significant proportion of patients also had depression at the baseline evaluation (43.3%), with the last month of suicidal behaviour at baseline independently predicting depression at this time. CONCLUSIONS: The findings of our study indicate that suicidal behaviour was prevalent on the year after FEP. Patients with a history of suicidal behaviour, depression at baseline and low levels of cholesterol should undergo close evaluation, monitoring and possible intervention in order to reduce suicide risk in the early phases of psychosis.