ABSTRACT
BRCA2 is an essential tumor suppressor protein involved in promoting faithful repair of DNA lesions. The activity of BRCA2 needs to be tuned precisely to be active when and where it is needed. Here, we quantified the spatio-temporal dynamics of BRCA2 in living cells using aberration-corrected multifocal microscopy (acMFM). Using multicolor imaging to identify DNA damage sites, we were able to quantify its dynamic motion patterns in the nucleus and at DNA damage sites. While a large fraction of BRCA2 molecules localized near DNA damage sites appear immobile, an additional fraction of molecules exhibits subdiffusive motion, providing a potential mechanism to retain an increased number of molecules at DNA lesions. Super-resolution microscopy revealed inhomogeneous localization of BRCA2 relative to other DNA repair factors at sites of DNA damage. This suggests the presence of multiple nanoscale compartments in the chromatin surrounding the DNA lesion, which could play an important role in the contribution of BRCA2 to the regulation of the repair process.
Subject(s)
BRCA2 Protein , DNA Damage , DNA Repair , BRCA2 Protein/metabolism , BRCA2 Protein/genetics , Humans , Chromatin/metabolism , Cell Nucleus/metabolism , Cell Nucleus/genetics , Cell Line, Tumor , DNA/metabolismABSTRACT
Spheroids derived from human mesenchymal stem cells (hMSCs) are of limited use for cartilage regeneration, as the viability of the cells progressively decreases during the period required for chondrogenic differentiation (21 days). In this work, spheroids based on hMSCs and a lactose-modified chitosan (CTL) were formed by seeding cells onto an air-dried coating of CTL. The polymer coating can inhibit cell adhesion and it is simultaneously incorporated into spheroid structure. CTL-spheroids were characterized from a morphological and biological perspective, and their properties were compared with those of spheroids obtained by seeding the cells onto a non-adherent surface (agar gel). Compared to the latter, smaller and more viable spheroids form in the presence of CTL as early as 4 days of culture. At this time point, analysis of stem cells differentiation in spheroids showed a remarkable increase in collagen type-2 (COL2A1) gene expression (~700-fold compared to day 0), whereas only a 2-fold increase was observed in the control spheroids at day 21. These results were confirmed by histological and transmission electron microscopy (TEM) analyses, which showed that in CTL-spheroids an early deposition of collagen with a banding structure already occurred at day 7. Overall, these results support the use of CTL-spheroids as a novel system for cartilage regeneration, characterized by increased cell viability and differentiation capacity within a short time-frame. This will pave the way for approaches aimed at increasing the success rate of procedures and reducing the time required for tissue regeneration.
Subject(s)
Cell Differentiation , Chitosan , Chondrogenesis , Lactose , Mesenchymal Stem Cells , Spheroids, Cellular , Chitosan/pharmacology , Chitosan/chemistry , Mesenchymal Stem Cells/drug effects , Mesenchymal Stem Cells/cytology , Humans , Cell Differentiation/drug effects , Chondrogenesis/drug effects , Spheroids, Cellular/drug effects , Spheroids, Cellular/cytology , Lactose/pharmacology , Lactose/chemistry , Cell Survival/drug effects , Cells, Cultured , Collagen Type II/metabolism , Collagen Type II/geneticsABSTRACT
In this review we focused on the putative therapeutic effect of Hericium erinaceus extract in the treatment of pathologic conditions of the lower urinary tract in which intestinal inflammation may play a role. To this aim we reviewed the available evidence on pelvic cross-organ sensitization as a possible mechanism through which intestinal inflammation and dysbiosis may affect the lower urinary tract. Also, we reviewed the clinical and experimental evidence supporting the role of Hericium erinaceus extract as an anti-inflammatory agent highlighting the role of a number of putative mediators and mechanisms which might make this nutraceutical suitable for the management of 'difficult to treat' lower urinary tract disorders.
Subject(s)
Basidiomycota , Hericium , Humans , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Inflammation , Plant Extracts/pharmacology , Plant Extracts/therapeutic useABSTRACT
BACKGROUND: In recent years, improvement of Health-Related Quality of Life (HRQoL) in Ulcerative colitis (UC) has become a relevant measure for treatment efficacy. METHODS: We report results from a multicenter prospective study in Italy investigating HRQoL in adult patients with UC treated with golimumab (GLM). Patients who had shown clinical response after a 6-week induction phase (w0), were followed for an additional 48 weeks (w48) (total 54-week treatment). RESULTS: Of the 159 patients enrolled 90 completed the study. Compared to values at the beginning of treatment (n = 137), significant improvements were observed for mean total Inflammatory Bowel Disease Questionnaire (IBDQ) scores at w0 (168.5) and w48 (181.7). Patients with baseline PMS above the median tended to have greater improvements in IBDQ at w0 (OR 2.037, p = 0.033) and w48 (OR 3.292, p = 0.027). Compared to beginning of GLM treatment, the mean Full Mayo Score (FMS) decreased by 5.9 points at w48, while mean Partial Mayo Score (PMS) decreased by 3.9 points at w0 and by 4.9 points at w48. CONCLUSIONS: GLM improved HRQoL, disease activity and inflammatory biomarkers in UC patients with moderate-to-severely active disease. The greater the burden of disease activity at baseline, the greater the improvement of HRQoL after 24 and 48 weeks of treatment.
Subject(s)
Colitis, Ulcerative , Inflammatory Bowel Diseases , Adult , Humans , Colitis, Ulcerative/drug therapy , Quality of Life , Prospective Studies , Antibodies, Monoclonal/therapeutic use , Treatment Outcome , Inflammatory Bowel Diseases/drug therapy , Severity of Illness IndexABSTRACT
In a previous study, we evaluated the degree of virulence of Mycobacterium avium subsp. paratuberculosis (Map) strains isolated from cattle in Argentina in a murine model. This assay allowed us to differentiate between high-virulent MapARG1347 and low-virulent MapARG1543 strains. To corroborate whether the differences in virulence could be attributed to genetic differences between the strains, we performed Whole Genome Sequencing and compared the genomes and gene content between them and determined the differences related to the reference strain MapK10. We found 233 SNPs/INDELS in one or both strains relative to Map K10. The two strains share most of the variations, but we found 15 mutations present in only one of the strains. Considering NS-SNP/INDELS that produced a severe effect in the coding sequence, we focus the analysis on four predicted proteins, putatively related to virulence. Survival of MapARG1347 strain in bMDM was higher than MapARG1543 and was more resistant to acidic pH and H2O2 stresses than MapK10. The genomic differences between the two strains found in genes MAP1203 (a putative peptidoglycan hydrolase), MAP0403 (a putative serine protease) MAP1003c (a member of the PE-PPE family) and MAP4152 (a putative mycofactocin binding protein) could contribute to explain the contrasting phenotype previously observed in mice models.
Subject(s)
Mycobacterium avium subsp. paratuberculosis , Mycobacterium tuberculosis , Animals , Cattle , Mice , Mycobacterium avium subsp. paratuberculosis/genetics , Hydrogen Peroxide , Genomics , PhenotypeABSTRACT
Among all the regulatory homeostatic networks in vertebrates, the activation of the hypothalamic-pituitaryadrenal axis during the stress response, has gained considerable attention, and the measurement of fecal glucocorticoids (FGC) has become an invaluable tool to assess adrenocortical activity related to stressful events in wild and captive animals. However, the use of FGC requires the validation of measurement techniques and the proper selection of the specific hormone according to the study species. The main objective of this study was to identify the dominant glucocorticoid (GC) hormone in the stress response of black-tailed prairie dogs (Cynomys ludovicianus) in an arid grassland of Chihuahua, Mexico. A capture stress challenge in the field was developed to determine if the levels of glucocorticoids (cortisol and corticosterone) both in serum and fecal samples could be attributed to stress in Cynomys ludovicianus. The samples were analysed with the technique of liquid phase radioimmunoassay , and this study showed that both cortisol and corticosterone are present at measurable levels in serum and fecal samples of black-tailed prairie dogs. We found that both GCs were present in similar concentrations in serum, however, corticosterone concentration in fecal samples was higher than cortisol. Likewise, biochemical validations performed in this study to test the assay reached acceptable levels of reliability. Therefore, we confirm that fecal analysis can be implemented as a method to measure stress responses in wild prairie dogs.
Subject(s)
Corticosterone , Glucocorticoids , Animals , Hydrocortisone , Mexico , Reproducibility of Results , SciuridaeABSTRACT
A 90-years-old man comes to the emergency department with diffuse abdominal pain and fever three days. On physical examination, the abdomen is soft and depressible with air-fluid sound present and bilateral reductible inguinal hernias. A mass is palpated at the level of the roght hypochondrium with a positive Murphy´s sign whitout signs of peritoneal irritation. An abdominal CT with intravenous contrast is requested due to the suspicion of acute abdominal pathology, observing cholelithiasis and a markedly distended gallbladder with torsion. (AU)
Subject(s)
Humans , Male , Aged, 80 and over , Hypertension , Anemia , Appendectomy , Nausea , GallbladderABSTRACT
A 79-year-old woman with multiple pathologies and multiple osteolytic lesions during the Covid-19 pandemic. Histopathological and radiological differential diagnosis of injuries. (AU)
Subject(s)
Humans , Female , Aged , Diabetes Mellitus , Hypertension , Heart Failure , Renal Insufficiency, ChronicABSTRACT
We extend the paradigmatic and versatile totally asymmetric simple exclusion process (TASEP) for stochastic 1D transport to allow for two different particle species, each having specific entry and exit rates. We offer a complete mean-field analysis, including a phase diagram, by mapping this model onto an effective one-species TASEP. Stochastic simulations confirm the results, but indicate deviations when the particle species have very different exit rates. We illustrate that this is due to a phenomenon of intermittency, and formulate a refined "intermittent" mean-field theory for this regime. We discuss how nonstationary effects may further enrich the phenomenology.
ABSTRACT
34-year-old woman, 16 weeks pregnant, with a personal history of cervical conization and previous cesarean section.Consultation for discomfort in the right groin region and signs of inflammation in the area, without a clear tumor. She refers that inthe first pregnancy she presented the same symptoms. In the intergestational period she has not noticed these symptoms. On physicalexamination, the 16-week pregnant abdomen did not reveal clear signs of hernia, although there was slight pain on palpation. It wasdecided to request a pelvic ultrasound to rule out the presence of hernia or other soft tissue lesions in the groin region. (AU)
Mujer de 34 años, gestante de 16 semanas, con antecedentes personales de conización cervical y cesárea anterior. Consultapor molestias en región inguinal derecha y signos de inflamación en la zona, sin clara tumoración. Refiere que en el primerembarazo presentaba la misma sintomatología. En el período intergestacional no ha notado estos síntomas. A la exploraciónfísica se palpa útero, con altura correspondiente a unas 16 semanas de gestación, no se demuestran signos evidentes de hernia,aunque si leve dolor a la palpación. Se decide solicitar ecografía pélvica para descartar la presencia de hernia u otras lesiones de partes blandas en región inguinal. (AU)
Subject(s)
Humans , Female , Adult , Cesarean Section , Pregnancy , Hernia , Wounds and Injuries , PatientsABSTRACT
Human gut microbiome is related to different clinical conditions and diseases. Recently several hypotheses have been theorized about a link between gut microbiota and genitourinary disease including urinary tract infections, and benign prostatic hyperplasia. Despite several data, underlying mechanisms still remain unclear. The aim of this review is to report the current state of knowledge in relation to urinary tract infections, benign prostatic hyperplasia and intestinal microbiota with a focus on its role in the development of disease and the underlying pathophysiologic mechanisms.
Subject(s)
Gastrointestinal Microbiome , Microbiota , Prostatic Hyperplasia , Male , Humans , Gastrointestinal Microbiome/physiology , PermeabilityABSTRACT
El SARS-CoV-2 es un virus emergente que se extendió por todo el mundo desde China a finales de 2019, generando en el individuo una respuesta inflamatoria sistémica con afectación severa de los principales órganos (pulmón, corazón, sistema nervioso central, etc.). A la enfermedad causada por el mismo, la denominamos COVID-19. Una de las complicaciones descritas en la literatura, más frecuentemente asociadas a la infección, es la enfermedad tromboembólica venosa (embolismo pulmonar y trombosis venosa profunda). Se cree que la causa es debida a la inflamación excesiva y un estado de hipercoagulabilidad, con activación plaquetaria, disfunción endotelial y estasis sanguínea, que favorece la adhesión plaquetaria y formación de trombos. Presentamos dos casos clínicos diagnosticados en nuestro centro durante el periodo de pandemia en los que se observó esta complicación (AU)
SARS-CoV-2 is an emerging virus spread over the world from China in late 2019, generating in the patient a systemic inflammatory response with severe involvement of the main organs (lung, heart, central nervous system, etc.). The disease produced by this virus is called COVID-19. Previous studies show that one of the most commonly associated complications with COVID-19 is venous thromboembolic disease (pulmonary embolism and deep vein thrombosis). The cause is thought to be due to excessive inflammation and hypercoagulability, with platelet activation, endothelial dysfunction and blood theses, which promotes platelet adhesion and thrombus formation. We present two clinical cases reported during the pandemic period in which this complication was reported (AU)
Subject(s)
Humans , Male , Middle Aged , /complications , /diagnosis , Pulmonary Embolism/diagnosis , Pulmonary Embolism/virology , Tomography, X-Ray ComputedABSTRACT
The chemical coupling of a protoplasmatic antigen from Mycobacterium avium subsp. paratubeculosis onto core-shell carboxylated particles was investigated with the aim of producing latex-protein complexes to be used in immunoagglutination assays capable of detecting bovine paratuberculosis disease. For this purpose, sensitizations were carried out using both colored and not colored carboxylated latexes as well as the protoplasmatic antigen at pH close to its isoelectric point to favor the antigenic protein to approach the particle surface. In all cases, higher fractions of proteins were chemically-bound to carboxyl groups on the surface of the particles. The assessment of the performance of the visual immunoagglutination assays consisted of evaluating 111 sera from healthy and infected bovines with Mycobacterium avium subsp. paratuberculosis. Complexes obtained from the colored latex allowed an acceptable visual discrimination between the studied positive and negative sera. Most of the positive samples showed strong to very strong agglutination and only a few samples reacted weakly, i.e. a sensitivity of 70%. The specificity of the assay, on the other hand, was 86%. Therefore, this rapid detection technique allows an easy and inexpensive identification of animals possibly infected with paratuberculosis "in situ" in the herds.
Subject(s)
Antigens, Bacterial/immunology , Latex Fixation Tests/veterinary , Latex/chemistry , Mycobacterium avium subsp. paratuberculosis/immunology , Paratuberculosis/diagnosis , Animals , Case-Control Studies , Cattle , Color , Microspheres , Paratuberculosis/immunology , Paratuberculosis/microbiology , Predictive Value of Tests , Time Factors , WorkflowABSTRACT
BACKGROUND: Percutaneous Endoscopic Gastrostomy (PEG) can involve some complications, despite the good safety of its track record. The Buried Bumper Syndrome (BBS) is a rare, late and dangerous complication that consists in the erosion of the internal bumper through the gastric wall. Case presentation We report the development of BBS in a man with chronic obstructive pulmonary disease (COPD) who had a persistent chronic cough which was prevalently but not solely in the morning and required placement of a PEG tube for continuous infusion of Levodopa/carbidopa intestinal gel for advanced Parkinson's disease. CONCLUSION: We believe that COPD with chronic cough while not representing an absolute contraindication to PEG placement, may potentially cause BBS and therefore an appropriate regimen of tube care by expert personnel is mandatory in this setting.
Subject(s)
Gastrostomy , Pulmonary Disease, Chronic Obstructive , Contraindications , Cough/etiology , Enteral Nutrition , Humans , Male , Pulmonary Disease, Chronic Obstructive/complications , Pulmonary Disease, Chronic Obstructive/therapyABSTRACT
Ustekinumab (UST), a human anti-IL12/23p40 monoclonal antibody, was approved by FDA and EMA for the treatment of moderate to severe Crohn's disease (CD). Whether UST is effective in inducing deep remission, including mucosal healing and transmural healing, in patients with CD in a real life setting is not completely clear. This study was performed on 92 subjects with confirmed diagnosis of moderate to severe Crohn's disease and no neoplasia. Before inclusion, all patients had been exposed and had failed to respond to conventional and/or at least one biological therapy. All patients underwent endoscopic examination and bowel MRI and ultrasonography at baseline (T0). At week 52 (T52), patients underwent colonoscopy for assessment of mucosal healing and MRI or ultrasonography for assessment of transmural healing. CDAI was used for the assessment of clinical response and clinical remission. SES-CD was used to assess endoscopic response and remission. Incidence of treatment-related adverse events (TRAEs) was recorded during the study period. Clinical response at week 52 was achieved in 38 (50.5%) patients and clinical remission in 29 (39%). Twenty-six (34%) patients showed mucosal healing, 34 (45%) showed partial endoscopic response. We observed a reduction in SES-CD of at least 50% in 34 (45%) patients as well as an SES-CD ≤ 2 in 26 (35%) patients. All patients with mucosal healing also showed transmural healing. No major TRAEs were observed during treatment. In this multicenter, real life study, we show that UST was well tolerated and effective in inducing clinical response and clinical remission in patients with moderate to severe CD who had previously failed to respond to conventional or biologic therapy. UST showed limited efficacy in inducing deep remission (i.e. mucosal+transmural healing).
Subject(s)
Crohn Disease , Ustekinumab , Biological Therapy , Crohn Disease/drug therapy , Humans , Prospective Studies , Remission Induction , Treatment Outcome , Ustekinumab/therapeutic useABSTRACT
Before there was use of ultrasonographic imaging, determination of the ratio of estrogens to androgens in the same individual was a technique used for differentiating the sex of monomorphic animals in captivity, with larger estrogen concentrations in the females. Due to species-specific differences in both concentration and changes throughout the year of these hormones, corroboration of the method is needed in each case. In this study, there was use of a chemo-immuno assay to quantify sex steroids in fecal samples collected from seven (five females and two males) Neotropical otters, Lontra longicaudis. The reproductive season for this species was determined to be between October and March, with increased estradiol in the females and relatively greater concentrations of testosterone in the males as compared with other seasons of the year. Results from utilization of a k-means analysis procedure indicated that the use of steroid ratios in fecal samples to differentiate otter sex is an effective technique when there are evaluations during the breeding season. The estrogen to androgen ratios during this period, however, are the inverse of what was expected, with there being larger testosterone concentrations in the female otters. The ratio of estrogens to androgens in feces of captive otters can be effectively used to determine the sex of otters in the field. We propose this method is reliable for sex determination in wild otter populations during the reproductive season.