Exosomal glypican-1 discriminates pancreatic ductal adenocarcinoma from chronic pancreatitis.

BACKGROUND AND AIMS: Pancreatic ductal adenocarcinoma (PDAC) diagnosis can be difficult in a chronic pancreatitis (CP) background, especially in its mass forming presentation. We aimed to assess the accuracy of glypican-1-positive circulating exosomes (GPC1+crExos) to distinguish PDAC from CP versus the state-of-the-art CA 19-9 biomarker. METHODS: This was a unicentric prospective cohort. Endoscopic ultrasound with fine-needle aspiration or biopsy and blood tests (GPC1+crExos and serum CA 19-9) were performed. RESULTS: The cohort comprised 60 PDAC and 29 CP (7 of which mass forming - MF) patients. Median levels of GPC1+crExos were significantly higher in PDAC (99.7%) versus CP (28.4%; p<0.0001) with an AUROC of 0.96 with 98.3% sensitivity and 86.2% specificity for a cut-off of 45.0% (p<0.0001); this outperforms CA 19-9 AUROC of 0.82 with 78.3% sensitivity and 65.5% specificity at a cut-off of 37 U/mL (p<0.0001). The superiority of% GPC1+crExos over CA 19-99 in differentiating PDAC from CP was observed in both early (stage I) and advanced tumors (stages II-IV). CONCLUSION: Levels of GPC1+crExos coupled to beads enable differential diagnosis between PDAC and CP including its mass-forming presentation.

Quality by design (QbD) optimization of diazepam-loaded nanostructured lipid carriers (NLC) for nose-to-brain delivery: Toxicological effect of surface charge on human neuronal cells.

Diazepam is commonly used in the management of epileptic seizures, although it has limitations that can be overcome by using formulations that are easier to administer and capable of directing the drug to the brain. In this field, it has been reported that the use of nanostructured lipid carriers (NLC) via intranasal (or via nose-to-brain) promotes the targeting of drugs to the brain, improving the effectiveness of therapy. The aim of this work was to optimize two diazepam-loaded NLC formulations for nose-to-brain delivery, one with positive surface charge and one with negative surface charge. The quality by design (QbD) approach was used to design the experiments, where the quality target product profile (QTPP), the risk assessment and the critical quality attributes (CQAs) were defined to ensure safety, efficacy and quality of the final formulations. The experiments started with the optimization of critical material attributes (CMAs), related to the ratios of lipids and emulsifiers, followed by the selection of critical process parameters (CPPs), related to the production methods of the diazepam-loaded NLC formulation (ultrasound technique and high-pressure homogenization - HPH). Afterwards, the positive surface charge of the diazepam-loaded NLC was optimized. Finally, the biocompatibility with human neuronal cells of the formulation with a negative surface charge and of the formulation with a positive surface charge was evaluated. The results of the optimization of the CMAs showed that the ratios of lipids and emulsifiers more adequate were 6.7:2.9 and 4.2:0.3 (% w,w), respectively. Regarding the CPPs, HPH was considered the most suitable production method, resulting in an optimized diazepam-loaded NLC formulation (F1C15) with negative surface charge, showing particle size of 69.59 ± 0.22 nm, polydispersity index (PDI) of 0.19 ± 0.00, zeta potential (ZP) of -23.50 ± 0.24 mV and encapsulation efficiency (EE) of 96.60 ± 0.03 %. The optimized diazepam-loaded NLC formulation (F2A8) with positive surface charge had particle size of 124.40 ± 0.84 nm, PDI of 0.17 ± 0.01, ZP of 32.60 ± 1.13 mV and EE of 95.76 ± 0.24 %. In addition, the incorporation of diazepam in NLC resulted in a sustained release of the drug. No significant changes in particle size, PDI, ZP and EE were observed for the formulation F1C15, after 3 months of storage, whereas for formulation F2A8, particle size increased significantly. Biocompatibility studies showed that the formulation F2A8 was more cytotoxic than the formulation F1C15. Thereby, we conclude that the formulation F1C15 is more suitable for targeting the brain, when compared with the formulation F2A8. From the results of these studies, it can be confirmed that the QbD approach is an adequate and central tool to optimize NLC formulations.

Applicability of lung ultrasound in COVID-19 diagnosis and evaluation of the disease progression: A systematic review.

INTRODUCTION: The COVID-19 pandemic originated in China and within about 4 months affected individuals all over the world. One of the limitations to the management of the COVID-19 is the diagnostic imaging to evaluate lung impairment and the patients' clinical evolution, mainly, in more severe cases that require admission into the intensive care unit. Among image examinations, lung ultrasound (LU) might be a useful tool to employ in the treatment of such patients. METHODS: A survey was carried out on PubMed to locate studies using the descriptors: ((Lung ultrasound OR ultrasound OR lung ultrasonography OR lung US) AND (coronavirus disease-19 OR coronavirus disease OR corona virus OR COVID-19 OR COVID19 OR SARS-CoV-2)). The period covered by the search was November 2019 to October 2020 and the papers selected reported LU in COVID-19. RESULTS: Forty-three studies were selected to produce this systematic review. The main LU findings referred to the presence of focal, multifocal and/or confluent B lines and the presence of pleural irregularities. CONCLUSIONS: The use of LU in the evaluation of patients with COVID-19 should be encouraged due to its intrinsic characteristics; a low cost, radiation free, practical method, with easy to sanitize equipment, which facilitates structural evaluation of lung damage caused by SARS-CoV-2. With the increase in the number of studies and the use of ultrasound scans, LU has been shown as a useful tool to evaluate progression, therapeutic response and follow-up of pulmonary disease in the patients with COVID-19.

[Frailty of the elderly and physical activity in general practice: A prospective study]. / Étude de la fragilité de la personne âgée et de l'activité physique en médecine générale : à propos d'une étude prospective.

INTRODUCTION: Frailty is a transient and reversible condition that can lead to significant morbidity and mortality and to the loss of autonomy. It is one of the key issues in public health and the prevention of addiction. Physical activity is often described as a protective factor against addiction and as a factor in limiting frailty. Our goal is to analyze the relationship between physical activity and frailty among the elderly METHODS: A cross-sectional observational study was carried out in three general practice clinics located in the French department of Eure. Patients aged 65 and over were included during a consultation with their GP. Dependent patients under the ADL scale were excluded. Level of physical activity was assessed by the Ricci and Gagnon questionnaire, which defines an active profile as a score≥18 points. Frailty was sought out by the Fried scale and the SEGA A grid. Multivariate analysis was performed to adjust frailty scores to age, gender, and level of physical activity. RESULTS: Out of the 70 patients included, 36 were active (51%) and 34 inactive (49%). They were predominantly female with 47 women (67%). Average age was 75.3years. Twelve patients were diagnosed as frail (17%) with the Fried scale and 24 (34%) with the SEGA A grid. Bivariate analysis revealed a greater frailty according to the Fried criteria in the inactive than in the active patients (mean for active patients 0.56 IC95 [0.31; 0.80], compared to 1.76 [1.21; 2.32] in the inactive patients, p<0.0001). The difference in mean was likewise significant regarding the SEGA A score (6.42 IC95 [5.34, 7.49] in the active population, as opposed to 8.65 IC95 [7.15, 10.15] among the inactive, p=0.017). In multivariate analysis, the Fried scale was primarily influenced by age and ADL, while the SEGA score was impacted by female gender and level of physical activity. CONCLUSION: Physical activity seems to have a positive effect on frailty. It would be interesting to propose systematic screening for frailty in general medicine and to institute preventive measures, including physical activity. Initiatives encouraging and promoting seniors' physical activity should be strengthened.

Comparison of obstetrical interventions in women with vaginal and cesarean section delivered: cross-sectional study in a reference tertiary center in the Northeast of Brazil.

OBJECTIVE: To compare the performance of obstetrical interventions and maternal and perinatal outcomes between vaginal and cesarean delivery routes in pregnant women at normal risk. Type of article: Original article. Desing: Cross-sectional study with 421 participants admitted for spontaneous or induced labor with full-term singleton gestations and fetuses weighing between 2,500 and 4,499 g. SETTING: Maternal Fetal-Medicine Service, Assis Chateaubriand Maternity, Federal University of Ceará (UFC), Fortaleza-CE, Brazil. METHODS: The instrument of data collection was divided into socio-demographic, clinical, and obstetric characteristics; data of labor and delivery; maternal morbidity; maternal outcome and perinatal outcomes. Pearsons chi-square test and Fishers exact test were used to verify associations between the groups. RESULTS: The mean age was 22.8 ± 6.0 (vaginal) and 22.9 ± 4.9 (cesarean section). Overall, 44.5% of vaginal deliveries and 85.5% of cesarean sections were monitored electronically (p < 0.001). Immediate skin-to-skin contact (84.1%) and first-hour breastfeeding (80.4%) were more frequent in vaginal deliveries compared with cesarean deliveries (27% vs. 61.0%, p < 0.001). The prevalence of puerperal infections was 1.2% (vaginal) and 5.0% (cesarean section) with a p value of 0.02; 40% of cesarean-delivered newborns and 9.7% of vaginally-delivered newborns were referred to the neonatal intensive care unit (p < 0.001). CONCLUSION: The cesarean section was associated with a lower frequency of useful practices, a higher frequency of harmful practices, worse neonatal outcomes, and a higher rate of postpartum infections.

Etanercept-induced granulomatous hepatitis as a rare cause of abnormal liver tests.

The authors report the case of a 76 year-old man with rheumatoid arthritis treated with prednisolone and etanercept. The patient was seen for persistent changes in liver tests lasting for six months, with a mixed pattern. The patient denied intake of new drugs or dietary/herbal supplements. Imaging studies showed mild steatosis. Additional study for chronic liver diseases only revealed positivity for anti-nuclear antibodies. Liver biopsy revealed noncaseating granulomas in some portal tracts. Consequent etiologic study for granulomatous diseases showed negative or normal results. So it was decided to suspend etanercept, with a subsequent gradual improvement on analytical parameters that normalized three months later. To date, only one case of granulomatous liver disease associated with an anti-TNF agent was described in the literature. This case also raises the question whether the development of granulomatous processes associated with anti-TNF agents has been underdiagnosed due to the presence of other concomitant immunosuppressant therapies.

Is a Single Preliminary Session of In-office Bleaching Beneficial for the Effectiveness of At-home Tooth Bleaching? A Randomized Controlled Clinical Trial.

OBJECTIVES: To evaluate the effect of combining in-office with at-home bleaching procedures in terms of the time required to obtain satisfactory tooth color, final color changes, and tooth sensitivity (TS) reported by patients. METHODS AND MATERIALS: Twenty-six patients enrolled in this study used 10% carbamide peroxide in a bleaching tray for 1 h/d until satisfactory tooth color was obtained. One-half of the participants underwent a preliminary session of in-office tooth bleaching with 35% hydrogen peroxide for 45 minutes. The time in days for the patients to obtain satisfactory tooth color by at-home bleaching procedures was recorded. The color change of the maxillary canines was assessed using the Vita Bleachedguide 3D Master scale and a spectrophotometer at 1 week and after the end of bleaching procedures. Participants' satisfaction with their smile was recorded using a visual analog scale, and TS was determined throughout the entire treatment. Data were analyzed by t-test, Mann-Whitney test, or Fisher exact test (α=0.05). RESULTS: The combined protocol reduced (by an average of 3.7 days) the time required to obtain satisfactory tooth color but increased the risk and level of TS. No difference in the final tooth color change (around 5.0 shade guide units; ΔE=11.6-14.9), or the level of patients' satisfaction with their smile, was observed. CONCLUSIONS: A preliminary session of in-office bleaching reduced the time necessary to obtain satisfactory tooth color with at-home bleaching but increased the risk and level of TS.

Kinetics of acid-base parameters in conventional hemodialysis

Details about the acid-base changes in hemodialysis are scarce in the literature but are potentially relevant to adequate management of patients. We addressed the acid-base kinetics during hemodialysis and throughout the interdialytic period in a cross-sectional study of adults undergoing conventional hemodialysis. Samples for blood gas analysis were obtained from the arterial limb of the arteriovenous fistula before the first session of the week (HD1), immediately at the end of HD1, and on sequential collections at 15, 30, 45, 60, and 120 min post-HD1. Additional blood samples were collected after ∼20 h following the end of the first dialysis and immediately prior to the initiation of the second dialysis of the week. Thirty adult patients were analyzed (55±15 years, 50% men, 23% diabetic; dialysis vintage 69±53 months). Mean serum bicarbonate levels increased at the end of HD1 (22.3±2.7 mEq/L vs 17.5±2.3 mEq/L, P<0.001) and remained stable until 20 h after the end of the session. The mean values of pCO2 before HD1 were below reference and at 60 and 120 min post-HD1 were significantly lower than at the start (31.3±2.7 mmHg and 30.9±3.7 mmHg vs 34.3±4.1 mmHg, P=0.041 and P=0.010, respectively). The only point of collection in which mean values of pCO2 were above 35 mmHg was 20 h post-dialysis. Serum bicarbonate levels remained stable for at least 20 h after the dialysis sessions, a finding that may have therapeutic implications. During dialysis, the respiratory response for correction of metabolic acidosis (i.e., pCO2 elevation) was impaired.

Kinetics of acid-base parameters in conventional hemodialysis.

Details about the acid-base changes in hemodialysis are scarce in the literature but are potentially relevant to adequate management of patients. We addressed the acid-base kinetics during hemodialysis and throughout the interdialytic period in a cross-sectional study of adults undergoing conventional hemodialysis. Samples for blood gas analysis were obtained from the arterial limb of the arteriovenous fistula before the first session of the week (HD1), immediately at the end of HD1, and on sequential collections at 15, 30, 45, 60, and 120 min post-HD1. Additional blood samples were collected after ∼20 h following the end of the first dialysis and immediately prior to the initiation of the second dialysis of the week. Thirty adult patients were analyzed (55±15 years, 50% men, 23% diabetic; dialysis vintage 69±53 months). Mean serum bicarbonate levels increased at the end of HD1 (22.3±2.7 mEq/L vs 17.5±2.3 mEq/L, P<0.001) and remained stable until 20 h after the end of the session. The mean values of pCO2 before HD1 were below reference and at 60 and 120 min post-HD1 were significantly lower than at the start (31.3±2.7 mmHg and 30.9±3.7 mmHg vs 34.3±4.1 mmHg, P=0.041 and P=0.010, respectively). The only point of collection in which mean values of pCO2 were above 35 mmHg was 20 h post-dialysis. Serum bicarbonate levels remained stable for at least 20 h after the dialysis sessions, a finding that may have therapeutic implications. During dialysis, the respiratory response for correction of metabolic acidosis (i.e., pCO2 elevation) was impaired.

Impact of cesarean section in a private health service in Brazil: indications and neonatal morbidity and mortality rates.

OBJECTIVE: To evaluate the incidence of, indications of, and maternal and neonatal morbidity and mortality rates in cesarean sections in a private health service in Brazil. DESIGN: Retrospective and observational study. SETTING: Private health service in Vitória, Espírito Santo, Brazil. METHODS: The patients were interviewed using a structured questionnaire to determine maternal age, gestational age at the time of delivery, number of previous deliveries, type of delivery performed, duration of labor, indications for cesarean delivery, point at which cesarean section was performed, physician responsible for delivery, and maternal morbidity, fetal morbidity, and fetal mortality rates. A descriptive analysis of the data was conducted. Students t-test was performed to compare quantitative variables, and Fishers exact test was performed for categorical variables. RESULTS: A total of 584 patients were evaluated. Of these, 91.8% (536/584) had cesarean sections, while only 8.2% (48/584) had vaginal deliveries. There were no reports of forceps-assisted vaginal deliveries. In 87.49% of the deliveries, the number of gestational weeks was more than 37. In terms of indications for performing cesarean section, 48.5% were for maternal causes, 30.41% were for fetal causes, and 17.17% were elective. Maternal re-hospitalization due to puerperal complications was necessary in 10.42% of the vaginal deliveries and in 0.93% of the cesarean deliveries (p<0.001). Complications were observed in 18.75% of the vaginally delivered newborns and in 17.16% of those delivered by cesarean section. Of the newborns with complications at birth, 40.59% (41/101) had to be admitted to the neonatal intensive care unit. There were no cases of maternal death. There were seven cases of fetal/neonatal death. CONCLUSION: We observed that the vast majority of deliveries in the private sector are performed by cesarean section, without labor, and by the patients obstetrician. We found no serious maternal complications or increased neonatal morbidity rates associated with cesarean section.

Effects of cervical cerclage on cervical length and the impact of changes in cervical length on pregnancy prognosis.

OBJECTIVE: To identify any cervix-related morphological and functional marker that can be correlated with pregnancy prognosis in patients who have undergone cerclage for cervical incompetence. DESIGN: An observational and prospective study. SETTING: Department of Obstetrics, Paulista School of Medicine - Federal University of São Paulo (UNIFESP-EPM). METHODS: Patients with cervical incompetence who underwent cervical cerclage using a modified version of the McDonald procedure during or before the 22nd week of pregnancy. The patients were examined by transvaginal ultrasound in the preoperative period, the immediate postoperative period, and between 20 and 24 weeks, 24 weeks + 1 day and 28 weeks, and 28 weeks + 1 day and 32 weeks. Cervical length and the presence of funneling were evaluated during all examinations. Changes in cervical length, presence or absence of funneling, percent increase or decrease in cervical length, and cervical length of less than established values (.

Biosafety Level 3 setup for multiphoton microscopy in vivo.

Multiphoton microscopy has revealed important insights into cellular behavior in vivo. However, its application in infectious settings often encounters technical, safety and regulatory limitations that prevent its wider use with highly virulent human pathogens. Herein, we present a method that renders multiphoton microscopy in vivo compatible with biosafety level 3 regulations and present an example of its application and potential to visualize a Mycobacterium tuberculosis infection of the mouse lung.

Expanding the view of Clock and cycle gene evolution in Diptera.

We expanded the view of Clock (Clk) and cycle (cyc) gene evolution in Diptera by studying the fruit fly Anastrepha fraterculus (Afra), a Brachycera. Despite the high conservation of clock genes amongst insect groups, striking structural and functional differences of some clocks have appeared throughout evolution. Clk and cyc nucleotide sequences and corresponding proteins were characterized, along with their mRNA expression data, to provide an evolutionary overview in the two major groups of Diptera: Lower Diptera and Higher Brachycera. We found that AfraCYC lacks the BMAL (Brain and muscle ARNT-like) C-terminus region (BCTR) domain and is constitutively expressed, suggesting that AfraCLK has the main transactivation function, which is corroborated by the presence of poly-Q repeats and an oscillatory pattern. Our analysis suggests that the loss of BCTR in CYC is not exclusive of drosophilids, as it also occurs in other Acalyptratae flies such as tephritids and drosophilids, however, but it is also present in some Calyptratae, such as Muscidae, Calliphoridae and Sarcophagidae. This indicates that BCTR is missing from CYC of all higher-level Brachycera and that it was lost during the evolution of Lower Brachycera. Thus, we can infer that CLK protein may play the main role in the CLK\CYC transcription complex in these flies, like in its Drosophila orthologues.