Keratin 17 modulates the immune topography of pancreatic cancer.

BACKGROUND: The immune microenvironment impacts tumor growth, invasion, metastasis, and patient survival and may provide opportunities for therapeutic intervention in pancreatic ductal adenocarcinoma (PDAC). Although never studied as a potential modulator of the immune response in most cancers, Keratin 17 (K17), a biomarker of the most aggressive (basal) molecular subtype of PDAC, is intimately involved in the histogenesis of the immune response in psoriasis, basal cell carcinoma, and cervical squamous cell carcinoma. Thus, we hypothesized that K17 expression could also impact the immune cell response in PDAC, and that uncovering this relationship could provide insight to guide the development of immunotherapeutic opportunities to extend patient survival. METHODS: Multiplex immunohistochemistry (mIHC) and automated image analysis based on novel computational imaging technology were used to decipher the abundance and spatial distribution of T cells, macrophages, and tumor cells, relative to K17 expression in 235 PDACs. RESULTS: K17 expression had profound effects on the exclusion of intratumoral CD8+ T cells and was also associated with decreased numbers of peritumoral CD8+ T cells, CD16+ macrophages, and CD163+ macrophages (p < 0.0001). The differences in the intratumor and peritumoral CD8+ T cell abundance were not impacted by neoadjuvant therapy, tumor stage, grade, lymph node status, histologic subtype, nor KRAS, p53, SMAD4, or CDKN2A mutations. CONCLUSIONS: Thus, K17 expression correlates with major differences in the immune microenvironment that are independent of any tested clinicopathologic or tumor intrinsic variables, suggesting that targeting K17-mediated immune effects on the immune system could restore the innate immunologic response to PDAC and might provide novel opportunities to restore immunotherapeutic approaches for this most deadly form of cancer.

Modeling Lysosomal Storage Disorders in an Innovative Way: Establishment and Characterization of Stem Cell Lines from Human Exfoliated Deciduous Teeth of Mucopolysaccharidosis Type II Patients.

Among the many lysosomal storage disorders (LSDs) that would benefit from the establishment of novel cell models, either patient-derived or genetically engineered, is mucopolysaccharidosis type II (MPS II). Here, we present our results on the establishment and characterization of two MPS II patient-derived stem cell line(s) from deciduous baby teeth. To the best of our knowledge, this is the first time a stem cell population has been isolated from LSD patient samples obtained from the dental pulp. Taking into account our results on the molecular and biochemical characterization of those cells and the fact that they exhibit visible and measurable disease phenotypes, we consider these cells may qualify as a valuable disease model, which may be useful for both pathophysiological assessments and in vitro screenings. Ultimately, we believe that patient-derived dental pulp stem cells (DPSCs), particularly those isolated from human exfoliated deciduous teeth (SHEDs), may represent a feasible alternative to induced pluripotent stem cells (iPSCs) in many labs with standard cell culture conditions and limited (human and economic) resources.

Trauma-focused treatments for victims of interpersonal violence: A comparison of treatment interventions and outcomes.

Background: Interpersonal violence represents a critical public health issue globally, with profound psychological impacts on victims. Objective: The main objective of this study was to analyze the effectiveness of different trauma-focused therapies on mental health outcomes of victims of interpersonal violence, at a community mental health clinic. Methods: Employing a secondary data methodology, the research involves 601 participants who reported being victims of sexual assault (49.1%), domestic violence (44.3%) or sexual trafficking (6.7%). The average age of the participants was 35.54 years, with a majority being female (89.8%). Results: Initial assessments revealed distinct symptomatology among the groups; however, by the ninth therapy session, symptom severity converged across the board, surpassing threshold levels for clinical concern. No significant interaction was observed between the type of trauma-focused therapy and the specific trauma encountered, suggesting a beneficial effect of trauma-focused therapies investigated. This uniformity in therapeutic outcomes underscores the potential of trauma-focused therapies to foster psychological healing in victims of diverse forms of interpersonal violence. Conclusions: The findings advocate for the widespread adoption of trauma-focused therapeutic interventions in community settings, emphasizing their role in the recovery of victims, independent of the nature of the trauma or the specific trauma-focused therapeutic model employed.

Development of Engineered-U1 snRNA Therapies: Current Status.

Splicing of pre-mRNA is a crucial regulatory stage in the pathway of gene expression. The majority of human genes that encode proteins undergo alternative pre-mRNA splicing and mutations that affect splicing are more prevalent than previously thought. Targeting aberrant RNA(s) may thus provide an opportunity to correct faulty splicing and potentially treat numerous genetic disorders. To that purpose, the use of engineered U1 snRNA (either modified U1 snRNAs or exon-specific U1s-ExSpeU1s) has been applied as a potentially therapeutic strategy to correct splicing mutations, particularly those affecting the 5' splice-site (5'ss). Here we review and summarize a vast panoply of studies that used either modified U1 snRNAs or ExSpeU1s to mediate gene therapeutic correction of splicing defects underlying a considerable number of genetic diseases. We also focus on the pre-clinical validation of these therapeutic approaches both in vitro and in vivo, and summarize the main obstacles that need to be overcome to allow for their successful translation to clinic practice in the future.

Dropout from evidence-based trauma treatment in a community mental health clinic serving victims of interpersonal violence.

Trauma-focused psychotherapies are increasingly offered in community-based mental health centers, but little is known about treatment dropout in these settings. The current study explored dropout at different stages of treatment in a treatment-seeking sample of 1,186 adults who experienced interpersonal violence and were offered trauma-focused and non-trauma-focused therapies. A total of 31.6% of participants dropped out before treatment initiation, 28.0% dropped out after treatment initiation and completed a mean of 4.02 (SD = 2.41) sessions, and 40.4% completed a full course of PTSD treatment. Being unemployed, p < .001, and scoring lower on measures of environment factors, p = .045, were significant predictors of pretreatment dropout. Being female, p < .001; Latinx, p = .032; and scoring higher on a measure of social relationships, p = .024, were independent predictors of postinitiation dropout. Individuals who completed nine sessions of treatment displayed significantly lower levels of posttraumatic stress disorder, depression, and anxiety symptoms. The present study provides preliminary evidence that survivors of interpersonal violence who seek therapy tend to drop out early during treatment, and most who complete treatment attain symptom reduction.

Post break-up stalking: Police-officers' perceptions.

Stalking is characterized by persistent unwanted behaviors and can disrupt the lives of victims. Its prevalence has been increasing with time. Due to the recent criminalization of stalking in Portugal and its being an understudied phenomenon, the objective of this study is to assess the perceptions of police officers (n = 1212) about stalking post intimate relationship breakup. We developed an adaptation of the questionnaire "Reasonable Perceptions of Stalking", using vignettes that describe stalking scenarios with the manipulation of severity, and victim and perpetrator sex. The main effects of severity, as well as the victim and the perpetrator's sex, were significant intrasubject variables in the perceptions of stalking and its severity, criminality, victim alert, fear of violence, victim-blaming, and the need for intervention. Participants' sex, previous training, and personal and/or professional experience with stalking were also significant variables concerning participants' perceptions. Our findings show that gender stereotypes are still perpetuated and have important implications for police officer training and intervention.

"It takes a community to save a child": Child trafficking knowledge of the Portuguese community.

This study aims to explore the knowledge of the Portuguese community on child trafficking. Through an online inventory, with a sample of 492 community members, with a mean age of 40.53 and mostly female (70.7%), participants were more knowledgeable about forms of exploitation, risk and vulnerability factors, the profile of the victims, and the situation in Portugal about child trafficking. On the contrary, they were less knowledgeable about the dynamics of child trafficking. Having a college education, and contacting with the subject through informal or/and formal sources were predictors of a higher level of knowledge of child trafficking. This study reinforces the importance of the community as a whole in fighting child trafficking. To protect children against child trafficking, public policies should prioritize the role of the community. An empowered community can actively contribute to the prevention, signaling, and (re)integration of victims of child trafficking.

Clinician-driven artificial intelligence in ophthalmology: resources enabling democratization.

PURPOSE OF REVIEW: This article aims to discuss the current state of resources enabling the democratization of artificial intelligence (AI) in ophthalmology. RECENT FINDINGS: Open datasets, efficient labeling techniques, code-free automated machine learning (AutoML) and cloud-based platforms for deployment are resources that enable clinicians with scarce resources to drive their own AI projects. SUMMARY: Clinicians are the use-case experts who are best suited to drive AI projects tackling patient-relevant outcome measures. Taken together, open datasets, efficient labeling techniques, code-free AutoML and cloud platforms break the barriers for clinician-driven AI. As AI becomes increasingly democratized through such tools, clinicians and patients stand to benefit greatly.

Nanostructured faujasite zeolite as metal ion adsorbent: kinetics, equilibrium adsorption and metal recovery studies.

The hydrothermal synthesis of nano-faujasite has been successfully performed and the effects of some crystallization parameters were investigated, along with the use of this material as a heavy-metal ion adsorbent. X-ray diffraction patterns have shown that the structure of the nano-faujasite is strongly dependent on both the crystallization time and the alkalinity of the synthesis medium. According to N2 physisorption, X-ray fluorescence, SEM/EDS, and solid state 29Si and 27Al NMR data, the produced nano-faujasite consists of a solid with low molar Si/Al ratio (1.7), with high availability of ion exchange sites and high surface area/small particle size, allowing easy diffusion of metal ions to adsorbent active sites. As a consequence, an excellent performance on removal of Cd2+, Zn2+ and Cu2+ ions was found for this solid. The adsorption capacity followed the order Cd2+ (133 mg·g-1) > Zn2+ (115 mg·g-1) > Cu2+ (99 mg·g-1), which agrees with the order of increasing absolute values of the hydration energy of the metal ions. Kinetic studies and adsorption isotherms showed that the metal ion removal takes place by ion exchange on the monolayer surface of the nano-faujasite. The electrochemical recovery of copper in metallic form exhibited an efficiency of 80.2% after 120 min, which suggests that this process can be adequately implemented for full-scale metal removal.

INTRAVITREAL DEXAMETHASONE IMPLANT MIGRATION INTO THE ANTERIOR CHAMBER: A Multicenter Study From the Pan-American Collaborative Retina Study Group.

PURPOSE: To establish the prevalence and risk factors for intravitreal dexamethasone implant migration into the anterior chamber in eyes with macular edema. METHODS: This was a multicenter, retrospective, observational chart review of data that included patients with macular edema who had been treated with at least one intravitreal dexamethasone injection. Patients with incomplete chart information during the follow-up period were excluded. RESULTS: The prevalence of implant migration in 468 patients, considering the number of injections, was 1.6%, with significant associations between implant migration and cataract surgery (P = 0.043) and intraocular lens status (P = 0.005) and a trend toward statistical significance (P = 0.057) with vitrectomy. A higher rate of implant migration into the anterior chamber was observed in vitrectomized eyes (4.8%) when compared with patients who did not undergo a vitrectomy (1.6%). The implants that migrated were removed with forceps with/without viscoelastic expression or with 20-gauge cannulas connected to the vitreous cutter machine. CONCLUSION: The risk of implant migration into the anterior chamber was 1.6%. Risk factors were a history of cataract surgery or vitrectomy and aphakia. When anterior migration occurs, rapid removal is advised, especially if corneal edema is present.

P-MAPA, a Fungi-Derived Immunomodulatory Compound, Induces a Proinflammatory Response in a Human Whole Blood Model.

P-MAPA is a complex compound, derived from Aspergillus oryzae cultures, that has shown immunomodulatory properties in infection and cancer animal models. Despite promising results in these models, the mechanisms of cellular activation by P-MAPA, suggested to be Toll-like receptor- (TLR-) dependent, and its effect on human immune cells, remain unclear. Using an ex vivo model of human whole blood, the effects of P-MAPA on complement system activation, production of cytokines, and the expression of complement receptors (CD11b, C5aR, and C3aR), TLR2, TLR4, and the coreceptor CD14 were analyzed in neutrophils and monocytes. P-MAPA induced complement activation in human blood, detected by increased levels of C3a, C5a, and SC5b-9 in plasma. As a consequence, CD11b expression increased and C5aR decreased upon activation, while C3aR expression remained unchanged in leukocytes. TLR2 and TLR4 expressions were not modulated by P-MAPA treatment on neutrophils, but TLR4 expression was reduced in monocytes, while CD14 expression increased in both cell types. P-MAPA also induced the production of TNF-α, IL-8, and IL-12 and oxidative burst, measured by peroxynitrite levels, in human leukocytes. Complement inhibition with compstatin showed that P-MAPA-induced complement activation drives modulation of C5aR, but not of CD11b, suggesting that P-MAPA acts through both complement-dependent and complement-independent mechanisms. Compstatin also significantly reduced the peroxynitrite generation. Altogether, our results show that P-MAPA induced proinflammatory response in human leukocytes, which is partially mediated by complement activation. Our data contribute to elucidate the complement-dependent and complement-independent mechanisms of P-MAPA, which ultimately result in immune cell activation and in its immunomodulatory properties in infection and cancer animal models.

Forced degradation studies of norepinephrine and epinephrine from dental anesthetics: Development of stability-indicating HPLC method and in silico toxicity evaluation.

Injectable solutions containing epinephrine (EPI) and norepinephrine (NE) are not stable, and their degradation is favored mainly by the oxidation of catechol moiety. As studies of these drugs under forced degradation conditions are scarce, herein, we report the identification of their degradation products (DP) in anesthetic formulations by the development of stability-indicating HPLC method. Finally, the risk assessment of the major degradation products was evaluated using in silico toxicity approach. HPLC method was developed to obtain a higher selectivity allowing adequate elution for both drugs and their DPs. The optimized conditions were developed using a C18 HPLC column, sodium 1-octanesulfonate, and methanol (80:20, v/v) as mobile phase, with a flow rate of 1.5 mL/min, UV detection at 199 nm. The analysis of standard solutions with these modifications resulted in greater retention time for EPI and NE, which allow the separation of these drugs from their respective DPs. Then, five DPs were identified and analyzed by in silico studies. Most of the DPs showed important alerts as hepatotoxicity and mutagenicity. To the best of our acknowledgment, this is the first report of a stability-indicating HPLC method that can be used with formulations containing catecholamines.

Development and validation of a stability-indicating HPLC method for topiramate using a mixed-mode column and charged aerosol detector.

The analysis of topiramate in the presence of its main degradation products is challenging due to the absence of chromophore moieties and their wide range of polarity. Mixed-mode chromatography has been used in such cases because it combines two or more modes of separation. Charged aerosol detector is also an alternative since its detection is independent of optical properties and analyte ionization. This study is aimed to develop and validate two new stability-indicating methods by high-performance liquid chromatography for the main degradation products of topiramate using mixed-mode chromatography and a charged aerosol detector. Method 1 employed an Acclaim Trinity P1® column (3.0 mm × 150 mm, 2.7 µm) with a mobile phase comprising of 80% ammonium acetate buffer (20 mM, pH 4.0) and 20% methanol at a flow rate of 0.5 mL/min at 35°C. Method 2 utilized a C18 Acclaim 120® column (4.6 mm × 250 mm; 5 µm) with ACN/water (50:50) at a flow rate of 0.6 mL/min at 50°C. Validation of the two methods demonstrated excellent performance with respect to linearity, precision, accuracy, and selectivity. The limits of detection for topiramate, fructose, sulfate, sulfamate, and compound A were 2.97, 12.08, 4.02, 13.91, and 3.94 µg/mL, respectively.

Blood Pressure Variability in Acute Ischemic Stroke: The Role of Early Recanalization.

We performed a retrospective study with the aim of investigating the association between blood pressure (BP) variability in the first 24 h after ischemic stroke and functional outcome, regarding arterial recanalization status. A total of 674 patients diagnosed with acute stroke and treated with revascularization therapies were enrolled. Systolic and diastolic BP values of the first 24 h after stroke were collected and their variation quantified through standard deviation. Recanalization state was evaluated at 6 h and clinical outcome at 3 months was assessed by modified Rankin Scale. In multivariate analyses systolic BP variability in the first 24 h post-stroke showed an association with 3 months clinical outcome in the whole population and non-recanalyzed patients. In recanalyzed patients, BP variability did not show a significant association with functional outcome.

Prenatal care: preparation for childbirth in primary healthcare in the south of Brazil. / Pré-natal: preparo para o parto na atenção primária à saúde no sul do Brasil.

OBJECTIVE: To evaluate the relationship between prenatal care and delivery guidelines In Primary Health Care. METHODS: This is a cross-sectional study, with 358 puerperal women of a public maternity from the south of Brazil. The data collection was performed from July to October of 2013, with prenatal card data transcription and a structured interview. The data has been analyzed through the use of the Chi-square test (p≤0.05). RESULTS: The prenatal care had a high coverage (85,5%) and early start by 71,8% of the women, however, 52% of them did not receive orientation for the childbirth. There was a statistical association between receiving orientation for the childbirth and fewer visits (p=0.028), longer interval between the last prenatal visit and the childbirth (p=0.002), and the classification of the prenatal care as intermediate and inadequate (p=0.024). CONCLUSIONS: Despite of the ideal number of visits, the quality of care has been classified as intermediate or inadequate, besides that, precarious access to the orientation for the childbirth during the prenatal care has been evidenced.

Effects of proanthocyanidin on oxidative stress biomarkers and adipokines in army cadets: a placebo-controlled, double-blind study.

PURPOSE: The relatively recent advent of polyphenol supplement for exercise studies has been tested in a variety of forms and doses. However, the dose-response on adipokines and oxidative stress biomarker effect remains unknown. The aim of the present study was to assess the effect of intense, long-duration (48-h) exercise, and a single dose of proanthocyanidin, on plasma leptin, adiponectin, and electronegative low-density lipoprotein (LDL(-)) concentrations. METHODS: Fifty-four healthy male army cadets (22 ± 2 years) participated in a double-blind, randomized, placebo-controlled study and were distributed between control (CG; n = 27) and supplemented groups (SG; n = 27). Immediately before the start of the exercise, both CG and SG groups received a capsule containing starch (200 mg) or proanthocyanidin (dry Vitis vinifera extract, 200 mg), respectively. Following a 12-h fasting period, the plasma adiponectin, leptin, and LDL(-) concentrations were measured prior to the start of the exercise after 24 and 48 h of military training, and after 24 h of rest. The effects of the proanthocyanidin (supplement), exercise (time), and their interaction were investigated using factorial two-way ANOVA. RESULTS: Plasma leptin concentration was only influenced by exercise (p = 0.001). Plasma adiponectin concentration was influenced by exercise (p = 0.037), and by the exercise x supplement interaction (p = 0.033). LDL(-) was influenced by the supplement (p = 0.001), exercise (p = 0.001), and their interaction (p = 0.001). CONCLUSIONS: A single dose of proanthocyanidin (200 mg) was able to reduce LDL(-) concentration and increase plasma adiponectin concentration after 24 h of rest in SG group, indicating its potential protective action.

Meta-analysis of effects of inoculation with homofermentative and facultative heterofermentative lactic acid bacteria on silage fermentation, aerobic stability, and the performance of dairy cows.

Forages are usually inoculated with homofermentative and facultative heterofermentative lactic acid bacteria (LAB) to enhance lactic acid fermentation of forages, but effects of such inoculants on silage quality and the performance of dairy cows are unclear. Therefore, we conducted a meta-analysis to examine the effects of LAB inoculation on silage quality and preservation and the performance of dairy cows. A second objective was to examine the factors affecting the response to silage inoculation with LAB. The studies that met the selection criteria included 130 articles that examined the effects of LAB inoculation on silage quality and 31 articles that investigated dairy cow performance responses. The magnitude of the effect (effect size) was evaluated using raw mean differences (RMD) between inoculated and uninoculated treatments. Heterogeneity was explored by meta-regression and subgroup analysis using forage type, LAB species, LAB application rate, and silo scale (laboratory or farm-scale) as covariates for the silage quality response and forage type, LAB species, diet type [total mixed ration (TMR) or non-TMR], and the level of milk yield of the control cows as covariates for the performance responses. Inoculation with LAB (≥105 cfu/g as fed) markedly increased silage fermentation and dry matter recovery in temperate and tropical grasses, alfalfa, and other legumes. However, inoculation did not improve the fermentation of corn, sorghum, or sugarcane silages. Inoculation with LAB reduced clostridia and mold growth, butyric acid production, and ammonia-nitrogen in all silages, but it had no effect on aerobic stability. Silage inoculation (≥105 cfu/g as fed) increased milk yield and the response had low heterogeneity. However, inoculation had no effect on diet digestibility and feed efficiency. Inoculation with LAB improved the fermentation of grass and legume silages and the performance of dairy cows but did not affect the fermentation of corn, sorghum, and sugar cane silages or the aerobic stability of any silage. Further research is needed to elucidate how silage inoculated with homofermentative and facultative heterofermentative LAB improves the performance of dairy cows.

Info-Gap robustness pathway method for transitioning of urban drainage systems under deep uncertainties.

In the urban water cycle, there are different ways of handling stormwater runoff. Traditional systems mainly rely on underground piped, sometimes named 'gray' infrastructure. New and so-called 'green/blue' ambitions aim for treating and conveying the runoff at the surface. Such concepts are mainly based on ground infiltration and temporal storage. In this work a methodology to create and compare different planning alternatives for stormwater handling on their pathways to a desired system state is presented. Investigations are made to assess the system performance and robustness when facing the deeply uncertain spatial and temporal developments in the future urban fabric, including impacts caused by climate change, urbanization and other disruptive events, like shifts in the network layout and interactions of 'gray' and 'green/blue' structures. With the Info-Gap robustness pathway method, three planning alternatives are evaluated to identify critical performance levels at different stages over time. This novel methodology is applied to a real case study problem where a city relocation process takes place during the upcoming decades. In this case study it is shown that hybrid systems including green infrastructures are more robust with respect to future uncertainties, compared to traditional network design.