Proteomics Analyses of Small Extracellular Vesicles of Aqueous Humor: Identification and Validation of GAS6 and SPP1 as Glaucoma Markers.

Cataracts and glaucoma account for a high percentage of vision loss and blindness worldwide. Small extracellular vesicles (sEVs) are released into different body fluids, including the eye's aqueous humor. Information about their proteome content and characterization in ocular pathologies is not yet well established. In this study, aqueous humor sEVs from healthy individuals, cataracts, and glaucoma patients were studied, and their specific protein profiles were characterized. Moreover, the potential of identified proteins as diagnostic glaucoma biomarkers was evaluated. The protein content of sEVs from patients' aqueous humor with cataracts and glaucoma compared to healthy individuals was analyzed by quantitative proteomics. Validation was performed by western blot (WB) and ELISA. A total of 828 peptides and 192 proteins were identified and quantified. After data analysis with the R program, 8 significantly dysregulated proteins from aqueous humor sEVs in cataracts and 16 in glaucoma showed an expression ratio ≥ 1.5. By WB and ELISA using directly aqueous humor samples, the dysregulation of 9 proteins was mostly confirmed. Importantly, GAS6 and SPP1 showed high diagnostic ability of glaucoma, which in combination allowed for discriminating glaucoma patients from control individuals with an area under the curve of 76.1% and a sensitivity of 65.6% and a specificity of 87.7%.

Capping of Mn-Doped ZnS Quantum Dots with DHLA for Their Stabilization in Aqueous Media: Determination of the Nanoparticle Number Concentration and Surface Ligand Density.

Colloidal Mn2+-doped ZnS quantum dots (QDs) were synthesized, surface modified, and thoroughly characterized using a pool of complementary techniques. Cap exchange of the native l-cysteine coating of the QDs with dihydrolipoic acid (DHLA) ligands is proposed as a strategy to produce nanocrystals with a strong phosphorescent-type emission and improved aqueous stability. Moreover, such a stable DHLA coating can facilitate further bioconjugation of these QDs to biomolecules using established reagents such as cross-linker molecules. First, a structural and morphological characterization of the l-cysteine QD core was performed by resorting to complementary techniques, including X-ray powder diffraction (XRD) and microscopy tools. XRD patterns provided information about the local structure of ions within the nanocrystal structure and the number of metal atoms constituting the core of a QD. The judicious combination of the data obtained from these complementary characterization tools with the analysis of the QDs using inductively coupled plasma-mass spectrometry (ICP-MS) allowed us to assess the number concentration of nanoparticles in an aqueous sample, a key parameter when such materials are going to be used in bioanalytical or toxicological studies. Asymmetric flow field-flow fractionation (AF4) coupled online to ICP-MS detection proved to be an invaluable tool to compute the number of DHLA molecules attached to the surface of a single QD, a key feature that is difficult to estimate in nanoparticles and that critically affects the behavior of nanoparticles when entering the biological media (e.g., cellular uptake, biodistribution, or protein corona formation). This hybrid technique also allowed us to demonstrate that the elemental composition of the nanoparticle core remains unaffected after the ligand exchange process. Finally, the photostability and robustness of the DHLA-capped QDs, critical parameters for bioanalytical applications, were assessed by molecular luminescence spectroscopy.

The talking card: Randomized controlled trial of a novel audio-recording tool for asthma control.

BACKGROUND: Asthma care plans typically include complicated written instructions. Customized, audio-recorded instructions may bridge health literacy gaps and improve treatment plan understanding. OBJECTIVE: To measure the effects of a recordable greeting card-style tool (Talking Card) on asthma control and parental care of children with asthma. METHODS: Multisite randomized trial in two primary care clinics, including children 4-11 years old with uncontrolled asthma and their parents. Parent-child dyads were randomized to usual care of asthma or usual care plus the Talking Card. Dyads completed three asthma-focused visits over 3 months. At the visit, card recipients received customized instructions recorded by the pediatrician onto an audio chip in the card. Asthma control was measured by using the Childhood Asthma Control Test. Card use and parental satisfaction were measured by parental survey (card arm only). Outcomes were analyzed by using generalized estimating equations and frequency distributions. RESULTS: Sixty-four dyads participated and attended 166 clinic visits. Card use was associated with a 1.6-point increase in Childhood Asthma Control Test score (p = 0.02) and a clinic visit regardless of card use with a three-point increase (p < 0.001). Satisfaction and self-efficacy were high among the card users. The mean satisfaction score was 8.9 of 10, with 96% agreeing or strongly agreeing that the card helped them take better care of asthma. CONCLUSIONS: The Talking Card, a novel audio communication tool, was associated with improved asthma control and deemed highly desirable by parents and children struggling to control asthma. This inexpensive portable tool may be useful in other chronic disorders and in locales with low literacy and poor access to digital technology.

Unraveling the nexus of oxidative stress, ocular diseases, and small extracellular vesicles to identify novel glaucoma biomarkers through in-depth proteomics.

Chronic ocular pathologies such as cataracts and glaucoma are emerging as an important problem for public health due to the changes in lifestyle and longevity. These age-related ocular diseases are largely mediated by oxidative stress. Small extracellular vesicles (sEVs) are involved in cell-to-cell communication and transport. There is an increasing interest about the function of small extracellular vesicles (sEVs) in the eye. However, the proteome content and characterization of sEVs released by ocular cells under pathological conditions are not yet well known. Here, we aimed to analyze the protein profile of sEVs and the intracellular protein content from two ocular cell lines (lens epithelial cells and retinal ganglion cells) exposed to oxidative stress to identify altered proteins that could serve as potential diagnostic biomarkers. The protein content was analyzed by quantitative mass spectrometry-based proteomics. Validation was performed by WB and ELISA using cell extracts and aqueous humor from cataract and glaucoma patients. After data analysis, 176 and 7 dysregulated proteins with an expression ratio≥1.5 were identified in lens epithelial cells' protein extract and sEVs, respectively, upon oxidative stress induction. In retinal ganglion cells, oxidative stress induction resulted in the dysregulation of 1033 proteins in cell extracts and 9 proteins in sEVs. In addition, by WB and ELISA, the dysregulation of proteins was mostly confirmed in aqueous humor samples from cataract or glaucoma patients in comparison to ICL individuals, with RAD23B showing high glaucoma diagnostic ability. Importantly, this work expands the knowledge of the proteome characterization of cataracts and glaucoma and provides new potential diagnostic glaucoma biomarkers.

Usefulness of aneurysm sac angiography in therapeutic management of an endoleak after endovascular aneurysm repair.

Endoleak treatment after endovascular aneurysm repair (EVAR) can be difficult and complex. The key to success lies mostly in the accurate interpretation of imaging tests. We describe the case of a patient who was urgently operated on due to an episode of acute arterial ischemia in the left lower limb for stent-graft iliac limb thrombosis and a proximal type I endoleak. We highlight the importance of performing aneurysm sac angiography as part of an effective therapeutic strategy.

Colloidal Physics Modeling Reveals How Per-Ribosome Productivity Increases with Growth Rate in Escherichia coli.

Faster-growing cells must synthesize proteins more quickly. Increased ribosome abundance only partly accounts for increases in total protein synthesis rates. The productivity of individual ribosomes must increase too, almost doubling by an unknown mechanism. Prior models point to diffusive transport as a limiting factor but raise a paradox: faster-growing cells are more crowded, yet crowding slows diffusion. We suspected that physical crowding, transport, and stoichiometry, considered together, might reveal a more nuanced explanation. To investigate, we built a first-principles physics-based model of Escherichia coli cytoplasm in which Brownian motion and diffusion arise directly from physical interactions between individual molecules of finite size, density, and physiological abundance. Using our microscopically detailed model, we predicted that physical transport of individual ternary complexes accounts for ~80% of translation elongation latency. We also found that volumetric crowding increases during faster growth even as cytoplasmic mass density remains relatively constant. Despite slowed diffusion, we predicted that improved proximity between ternary complexes and ribosomes wins out, illustrating a simple physics-based mechanism for how individual elongating ribosomes become more productive. We speculate that crowding imposes a physical limit on growth rate and undergirds cellular behavior more broadly. Unfitted colloidal-scale modeling offers systems biology a complementary "physics engine" for exploring how cellular-scale behaviors arise from physical transport and reactions among individual molecules. IMPORTANCE Ribosomes are the factories in cells that synthesize proteins. When cells grow faster, there are not enough ribosomes to keep up with the demand for faster protein synthesis without individual ribosomes becoming more productive. Yet, faster-growing cells are more crowded, seemingly making it harder for each ribosome to do its work. Our computational model of the physics of translation elongation reveals the underlying mechanism for how individual ribosomes become more productive: proximity and stoichiometry of translation molecules overcome crowding. Our model also suggests a universal physical limitation of cell growth rates.

Fungal Diseases in Elasmobranchs and Their Possible Treatment with a Special Mention to Azole Antifungal Agents.

INTRODUCTION: Elasmobranchs currently constitute an important part of the animal collection of many aquariums worldwide. Their maintenance under human care has allowed us to describe and identify new pathogens and diseases affecting them, as well as to determine different treatments for these diseases. Great advances in elasmobranch husbandry have been developed. METHODS: A search was performed on scientific databases as PubMed and other specialized sources (IAAAM archive). RESULTS: Little information on pharmacotherapeutics is available in this taxonomic group, and treatments lack a scientific base and instead are frequently dependent on empirical knowledge. Pharmacokinetic studies are the first step to determining therapeutic protocols that are safe and effective. The available bibliography shows that a majority of the mycoses recorded in cartilaginous fish are severe, aggravated by the fact that the antifungal treatments administered, following the guidelines used for teleost species, are ineffective in elasmobranchs. Azoles appear to be a promising group of antifungals for use in treating systemic mycoses in sharks and rays. CONCLUSIONS: Based on the findings of this review, it is essential to investigate the pharmacokinetics of the different antifungals in these species in order to provide therapeutic options for fungal infections in cartilaginous fish.

Leptospirosis: Epidemiological and Clinical Characteristics at the National Reference Hospital in Panama.

Leptospirosis represents a public health problem in Panama, with an incidence rate of 1 in 100,000 inhabitants in 2014. Despite active surveillance and reports of outbreaks in the news, publications about human leptospirosis in Panama are scarce. The objective of this study was to describe the epidemiological and clinical features of leptospirosis in a cohort of patients admitted to the national reference hospital from January 2013 to December 2018. A total of 188 patients with suspected leptospirosis were identified, but only 56.9% (107 of 188) of the medical records could be retrieved. Microagglutination assays were completed in 45% (48 of 107) of the patients, confirming leptospirosis in 29.2% (14 of 48) of the patients. The most prevalent serogroup identified was Leptospira interrogans icterohemorrhagiae (4 of 14, 28.6%). The majority of patients with confirmed disease were middle-aged (36.4 ± 15.7 years), male (11 of 14, 78.6%), and symptomatic for 6.8 ± 0.7 days before admission. The predominant clinical presentation was fever (13 of 14, 92.9%), abdominal pain (7 of 14, 50%), and jaundice (8 of 14, 57.1%). Respiratory failure (8 of 14, 57.1%), elevated creatinine levels on admission (8 of 14, 57.1%), transfusion of blood-derived products (6 of 14, 42.9%), and required use of vasopressors (4 of 14, 28.6%) were common complications. Mortality was 28.6% (4 of 14). Empiric antibiotic therapy was initiated in almost all patients (10 of 12, 83.3%), and was appropriate in 90% (9 of 10) of them. Our study highlights the high prevalence of severe disease and reveals the diagnostic challenges concealing the true burden of leptospirosis in Panama. However, the small number of confirmed patients limits the generalization of these findings.

Carcinoid Heart Disease From a Primary Liver Carcinoid Tumor.

Carcinoid heart disease (CaHD) is part of the carcinoid syndrome. Cardiac involvement is present in 20% to 60% of patients with carcinoid syndrome and is normally from liver metastasis. We report the case of a patient who presented with CaHD disease with an undiagnosed primary tumor or a possible primary liver carcinoid tumor. (Level of Difficulty: Advanced.).

Nanoparticles as fluorescent labels for optical imaging and sensing in genomics and proteomics.

Optical labelling reagents (dyes and fluorophores) are an essential component of probe-based biomolecule detection, an approach widely employed in a variety of areas including environmental analysis, disease diagnostics, pharmaceutical screening, and proteomic and genomic studies. Recently, functional nanomaterials, as a new generation of high-value optical labels, have been applied to molecular detection. The great potential of such recent optical labels has paved the way for the development of new biomolecule assays with unprecedented analytical performance characteristics, related to sensitivity, multiplexing capability, sample throughput, cost-effectiveness and ease of use. This review aims to provide an overview of recent advances using different nanoparticles (such as quantum dots, rare earth doped nanoparticles or gold nanoparticles) for analytical genomics and proteomics, with particular emphasis on the outlook for different strategies of using nanoparticles for bioimaging and quantitative bioanalytical applications, as well as possibilities and limitations of nanoparticles in such a growing field.

Predictive markers of nonalcoholic fatty liver disease in lean patients. A multinomial regression model and a 2k factorial analysis.

OBJECTIVE: Nonalcoholic fatty liver disease (NAFLD) is associated with obesity and insulin resistance; however, there is a group of non-obese patients with NAFLD that need to be characterized. Our aim was to evaluate the factors associated with NAFLD in non-obese subjects in a third-level hospital. METHODS: A comparative cross-sectional study was performed. Participants were divided into four groups: non-obese without NAFLD (group 1), non-obese with NAFLD (group 2), obese without NAFLD (group 3), and obese with NAFLD (group 4). We evaluated the effect of clinical and biochemical characteristics with the disease by groups using a multinomial regression model and a 2K factorial analysis. RESULTS: We included 278 participants. Low platelet-lymphocyte ratio (PLR) as a novel parameter associated with NAFLD in non-obese subjects. Age, uric acid, alanine transaminase (ALT), high-density lipoprotein (HDL)-cholesterol, and neutrophil-lymphocyte ratio (NLR) were other related parameters (akaike information criterion = 557). NLR had the larger OR in groups with NAFLD (lean with NAFLD 7.12, obese with NAFLD 13.02). The 2k factorial design found inverse effect on PLR by NAFLD (effect -21.89, P < 0.001), which was higher than BMI (effect -1.33, P < 0.045). CONCLUSION: Our study found that PLR is a novel parameter with inverse correlation with NAFLD in non-obese patients. Other related parameters are age, hyperuricemia, elevation of ALT and NLR, and low HDL-cholesterol.

Fatal vascular complication due to metallosis.

Metallosis is a well-known complication at the site of total hip and knee arthroplasty. Vascular involvement of this complication is rare and generally results from vascular compression by a mass or pseudotumor. In the present report, we have described a case of lower limb acute arterial ischemia due to arterial injury as a complication of metallosis with a fatal outcome.

Dyslipidemia as a risk factor for liver fibrosis progression in a multicentric population with non-alcoholic steatohepatitis.

Background: Nonalcoholic fatty liver disease (NAFLD) is a serious worldwide health problem, with an estimated global prevalence of 24%; it has a notable relationship with other metabolic disorders, like obesity and type 2 diabetes mellitus (T2DM). Nonalcoholic steatohepatitis (NASH) is one of the most important clinical entities of NAFLD, which is associated with an increased risk of progression to liver cirrhosis and hepatocellular carcinoma (HCC). Mexico is one of the countries with the highest prevalence of metabolic diseases; therefore, we sought to investigate the impact that these clinical entities have in the progression to advanced fibrosis in Mexican patients with NASH. Methods: We performed a multicenter retrospective cross-sectional study, from January 2012 to December 2017. A total of 215 patients with biopsy-proven NASH and fibrosis were enrolled. NASH was diagnosed according NAS score and liver fibrosis was staged by the Kleiner scoring system. For comparing the risk of liver fibrosis progression, we divided our sample into two groups. Those patients with stage F0-F2 liver fibrosis were included in the group with non-significant liver fibrosis (n=178) and those individuals with F3-F4 fibrosis were included in the significant fibrosis group (n=37). We carried out a multivariate analysis to find risk factors associated with liver fibrosis progression. Results: From the 215 patients included, 37 had significant liver fibrosis (F3-4). After logistic regression analysis T2DM (p=0.044), systemic arterial hypertension (p=0.014), cholesterol (p=0.041) and triglycerides (p=0.015) were the main predictor of advanced liver fibrosis. Conclusions: In a Mexican population, dyslipidemia was the most important risk factor associated with advanced liver fibrosis and cirrhosis.

Duplex mapping of 2036 primary varicose veins.

OBJECTIVE: To produce a comprehensive anatomical and hemodynamic ultrasound scan mapping of the whole limb in patients with primary varicose veins (VVs). DESIGN: An analysis of venous duplex scans performed on patients referred for treatment of primary VVs. METHODS: A total of 2036 limbs were evaluated, looking for the origin of VVs in the saphenous systems and in the perforating vein (PV) systems, as well as for the presence of non-saphenous reflux. RESULTS: The sapheno-femoral junction (SFJ) of the great saphenous vein (GSV) was involved in 1330 limbs (65.3%). We have noted that finding reflux in the groin does not imply that it originates at that point necessarily, as reflux from the pelvis or abdominal wall can also cause primary VVs (SFJ reflux, 41.9% and competent SFJ with reflux from proximal veins, 35.4%). We also noted that analyzing only the presence of reflux in the SFJ of the GSV would miss 10.9% of limbs of reflux in the SFJ of the anterior accessory GSV. In 237 limbs (11.6%), reflux was observed in the popliteal fossa. In the PV system, we distinguished those PVs with retrograde flow that acted as an origin of the VVs, and other PVs that acted as re-entry points. Based on this difference, the location identified as the most frequent origin of VVs in the PV system was the thigh, specifically in the group of PVs of the medial thigh of the femoral canal, with 85 PVs with a total of 238 incompetent PVs identified. Pure non-saphenous reflux was observed in 162 limbs (8%). CONCLUSION: The assumption that the origin of VVs would be exclusively in the sapheno-femoral or sapheno-popliteal junction, is a mistaken attitude and a comprehensive duplex scan mapping is recommended.

Acute pancreatitis in children with acute lymphoblastic leukemia treated with L-asparaginase.

BACKGROUND: L-asparaginase (L-asp) is used as part of the initial treatment in children with acute lymphoblastic leukemia (ALL), inducing remission in 83% to 95% of the treated patients. Major toxicity effects reported are hypersensitivity reactions and dysfunctions of the liver and pancreas. Acute pancreatitis (AP) induced by L-asp has been noted in 2.5% to 16% of the treated patients. The purpose of this study was to determine the frequency and outcome of AP in children with ALL treated with L-asp in a tertiary care pediatric hospital. METHODS: From January 1999 to June 2005, the charts of children with ALL admitted for L-asp treatment were reviewed. Data from children who developed AP were analyzed retrospectively. AP was defined as the presence of clinical data (nausea, vomiting, and abdominal pain), elevated pancreatic enzymes, and changes in the abdominal ultrasound and/or computed tomography (CT) scan. Clinical and biochemical data, abdominal ultrasound, and CT scan findings, complications, treatment, and outcome were analyzed retrospectively. RESULTS: During the last 6 years, 266 ALL new cases were started on chemotherapy including L-asp, of which 18 of 266 (6.7%) developed AP. Pancreatic necrosis by CT scan was found in 10 patients, peripancreatic collections and pseudocyst formation were detected in 5 and 3 cases, respectively, and resolved by cystogastrostomy or drainage. Two patients developed chronic pancreatitis and 3 diabetes. There was no relationship between number of doses and pancreatic toxicity. None of the patient died due to pancreatic toxicity. CONCLUSIONS: L-asp is an effective drug to treat ALL, the administration of L-asp requires the monitoring of pancreatic toxicity to detect AP and have treatment initiated as early as possible. Chronic complications after AP occur in almost one third of cases.

Celiac disease pathogenesis: the proinflammatory cytokine network.

In susceptible individuals, the adaptive response, mediated by the activation of antigen-specific T lymphocytes, drives a proinflammatory response, which ends in an immune-mediated enteropathy characterized by villous atrophy, crypt hyperplasia, and recruitment of intraepithelial lymphocytes. In addition, some gluten peptides are able to induce an innate immune response in intestinal mucosa. The molecular mechanisms and the cells involved in the initial stages of the gluten-intestinal mucosa interaction are poorly understood to date. There is evidence of a direct toxic effect of gluten peptides in several biological models. However, the failure to control the inflammatory response may be one of the factors underlying gluten intolerance in these individuals. The cytokine network involved in celiac disease is characterized by abundant interferon-gamma in the intestinal mucosa. In addition, the production of interleukin (IL)-15, IL-18, and IL-21 is linked to gluten intake, which can drive the inflammatory response probably sustained by IL-18, IL-21, and perhaps IL-27 through STAT1 and STAT5 pathways, whereas neither IL-12 nor IL-23 plays a significant role in pathogenic mechanisms. Herein we describe the involvement of these activation pathways in the context of the pathogenesis of celiac disease.

Epidemiology of home enteral nutrition: an approximation to reality.

INTRODUCTION: home enteral nutrition (HEN) has undergone an important development; however, there is a notable lack of information with regard to its incidence and characteristics. OBJECTIVES: our aim was to assess the state of HEN in our area. METHODS: an observational, prospective study, involving all patients who had initiated HEN in the Nutrition Unit during a year. Epidemiological, functional, and nutritional evolution of the patients was described and incidence of HEN was calculated. RESULTS: HEN incidences totalled 229/100,000 inhabitants/year. The HEN population in our area was characterized by the aged and a high frequency of comorbidity and functional limitations. Neurological and oncological diseases accounted for 50% of indications. The remaining cases were malnourished patients who had received short periods of HEN after hospitalization or a hip fracture. Oral supplements (60%) with standard and hypercaloric formulas were used the most. At baseline, 75% of the patients suffered from malnutrition. During the follow-up, patients showed weight gain (1.6%), an increase in the percentage of normal weight and overweight (from 74% to 82.7%, p = 0.001) and a reduction in pressure ulcers (15.7% vs10.3%, p < 0.001). The median duration of HEN was 8.5 months. Only a quarter of the patients experienced complications (mostly mild gastrointestinal complications); 43.1% had died at the end of the follow-up. CONCLUSIONS: in our area, the HEN incidence was much higher than those described in the literature. HEN appears to be a safe therapy with few complications that improves the nutritional status of the patients, even with short periods of administration.

Current trends of liver cirrhosis in Mexico: Similitudes and differences with other world regions.

AIM: To investigate the main current etiologies of cirrhosis in Mexico. METHODS: We performed a cross-sectional retrospective multicenter study that included eight hospitals in different areas of Mexico. These hospitals provide health care to people of diverse social classes. The inclusion criteria were a histological, clinical, biochemical, endoscopic, or imaging diagnosis of liver cirrhosis. Data were obtained during a 5-year period (January 2012-December 2017). RESULTS: A total of 1210 patients were included. The mean age was 62.5 years (SD = 12.1), and the percentages of men and women were similar (52.0% vs 48.0%). The most frequent causes of liver cirrhosis were hepatitis C virus (HCV) (36.2%), alcoholic liver disease (ALD) (31.2%), and nonalcoholic steatohepatitis (23.2%), and the least frequent were hepatitis B virus (1.1%), autoimmune disorders (7.3%), and other conditions (1.0%). CONCLUSION: HCV and ALD are the most frequent causes of cirrhosis in Mexico. However, we note that non-alcoholic fatty liver disease (NAFLD) as an etiology of cirrhosis increased by 100% compared with the rate noted previously. We conclude that NAFLD will soon become one of the most frequent etiologies of liver cirrhosis in Mexico.

Papel del cirujano vascular en la fijación anterior de columna / Role of the vascular surgeon in anterior spine fixation

La fijación anterior de columna es una técnica bien implantada en la cirugía espinal. La presencia y necesidad de manipular los vasos principales, la aorta y las ilíacas y el riesgo hemorrágico que ello presenta hacen recomendable la colaboración de un cirujano vascular como cirujano de abordaje durante los procedimientos. Presentamos la experiencia de nuestro hospital en el tratamiento de fijación anterior de columna entre los servicios de traumatología y de cirugía vascular con una serie de 28 casos comprendidos entre el 2017 y el 2021.(AU)

Anterior spinal fixation is a well-established technique in spinal surgery: The presence and need for manipulation of the main vessels, aorta and iliac, and the risk of bleeding that presents, makes it advisable to have a Vascular Surgeon as an approach surgeon during the procedures. We present the experience of our Hospital in the treatment of anterior fixation of the experience of our Hospital in the treatment of anterior of the spine with the traumatology and vascular surgery service with a series of 28 cases between 2017 and 2021.(AU)