Study protocol for a randomized, controlled trial using a novel, family-centered diet treatment to prevent nonalcoholic fatty liver disease in Hispanic children.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is the leading liver disorder among U.S. children and is most prevalent among Hispanic children with obesity. Previous research has shown that reducing the consumption of free sugars (added sugars + naturally occurring sugars in fruit juice) can reverse liver steatosis in adolescents with NAFLD. This study aims to determine if a low-free sugar diet (LFSD) can prevent liver fat accumulation and NAFLD in high-risk children. METHODS: In this randomized controlled trial, we will enroll 140 Hispanic children aged 6 to 9 years who are ≥50th percentile BMI and without a previous diagnosis of NAFLD. Participants will be randomly assigned to either an experimental (LFSD) or a control (usual diet + educational materials) group. The one-year intervention includes removal of foods high in free sugars from the home at baseline, provision of LFSD household groceries for the entire family (weeks 1-4, 12, 24, and 36), dietitian-guided family grocery shopping sessions (weeks 12, 24, and 36), and ongoing education and motivational interviewing to promote LFSD. Both groups complete assessment measures at baseline, 6, 12, 18, and 24 months. Primary study outcomes are percent hepatic fat at 12 months and incidence of clinically significant hepatic steatosis (>5%) + elevated liver enzymes at 24 months. Secondary outcomes include metabolic markers potentially mediating or moderating NAFLD pathogenesis. DISCUSSION: This protocol describes the rationale, eligibility criteria, recruitment strategies, analysis plan as well as a novel dietary intervention design. Study results will inform future dietary guidelines for pediatric NAFLD prevention. TRIAL REGISTRATION: ClinicalTrials.gov, NCT05292352.

The influence of acetylsalicylic acid on bone regeneration: systematic review and meta-analysis.

Acetylsalicylic acid (ASA) is commonly used as a non-steroidal anti-inflammatory drug that interferes with multiple biological pathways. ASA acts by stimulating osteogenesis and inhibiting osteoclastogenesis. Thus, the objective of this study was to perform a systematic review and meta-analysis to evaluate the effectiveness of the use of ASA in the bone regeneration in animal models. This review was structured based on the PRISMA Statement and registered on PROSPERO database according to protocol number #CDR42018111403. The quality of evidence was assessed by using the Grades of Recommendation, Assessment, Development, and Evaluation (GRADE). With the development of search strategies, we identified studies on the use of ASA from the following databases: 1- Medline (via PubMed); 2 - Web of Science; 3 - Scopus; and 4 - EMBASE. A total of 296 articles were identified and after screening the title, abstract, and full text, only 18 studies were selected for qualitative analysis and 12 were selected for performance of the quantitative analysis (meta-analysis). A meta-analysis of the amount of bone tissue formed showed a significant advantage when ASA was locally used, revealing a mean difference (MD) of 22.75% (95% CI: 15.39-30.12) p < 0.00001. Within the limitations of the available data, the results were promising and showed that ASA can be effective in bone formation in animal models.

Permeation of beta-defensin-3 encapsulated with polyethylene glycol in lung surfactant models at air-water interface.

Coarse grained molecular dynamics of the permeation of the peptide human beta-defensin-3 (HBD3) in two different lung surfactant models (BLES and CUROSURF) at surface tension of 20 mN m-1 was performed in order to simulate the effect of charge distribution on the permeation process. We found that HBD3 permeates in lung surfactant models at surface tension of 20 mN m-1. However, it sticks to the phospholipid polar heads, possibly having its bactericidal properties inactivated by electrostatic interaction. The HBD3 encapsulated with polyethylene glycol (PEG) molecules does permeate, but it does not interact with the polar heads. The Gibbs free energies of this process in both models were estimated to be -39 and -64 Kcal mol-1 for HBD3 and PEG-encapsulated HBD3, respectively. The peptide HBD3 was pulled from the polar heads to the water phase overcoming an energy barrier of around 17 and 22 Kcal mol-1, respectively. The peptide was encapsulated with PEG molecules in order to reduce this barrier, thereby decreasing to roughly 12 and 0 Kcal mol-1 in BLES and CUROSURF models, respectively. After permeation of the HBD3 encapsulated with PEG, it freely moves in the water phase with little or none interaction with the phospholipid head groups. Lung surfactant with low charge distribution seems to have less or none electrostatic interaction with HBD3, possibly preserving its antimicrobial activity.

IL-4 receptor α blockade prevents sensitization and alters acute and long-lasting effects of allergen-specific immunotherapy of murine allergic asthma.

BACKGROUND: Allergen-specific immunotherapy (AIT) is the only causal treatment for allergy. However, success rates vary depending on the type of allergy and disease background of the patient. Hence, strategies targeting an increased therapeutic efficacy are urgently needed. Here, the effects of blockade of IL-4 and IL-13 signaling on different phases of AIT were addressed. METHODS: The impact of the recombinantly produced IL-4 and IL-13 antagonist IL-4 mutein (IL-4M) on allergic sensitization and AIT outcome in experimental allergic asthma were analyzed in a murine model. The effects of IL-4M administration were assessed prior/during sensitization, immediately after AIT under allergen challenge, and two weeks post-treatment. RESULTS: Intervention with IL-4M prior/during sensitization led to strong induction of IgG1, IgG2a, IgG2b, and IgG3, decrease of specific and total IgE, as well as of IL-5 in serum. Similar effects on the serum immunoglobulin levels were observed immediately after IL4M-supplemented AIT during allergen challenge. Additionally, IL4M markedly suppressed type-2 cytokine secretion of splenocytes beyond the effect of AIT alone. These effects were equaled to those of AIT alone two weeks post-treatment. Intriguingly, here, IL-4M induced a sustained decrease of Th2-biased Tregs (ST2+ FOXP3+ GATA3intermediate ). CONCLUSIONS: IL-4 and IL-13 blockade during experimental AIT demonstrates beneficial effects on immunological key parameters such as immunoglobulin and cytokine secretion immediately after AIT. Although two weeks later these effects were dropped to those of AIT alone, the number of potentially disease-triggering Th2-biased Tregs was further significantly decreased by IL-4M treatment. Hence, IL-4/IL13-targeting therapies prime the immune memory in therapy success-favoring manner.

Penetration of antimicrobial peptides in a lung surfactant model.

Molecular dynamics simulations were successfully performed to understand the absorption mechanism of antimicrobial peptides LL-37, CATH-2, and SMAP-29 in a lung surfactant model. The antimicrobial peptides quickly penetrate in the lung surfactant model in dozens or hundreds nanoseconds, but they electrostatically interact with the lipid polar heads during the simulation time of 2 µs. This electrostatic interaction should be the explanation for the inactivation of the antimicrobial peptides when co-administrated with lung surfactant. As they strongly interact with the lipid polar heads of the lung surfactant, there is no positive charge available on the antimicrobial peptide to attack the negatively charged bacteria membrane. In order to avoid the interaction of peptides with the lipid polar heads, sodium cholate was used to form nanoparticles which act as an absorption enhancer of all antimicrobial peptides used in this investigation. The nanoparticles of 150 molecules of sodium cholate with one peptide were inserted on the top of the lung surfactant model. The nanoparticles penetrated into the lung surfactant model, spreading the sodium cholate molecules around the lipid polar heads. The sodium cholate molecules seem to protect the peptides from the interaction with the lipid polar heads, leaving them free to be delivered to the water phase. The penetration of peptides alone or even the peptide nanoparticles with sodium cholate do not collapse the lung surfactant model, indicating to be a promisor drug delivery system to the lung. The implications of this finding are that antimicrobial peptides may only be co-administered with an absorption enhancer such as sodium cholate into lung surfactant in order to avoid inactivation of their antimicrobial activity.

The roles of mucus-forming mucins, peritrophins and peritrophins with mucin domains in the insect midgut.

Most insects have a gut lined with a peritrophic membrane (PM) consisting of chitin and proteins, mainly peritrophins that have chitin-binding domains. The PM is proposed to originate from mucus-forming mucins (Mf-mucins), which acquired a chitin-binding domain that interlocked with chitin, replacing mucus in function. We evaluated the expression of Mf-mucins and peritrophins by RNA-sequencing (RNA-seq) throughout the midgut of four distantly related insects. Mf-mucins were identified as proteins with high o-glycosylation and a series of uninterrupted Pro/Thr/Ser residues. The results demonstrate that the mucus layer is widespread in insects, and suggest that insect Mf-mucins are derived from those found in other animals by the loss of the cysteine knot and von Willebrand domains. The data also support a role of Mf-mucins in protecting the middle midgut of Musca domestica against acidic buffers. Mf-mucins may also produce a jelly-like material associated with the PM that immobilizes digestive enzymes in Spodoptera frugiperda. Peritrophins with a domain similar to Mf-mucins may be close to the ancestor of peritrophins. Expression data of peritrophins and chitin synthase genes throughout the midgut of M. domestica, S. frugiperda and Tenebrio molitor indicated that peritrophins were incorporated along the PM, according to their preferential sites of formation. Finally, the data support the view that mucus has functions distinct from the PM.

MALDI MS imaging investigation of the host response to visceral leishmaniasis.

Mass spectrometry imaging (MSI) of animal tissues has become an important tool for in situ molecular analyses and biomarker studies in several clinical areas, but there are few applications in parasitological studies. Leishmaniasis is a neglected tropical disease, and experimental mouse models have been essential to evaluate pathological and immunological processes and to develop diagnostic methods. Herein we have employed MALDI MSI to examine peptides and low molecular weight proteins (2 to 20 kDa) differentially expressed in the liver during visceral leishmaniasis in mice models. We analyzed liver sections of Balb/c mice infected with Leishmania infantum using the SCiLS Lab software for statistical analysis, which facilitated data interpretation and thus highlighted several key proteins and/or peptides. We proposed a decision tree classification for visceral leishmaniasis with distinct phases of the disease, which are named here as healthy, acute infection and chronic infection. Among others, the ion of m/z 4963 was the most important to identify acute infection and was tentatively identified as Thymosin ß4. This peptide was previously established as a recovery factor in the human liver and might participate in the response of mice to Leishmania infection. This preliminary investigation shows the potential of MALDI MSI to complement classical compound selective imaging techniques and to explore new features not yet recognized by these approaches.

Masking Colored Substrates Using Monolithic and Bilayer CAD-CAM Ceramic Structures.

OBJECTIVE: To evaluate the masking ability and translucency of monolithic and bilayer CAD-CAM ceramic structures. METHODS: Discs of high translucency (HT) and low translucency (LT) lithium disilicate-based ceramic (IPS e.max CAD) with different thicknesses (0.7, 1, 1.5, and 2 mm) were evaluated as a monolithic structure or combined (bilayer) with a 0.5-mm-thick zirconia framework (IPS e.max ZirCAD). The masking ability and translucency were calculated based on CIE L*a*b* color coordinates measured with a spectrophotometer (SP60, X-Rite). The translucency parameter (TP) was calculated using color coordinates measured over standard white-and-black backgrounds. The masking ability was calculated by CIEDE2000 color difference metric (ΔE00) for each specimen measured over a tooth-colored substrate (shade A2) compared to three darker backgrounds (shade C4 and two metal substrates). Confidence intervals (CI) for the means (95% CI) were calculated for TP and ΔE00. The Pearson correlation between ΔE00 and TP was investigated for monolithic and bilayer structures over all backgrounds. RESULTS: The thinner the lithium disilicate layer, the greater the translucency and the higher the ΔE00 values. The effect of ceramic thickness on both translucency and masking ability was more pronounced for the monolithic structures. In addition, monolayers always presented a greater color variation than their bilayer counterparts. The metallic background produced greater ΔE00 than the C4-shaded substrate. CONCLUSION: Monolithic veneers were able to mask C4-shaded background but did not mask metallic backgrounds. Bilayer structures showed greater shade masking ability than monolithic structures.

Management Strategies Aiming to Improve Horse Welfare Reduce Embryonic Death Rates in Mares.

The objective of this retrospective study was to evaluate the effect of management strategies aiming to improve animal well-being on pregnancy and embryonic death (ED) rates. Breeding records of a cohort of 1206 Thoroughbred mares brought to a stallion station facility, to be bred with the stallions housed there, were evaluated during ten breeding seasons. Mares were blocked according to management strategies in two groups: Stress and Relax. Strategies used to improve animal well-being (Relax group) were as follows: stopping the teasing routine, reducing or eliminating stall confinement, reducing the number of mares per group and maintaining herd stability during the breeding season. In barren mares, the pregnancy rate was higher in the Relax group (91.8%) when compared to the observed in Stress group (84.7%). However, no difference in pregnancy rates were observed (Stress = 85.2% vs. Relax = 86.2) in foaling mares. ED rate was higher in barren and foaling mares of the Stress group mares (25.5% and 26.8%, respectively) compared with the Relax group (16.1% and 14.7%, respectively). No significant differences were observed on foal heat pregnancy rate between groups; yet, the embryo loss on foal heat was significant reduced in Relax mares (Relax = 8.7% vs Stress = 24.5%). In conclusion, management strategies aimed to reduce social stress can reduce early pregnancy losses and the average cycles per pregnancy, improving reproductive performance in mares.

Histological correlates of N40 auditory evoked potentials in adult rats after neonatal ventral hippocampal lesion: animal model of schizophrenia.

The neonatal ventral hippocampal lesion (NVHL) is an established neurodevelopmental rat model of schizophrenia. Rats with NVHL exhibit several behavioral, molecular and physiological abnormalities that are similar to those found in schizophrenics. Schizophrenia is a severe psychiatric illness characterized by profound disturbances of mental functions including neurophysiological deficits in brain information processing. These deficits can be assessed by auditory evoked potentials (AEPs), where schizophrenics exhibit abnormalities in amplitude, duration and latency of such AEPs. The aim of the present study was to compare the density of cells in the temporal cerebral cortex and the N40-AEP of adult NVHL rats versus adult sham rats. We found that rats with NVHL exhibit significant lower amplitude of the N40-AEP and a significant lower number of cells in bilateral regions of the temporal cerebral cortex compared to sham rats. Because the AEP recordings were obtained from anesthetized rats, we suggest that NVHL leads to inappropriate innervation in thalamic-cortical pathways in the adult rat, leading to altered function of cortical networks involved in processing of primary auditory information.

Hidradenitis suppurativa: retrospective study of 20 cases.

In this study, twenty cases of severe hidradenitis suppurativa are reported, mainly in non-white people and in axillary areas. Wide surgical excision has offered good results, although relapses have occurred at variable intervals in the follow-up period.

Hidradenitis suppurativa: retrospective study of 20 cases / Hidradenite supurativa: estudo retrospectivo de 20 casos

In this study, twenty cases of severe hidradenitis suppurativa are reported, mainly in non-white people and in axillary areas. Wide surgical excision has offered good results, although relapses have occurred at variable intervals in the follow-up period.

São relatados 20 casos de hidradenite supurativa grave, com predomínio de não brancos e regiões axilares. A excisão cirúrgica ampla trouxe bons resultados, embora a recidiva ocorresse em intervalos variáveis no acompanhamento.

Influence of the layer thickness in plasmonic gold nanoparticles produced by thermal evaporation.

Metallic nanoparticles (NPs) have received recently considerable interest of photonic and photovoltaic communities. In this work, we report the optoelectronic properties of gold NPs (Au-NPs) obtained by depositing very thin gold layers on glass substrates through thermal evaporation electron-beam assisted process. The effect of mass thickness of the layer was evaluated. The polycrystalline Au-NPs, with grain sizes of 14 and 19 nm tend to be elongated in one direction as the mass thickness increase. A 2 nm layer deposited at 250°C led to the formation of Au-NPs with 10-20 nm average size, obtained by SEM images, while for a 5 nm layer the wide size elongates from 25 to 150 nm with a mean at 75 nm. In the near infrared region was observed an absorption enhancement of amorphous silicon films deposited onto the Au-NPs layers with a corresponding increase in the PL peak for the same wavelength region.

Delayed reconstruction after eyelid tumor excision.

BACKGROUND: The aim of this study is to report the feasibility of one to two weeks delayed reconstruction after eyelid tumor excision. PATIENTS AND METHODS: A retrospective case-series study was designed. Fourteen consecutive outpatients referred to a tertiary referral eye center for eyelid tumor management were enrolled. The intervention imvolved eyelid tumor excision (with four millimeters surrounding clinically tumor-free tissue), compressive patching of the excised area until reconstruction (with or without a therapeutic contact lens), extensive histological work-up to assess histological tumor clearance, and reconstruction one week after excision (in a couple of patients after two weeks when further excision was necessary to achieve tumor clearance). Excision and closure were performed by a single external trained oculoplastic surgeon (IOH) with a day per week (Fridays) operating slot. As outcome we assessed whether such an approach has led before, during, or after eyelid reconstruction to unusual major adverse events (follow-up time: 18 - 36 months). RESULTS: Delaying up to one week, in some cases even two weeks, reconstruction after eyelid tumor excision was not associated with any unusual major adverse events before, during, or after surgery. CONCLUSIONS: When necessary, after eyelid tumor excision, it is possible to delay for one week or even for two weeks reconstruction, apparently without experiencing major unusual adverse events.

Mucosal excision instead of fashioning nasolacrimal mucosae flaps during external dacryocystorhinostomy: a pilot study.

BACKGROUND: To report results of a simplified external dacryocytorhinostomy procedure in which nasal and lacrimal sac mucosal flaps are simply removed instead of being sutured together. DESIGN: Retrospective non-comparative case-series study. PARTICIPANTS: Fourteen consecutive outpatients patients undergoing external dacryocytorhinostomy. INTERVENTION: Modified and simplified transcutaneous external dacryocytorhinostomy where basically the lacrimal sac and the nasal mucosae are widely excised in front of DCR's osteotomy instead of being used to fashion nasolacrimal flaps. OUTCOME: Assessment of dacryocytorhinostomy anatomical patency by syringing and patient's self-perception of epiphora symptoms improvement about twelve weeks and one year after surgery, as well as patient's report of ocular air-reflux during Valsalva maneuver a year after surgery. RESULTS: Syringing showed anatomic patency in 93 % and 92 % of patients twelve weeks (mean ± SD: 12 weeks ± 6 weeks) and one year (15 months ± 3 months) after surgery, respectively. One year after surgery, 62 % of patients reported ocular air-reflux. Twelve weeks and one year after surgery, mean symptom improvement was 82 % ± 17 % and 79 % ± 29 %, respectively. Furthermore, 79 % and 85 % of patients reported a postoperative improvement of their symptoms greater or equal to 80 % after twelve weeks and one year, respectively. CONCLUSIONS: Removing lacrimal sac and nasal mucosae in front of the osteotomy did not appear to have a major negative impact on the outcome of the external dacryocytorhinostomy. The findings of this pilot study need to be confirmed by a larger prospective trial.

Improvement of chronic epiphora symptoms after surgery in patients with different preoperative schirmer-test values.

BACKGROUND: To compare improvement of epiphora symptoms after surgery between patients who had different preoperative basal Schirmer test values. DESIGN: Retrospective comparative case-series study. PARTICIPANTS: Twenty-two consecutive patients (one eye/patient) undergoing surgery for chronic epiphora symptoms resistant to ≥ 6 months of medical treatments (who incidentally had a preoperative basal [oxybuprocain-HCL 0.4 %] Schirmer-test and a postoperative one-year follow-up) were ranked according to their preoperative basal Schirmer test values and divided into two groups of equal size: group I, patients with lower (0 to 6 mm) and group II, patients with higher (7 to 25 mm) Schirmer test values. INTERVENTION: Combination of Piffaretti's endocanlicular endoscopic lacrimal drainage (22 patients), lacrimal intubation (8 patients), lacrimal punctoplasty (22 patients), lateral canthoplasty (18 patients), and/or conjunctivochalasis (8 patients) surgical procedures. OUTCOME: Subjective self-perceptions of improvement of their epiphora symptoms was made by the patients one-year after surgery. RESULTS: While between groups I and II mean (± SD) preoperative basal Schirmer test values significantly differed (4 ± 3 mm/ 5 min vs. 10 ± 6 mm/ 5 min, p ≤ 0.001, Mann-Whitney rank sum test), the mean postoperative improvement of epiphora symptoms did not (82 ± 30 % vs. 76 ± 34 %, p = 0.92) and no significant (rho = -0.11, p = 0.62, Spearman rank correlation coefficient) correlation between Schirmer test values and symptoms improvement was found. In both groups > 70 % of patients reported ≥ 70 % (satisfactory) postoperative improvement. CONCLUSIONS: In the present study, patients who had different preoperative basal Schirmer test values reported having very similar epiphora symptoms improvement one-year after surgery.

Sentinel node biopsy in breast cancer: results in a large series.

Sentinel lymph node biopsy (SLNB) is an appropriate method for the evaluation of axillary status in cases of early breast cancer. We report our experience in treating cases evaluated using SLNB. We analyzed a total of 1192 cases assessed by means of SLNB from July 1999 to December 2007. SLNB processing was successfully completed in 1154 cases with the use of blue dye or radiolabeled 99mTc-Dextran-500, or both. Of these 1154 patients, 857 were N0(i-) (no regional lymph node metastasis, negative immunohistochemistry, IHC), 96 were N0(i+) (no regional lymph node metastasis histologically, positive IHC, no IHC cluster greater than 0.2 mm) and 201 were N1mi (greater than 0.2 mm, none greater than 2.0 mm). Most of the tumors (70%) were invasive ductal carcinomas and tumors were staged as T1 in 770 patients (65%). A total of 274 patients underwent SLNB and axillary dissections up to April 2003. The inclusion criteria were tumor size equal to or less than 3 cm in diameter, no clinically palpable axillary lymph nodes, no neoadjuvant therapy. In 19 cases, the SLN could not be identified intraoperatively. A false-negative rate of 11% and a negative predictive value of 88.2% were obtained for the 255 assessable patients. The overall concordance between SLNB and axillary lymph node status was 92%. SLNB sensitivity for nodes was 81% and specificity was 100%. The higher sensitivity, specificity, accuracy, and lower false-negative rates of SLNB suggest that this method may be an appropriate alternative to total axillary dissection in early breast cancer patients.

Sentinel node biopsy in breast cancer: results in a large series

Sentinel lymph node biopsy (SLNB) is an appropriate method for the evaluation of axillary status in cases of early breast cancer. We report our experience in treating cases evaluated using SLNB. We analyzed a total of 1192 cases assessed by means of SLNB from July 1999 to December 2007. SLNB processing was successfully completed in 1154 cases with the use of blue dye or radiolabeled 99mTc-Dextran-500, or both. Of these 1154 patients, 857 were N0(i-) (no regional lymph node metastasis, negative immunohistochemistry, IHC), 96 were N0(i+) (no regional lymph node metastasis histologically, positive IHC, no IHC cluster greater than 0.2 mm) and 201 were N1mi (greater than 0.2 mm, none greater than 2.0 mm). Most of the tumors (70 percent) were invasive ductal carcinomas and tumors were staged as T1 in 770 patients (65 percent). A total of 274 patients underwent SLNB and axillary dissections up to April 2003. The inclusion criteria were tumor size equal to or less than 3 cm in diameter, no clinically palpable axillary lymph nodes, no neoadjuvant therapy. In 19 cases, the SLN could not be identified intraoperatively. A false-negative rate of 11 percent and a negative predictive value of 88.2 percent were obtained for the 255 assessable patients. The overall concordance between SLNB and axillary lymph node status was 92 percent. SLNB sensitivity for nodes was 81 percent and specificity was 100 percent. The higher sensitivity, specificity, accuracy, and lower false-negative rates of SLNB suggest that this method may be an appropriate alternative to total axillary dissection in early breast cancer patients.