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Mariana Ramírez Mejía,1 Leticia Arroyo Muñoz,1 Ana Beatriz Medina Perez,1 Cristina Mendoza Velasquez,1 Jimena Ceja Martínez,1 Azyadeh Camacho Ordonez,1 Oscar Guerrero-Berger1,2 1Department of Anterior Segment Surgery, Fundación Hospital Nuestra Señora de la Luz, Mexico City, Mexico; 2Centro Oftalmológico Mira, Mexico City, MéxicoCorrespondence: Oscar Guerrero-Berger, Department of Anterior Segment Surgery, Fundación Hospital Nuestra Señora de la Luz, Ezequiel Montes 135, Tabacalera, Cuauhtémoc, Mexico City, 06030, Mexico, Tel +525551281140, Email droscarguerrero@gmail.comPurpose: To compare magnification and refocusing during phacoemulsification with the NGENUITY® 3-D Visualization System (3-D) versus the conventional microscope (CM) OPMI LUMERA 700.Setting: This study was performed in the Department of Anterior Segment of the Fundación Hospital Nuestra Señora de la Luz.Design: Prospective, randomized, cross-sectional, multi-surgeon, and comparative study.Methods: This study enrolled 100 patients (eyes) scheduled for phacoemulsification to measure the number of times changes in focusing and magnification were needed during cataract surgery.Results: Our study included 100 patients. From the endpoints evaluated, “zoom-in” showed statistically significant differences for all of the four predefined cataract surgery steps (means: Step 1, 0.38 (CM) vs 0.08 (3-D); Step 2, 0.36 (CM) vs 0.06 (3-D); Step 3, 0.54 (CM) vs 0.22 (3-D); Step 4, 0.56 (CM) vs 0.24 (3-D); all comparisons, p < 0.05). In Step 4, there was a statistically significant increased use of “focus-out” for the 3-D system (mean 0.16 (CM) vs 0.58 (3-D); p < 0.05). “Focus-in” and “zoom-out” showed no group differences for all steps. The duration of surgery with the 3-D system was longer at each step and overall. The percentage of light intensity did not show a statistically significant difference between both systems, with a mean of 99.45 for CM vs 98.43% for the heads-up system.Conclusion: The heads-up 3-D system is a safe option that offers excellent magnification for anterior segment visualization. The surgical time is longer, but adjusting settings like light intensity and brightness may facilitate some surgical steps early in the learning curve.Keywords: three-dimensional visualization, cataract surgery, heads-up surgery, visualization systems

Schizymenia dubyi (Chauvin ex Duby) J. Agardh is a red alga of the order Nemastomatales that is native to Europe, Asia and Oceania. The first specimen of this seaweed from the Argentinian coast was collected in the Chapadmalal intertidal wave-cut platform (38°12′S, 57°40′W) in September 2008. Taxonomic identification of the species was made on the basis of vegetative and reproductive characters and confirmed by DNA sequence analysis of the rbcL gene. The S. dubyi samples collected in the field were represented by foliose gametophytes as well as crustose tetrasporophytes. The gametophytes were distributed in patches of c. 50% coverage of the analysed quadrants (n = 20). Gametophyte abundance was 3.1±2.5 (mean±SD) plants per quadrant (0.04 m2). All these were fertile females. Molecular results placed the specimens from Argentina within the S. dubyi clade, closely related to S. dubyi from Japan and France. [ABSTRACT FROM AUTHOR]

It is commonly believed that near-field head-related transfer functions (HRTFs) provide perceptual benefits over far-field HRTFs that enhance the plausibility of binaural rendering of nearby sound sources. However, to the best of our knowledge, no study has systematically investigated whether using near-field HRTFs actually provides a perceptually more plausible virtual acoustic environment. To assess this question, we conducted two experiments in a six-degrees-of-freedom multimodal augmented reality experience where participants had to compare non-individual anechoic binaural renderings based on either synthesized near-field HRTFs or intensity-scaled far-field HRTFs and judge which of the two rendering methods led to a more plausible representation. Participants controlled the virtual sound source position by moving a small handheld loudspeaker along a prescribed trajectory laterally and frontally near the head, which provided visual and proprioceptive cues in addition to the auditory cues. The results of both experiments show no evidence that near-field cues enhance the plausibility of non-individual binaural rendering of nearby anechoic sound sources in a dynamic multimodal virtual acoustic scene as examined in this study. These findings suggest that, at least in terms of plausibility, the additional effort of including near-field cues in binaural rendering may not always be worthwhile for virtual or augmented reality applications.

Pharmacological management of insulin resistance and dyslipidaemias in children and adolescents is required to prevent the development of metabolic syndrome (MS) and type II diabetes mellitus (DM2). Material and method: we developed extemporaneous formulations from 500 mg-tablets from three generic brands, commercially available in Mexican drugstores: MedimartTM, Farmacias del AhorroTM and Primer NivelTM, dissolved in water sweetened with Splenda, which made them palatable and allowed customised dose adjustment. Solutions were stored at four environmental conditions: 25oC exposed to light, 25oC protected from light, 4oC and 40oC, and their physicochemical stability was assayed. The stability of the drug was determined by ultra performance liquid chromatography and ultraviolet detection (UPLC-UV) and by measuring the pH of the stock solution. The mobile phase consisted of (KH2PO4) 0.1 M, pH = 6.5, 4.6 mM sodium dodecyl sulphate (SDS) and acetonitrile (63:7:30) at 0.8 mL/min, column VARIAN Pursuit C8 150 × 3.9 mm tempered at 40oC, with detection at 236 nm. Results: Metformin from all trademarks was stable at all storage conditions for up to 30 days, retaining more than 90% of the initial amount of active drug, with a pH of 7.4 ± 0.3. Conclusion: Metformin extemporaneous formulations may be developed from either the innovator or generic brands, having the advantage of saving money and conserving the stability of its physicochemical properties. Keywords: Generic metformin, Innovator metformin, Extemporaneous formulation, Physicochemical stability, UPLC-UV.

Abstract Background The flavonoid pathway is a long-standing and important tool for plant genetics, biochemistry, and molecular biology. Numerous flavonoid mutants have been identified in Arabidopsis over the past several decades in a variety of ecotypes. Here we present an analysis of Arabidopsis lines of ecotype Columbia carrying T-DNA insertions in genes encoding enzymes of the central flavonoid pathway. We also provide a comprehensive summary of various mutant alleles for these structural genes that have been described in the literature to date in a wide variety of ecotypes. Findings The confirmed knockout lines present easily-scorable phenotypes due to altered pigmentation of the seed coat (or testa). Knockouts for seven alleles for six flavonoid biosynthetic genes were confirmed by PCR and characterized by UPLC for altered flavonol content. Conclusion Seven mutant lines for six genes of the central flavonoid pathway were characterized in ecotype, Columbia. These lines represent a useful resource for integrating biochemical and physiological studies with genomic, transcriptomic, and proteomic data, much of which has been, and continues to be, generated in the Columbia background.

Abstract Background We compared the diagnostic accuracy and reproducibility of commercially available NS1-based dengue tests and explored factors influencing their sensitivities. Methods Paired analysis of 310 samples previously characterized as positive (n = 218) and negative (n = 92) for viral isolation and/or RT-PCR and/or IgM seroconversion. Masked samples were tested by two observers with Platelia™ Dengue NS1 Ag, second generation Pan-E™ Dengue Early ELISA, SD Dengue NS1 Ag ELISA, Dengue NS1 Ag STRIP™, and SD BIOLINE™ Dengue Duo (NS1/IgM/IgG). Results SD BIOLINE™ NS1/IgM/IgG had the highest sensitivity (80.7% 95%CI 75-85.7) with likelihood ratios of 7.4 (95%CI 4.1-13.8) and 0.21 (95%CI 0.16-0.28). The ELISA-format tests showed comparable sensitivities; all below 75%. STRIP™ and SD NS1 had even lower sensitivities ( Conclusions The simultaneous detection of NS1/IgM/IgG would be potentially useful for dengue diagnosis in both endemic and non endemic areas. A negative result does not rule out dengue. Further studies are required to assess the performance and impact of early laboratory diagnosis of dengue in the routine clinical setting.

T1 ρ and Quantitative Susceptibility Mapping (QSM) are evolving as substrates for quantifying the progressive nature of knee osteoarthritis., Objective: To evaluate the effects of spin lock time combinations on depth-dependent T1 ρ estimation, in adjunct to QSM, and characterize the degree of shared variance in QSM and T1 ρ for the quantitative measurement of articular cartilage., Design: Twenty healthy participants (10 ​M/10F, 22.2 ​± ​3.4 years) underwent bilateral knee MRI using T1 ρ MAPPS sequences with varying TSLs ([0-120] ms), along with a 3D spoiled gradient echo for QSM. Five total TSL combinations were used for T1 ρ computation, and direct depth-based comparison. Depth-wide variance was assessed in comparison to QSM as a basis to assess for depth-specific variation in T1 ρ computations across healthy cartilage., Results: Longer T1 ρ relaxation times were observed for TSL combinations with higher spin lock times. Depth-specific differences were documented for both QSM and T1 ρ , with most change found at ∼60% depth of the cartilage, relative to the surface. Direct squared linear correlation revealed that most T1 ρ TSL combinations can explain over 30% of the variability in QSM, suggesting inherent shared sensitivity to cartilage microstructure., Conclusions: T1 ρ mapping is subjective to the spin lock time combinations used for computation of relaxation times. When paired with QSM, both similarities and differences in signal sensitivity may be complementary to capture depth-wide changes in articular cartilage., Competing Interests: Dr. Myer consults with commercial entities to support commercialization strategies and applications to the US Food and Drug Administration but has no direct financial interest in the products. Dr. Myer's institution receives current and ongoing grant funding from National Institutes of Health/NIAMS Grants U01AR067997, R01 AR070474, R01AR055563, R01AR076153, R01 AR077248 and industry sponsored research funding related to injury prevention and sport performance to his institution. Dr. Myer receives author royalties from Human Kinetics and Wolters Kluwer. Dr. Myer is an inventor of biofeedback technologies (Patent No: US11350854B2, Augmented and Virtual reality for Sport Performance and Injury Prevention Application, Approval Date: July 06, 2022, Software Copyrighted) designed to enhance rehabilitation and prevent injuries and receives licensing royalties. Dr. Myer and Dr. Diekfuss receive inventor-related royalties resultant from biofeedback technologies (Include Health: LIC1907082014-0706). Dr. Diekfuss also receives author royalties from Kendall Hunt Publishing Company. Dr. Mandava is an employee of GE HealthCare., (© 2024 The Authors.)

Background: The main objectives of this paper are to outline the essential tools, instruments, and equipment needed to set up a functional microsurgery laboratory that is affordable for low-income hospitals and to identify cost-effective alternatives for acquiring microsurgical equipment, such as refurbished or donated instruments, collaborating with medical device manufacturers for discounted rates, or exploring local suppliers., Methods: Step-by-step instructions were provided on setting up the microsurgery laboratory, including recommendations for the layout, ergonomic considerations, lighting, and sterilization processes while ensuring cost-effectiveness, as well as comprehensive training protocols and a curriculum specifically tailored to enhance microsurgical skills in neurosurgery residents., Results: We explored cost-effective options for obtaining microsurgery simulators and utilizing open-source or low-cost virtual training platforms. We also included guidelines for regular equipment maintenance, instrument sterilization, and establishing protocols for infection control to ensure a safe and hygienic learning environment. To foster collaboration between low-income hospitals and external organizations or institutions that can provide support, resources, or mentorship, this paper shows strategies for networking, knowledge exchange, and establishing partnerships to enhance microsurgical training opportunities further. We evaluated the impact and effectiveness of the low-cost microsurgery laboratory by assessing the impact and effectiveness of the established microsurgery laboratory in improving the microsurgical skills of neurosurgery residents. About microsutures and microanastomosis, after three weeks of training, residents showed improvement in "surgical time" for ten separate simple stitches (30.06 vs. 8.65 min) and ten continuous single stitches (19.84 vs. 6.51 min). Similarly, there was an increase in the "good quality" of the stitches and the suture pattern from 36.36% to 63.63%., Conclusion: By achieving these objectives, this guide aims to empower low-income hospitals and neurosurgery residents with the necessary resources and knowledge to establish and operate an affordable microsurgery laboratory, ultimately enhancing the quality of microsurgical training and patient care in low-income countries., Competing Interests: There are no conflicts of interest., (Copyright: © 2023 Surgical Neurology International.)

Background: This study evaluates the factors affecting the return to work of endoscopic surgery for lumbar foraminal stenosis (LFS), including symptoms, functional status, complications, and reoperation rates., Methods: The authors' retrospective cohort study included 100 consecutive patients (50 males and 50 females) diagnosed with LFS who underwent endoscopic surgery at Trotsky National Research Center of Surgery between January 2018 and December 2021., Results: There were no significant differences in age and preoperative visual analog scale and Oswestry disability index scores between the male and female groups, time to return to work for different patient groups after undergoing endoscopic lumbar foraminotomy (ELF). However, patients with more severe stenosis and comorbidities may take longer to recover. Confounding factors were patient age, preoperative physical function, and job requirements., Conclusion: This study confirms that study ELF can effectively improve symptoms associated with lumbar radiculopathy, as well as back pain, and improve patients' quality of life. Comorbidity, smoking status, and complications prolong the time to return to work following ELF surgery compared to healthy subjects., Competing Interests: There are no conflicts of interest., (Copyright: © 2023 Surgical Neurology International.)

Background and Objectives: Fibrin sheath (FS) formation around tunneled central venous catheters (CVC) increases the risk of catheter-related bloodstream infections due to bacterial adherence to a biofilm. We sought to investigate whether FS disruption (FSD) at the time of CVC removal or exchange affects infectious outcomes in patients with CVC-related infections., Design, Setting, Participants, and Measurements: Retrospective cohort study of 307 adult maintenance hemodialysis patients aged 18 years or older at a single center academic-based hemodialysis program (UHN, Toronto) who developed CVC-related infections requiring CVC removal or exchange between January 2000 and January 2019. Exposure was FSD at the time of CVC removal or exchange. Outcomes were infectious metastatic complications, recurrent infection with the same organism within 1 year, or death due to infection. We created a Markov Multi-State Model (MMSM) to assess patients' trajectories through time as they transitioned between states. A time-to-event analysis was performed, adjusted for clinically relevant factors., Results: There was no significant relationship between FSD status at the time of CVC removal, the development of infectious complications in the multivariable model (adjusted HR = 0.71, 95% CI 0.09-5.80, p  = 0.76), or mortality from infection (HR = 0.84, 95% CI 0.34-2.11, p  = 0.73)., Conclusions: FSD at the time of CVC removal was not associated with increased risk of infectious complications or death due to infection. Further prospective study is needed to determine whether FSD contributes to reducing CVC infectious related complications., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Objectives: Less than a quarter of the world population has access to microneurosurgical care within a range of 2 h. We introduce a simplified exoscopic visualization system for low-resource settings., Materials and Methods: We purchased a 48 megapixels microscope camera with a c-mount lens and a ring light at a total cost of US$ 125. Sixteen patients with lumbar degenerative disk disease were divided into an exoscope group and a microscope group. In each group, we performed four open and four minimally invasive transforaminal lumbar interbody fusions (TLIF). We conducted a questionnaire-based assessment of the user experience., Results: The exoscope achieved similar outcomes with comparable blood loss and operating time as the microscope. It provided similar image quality and magnification. Yet, it lacked stereoscopic perception and the adjustability of the camera position was cumbersome. Most users strongly agreed the exoscope would significantly improve surgical teaching. Over 75% reported that they would recommend the exoscope to colleagues and all users saw its great potential for low-resource environments., Conclusion: Our low-budget exoscope is safe and feasible for TLIF and purchasable at a fraction of the cost of conventional microscopes. It may thus help expand access to neurosurgical care and training worldwide., Competing Interests: There are no conflicts of interest., (© 2023 Published by Scientific Scholar on behalf of Journal of Neurosciences in Rural Practice.)

Introduction: The stromal vascular fraction (SVF) is a heterogeneous population of cells that, interacting with each other, can affect the processes of regeneration, angiogenesis, and immunomodulation. Over the past 20 years, there has been a trend towards an increase in the number of clinical studies on the therapeutic use of SVF. MicroRNAs (miRNAs) are also important regulators of cellular function and they have been shown to be involved in SVF cellular component function. The purpose of this study was to analyze existing clinical studies on the therapeutic use of SVF including the role of miRNAs in the regulation of the function of the cellular component of SVF as an anti-inflammatory, pro-angiogenic and cell differentiation activity., Methods: The search strategy was to use material from the clinicaltrials.gov website, which focused on the key term "Stromal vascular fraction", and the inclusion and exclusion criteria were divided into two stages., Results: By August 2022, there were 149 registered clinical trials. Most studies belong to either Phase 1-2 (49.37%), Phase 1 (25.32%) or Phase 2 (22.78%). Most of them focused in the fields of traumatology, neurology/neurosurgery, endocrinology, vascular surgery, and immunology. However, only 8 clinical trials had published results. All of clinical trials have similar preparation methods and 8 clinical trials have positive results with no serious adverse effects., Conclusions: There appears to be a wide potential for the clinical use of SVF without reports of serious side effects. Many preclinical and clinical studies are currently underway on the use of SVF, and their future results will help to further explore their therapeutic potential. Nevertheless, there are not many studies on the role of miRNAs in the SVF microenvironment; however, this topic is very important for further study of the clinical application of SVF, including safety, in various human diseases., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2022 The Authors.)

A series of fourteen 2-aryl-3-phenyl-2,3-dihydro-4 H -pyrido[3,2- e ][1,3]thiazin-4-ones was prepared at room temperature by T3P-mediated cyclization of N -phenyl- C -aryl imines with thionicotinic acid, two difficult substrates. The reactions were operationally simple, did not require specialized equipment or anhydrous solvents, could be performed as either two or three component reactions, and gave moderate-good yields as high as 63%. This provides ready access to N -phenyl compounds in this family, which have been generally difficult to prepare. As part of the study, the first crystal structure of neutral thionicotinic acid is also reported, and showed the molecule to be in the form of the thione tautomer. Additionally, the synthesized compounds were tested against T. brucei, the causative agent of Human African Sleeping Sickness. Screening at 50 µM concentration showed that five of the compounds strongly inhibited growth and killed parasites.

The native flora of Chile has unique characteristics due to the geographical situation of the country, with the vast desert in the North, Patagonia in the South, the Andean Mountains on the east and the Pacific Ocean on the west. This exclusivity is reflected in high concentrations of phytochemicals in the fruits and leaves of its native plants. Some examples are Aristotelia chilensis (Molina), Stuntz (maqui), Berberis microphylla G. Forst. (calafate), Peumus boldus Molina (boldo), Ribes magellanicum Poir. (Magellan currant), Ugni molinae Turcz. (murtilla), Rubus geoides Sm. (miñe miñe), Drimys winteri J.R.Forst. & G.Forst. (canelo), Luma apiculata (DC.) Burret (arrayán) distributed throughout the entire Chilean territory. Some of these Chilean plants have been used for centuries in the country's traditional medicine. The most recent studies of phytochemical characterization of parts of Chilean plants show a wide spectrum of antioxidant compounds, phenolic components, terpenoids and alkaloids, which have shown biological activity in both in vitro and in vivo studies. This manuscript covers the entire Chilean territory characterizing the phytochemical profile and reporting some of its biological properties, focusing mainly on antioxidant, anti-inflammatory, antimicrobial, chemopreventive and cytotoxic activity, and potential against diabetes, metabolic syndrome and gastrointestinal disorders., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Here we report de novo macrocyclic peptide binders to Wnt3a, a member of the Wnt protein family. By means of the Random non-standard Peptides Integrated Discovery (RaPID) system, we have performed in vitro selection against the complex of mouse Wnt3a (mWnt3a) with human afamin (hAFM) to discover macrocyclic peptides that bind mWnt3a with K D values as tight as 110 nM. One of these peptides, WAp-D04 (Wnt-AFM-peptide-D04), was able to inhibit the receptor-mediated signaling process, which was demonstrated in a Wnt3a-dependent reporter cell-line. Based on this initial hit, we applied a block-mutagenesis scanning display to identify a mutant inhibitor, WAp-D04-W10P, with 5-fold greater potency in a reporter assay. This work represents the first instance of molecules capable of inhibiting Wnt signaling through direct interaction with a Wnt protein, a molecular class for which targeting has been challenging due its highly hydrophobic nature., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Background: Adequate phonation is self-regulated by auditory feedback. Children with bilateral profound hearing loss (PHL) lack this feedback resulting in abnormal voice. Adequate hearing aid use and auditory-verbal therapy (AVT) may improve voice quality in deaf children., Objective: To study whether hearing aid use and AVT approach improve acoustic parameters of voice of children with bilateral PHL., Materials and Methods: Nineteen children with bilateral PHL were studied. Age range 2-5 years (X = 53.04 months; SD = 9.54). All children were fitted with hearing aids according to auditory testing and they underwent a 1-year auditory habilitation period using the AVT approach. Acoustic analysis of voice including F0, shimmer, and jitter was performed at the onset and at the end of the auditory habilitation period. Final acoustic data were compared to a matched control group of 19 children, age range 2-5 years (X = 52.85; SD = 9.74) with normal hearing., Results: Mean fundamental frequency (F0) was significantly increased after AVT intervention. Shimmer and jitter significantly (P < 0.05) improved after the intervention period. However, despite the improvements, mean F0 at the end of the intervention period was still significantly (P < 0.05) decreased as compared to controls. Also, mean shimmer and jitter at the end of the habilitation period were still significantly (P < 0.05) higher as compared to controls., Conclusions: The results of this preliminary study suggest that hearing aid use and auditory habilitation with AVT approach improved acoustic voice parameters of children with PHL. However, acoustic parameters persisted abnormal as compared to matched normal hearing controls. AVT approach and regular hearing aid use seem to be safe and reliable clinical tools for improving voice quality of children with PFL., (Copyright © 2018 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

The Report on Carcinogens (RoC), from the National Toxicology Program of the USA, is one of the world-leading programs for the identification and acknowledgment of substances that represent a hazard of cancer to humans. RoC covers several essential topics concerning environmental, occupational, and pharmaceutical agents that are known to be, or reasonably anticipated to be carcinogenic to humans. To promote the highest exploitation by its potential users, several RoC aspects and features were put together into one article. For doing so, a comprehensive description is provided regarding RoC history, scope, general features, listing criteria, contents, handbook, and website. Secondary and tertiary aims for this work were (a) to point out some improvement opportunities for the RoC, and (b) to discuss pending issues in regulatory science and cancer hazard assessments. In this regard, for agents classified as probably, likely, reasonably anticipated, possibly or suspected to be a human carcinogen, there is a lack of quantitative knowledge concerning the likelihood of those agents actually being carcinogenic to humans. Elucidating these probabilities is necessary, because the duration of current regulations and the arrival of new acts may depend on it. On the other hand, there is a dramatic imbalance in priorities toward carcinogens, compared with non-carcinogens, in current cancer hazard identification programs. That vision may ignore that the availability on the market of chemicals classified as probably not carcinogenic to humans can also be important for the employment, alimentation, economy, quality of life of consumers, and human health., (© 2019 John Wiley & Sons, Ltd.)

Flexizymes, highly flexible tRNA aminoacylation ribozymes, have enabled charging of virtually any amino acid (including non-proteogenic ones) onto tRNA molecules. Coupling to a custom-made in vitro translation system, namely the flexible in vitro translation (FIT) system, has unveiled the remarkable tolerance of the ribosome toward molecules, remote from what nature has selected to carry out its elaborate functions. Among the very diverse molecules and chemistries that have been ribosomally incorporated, a plethora of entities capable of mediating intramolecular cyclization have revolutionized the design and discovery of macrocyclic peptides. These macrocyclization reactions (which can be spontaneous, chemical, or enzymatic) have all served as tools for the discovery of peptides with natural-like structures and properties. Coupling of the FIT system and mRNA display techniques, known as the random non-standard peptide integrated discovery (RaPID) system, has in turn allowed for the simultaneous screening of trillions of macrocyclic peptides against challenging biological targets. The macrocyclization methodologies are chosen depending on the structural and functional characteristics of the desired molecule. Thus, they can emanate from the peptide's N-terminus or its side chains, attributing flexibility or rigidity, or even result in the installation of fluorescent probes.

Macrocyclic peptides are an emerging class of bioactive compounds for therapeutic use. In part, this is because they are capable of high potency and excellent target affinity and selectivity. Over the last decade, several biochemical techniques have been developed for the identification of bioactive macrocyclic peptides, allowing for the rapid isolation of high affinity ligands to a target of interest. A common feature of these techniques is a general reliance on thioether formation to effect macrocyclization. Increasingly, the compounds identified using these approaches have been subjected to x-ray crystallographic analysis bound to their respective targets, providing detailed structural information about their conformation and mechanism of target binding. The present review provides an overview of the target bound thioether-closed macrocyclic peptide structures that have been obtained to date.

Calreticulin (CRT) is a highly conserved protein in the endoplasmic reticulum that plays important roles in the regulation of key cellular functions. Little is known about the participation of E. histolytica CRT ( Eh CRT) in the processes of pathogenicity or in the modulation of the host immune response. The aim of this study was to evaluate the role of CRT in the proliferation and the cytokine profile in peripheral blood mononuclear cells (PBMCs) from patients with amebic liver abscess (ALA) during the acute phase (AP-ALA) of the disease compared to patients during the resolution phase (R-ALA). The PBMCs from each participant were cocultured with Eh CRT and tested by the colorimetric method to evaluate their proliferation index (PI). The supernatants were subjected to an enzyme-linked immunosorbent assay (ELISA) to evaluate the concentration of cytokines. The mean values of all groups were compared using the independent t -test. When the PIs of individuals without diagnosis of liver abscess (NEG) were compared, there were no statistically significant differences in the proliferation of PBMCs between patients with AP-ALA and R-ALA when stimulated with Eh CRT or concanavalin A (ConA). However, the levels of interleukins [IL-6, IL-10, granulocyte colony stimulating factor (GCSF), and transforming growth factor β1 (TGFβ1)] were higher in patients with AP-ALA, whereas in patients with R-ALA, higher levels of interferon gamma (IFNγ) were detected. These results suggest that Eh CRT acts as a mitogen very similar to the activity of ConA. In addition, Eh CRT is an excellent immunogen for the specific activation of PBMCs, inducing the differential expression of ILs depending on the outcome of disease, determining the type of immune response: a Th2 cytokine profile during the acute phase and a Th1 profile during the resolution phase.

Cobalamin C (cblC) deficiency is the most commonly inherited inborn error of vitamin B12 metabolism. It is characterized by multisystem involvement with severe neurological, hematological, renal and cardiopulmonary manifestations. Disease is most commonly diagnosed early in the first decade of life. We report a case of a 20-year-old woman who developed severe pulmonary arterial hypertension while under nephrologic follow-up for chronic kidney disease. She had initially presented at 14 years of age with visual disturbance and acute renal failure and been diagnosed with thrombotic thrombocytopenic purpura on the basis of kidney biopsy findings of thrombotic microangiopathy and compatible ADAMTS13 (a disentegrin and metalloproteinase with a thrombospondin type 1 motif member 13). When cblC deficiency was eventually diagnosed, remarkable improvement in cardiopulmonary function was evident upon initiation of treatment. This case highlights the importance of a timely diagnosis and initiation of treatment for cblC deficiency. Clinical diagnosis may be challenged by asynchronous organ symptom presentation and by misleading laboratory tests, in this case: an initial low ADAMTS13. A simple test of plasma homocysteine level should be encouraged in cases of thrombotic microangiopathy and/or pulmonary artery hypertension.

Teaching models in endoscopy are important tools to minimize risks derived from endoscopic procedures, taking into account that therapeutic endoscopy, also known as surgical endoscopy, has greatly developed during the last decade. This results from the fact that minimally invasive procedures present relevant contributions and promote more comfort to patients. In this context, ex vivo teaching models and virtual simulators are important tools to the safe acquisition of abilities. In this article, the Brazilian Society of Digestive Endoscopy presents and describes its first course of therapeutic ERCP and EUS in models of laboratory teaching.

The main objective of this study is to establish potential neuromorphometric differences which might act as markers of genetic risk for bipolar disorder and therefore serve as endophenotypes for discovery of genes that contribute to bipolar disorder. Magnetic resonance imaging (MRI) was used to assess structural brain volumes of 49 subjects. Volumetric analyses were first performed to test possible differences in the volume of brain structures between subjects with bipolar disorder type I (BPI) and control subjects in a new sample, based on regions previously reported in the literature as being either increased or decreased in size in bipolar patients. Subsequently, for those brain regions showing statistical difference between subjects with BPI and control subjects in our new sample, we tested whether unaffected first degree relatives (UFRs) of the BPI subjects also showed similar differences compared with controls. Four specific regions (right prefrontal, right middle prefrontal, right globus pallidus and left globus pallidus) met criteria for being possible endophenotypes for BPI in this sample., (Copyright © 2015 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.)

Enzymatic depletion of the nonessential amino acid l-Arginine (l-Arg) in patients with cancer by the administration of a pegylated form of the catabolic enzyme arginase I (peg-Arg I) has shown some promise as a therapeutic approach. However, l-Arg deprivation also suppresses T-cell responses in tumors. In this study, we sought to reconcile these observations by conducting a detailed analysis of the effects of peg-Arg I on normal T cells. Strikingly, we found that peg-Arg I blocked proliferation and cell-cycle progression in normal activated T cells without triggering apoptosis or blunting T-cell activation. These effects were associated with an inhibition of aerobic glycolysis in activated T cells, but not with significant alterations in mitochondrial oxidative respiration, which thereby regulated survival of T cells exposed to peg-Arg I. Further mechanistic investigations showed that the addition of citrulline, a metabolic precursor for l-Arg, rescued the antiproliferative effects of peg-Arg I on T cells in vitro. Moreover, serum levels of citrulline increased after in vivo administration of peg-Arg I. In support of the hypothesis that peg-Arg I acted indirectly to block T-cell responses in vivo, peg-Arg I inhibited T-cell proliferation in mice by inducing accumulation of myeloid-derived suppressor cells (MDSC). MDSC induction by peg-Arg I occurred through the general control nonrepressed-2 eIF2α kinase. Moreover, we found that peg-Arg I enhanced the growth of tumors in mice in a manner that correlated with higher MDSC numbers. Taken together, our results highlight the risks of the l-Arg-depleting therapy for cancer treatment and suggest a need for cotargeting MDSC in such therapeutic settings., (©2014 American Association for Cancer Research.)

The accumulation of myeloid-derived suppressor cells (MDSC) in tumor-bearing hosts is a hallmark of malignancy-associated inflammation and a major mediator for the induction of T cell suppression in cancer. MDSC can be divided phenotypically into granulocytic (G-MDSC) and monocytic (Mo-MDSC) subgroups. Several mechanisms mediate the induction of T cell anergy by MDSC; however, the specific role of these pathways in the inhibitory activity of MDSC subpopulations remains unclear. Therefore, we aimed to determine the effector mechanisms by which subsets of tumor-infiltrating MDSC block T cell function. We found that G-MDSC had a higher ability to impair proliferation and expression of effector molecules in activated T cells, as compared to Mo-MDSC. Interestingly, both MDSC subgroups inhibited T cells through nitric oxide (NO)-related pathways, but expressed different effector inhibitory mechanisms. Specifically, G-MDSC impaired T cells through the production of peroxynitrites (PNT), while Mo-MDSC suppressed by the release of NO. The production of PNT in G-MDSC depended on the expression of gp91(phox) and endothelial NO synthase (eNOS), while inducible NO synthase (iNOS) mediated the generation of NO in Mo-MDSC. Deletion of eNOS and gp91(phox) or scavenging of PNT blocked the suppressive function of G-MDSC and induced anti-tumoral effects, without altering Mo-MDSC inhibitory activity. Furthermore, NO-scavenging or iNOS knockdown prevented Mo-MDSC function, but did not affect PNT production or suppression by G-MDSC. These results suggest that MDSC subpopulations utilize independent effector mechanisms to regulate T cell function. Inhibition of these pathways is expected to specifically block MDSC subsets and overcome immune suppression in cancer., (© 2013 UICC.)

Tumor necrosis factor (TNF)-α promotes tumor development under chronic inflammation. Because TNF also activates caspase-8, selective inhibition of TNF-induced extrinsic apoptosis would be required for inflammation-associated tumor growth. In a mouse model of inflammation-associated colon carcinogenesis, we found nuclear expression of β-catenin in tumors of wild-type, but not mutant, mice that were made resistant to TNF-induced apoptosis by a germline mutation blocking caspase cleavage of the retinoblastoma (RB) protein, despite similar frequencies of β-catenin exon-3 mutations in these two genetic backgrounds. TNF-induced apoptosis was also attenuated in human colon cancer cell lines with genetically activated β-catenin. However, we found that HCT116 cells, which contain an activated allele of β-catenin but do not express nuclear β-catenin, were sensitive to TNF-induced apoptosis. In HCT116 cells, TNF stimulated efficient RB cleavage that preceded chromatin condensation. In contrast, TNF did not induce RB cleavage in colon cancer cells expressing nuclear β-catenin and these cells could be sensitized to basal and/or TNF-induced apoptosis by the knockdown of β-catenin or RB. In the apoptosis-resistant colon cancer cells, knockdown of β-catenin led to a reduction in the RB protein without affecting RB mRNA. Furthermore, ectopic expression of the caspase-resistant, but not the wild-type, RB re-established resistance to TNF-induced caspase activation in colon cancer cells without β-catenin. Together, these results suggest that nuclear β-catenin-dependent RB stabilization suppresses TNF-induced apoptosis in caspase-8-positive colon cancer cells.

Development of cervical cancer is a long process of abnormal cancerous cell growth in the cervix and is primarily the result of infection with specific high-risk types of human papillomavirus (HPV). The cytokines tumor necrosis factor α (TNFα) and lymphotoxin α (LTA) have an important role in all stages of cervical cancer and have the ability to induce the regression or promote the development of human tumors. Biologically important single-nucleotide polymorphisms (SNPs) occur within the TNFα and LTA genes. Therefore, the purpose of this study was to investigate the SNPs in the TNFα promoter region (-163, -238, -244, -308, -376, -857, -863, and -1031) and in the first intron of LTA (+252) in women with precursor lesions of cervical cancer. Overall, we studied 396 women from Mexico City. A total of 191 patients with HPV infection and precursor cervical lesions were subdivided in two groups: those with low-grade squamous intraepithelial lesions (n = 132) and those with high-grade squamous intraepithelial lesions (n = 59). Women (n = 205) negative for HPV and without cervical lesions were also included in the study. DNA was extracted from peripheral white blood cells and from cervical samples, and detection of biallelic polymorphisms of TNFα and LTA was performed using the polymerase chain reaction-sequence-specific oligonucleotide probe and restriction fragment length polymorphism techniques, respectively. We demonstrated that risk is associated with the genotype G/A (odds ratio = 2.48) and that protection is associated with the genotype G/G of SNP TNFα -376 (odds ratio = 0.37).

Soybean [Glycine max (L.) Merr] plants were exposed to three temperature regimens during seed development to investigate the effect of temperature on the expression of eight defense-related genes and the accumulation of two fungal pathogens in inoculated seeds. In seeds prior to inoculation, either a day/night warm (34/26 °C) or a cool temperature (22/18 °C) relative to normal (26/22 °C) resulted in altered patterns of gene expression including substantially lower expression of PR1, PR3 and PR10. After seed inoculation with Cercospora kikuchii, pathogen accumulation was lowest in seeds produced at 22/18 °C in which of all defense genes, MMP2 was uniquely most highly induced. For seeds inoculated with Diaporthe phaseolorum, pathogen accumulation was lowest in seeds produced at 34/26 °C in which of all defense genes, PR10 was uniquely most highly induced. Our detached seed assays clearly demonstrated that the temperature regimens we applied during seed development produced significant changes in seed defense-related gene expression both pre- and post inoculation and our findings support the hypothesis that global climate change may alter plant-pathogen interactions and thereby potentially crop productivity., (© Springer Science+Business Media B.V. 2011)

Prophylaxis is increasingly prescribed in treatment of haemophilia and its benefit is believed to be most significant for the youngest patients as haemophilic arthropathy may be prevented if prophylaxis is initiated prior to recurrent haemarthroses. While clinical prophylaxis data are readily available for haemophilia A, analogous data for haemophilia B are relatively limited. A prospective clinical study of recombinant factor IX (BeneFIX; rFIX), designed to allow investigator prescribed prophylaxis according to customary practices, was conducted in children <6 years old with severe haemophilia B. Nearly all children were prescribed prophylaxis (22/25; 88%) for all or part of their study participation. Favourable efficacy and safety profiles were reported. Routine prophylaxis with 1 or 2 rFIX infusions per week over an average of greater than 6 months of therapy resulted in near complete prevention of spontaneous breakthrough haemorrhages (<1 per year), with most children (77%) having none, including seven patients (32%) who had no bleeding episodes at all. Haemorrhages in joints were less common than those outside joints (27% vs. 73% of haemorrhages). In a patient population that included children with multiple prior haemarthroses, 68% of children had no joint bleeding. Breakthrough haemorrhages resolved with 1 or 2 infusions in 89% of episodes. The absence of changes in prophylaxis infusion schedules suggests that 1 or 2 rFIX infusion(s) per week were well-tolerated by these young patients, including those with (41%) and without (59%) central venous access devices. Safety was established by the low incidence of treatment-related adverse events.

Objectives: The forkhead box P3 (FOXP3) gene is considered to be the master gene of regulatory T cells. The significance of regulatory T cells in liver transplant has been investigated in previous reports, but quantitative FOXP3 messenger RNA (mRNA) expression after living-donor liver transplant has not been assessed. The objective of this study was to determine whether the human FOXP3 gene is a good marker for regulatory activity in T cells in living-donor liver transplant recipients during the immediate posttransplant period., Materials and Methods: In peripheral blood mononuclear cells of 15 living-donor liver transplant recipients during the first month after transplant; we measured the population of CD4+CD25+ T cells using flow-assisted cell sorting and the expression of FOXP3 mRNA using real-time polymerase chain reaction., Results: Fold induction of FOXP3 mRNA significantly increased on postoperative day 7 (3.3-fold) compared with the reference preoperative value (P < .01) but returned to baseline by 28 days after transplant. The population of CD4+CD25+ T cells did not change significantly. Expression of FOXP3 mRNA on days 14, 21, and 28 were lower in recipients with acute cellular rejection within 60 days after living-donor liver transplant., Conclusions: Increased expression of FOXP3 mRNA immediately after living-donor liver transplant might be influenced by activation of T cells including regulatory T cells and other T cells. However, after stabilization of these activation profiles, it seems likely that FOXP3mRNA expression is associated with graft acceptance. Further studies are necessary with measurement of FOXP3 mRNA expression at appropriate sampling points.

This study attempted to replicate evidence for association of the Epsin 4 gene (which encodes enthoprotin, a protein involved in vesicular transport) to schizophrenia in a new sample of families segregating schizophrenia drawn from the Latin American population. 1,423 subjects (767 with a history of psychosis) from 337 Latino families were genotyped using three single nucleotide polymorphisms (SNPs) spanning the Epsin 4 gene. A family based association test was utilized to test for association of these SNPs to the phenotypes of psychosis and schizophrenia. Haplotypes defined by these three SNPs showed significant association to the phenotype of psychosis in this sample (global p value=0.014, bi-allelic p value=0.047). Variation in the Epsin 4 gene is significantly associated with psychotic disorder in this Latino population. This provides additional support for the involvement of enthoprotin in the pathogenesis of schizophrenia and other psychotic disorders.

It has been well documented that transthyretin (TTR) shows an affinity for lipoproteins and amyloid is deposited around adipocytes in patients with familial amyloidotic polyneuropathy (FAP). We examined the involvement of lipids in amyloid fibrils in the tissues by histopathologic methods. Sudan black B staining for frozen tissues of autopsied FAP patients and biospied dialysis related amyloidosis (DRA) patients revealed colocalization of lipids in the tissue sections where Congo red staining was positive, while no such positive staining was observed in paraffin embedded tissues. Immunohistochemical study using lipoprotein antibodies revealed that only anti-high density lipoprotein (HDL) antibody showed immunoreactivity in both types of amyloid specimens where Congo red was positively stained. Measurement of the components of lipids in the frozen cardiac samples from an FAP patient and an amyotrophic lateral sclerosis (ALS) patient revealed that concentrations of triglyceride and cholesterol in each lipoprotein, except for HDL-triglyceride, were markedly elevated in the FAP patient's material, compared to that of the ALS patient. These results suggest that interaction of amyloid fibrils with HDL may play an important role in amyloid formation in FAP.

Duct-to-duct biliary reconstruction has been introduced in adult living donor liver transplantation (LDLT). In right-lobe grafts, however, the presence of two or three separated bile duct orifices is not rare and makes an alternative approach for reconstruction necessary. We used the cystic duct for one of the anastomoses in biliary reconstruction for 5 right-lobe living donor liver transplants with two separated ducts. Before the anastomosis, the inside lumen of the cystic duct was straightened with a metal probe. Two external drainage tubes were placed in all recipients, and posttransplant cholangiography through the tubes approximately one month after transplantation showed no leakage or stricture at any of the anastomotic sites. The drainage tubes were removed between 17 and 37 weeks after transplantation. All of the patients except one who died of chronic rejection have been doing well without any late biliary complications during follow-up periods ranging from 10 to 28 months after transplantation. In conclusion, our results indicate that biliary reconstruction using the cystic duct is feasible and safe for living donor liver transplantation and that external drainage tubes may be effective for prevention of complications.

The multidrug resistance EhPgp1 gene is constitutively expressed in drug resistant trophozoites from Entamoeba histolytica. It has been demonstrated that two CCAAT/enhancer binding sites located in the EhPgp1 gene promoter control its transcriptional activation. However, functional assays of the 5' end of its promoter showed that region from -234 to -196 bp (38 bp) is also important for the EhPgp1 gene transcription. Here, we demonstrated that in the 38 bp region putative cis-activator sequences are located. In silico analysis showed the presence of GATA1, Gal4, Nit-2, and C/EBP consensus sequences. Additionally, we identified three specific DNA-protein complexes, which were competed by a C/EBP, GATA1, and HOX oligonucleotides. Finally, we partially purified three proteins of 64.4, 56.7, and 27.4 kDa. Further investigations are currently in progress to determine the identity of these nuclear factors and how they are interacting with the EhPgp1 gene promoter.

This open-label, randomised, two-way crossover study compared the steady-state bioavailability of oestradiol administered by way of a new oestradiol matrix transdermal delivery system (Alora 0.1 mg/day) with that of Estraderm (0.1 mg/day) in 24 subjects. Serum oestradiol, oestrone and oestrone sulphate concentrations were determined by measurement of blood samples. Mean SD pre-dosing, nonadjusted oestradiol levels for Alora (71.9 27.0 pg/ml) were substantially higher than those for Estraderm (26.7 9.7 pg/ml), while peak oestradiol concentrations were comparable. Consequently, fluctuations in steady-state levels were substantially smaller for Alora than for Estraderm; the fluctuation index values (\[C- C ])/ max min C ) were significantly lower for Alora (0.97 0.23) than av for Estraderm (1.68 0.45). Oestradiol levels remained constant over the dosing interval with Alora but decreased significantly after 48 hours with Estraderm. The bioavailability of oestradiol with Alora was 127 56% that of Estraderm. Oestrone and oestrone sulphate data showed the same qualitative and quantitative differences between the two systems. Both systems were well tolerated. In summary, Alora delivered more oestradiol to the systemic circulation with greater consistency and over a longer time than did Estraderm.

Keratinases can be used for the production of potentially important hydrolyzed proteins and chemicals. This study investigated the keratinolytic activity of Streptomyces sp on keratinaceous materials like wool. High levels of proteolytic and keratinolytic activity were obtained after 96 h of culture when two Streptomyces sp strains were grown on basal medium containing mineral salts and 3% (w/v) of defatted wool as a source of energy, carbon, and nitrogen. The cell-free culture filtrates exhibited rapid proteolytic digestion of keratin powder. Currently, the authors are testing whether the enzymatic activity obtained is in fact keratinolytic, and not only an alkaline protease activity.

Plasma estradiol has been suggested to suppress adipose tissue lipoprotein lipase (LPL) activity in women. The present study explores the regulation of LPL by sex steroids in sedentary obese men (N = 24) at their usual weight. Femoral adipose tissue LPL activity, eluted with serum and heparin or extracted with detergent, showed significant inverse correlations with plasma levels of testosterone, bioavailable testosterone, dihydrotestosterone, and estradiol. Both measures of femoral LPL activity were also correlated with the weight change occurring despite efforts to maintain a constant weight. Abdominal LPL activity showed significant but weaker inverse correlations with bioavailable testosterone only. Multivariate analysis of potential predictors for eluted femoral LPL activity showed that plasma testosterone, dihydrotestosterone, and estradiol were interdependent, whereas the rate of weight change was an independent variable. In the regression equation, only bioavailable testosterone and weight change were retained, explaining 63% of the variability (R = .79, P = .0002). These results suggest that sex steroids suppress adipose tissue LPL activity in men, and more so in the thigh than in the abdomen, thereby possibly contributing to a central fat accumulation. The data are compatible with a model from male animals suggesting that testosterone effects on adipose tissue LPL are mediated by estradiol formed locally.

We describe an outbreak of skin lesions due to Mycobacterium chelonae subsp. abscessus associated with injections of lidocaine (lignocaine) given by a 'bioenergetic' (a practitioner of alternative medicine) in Colombia. The lidocaine carpules and the lesions of the patients yielded mycobacteria with identical biochemical characteristics. Using the methodology of Sartwell and a case control design we examined the incubation period and assessed risk factors. Of 667 potentially exposed individuals, a total of 298 patients were interviewed, of whom 232 had skin lesions. The median incubation period was 30.5 days (range 15-59 days). Male sex (OR 2.85, 95% CI 1.26-6.51), increasing age (OR 1.25, 95% CI 1.03-1.53), subcutaneous injection route (OR 3.72, 95% CI 1.09-12.7) and number of injections (OR 1.01, 95% CI 1.00-1.03) were risk factors for disease. To our knowledge, this is the largest reported outbreak of M. chelonae infection, the first in which the organism has been isolated from the putative vehicle of infection, and the first in which the incubation period could be determined.

Sibling resemblance in fatness and subcutaneous fat distribution is addressed in a sample of healthy adolescents who belong to 122 nuclear families. Relative fatness and fat distribution were estimated using anthropometric measurements, bioelectric impedance, and ultrasound images. The patterns of fat distribution were defined by indexes constructed using combinations of subcutaneous fat measurements. Adjusted Z scores were used to analyze age- and sex-related changes on fatness and fat distribution. An analysis of variance was used to calculate the resemblance within and between families. The results show that there is significant family resemblance for the level of fatness. There is also significant resemblance between sibs of like sex for the fat distribution patterns. The results also show that subcutaneous fat thickness has a strong genetic component (60-65% of the variance) and that the patterns of fat distribution are apparent from early adolescence.

Recent studies have shown major gene effects for obesity in randomly ascertained families. To investigate the familial aggregation of a specific subset of obesity, which is particularly prone to medical complications, families with morbid obesity were studied. This condition occurs in 1%-2% of the population and is defined as 45.5 kg (100 pounds) or more over ideal weight. First-degree relatives of 221 morbidly obese probands (1560 adults) were identified, and height and weight (current and greatest) were obtained from each family member. Morbid obesity occurred in the family members of the probands 8 times more often than in the general population. Of the morbidly obese probands, 48% had one or more first-degree relatives who were also morbidly obese compared to a 6% population estimate. By the ages of 20-24, 12% of the morbidly obese probands were already 45.5 kg or more overweight, and 45% were 22.7 kg (50 pounds) or more overweight. There was little difference in the prevalence of familial morbid obesity by the gender of the probands: 47% of the male probands and 48% of the female probands had another morbidly obese relative, while 67% and 53% of the early onset (before age 25) male and female probands, respectively, had one or more first-degree relatives who were also morbidly obese. In addition to the extreme degree of familial aggregation, the prevalence of morbid obesity in parent-offspring sets was calculated within the morbidly obese families. Morbidly obese families who have one or two morbidly obese parents have a 2.6 times increased risk (p<0.002) of having one or more morbidly obese adult offspring, compared to families who have neither parent morbidly obese. Evidence for trimodality of the body mass index distribution was found for each gender (p = 0.0006 for male relatives and p = 0.075 for female relatives). The strong familial aggregation of morbid obesity indicates the need for further understanding of the genetic determinants of this extreme clinical disorder and how environmental factors affect the genetic expression of the trait.