No description available.

Otitis Media is one of the most common childhood diseases. Recurrent acute otitis media RAOM is characterized by repeated episodes of inflammation of the middle ear in conjunction with middle ear fluid, and often with an inflamed or bulging eardrum. Defective clearance by the Eustachian tube results in mucus build-up and is characteristic of otitis media with effusion (OME). Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, respiratory syncytial virus, and rhinovirus are the most common contributors to otitis media pathogenesis. In New Zealand, OME has been implicated with conductive hearing loss in childhood and has been shown to significantly impact on speech and language development. New Zealand Māori and Polynesian children have displayed significantly higher hearing test failure rates than European-Caucasian children. The collectins, Surfactant Protein (SP)-A and -D are encoded by three genes (SP-A1, SP-A2, and SP-D) and are host defense proteins present in the middle ear and Eustachian tube. Single nucleotide polymorphisms (SNPs) in SP-A1 and SP-A2 have been associated with increased or decreased susceptibility to otitis media, meningococcal disease, and range of respiratory diseases. Using allele-specific primers and real-time PCR with SYBR Green I melting curve analysis, four groups of individuals were genotyped for eleven SP-A1, SP-A2, and SP-D SNPs: European-Caucasian individuals with RAOM/OME; New Zealand Māori/Polynesian individuals with RAOM/OME; individuals with meningococcal disease; and a control group. The computer program, Haploview, was employed to perform χ2 analyses and identify statistically significant associations of alleles/haplotypes with RAOM/OME or meningococcal disease. In the European-Caucasian population, two SP-A1 alleles, one SP-A2 allele, and four haplotypes (CGAGC, 1A3, 1A9, and 1A10) were found to be associated with increased risk of RAOM/OME (P lt; 0.05). Conversely, haplotypes 6A2 and 1A2 were found to be protective against susceptibility to RAOM/OME (P lt; 0.05). In New Zealand Māori and Polynesian individuals, two SP-A1 alleles, three SP-A2 alleles, one SP-D allele, and four haplotypes (6A8, 6A10, 1A3, and 1A10) were found to be associated with increased risk of RAOM/OME (P lt; 0.05). An additional four haplotypes (6A2, 1A0, 1A2, and TA) were determined to be protective against susceptibility to RAOM/OME (P lt; 0.05). However, protective SPA1/SPA2/SPD haplotype 6A2-1A0-TA was significantly under-represented in the New Zealand Māori and Polynesian population (P lt; 0.05). A single allele and haplotype were associated with increased risk of meningococcal disease (P lt; 0.05). The findings of this study confirm that specific genetic variants of SP-A and SP-D are associated with either increased or decreased risk of developing RAOM and/or OME. Furthermore, it was demonstrated that New Zealand Māori and Polynesian individuals appear to exhibit more haplotypes susceptible to RAOM/OME. This may provide a partial explanation for the higher RAOM/OME-related failure rates of hearing tests in New Zealand Māori and Polynesian children. However, there are numerous socio-economic and environmental factors that also contribute to otitis media pathogenesis which were not considered in this study. The effects of the SP-A1, SP-A2, and SP-D alleles and haplotypes on the bacterial/viral binding efficiencies of SP-A and SP-D need to be investigated by further research, using a large population, to confirm the association with susceptibility or resistance with RAOM/OME.

Iron is essential in nearly all organisms. It is a cofactor in many proteins and enzymes. This transition metal can also be toxic because it participates in reactions which produce reactive oxygen species. To avoid these toxic effects while still being used for essential processes, the cell must regulate tightly iron import, metabolism, trafficking, and homeostasis. These processes were studied using biophysical methods centered on Mossbauer spectroscopy supplemented by electron paramagnetic resonance, electronic absorption spectroscopy, and inductively coupled plasma mass spectrometry. This integrated biophysical approach was applied to yeast cells, isolated yeast mitochondria, and mouse brains. We determined the concentration of Fe, and the proportion of that Fe present as iron-sulfur clusters, heme centers, mononuclear nonheme centers, and as Fe3+ oxyhydroxide (phosphate) nanoparticles for each system. In yeast, the dependence of metabolic mode of growth and iron in the growth medium on this distribution was studied. Approximately three-quarters of the iron in fermenting cells was located in vacuoles, where it was present as high-spin mononuclear Fe3+ species with rhombic symmetry. The remaining quarter was present in the mitochondria. In fermenting mitochondria 4 distinct species of iron were observed, including [Fe4S4]2+ clusters and low-spin Fe2+ hemes arising from respiratory complexes, non-heme high spin (NHHS) Fe2+ species, high spin nonheme Fe3+ species, and nanoparticles. These distributions (in both the cells and mitochondria) change when the cells are grown on iron deficient medium but remained relatively unaltered as iron in the growth medium was increased. Respiring cells had less Fe associated with vacuoles, and more Fe present as HS Fe2+. Respiring mitochondria contain more [Fe4S4]2+ clusters and low-spin Fe2+ hemes, more S = 1/2 [Fe2S2]1+ clusters, and less NHHS Fe2+, HS Fe3+ species and Fe3+ nanoparticles. These changes were rationalized by assuming that the NHHS Fe2+ and Fe3+ species, and the nanoparticles were in equilibrium within the matrix of the mitochondria, and that the Fe2+ species served as feedstock for the synthesis of iron-sulfur clusters and heme centers. The iron in the mouse brain consisted mostly of [Fe4S4]2+ clusters and Fe2+ hemes from mitochondria respiratory complexes, and of ferritin, an Fe storage protein complex. NHHS Fe2+ and Fe3+ species were also observed. The ratio of stored Fe to mitochondrial Fe was sensitive to age. The brains of prenatal animals were dominated by ferritin. Following birth up to the first 4 weeks of life, there was an increase in mitochondrial Fe and a decline of ferritin Fe. Beyond 4 weeks up to 58 weeks, levels of ferritin increased and mitochondrial Fe remained constant. The brains of mice fed an Fe-deficient diet were also studied; most of the Fe in these brains was present as mitochondrial Fe, with little stored as ferritin. A model was developed to explain these changes.

No description available.

No description available.

O OME é empregado no tratamento de curto e longo prazo em diferentes desordens gastrointestinais. É um inibidor da bomba de prótons, que atua seletivamente na enzima H+/K+ ATPase. É completamente metabolizado pelas enzimas do citocromo P450 gerando três metabólitos principais: HOME, OMES e OS. É utilizado no tratamento de refluxo gastroesofágico em pacientes que sofrem de obesidade e sobrepeso no período anterior e posterior à cirurgia bariátrica. Entretanto, a realização deste procedimento cirúrgico pode prejudicar a absorção de nutrientes e medicamentos. Assim, a determinação das concentrações plasmáticas do OME é de grande importância clínica para o ajuste de dose evitando-se o comprometimento do tratamento. Portanto, foram desenvolvidos e validados dois métodos para quantificação do OME e metabólitos em amostras de plasma: LLE, método padrão e OS-DLLME e avaliada a IL-DLLME como método alternativo de extração. O método padrão, LLE, foi desenvolvido para quantificação do OME e metabólitos em amostras de plasma por HPLC-UV. A análise foi feita em uma coluna cromatográfica Zorbax Eclipse XDB - C18 (25 cm x 4,6 mm, partículas de 5 ?m) (Agilent Technologies, Palo Alto, CA, EUA) e coluna de guarda Zorbax Eclipse XDB - C18 (12,5 mm x 4,6 mm, partículas de 5 ?m) e FM composta por ACN: água (30:70, v/v), na vazão de 1 mL/min, detecção em 302 nm. Nestas condições foi possível a separação do OME e metabólitos em menos de 20 minutos. O método apresentou-se linear no intervalo de concentração plasmática de 20- 1000 ng/mL (r>0,99). Os parâmetros de precisão, exatidão, efeito carry-over e estabilidade estão em conformidade com o guia de validação da ANVISA (2012). O método foi satisfatoriamente aplicado em amostras de plasma de 20 pacientes submetidos à cirurgia bariátrica, no período pré e pós-cirúrgico. Outros dois métodos foram avaliados utilizando a técnica de microextração em fase líquida (DLLME). O método de OS-DLLME foi desenvolvido e validado para quantificação do OME e metabólitos em amostras de plasma por LCMS/ MS. A análise foi realizada na mesma coluna cromatográfica descrita anteriormente com FM composta por ACN:tampão formiato de amônio 10 mM pH 8,5 (50:50, v/v) vazão de 0,4 mL/min. Foram monitorados os íons precursores do OME, HOME, OMES e PI. As amostras de plasma foram pré-tratadas com ISO e com sobrenadante foi desenvolvida a OS-DLLME. Os parâmetros da OS-DLLME (tipo e volume de solvente extrator e dispersor, pH, força iônica do meio, tempo de agitação, tempo de centrifugação, volume de diluição da amostra e volume de amostra) foram otimizados com o auxílio de um delineamento experimental Plackett Burmann seguido por um CCD. Ao final uma metodologia de superfície de resposta facilitou a determinação da condição ótima de extração utilizando a ferramenta desejabilidade (D) na qual as condições otimizadas foram: 400?L de plasma, 2,5 mL de solução tampão tetraborato de sódio 0,1 mM pH 9,5, 120 ?L de CHCl3, 500 ?L de ISO, 0% de NaCl, 5 minutos de centrifugação, sem agitação das amostras. Posteriormente, o método foi aplicado com sucesso para análise de amostras de plasma de 5 pacientes submetidos à cirurgia bariátrica, no período pré e pós-cirúrgico.A IL-DLLME foi avaliada como técnica de extração utilizando ILs como solventes extratores alternativos. A análise foi realizada na mesma coluna cromatográfica com fase móvel no modo eluição por gradiente composta por ACN:água: 0-10 min, 5:95; 10,0-15 min, 10:90; 15,01-40 min, 30:70, v/v, vazão de 1 mL/min e detecção em 302 nm. Foram avaliados três ILs comerciais e oito ILs sintetizados no laboratório (NPPNS). Foram avaliados os parâmetros da IL-DLLME: solvente dispersor, IL extrator, pH e composição da solução tampão. Também foi realizada a avaliação por RMN dos IL utilizados. Dentre os ILs comerciais foi selecionado o OMImPF6, e dentre os ILs sintetizados foi selecionado o C8PF6. Entretanto, não foi possível o desenvolvimento do método devido a escassa quantidade de IL comercial. Em relação aos ILs sintetizados foram observados problemas na síntese e purificação, como presença de interferentes que coeluiam juntamente com os analitos avaliados. Entretanto, apesar deste método não ter sido validado foi possível obter valores de recuperação, para ambos os IL selecionados, superiores a 40%. / OME is used in the short- and long-term treatment of different gastrointestinal disorders. It is a proton pump inhibitor, which selectively acts on the enzyme H + / K + ATPase. It is completely metabolized by the cytochrome P450 enzymes generating three main metabolites: HOME, OMES and OS. It is used in the treatment of gastroesophageal reflux in patients who are obese and overweight in the period before and after bariatric surgery. However, performing this surgical procedure may impair the absorption of nutrients and medications. Thus, the determination of plasma concentrations of OME is of great clinical importance for dose adjustment, avoiding compromised treatment. Thus, two methods were developed for the quantification of OME and metabolites in plasma samples: LLE, standard method and OS-DLLME and IL-DLLME as an alternative method of extraction. The standard method, LLE, was developed for quantification of OME and metabolites in plasma samples by HPLC-UV. The analysis was done on a Zorbax Eclipse XDB-C18 (25 cm x 4.6 mm, 5 ?m particles) (Agilent Technologies, Palo Alto, CA, USA) and Zorbax Eclipse XDB-C18 (12, 5 mm x 4.6 mm, 5 ?m particles) and mobile phase composed of ACN: water (30:70, v / v) at the flow rate of 1 mL / min, detection at 302 nm. Under these conditions it was possible to separate OME and metabolites in less than 20 minutes. The method was linear in the plasma concentration range of 20-1000 ng / mL (r> 0.99). The parameters of accuracy, accuracy, carry-over effect and stability are in accordance with ANVISA\'s validation guide (2012). The method was satisfactorily applied in plasma samples from 20 patients submitted to bariatric surgery in the pre- and post-surgery period. Two other methods were evaluated using the liquid phase microextraction technique (DLLME). The OS-DLLME method was developed and validated for quantification of OME and metabolites in plasma samples by LC-MS / MS. The analysis was performed in the same chromatographic column described above with mobile phase composed of ACN: 10 mM ammonium formate buffer pH 8.5 (50:50, v / v) flow rate of 0.4 mL / min. The precursor ions of OME, HOME, OMES and PI were monitored. Plasma samples were pre-treated with ISO and the supernatant was grown to OS-DLLME. The parameters of OS-DLLME (type and volume of solvent extractor and dispersant, pH, ionic strength of medium, agitation time, centrifugation time, sample dilution volume and sample volume) were optimized with the aid of an experimental design Plackett Burmann followed by a central composite outline. At the end, a response surface methodology facilitated the determination of the optimal extraction condition using the desirability tool (D) in which optimized conditions were: 400?L of plasma, 2.5 mL of buffer solution 0.1 mM sodium tetraborate pH 9,5, 120 ?L CHCl3, 500 ?l ISO, 0% NaCl, 5 minutes centrifugation, without sample shaking. Subsequently, the method was validated and successfully applied for analysis of plasma samples from 5 patients submitted to bariatric surgery in the pre and post-surgical period. IL-DLLME was evaluated as an extraction technique using ILs as alternative extractive solvents. The analysis was performed on the iv same mobile phase chromatographic column in the gradient elution mode composed of ACN: water: 0-10 min, 5:95; 10.0-15 min, 10:90; 15.01 - 40 min, 30:70, v/v, flow rate of 1 mL / min and detection at 302 nm. Three commercial ILs and eight ILs synthesized in the laboratory (NPPNS) were evaluated. The parameters of IL-DLLME were evaluated: dispersing solvent, IL extractor, pH and buffer composition. NMR evaluation of the ILs used was also performed. Among the commercial ILs, OMImPF6 was selected, and C8PF6 was selected among the ILs synthesized. However, the development of the method was not possible due to the small amount of commercial IL. Concerning ILs synthesized, problems were observed in the synthesis and purification, as presence of interfering factors that coeluted together with the analyzed analytes. Thus, although this method was not validated, it was possible to obtain recovery values, for both ILs selected, higher than 40%.

OBJECTIVES: Bacterial infections in the normally sterile environment of the middle ear cavity usually trigger host immune response, whereby the innate immune system plays a dominant role as the host’s first line of defense. In this study we evaluated the expression levels of Toll-like receptors (TLRs) -2, -4, -5, -9, and nucleotidebinding oligomerization domain-containing proteins (NODs) -1 and -2, all of which are related to bacterial infection in pediatric patients with otitis media with effusion (OME). METHODS: The study sample consisted of 46 pediatric patients with OME, all of whom had ventilation tubes inserted. The expression levels of TLR-2, -4, -5, -9, NOD-1 and -2 mRNA in middle ear effusion were assessed by polymerase chain reaction. RESULTS: All effusion fluid samples collected from patients with OME showed expression of TLR-2, -4, -5, -9, NOD-1, and -2 mRNA. However, we found no correlations among expression levels of pattern recognition receptors (PRRs) in relation to characteristics of exudates, presence of bacteria, or frequencies of ventilation tube insertion (p>0.05). CONCLUSION: Our findings suggest that exudates of OME patients show PRR expressions that are related to the innate immune response regardless of the characteristics of effusion fluid, presence of bacteria in exudates, or frequency of ventilation tube insertion.

Introduktion: Sekretorisk media otit (SOM) är den vanligaste öronsjukdomen och uppkommer oftast i efterförloppet till akut media otit (AOM). Om barnet får SOM leder det till hörselnedsättning som kan påverka språklig utveckling och ge beteendeproblem. Etiologin bakom SOM anses vara multifaktoriell. Barnets tryck i mellanörat kan påverkas vid flaskmatning. Allergier, bakteriella/virus infektioner och inflammationer har också betydelse för utvecklandet av SOM. Syfte: att studera frekvensen av att använda nappflaska och liggande position vid flaskmatning relaterat till sekretorisk media otit och akut media otit i samband med rörbehandling. Metod: En deskriptiv tvärsnittsstudie med kvantitativ ansats. Datainsamling skedde via enkäter som besvarades av närstående till barn 2- 5 år som genomgått rörinsättning på öronoperation. Resultat: 103 barn med närstående deltog i studien. 87% av barnen som genomgick rörinsättning använde eller hade använt nappflaska och av dessa barn låg 55% i planläge. Hereditet gällande SOM kunde skönjas. Konklusion: Studien visade hög frekvens av flaskmatning där lite mer än hälften, 55% låg i planläge men studiens begränsning kunde inte åskådligöra samband med planläge och SOM. Det är av vikt att belysa faktorer som har betydelse för utveckling av SOM för att minska behov av rörbehandling. Dessa faktorer kan sjuksköterskan informera närstående om via hälsoundervisning för att öka barn och närståendes empowerment. / Introduction: Otitis Media with Effusion (OME) is the most common ear disease and arises mostly in the end of akut media otit (AOM). If a child contracts OME, it will result in a hearing loss which can affect the verbal development and may lead to behavior difficulties. The etiology behind OME is considered to be multifactorial. A child’s middle ear pressure may be affected by bottle nursing. Furthermore, the development of OME may be caused by allergies, bacterial/virus infections and inflammations. Purpose: To study the frequency of using a feeding bottle while the child is lying down related to OME and AOM when tube insertion. Method: A descriptive cross-sectional study with a quantitative approach. A survey data collection was used wherein the people who were surveyed were close family to two to five year old children, who have had ear tubes inserted. Result: 103 children with their close family participated in the study. 90% of these children who went through an ear tube insertion used or had used a feeding bottle and 50% of them lied down as they were fed. The heredity regarding SOM could be picked out. Conclusion: The study showed a high frequency of bottle nursing where just over the half (55%) lied down, but the restriction of the study could however not visualize the relation between lying down and SOM. Moreover, it is important to bring up the factors that are significant for the development of SOM to decrease the need of ear tube treatment/insertion. The nurse can inform close family about these factors through health education in order to increase children and close family´s empowerment.

This Master Thesis, inspired by the relevance that home-offices have assumed as a consequence of the COVID-19 pandemic, originated from the architectural, quantitative, and qualitative analysis of lighting within the author’s student accommodation on KTH Main Campus, in Stockholm (Sweden). As a matter of fact, this background study proved that recommended lighting levels (Illuminance, Contrast Ratio, CCT, CRI, M-EDI, and DER values) were not met, making the space unsuitable and unhealthy to both live and work in. Moreover, evaluating individual lighting preferences, it was demonstrated that flexibility, customizability, and adaptability to daylighting and tasks are fundamental requirements for working from home.Considering the temporary nature of students’ leases, the idea of developing an inexpensive and easily assemblable kit of smart products, aimed at improving lighting in dormitories, came quite naturally.To study the implementation of visual comfort and focus on tasks, literature review, market analysis, quantitative measurements, and hands-on experiments were carried on.Thus, with the gained knowledge and experience, the Home-Office Lighting Kit, an array of commercially available sockets, shades, and smart sources, supporting circadian rhythm and individual preferences, was proposed. Along with it, instructions on set-up and scenarios were provided.Although the overall conclusion was that the market needs products with a better balance between price, sturdiness, and reliability, the lighting conditions within the case study space were substantially improved, making other students interested in doing the same.Eventually, interesting cues for the sector’s development were proposed.