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BACKGROUND: Hallux rigidus is a common condition characterised by first metatarsophalangeal joint (MTPJ) degeneration, pain and limited range of motion (ROM). The gold standard surgical treatment is arthrodesis, providing good pain relief, but sacrifices ROM. The Cartiva synthetic cartilage implant (SCI) has been utilised as an interpositional arthroplasty, aiming to reduce pain whilst preserving range of motion. Current evidence for Cartiva SCI is limited. The aim was to evaluate the clinical outcomes of Cartiva SCI compared to arthrodesis undertaken in our centre. METHODS: A retrospective cohort study was conducted, enrolling all adult patients who underwent primary first MTPJ SCI arthroplasty or arthrodesis for the treatment hallux rigidus. The primary outcome was a validated patient-reported outcome measure (PROM), the Manchester-Oxford Foot Questionnaire (MOXFQ). Secondary outcomes included EQ-5D, complication rates, VAS Pain and FAAM (ADL). RESULTS: Between 2017 and 2020 there were 33 cases divided into two groups (17 Cartiva SCI, 16 arthrodesis, mean age 59.0 ± 9.9 years) with a mean follow up of 2.3 years. There was no statistically significant difference in any of the MOXFQ, EQ-5D, VAS Pain or FAAM (ADL) outcome scores between the Arthrodesis and SCI groups (p > 0.05). The mean MOXFQ Index score was 7.2 ± 6.4 for the SCI group and 3.9 ± 5.8 for the Arthrodesis group at final follow up (p = 0.15). Although complications were high in both groups, the overall hallux reoperation rate was 29.4 % in the SCI cohort and 0.0 % for arthrodesis. CONCLUSION: This retrospective comparative study found no significant superiority of Cartiva SCI over arthrodesis in terms of PROMs. Due to the higher rate of further surgical intervention in the SCI cohort, we recommend arthrodesis as the preferred surgical option for hallux rigidus. LEVEL OF EVIDENCE: III.
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Myopia is becoming increasingly common in young generations all over the world, and it is predicted to become the most common cause of blindness and visual impairment in later life in the near future. Because myopia can cause serious complications and vision loss, it is critical to create and prescribe effective myopia treatment solutions that can help prevent or delay the onset and progression of myopia. The scientific understanding of myopia's causes, genetic background, environmental conditions, and various management techniques, including therapies to prevent or postpone its development and slow its progression, is rapidly expanding. However, some significant information gaps exist on this subject, making it difficult to develop an effective intervention plan. As with the creation of this present algorithm, a compromise is to work on best practices and reach consensus among a wide number of specialists. The quick rise in information regarding myopia management may be difficult for the busy eye care provider, but it necessitates a continuing need to evaluate new research and implement it into daily practice. To assist eye care providers in developing these strategies, an algorithm has been proposed that covers all aspects of myopia mitigation and management. The algorithm aims to provide practical assistance in choosing and developing an effective myopia management strategy tailored to the individual child. It incorporates the latest research findings and covers a wide range of modalities, from primary, secondary, and tertiary myopia prevention to interventions that reduce the progression of myopia.
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AIM: The aim of the study was to determine whether ocular movement can affect the shape of the globe and lead to measurable change in axial and peripheral eye length. METHODS: Ten subjects aged 18-30 years (6 M/4 F) participated with informed consent. The mean spherical equivalent refractive error was ≤-1.00 DS with cylindrical refraction <-1.25 DC. One drop of tropicamide hydrochloride 1% was instilled 20 min before measurement to induce mydriasis and mild cycloplegia. Using IOLMaster, eye length was measured centrally and temporally (25° off-axis) in four different positions. Subjects then rotated their eyes 25° in the temporal direction to fixate on a target for 10 min. After that, the same measurements were repeated. RESULTS: Before rotation, the group mean peripheral eye length was significantly shorter than the central eye length (P < 0.05). There was no significant variation in central or peripheral eye length due to off-axis fixation, either after the initial eye rotation or after fixation for 10 min at the off-axis point. The difference between central and peripheral eye lengths was maintained after 10 min of temporal fixation (P < 0.05). CONCLUSION: Peripheral eye length was shorter than central eye length showing the prolate shape associated with myopia. The action of the extraocular muscles on the globe has no significant effect upon the retinal shape assessed by off-axis eye length measurement in myopic subjects.
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CLINICAL RELEVANCE: Swept-source optical coherence tomography may not provide reliable measurements of choroidal thickness for pigmented subjects. BACKGROUND: The repeatability of choroidal thickness measurements in young healthy adults was investigated using swept source optical coherence tomography. METHODS: Choroidal thickness was measured using swept source optical coherence tomography in 98 healthy subjects (49 South Asian and 49 Caucasian) aged between 19 and 28 years old. Repeated radial scan images centred on the fovea were obtained from the right eye with 5 min between measurements. Choroidal thickness values were obtained from the built-in automated segmentation algorithm for the 9 subfields defined by the Early Treatment Diabetic Retinopathy Study. Interclass correlation coefficients and Bland-Altman methods were used to assess test-retest repeatability. RESULTS: The mean ± standard deviation sub-foveal choroidal thickness was 316.53 ± 63.36 µm for the Caucasian group and 292.70 ± 60.13 µm for the South Asian group. Bland-Altman analysis showed smaller test-retest variability for choroidal thickness measurements in the Caucasian group (mean difference ± SD = -0.01 ± 6.59 µm) compared to the South Asian group (mean difference ± SD = -2.38 ± 12.20 µm). No relationship was found between the mean choroidal thickness and mean spherical equivalent of refractive error for both subject groups. CONCLUSION: Swept source optical coherence tomography is capable of obtaining choroidal thickness measurements with good repeatability. However, test-retest limits of agreement are substantially greater in South Asian subjects, suggesting that successful identification of the choroidal scleral interface may be affected by the amount of pigmentation present in the retinal and choroidal tissues.
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Retinopatia Diabética , Tomografia de Coerência Óptica , Adulto , Corioide/diagnóstico por imagem , Humanos , Retina , Esclera , Tomografia de Coerência Óptica/métodos , Adulto JovemRESUMO
The prevalence of myopia is increasing extensively worldwide. The number of people with myopia in 2020 is predicted to be 2.6 billion globally, which is expected to rise up to 4.9 billion by 2050, unless preventive actions and interventions are taken. The number of individuals with high myopia is also increasing substantially and pathological myopia is predicted to become the most common cause of irreversible vision impairment and blindness worldwide and also in Europe. These prevalence estimates indicate the importance of reducing the burden of myopia by means of myopia control interventions to prevent myopia onset and to slow down myopia progression. Due to the urgency of the situation, the European Society of Ophthalmology decided to publish this update of the current information and guidance on management of myopia. The pathogenesis and genetics of myopia are also summarized and epidemiology, risk factors, preventive and treatment options are discussed in details.
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Miopia Degenerativa , Oftalmologia , Procedimentos Ortoceratológicos , Progressão da Doença , Humanos , Miopia Degenerativa/epidemiologia , Miopia Degenerativa/prevenção & controle , PrevalênciaRESUMO
INTRODUCTION: Body height and axial length (AL) increase during childhood with excessive axial elongation resulting in myopia. There is no consensus regarding the association between body growth and AL during refractive development. This study explored the association between change in body height, AL and refractive status over 4-years in children and young adults. MATERIAL AND METHODS: Measures were collected biennially (timepoints: t1, t2, t3) (t1 nâ¯=â¯140, aged 5-20years). Non-cycloplegic autorefraction was obtained using the Shin-Nippon openfield autorefractor. AL, corneal curvature (CC) and anterior chamber depth (ACD) were measured by IOL Master. Body height (cm) was measured using a wall mounted tape measure. Refractive status was classified using spherical equivalent refraction (SER): persistent emmetropes (PE) (-0.50D to +1.00D), persistent myopes (PM) (≤-0.50D), progressing myopes (PrM) (increase of ≤-0.50D between timepoints), incident myopes (IM) (subsequent SER≤-0.50D) and persistent hyperopes (PH) (>+1.00D). RESULTS: Change in AL and change in height were correlated in the PE (all t:pâ¯≤â¯0.003) and the IM (t1-t2 pâ¯=â¯0.04). For every increase in body height of 1â¯cm: t1-t2: AL increased by 0.03â¯mm in the PE, 0.15 in the PM, 0.11â¯mm in the IM, 0.14â¯mm in the PrM, -0.006â¯mm in the PH. T2-t3: AL increased by 0.02â¯mm in the PE, 0.06 in the PM, 0.16â¯mm in the PrM, 0.12â¯mm in the IM and -0.03â¯mm in the PH. CONCLUSIONS: In emmetropia body growth and axial elongation are correlated. In participants with myopia, body growth appears to stabilise whilst axial elongation continues at a much faster rate indicating dysregulation of normal ocular growth.
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Comprimento Axial do Olho/fisiologia , Estatura/fisiologia , Emetropia/fisiologia , Hiperopia/fisiopatologia , Miopia/fisiopatologia , Refração Ocular/fisiologia , População Branca , Adolescente , Câmara Anterior/anatomia & histologia , Biometria , Estatura/etnologia , Criança , Pré-Escolar , Córnea/anatomia & histologia , Feminino , Humanos , Masculino , Acuidade Visual/fisiologia , Adulto JovemRESUMO
BACKGROUND: The authors assess the impact of rhinoplasty on public perception of a patient's appearance and personality. METHODS: A survey was created using standardized before-and-after photographs of 10 Caucasian women who had undergone primary rhinoplasty. Photographs of two additional women who had not undergone facial surgery were randomly included as controls, for a total of 12 survey items. Preoperative and postoperative photographs were placed side by side. The survey was administered by means of crowd-sourcing. Respondents were asked to evaluate which photograph better represented 11 traits of appearance or personality, according to a seven-point Likert scale. A score of 1 meant the preoperative photograph was much better, 7 meant the postoperative photograph was much better, and 4 meant no difference. T tests and analyses of variance were used to evaluate rating changes for each trait and differences between demographic groups. RESULTS: There were 264 responses received. Averaged scores across the 10 survey patients produced a value for each appearance or personality trait. In 10 of 11 categories (i.e., symmetry, youthfulness, facial harmony, likeability, trustworthiness, confidence, femininity, attractiveness, approachability, and intelligence), the postoperative photograph was significantly favorable compared with the preoperative photograph (p < 0.00001). The preoperative photograph was rated higher only in aggressiveness (p < 0.001). The same scores were calculated for the controls; no significant difference in any category was seen except confidence, where the right image was viewed as more confident (mean, 4.19; p < 0.005). CONCLUSION: Aesthetic rhinoplasty improves the public perception of a person's appearance and personality in multiple aspects.
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Beleza , Face , Rinoplastia/psicologia , Adulto , Agressão , Estudos Controlados Antes e Depois , Estética , Expressão Facial , Feminino , Humanos , Percepção , Personalidade , Fotografação , Período Pós-Operatório , Período Pré-OperatórioRESUMO
As the US population continues to age, new cases of end-stage renal disease (ESRD) in individuals, aged 85 years or older (the oldest old), are increasing. Many patients who begin hemodialysis despite questionable benefit may struggle with high symptom burden and rapid functional decline. This article reviews the history regarding the funding and development of the Medicare ESRD program, reviews current approaches to the oldest old with ESRD, and considers strategies to improve the management approach of this vulnerable population.
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BACKGROUND: Broader use of value-based reimbursement models will require providers to transparently demonstrate health care value. We sought to determine and report cost and quality data for episodes of hip and knee arthroplasty surgery among 13 members of the High Value Healthcare Collaborative (HVHC), a consortium of health care systems interested in improving health care value. METHODS: We conducted a retrospective, cross-sectional observational cohort study of 30-day episodes of care for hip and knee arthroplasty in fee-for-service Medicare beneficiaries aged 65 or older who had hip or knee osteoarthritis and used 1 of 13 HVHC member systems for uncomplicated primary hip arthroplasty (N = 8853) or knee arthroplasty (N = 16,434), respectively, in 2012 or 2013. At the system level, we calculated: per-capita utilization rates; postoperative complication rates; standardized total, acute, and postacute care Medicare expenditures for 30-day episodes of care; and the modeled impact of reducing episode expenditures or per-capita utilization rates. RESULTS: Adjusted per-capita utilization rates varied across HVHC systems and postacute care reimbursements varied more than 3-fold for both types of arthroplasty in both years. Regression analysis confirmed that total episode and postacute care reimbursements significantly differed across HVHC members after considering patient demographic differences. Potential Medicare cost savings were greatest for knee arthroplasty surgery and when lower total reimbursement targets were achieved. CONCLUSION: The substantial variation that we found offers opportunities for learning and collaboration to collectively improve outcomes, reduce costs, and enhance value. Ceteris paribus, reducing per-episode reimbursements would achieve greater Medicare cost savings than reducing per-capita rates.
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Artroplastia de Quadril/economia , Artroplastia do Joelho/economia , Complicações Pós-Operatórias/epidemiologia , Idoso , Artroplastia de Quadril/estatística & dados numéricos , Artroplastia do Joelho/estatística & dados numéricos , Redução de Custos , Estudos Transversais , Atenção à Saúde , Cuidado Periódico , Planos de Pagamento por Serviço Prestado , Feminino , Gastos em Saúde , Humanos , Tempo de Internação , Masculino , Medicare/economia , Pessoa de Meia-Idade , Osteoartrite do Joelho , Análise de Regressão , Estudos Retrospectivos , Cuidados Semi-Intensivos , Estados Unidos/epidemiologiaAssuntos
Hiperopia/fisiopatologia , Miopia/fisiopatologia , Pupila/fisiologia , Feminino , Humanos , MasculinoRESUMO
PURPOSE: To investigate the relationship between pupil diameter and refractive error and how refractive correction, target luminance, and accommodation modulate this relationship. METHODS: Sixty emmetropic, myopic, and hyperopic subjects (age range, 18 to 35 years) viewed an illuminated target (luminance: 10, 100, 200, 400, 1000, 2000, and 4100 cd/m) within a Badal optical system, at 0 diopters (D) and -3 D vergence, with and without refractive correction. Refractive error was corrected using daily disposable contact lenses. Pupil diameter and accommodation were recorded continuously using a commercially available photorefractor. RESULTS: No significant difference in pupil diameter was found between the refractive groups at 0 D or -3 D target vergence, in the corrected or uncorrected conditions. As expected, pupil diameter decreased with increasing luminance. Target vergence had no significant influence on pupil diameter. In the corrected condition, at 0 D target vergence, the accommodation response was similar in all refractive groups. At -3 D target vergence, the emmetropic and myopic groups accommodated significantly more than the hyperopic group at all luminance levels. There was no correlation between accommodation response and pupil diameter or refractive error in any refractive group. In the uncorrected condition, the accommodation response was significantly greater in the hyperopic group than in the myopic group at all luminance levels, particularly for near viewing. In the hyperopic group, the accommodation response was significantly correlated with refractive error but not pupil diameter. In the myopic group, accommodation response level was not correlated with refractive error or pupil diameter. CONCLUSIONS: Refractive error has no influence on pupil diameter, irrespective of refractive correction or accommodative demand. This suggests that the pupil is controlled by the pupillary light reflex and is not driven by retinal blur.
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Hiperopia/fisiopatologia , Miopia/fisiopatologia , Pupila/fisiologia , Acomodação Ocular/fisiologia , Adolescente , Adulto , Emetropia/fisiologia , Feminino , Humanos , Luz , Masculino , Estimulação Luminosa , Refração Ocular , Adulto JovemRESUMO
The abscopal effect is an extremely rare phenomenon occurring when irradiation or treatment of a primary tumor burden not only results in debulking of the targeted site but also reduces tumor size at distant sites from the intended treatment area. We present the abscopal effect occurring in a patient with low-grade marginal zone lymphoma who subsequently received radioactive iodine therapy for papillary thyroid carcinoma. She was 67 years old when a routine complete blood count at her primary care physician's office yielded a persistent leukocytosis of 14,500/µL with lymphocytosis of 9,870/µL. Immunophenotyping and fluorescence in situ hybridization (FISH) analysis confirmed low-grade marginal zone lymphoma. Over eight years, her peak leukocyte and lymphocyte counts were 24,100/µL and 18,100/µL, respectively. Subsequently, she was diagnosed with papillary thyroid carcinoma after presenting with a new complaint of dysphagia. A total thyroidectomy was performed, followed by 172.1 millicuries of oral I-131 sodium iodine radioactive ablation therapy. Following treatment, her leukocyte and lymphocyte counts were 3,100/µL and 1,100/µL, respectively. Over the next four years, her leukocyte and lymphocyte counts remained within normal limits and she remained symptom free. To our knowledge, there has never been a published report describing the use of radioactive iodine causing abscopal effect benefits for patients with underlying lymphoproliferative diseases.
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Stimulation of the accommodation system results in a response in the vergence system via accommodative vergence cross-link interactions, and stimulation of the vergence system results in an accommodation response via vergence accommodation cross-link interactions. Cross-link interactions are necessary in order to ensure simultaneous responses in the accommodation and vergence systems. The crosslink interactions are represented most comprehensively by the response AC/A (accommodative vergence) and CA/C (vergence accommodation) ratios, although the stimulus AC/A ratio is measured clinically, and the stimulus CA/C ratio is seldom measured in clinical practice. The present study aims to quantify both stimulus and response AC/A and CA/C ratios in a binocularly normal population, and determine the relationship between them. 25 Subjects (mean ± SD age 21.0 ± 1.9 years) were recruited from the university population. A significant linear relationship was found between the stimulus and response ratios, for both AC/A (r² = 0.96, p < 0.001) and CA/C ratios (r² = 0.40, p < 0.05). Good agreement was found between the stimulus and response AC/A ratios (95% CI -0.06 to 0.24 MA/D). Stimulus and response CA/C ratios are linearly related. Stimulus CA/C ratios were higher than response ratios at low values, and lower than response ratios at high values (95% CI -0.46 to 0.42 D/MA). Agreement between stimulus and response CA/C ratios is poorer than that found for AC/A ratios due to increased variability in vergence responses when viewing the Gaussian blurred target. This study has shown that more work is needed to refine the methodology of CA/C ratio measurement.
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Acomodação Ocular/fisiologia , Convergência Ocular/fisiologia , Visão Binocular/fisiologia , Adulto , Feminino , Humanos , Masculino , Estimulação Luminosa/métodos , Análise de Regressão , Adulto JovemRESUMO
Placentitis is reported to be the cause of 9.8-33.5% of abortions, stillbirths and perinatal losses in horses. Bacterial infections are responsible for 53% of placentitis cases with Streptococcus equi ssp. zooepidemicus being isolated in 28% of these cases. Clinically, mares may have a vaginal discharge, show udder development, lactate prenatally and deliver a premature or dead foal. Major aspects of the pathogenesis of infectious preterm delivery that may require more effective therapeutic targeting are myometrial contraction, immunological aspects of preterm delivery, and the effects of proinflammatory cytokine signalling on activation of the fetal hypothalamic-pituitary-adrenal axis. This article focuses on current knowledge of inflammatory signalling secondary to equine placentitis, and the interplay among inflammation, loss of myometrial quiescence and activation of the fetal hypothalamic-pituitary-adrenal axis.
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Doenças dos Cavalos/imunologia , Trabalho de Parto Prematuro/veterinária , Doenças Placentárias/veterinária , Nascimento Prematuro , Animais , Feminino , Doenças dos Cavalos/microbiologia , Doenças dos Cavalos/patologia , Cavalos , Doenças Placentárias/microbiologia , GravidezRESUMO
PURPOSE: To determine whether variation in ocular rigidity (a quantity that describes the elastic properties of the globe) affects the characteristics of horizontal saccadic eye movements. METHODS: Thirty-three young, visually healthy subjects participated with informed consent in the study. Axial length was measured using the IOLMaster ocular biometer. Ocular rigidity coefficients were determined using Schiotz tonometry. Horizontal saccades were stimulated randomly to 40° in 10° steps. Eye movements were recorded continuously at a sampling rate of 60 Hz using the Viewpoint video-eyetracker. RESULTS: Peak velocity increased significantly with increasing ocular rigidity (F [2,263] = 30.635, P < 0.001). Time to peak velocity (F [2,263] = 27.723, P < 0.001) and total response time (F [2,263] = 21.133, P < 0.001) decreased significantly with increasing ocular rigidity. Ocular rigidity was significantly positively correlated with peak velocity (R(2) = 0.67, P < 0.001), and significantly negatively correlated with time to peak velocity (R(2) = 0.64, P < 0.001), and total response time (R(2) = 0.62, P < 0.001). CONCLUSIONS: The known relationship of ocular rigidity with myopia can be extended to shorter hyperopic eyes, which are found to have higher ocular rigidity. The dynamic characteristics of saccadic eye movements are found to vary systematically with ocular rigidity. These findings suggest that the structural characteristics of the eye are an important factor in determining dynamic characteristics of eye movements.
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Pressão Intraocular/fisiologia , Movimentos Sacádicos/fisiologia , Adolescente , Adulto , Comprimento Axial do Olho , Elasticidade , Feminino , Humanos , Masculino , Valores de Referência , Adulto JovemRESUMO
It was hypothesized that optimal sperm recovery rate (RR) without damage to the sperm would be obtained after centrifugation without a cushion solution. Semen collected three times from six light breed stallions was extended to 25 × 10(6) sperm/mL and centrifuged at CON (noncentrifuged), 900NC (no-cushion), 900C (cushion), 1800NC, and 1800C × g for 10 minutes. Sperm concentration, motility (TM and PM), and intact plasma membranes (PLM) and acrosomes (ACR) pre- and postcentrifugation (D0) and after 24 hours (D1) of cooling were evaluated. The RR in the CON (100 ± 0.0), 900NC (93.7 ± 2.9), and 1800NC (96.7 ± 2.6) groups was significantly higher than the 900C (68.7 ± 4.6) and 1800C (79.6 ± 3.5) groups. The D0 TM and PM were not different between the CON, 900NC, 900C, and 1800C, but were lower for the 1800NC group. The D1 TM and PM of the 900NC (75.2 ± 3.8 and 71.1 ± 4.1) and 900C (76.2 ± 3.7 and 72.4 ± 4.0) groups were significantly higher than the 1800NC (71.7 ± 4.1 and 67.3 ± 4.4) and 1800C (71.6 ± 4.1 and 67.2 ± 4.4) groups, and the CON (66.2 ± 4.5 and 60.0 ± 4.8) group was significantly lower than the other groups. The D1 PLM of the CON, 900NC, 900C, 1800NC, and 1800C groups were not different. The ACR on D1 was significantly lower for the CON (93.0 ± 2.4) group compared with all other groups. Optimal RR preserving sperm integrity was obtained in the 900NC group.
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Análise do Sêmen/métodos , Sêmen/citologia , Acrossomo/fisiologia , Animais , Membrana Celular/fisiologia , Separação Celular/métodos , Separação Celular/estatística & dados numéricos , Separação Celular/veterinária , Centrifugação/métodos , Centrifugação/veterinária , Cavalos , Masculino , Análise do Sêmen/estatística & dados numéricos , Análise do Sêmen/veterinária , Contagem de Espermatozoides/estatística & dados numéricosRESUMO
PURPOSE: To characterize corneal, internal, and total ocular monochromatic higher-order aberration (MHOA) changes that occur in the aging eye. METHODS: Prospective observational case series including 300 eyes of 167 patients (mean age = 63.8 years) attending the ophthalmology service at University Hospital Ayr, Scotland. Corneal, internal, and total ocular aberrations were measured over a 6-mm dilated pupil. Zernike coefficients were obtained to the sixth order. Changes in MHOA between age groups and inter-eye correlations between right and left eyes were analyzed. RESULTS: A significant inter-eye correlation was found for refractive mean spherical equivalent and cylinder. A significant inter-eye correlation for the whole eye, corneal, and internal MHOA was found (p < 0.001). Right eye analysis found a significant positive correlation between age and the root mean square of whole eye MHOA (p = 0.012), with an increase from 0.517 µm in the fifth decade to 0.824 µm in the ninth. Total internal MHOA increased from 0.411 to 0.704 µm. A significant positive correlation was found between age and internal fourth- (p = 0.007), fifth- (p = 0.029), and sixth-order (p = 0.025) root mean square aberrations. There were no significant age-related changes in corneal MHOA or corneal spherical aberration. Overall mean (SD) corneal SA was 0.203 (0.082) µm. CONCLUSIONS: A strong correlation between the right and left eyes exists for MHOA. Whole eye MHOA increases with age. Such changes can be attributed to age-related changes in the internal optical quality of the eye. Such normative data are useful to the cataract surgeon when considering the use of an aspherical IOL to counteract corneal-induced SA during cataract surgery.
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Envelhecimento/fisiologia , Córnea/fisiopatologia , Aberrações de Frente de Onda da Córnea/fisiopatologia , Aberrometria , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Topografia da Córnea , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Erros de Refração/fisiopatologia , Adulto JovemRESUMO
Antibody arrays have been used as an effective method for simultaneously detecting multiple proteins such as cytokines. However, their use in quantifying a large number of cellular proteins has the following limitations: (i) unsuitable for simultaneously detecting proteins that may form complexes with each other; (ii) incapable of quantitatively detecting more than one epitope of each protein such as phospho- and nonphospho-epitopes; and (iii) incapable of simultaneously detecting multiple biomarkers on solid surfaces such as formalin-fixed tissue sections. Using a novel multiple epitope detection (MED) technique, we have overcome these limitations and have improved upon currently available antibody-based protein detection technologies. The MED technique employs primary antibody detection of epitopes within fixed cells or tissue, followed by elution of bound antibodies, and subsequent quantification of the eluted antibodies by an epitope array. Using the MED method, we demonstrate accurate detection of individual proteins even in complex with each other, simultaneous quantitative detection of phospho- and nonphospho-epitopes on a protein, and sensitive detection of multiple biomarkers on formalin-fixed tissue sections.
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Anticorpos/química , Epitopos/metabolismo , Análise Serial de Proteínas/métodos , Anticorpos/isolamento & purificação , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/metabolismo , Epitopos/química , Feminino , Fixadores/química , Formaldeído/química , Humanos , Proteínas Imobilizadas/química , Imuno-Histoquímica , Placenta/metabolismo , Gravidez , Ligação Proteica , Proteoma/metabolismo , Fixação de TecidosRESUMO
Conventional centrifugation protocols result in important sperm losses during removal of the supernatant. In this study, the effect of centrifugation force (400 or 900 × g), duration (5 or 10 min), and column height (20 or 40 mL; Experiment 1); sperm concentration (25, 50, and 100 × 10(6)/mL; Experiment 2), and centrifugation medium (EZ-Mixin CST [Animal Reproduction Systems, Chino, CA, USA], INRA96 [IMV Technologies, Maple Grove, MN, USA], or VMDZ [Partnar Animal Health, Port Huron, MI, USA]; Experiment 3) on sperm recovery and survival after centrifugation and cooling and storage were evaluated. Overall, sperm survival was not affected by the combination of centrifugation protocol and cooling. Total sperm yield was highest after centrifugation for 10 min at 400 × g in 20-mL columns (95.6 ± 5%, mean ± SD) or 900 × g in 20-mL (99.2 ± 0.8%) or 40-mL (91.4 ± 4.5%) columns, and at 900 × g for 5 min in 20-mL columns (93.8 ± 8.9%; P < 0.0001). Total (TMY) and progressively motile sperm yield followed a similar pattern (P < 0.0001). Sperm yields were not significantly different among samples centrifuged at various sperm concentrations. However, centrifugation at 100 × 10(6)/mL resulted in significantly lower total sperm yield (83.8 ± 10.7%) and TMY (81.7 ± 6.8%) compared with noncentrifuged semen. Centrifugation in VMDZ resulted in significantly lower TMY (69.3 ± 22.6%), progressively motile sperm yield (63.5 ± 18.2%), viable yield (60.9 ± 36.5%), and survival of progressively motile sperm after cooling (21 ± 10.8%) compared with noncentrifuged semen. In conclusion, centrifuging volumes of ≤ 20 mL minimized sperm losses with conventional protocols. With 40-mL columns, it may be recommended to increase the centrifugal force to 900 × g for 10 min and dilute the semen to a sperm concentration of 25 to 50 × 10(6)/mL in a milk- or fractionated milk-based medium. The semen extender VMDZ did not seem well suited for centrifugation of equine semen.