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Treatment of urethral stricture is a complex reconstructive urology treatment. Previous surgery in the treatment of urethral stricture increases the risk of urinary incontinence. Therefore, this study aimed to assess the outcome of comparing two preservative or nonsurgical methods, clean intermittent catheterization (CIC) or hydraulic urethral dilatation (HUD). Forty-eight patients diagnosed with symptomatic urethral stricture were enrolled in the study. Patients were randomly divided into two groups: Nelaton-based dilatation and HUD treatment. Patients were assessed using patient-reported outcome measure for urethral stricture surgery (USS-PROM) through direct clinical interview and were followed up for 12 months. The rate of urinary infection, the need for surgical intervention, and the patient's quality of life were considered as outcomes of the interventions. SPSS software, version 20.0, was used for statistical analysis. Overall, 23 (47.9%) patients underwent HUD of stricture and Nelaton-based dilatation of stricture was done in 25 (52.1%) patients. We found that surgical intervention was required in four patients in the HUD group, but 15 patients in the CIC group required surgical intervention. Also, the need for surgical intervention and urinary tract infection (UTI) was significantly greater in the CIC group compared with the HUD group. With respect to the patient-reported outcomes, the mean Peeling's voiding picture (Q8) and EQ-VAS score (Q16) in the HUD group (p = .02) were significantly greater than those in the CIC group (p = .02). We obtained excellent results by using an HUD catheter. This safe, simple, well-tolerated, home-based procedure reduced the need for hospital admission to operate for repeat.
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Estrechez Uretral , Humanos , Estrechez Uretral/cirugía , Constricción Patológica , Dilatación/métodos , Calidad de Vida , Uretra/cirugía , Resultado del TratamientoRESUMEN
The maternal cardiovascular-circulatory system undergoes profound changes almost from the conception of a pregnancy until the postpartum period to support the maternal adaptions required for pregnancy and lactation. Maintenance of cardiovascular homeostasis requires changes in the cardiovascular autonomic responses. Here, we present a longitudinal study of the maternal cardiovascular autonomic responses to pregnancy and maternal position. Over a normal gestation, in the left lateral position there are significant changes in both time and frequency domain parameters reflecting heart rate variability. We show that cardiovascular autonomic responses to physiological stressors (standing and supine positions in late pregnancy) became significantly different with advancing gestation. In the third trimester, 60% of the subjects had an unstable heart rate response on standing, and these subjects had a significantly reduced sample entropy evident in their heart rate variability data. By 6 weeks, postpartum function returned to near the non-pregnant state, but there were consistent differences in high-frequency power when compared to nulligravid cases. Finally, we review complementary evidence, in particular from magnetic resonance imaging, that provides insights into the maternal and fetal impacts of positioning in pregnancy. This demonstrates a clear relationship between supine position and maternal hemodynamic parameters, which relates to compression of the inferior vena cava (p = 0.05). Together, these studies demonstrate new understanding of the physiology of physiological stressors related to position.
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Fetal growth restriction (FGR) and maternal supine going-to-sleep position are both risk factors for late stillbirth. This study aimed to use magnetic resonance imaging (MRI) to quantify the effect of maternal supine position on maternal-placental and fetoplacental blood flow, placental oxygen transfer and fetal oxygenation in FGR and healthy pregnancies. Twelve women with FGR and 27 women with healthy pregnancies at 34-38 weeks' gestation underwent MRI in both left lateral and supine positions. Phase-contrast MRI and a functional MRI technique (DECIDE) were used to measure blood flow in the maternal internal iliac arteries (IIAs) and umbilical vein (UV), placental oxygen transfer (placental flux), fetal oxygen saturation (FO2 ), and fetal oxygen delivery (delivery flux). The presence of FGR, compared to healthy pregnancies, was associated with a 7.8% lower FO2 (P = 0.02), reduced placental flux, and reduced delivery flux. Maternal supine positioning caused a 3.8% reduction in FO2 (P = 0.001), and significant reductions in total IIA flow, placental flux, UV flow and delivery flux compared to maternal left lateral position. The effect of maternal supine position on fetal oxygen delivery was independent of FGR pregnancy, meaning that supine positioning has an additive effect of reducing fetal oxygenation further in women with FGR, compared to women with appropriately grown for age pregnancies. Meanwhile, the effect of maternal supine positioning on placental oxygen transfer was not independent of the effect of FGR. Therefore, growth-restricted fetuses, which are chronically hypoxaemic, experience a relatively greater decline in oxygen transfer when mothers lie supine in late gestation compared to appropriately growing fetuses. KEY POINTS: Fetal growth restriction (FGR) is the most common risk factor associated with stillbirth, and early recognition and timely delivery is vital to reduce this risk. Maternal supine going-to-sleep position is found to increase the risk of late stillbirth but when combined with having a FGR pregnancy, maternal supine position leads to 15 times greater odds of stillbirth compared to supine sleeping with appropriately grown for age (AGA) pregnancies. Using MRI, this study quantifies the chronic hypoxaemia experienced by growth-restricted fetuses due to 13.5% lower placental oxygen transfer and 26% lower fetal oxygen delivery compared to AGA fetuses. With maternal supine positioning, there is a 23% reduction in maternal-placental blood flow and a further 14% reduction in fetal oxygen delivery for both FGR and AGA pregnancies, but this effect is proportionally greater for growth-restricted fetuses. This knowledge emphasises the importance of avoiding supine positioning in late pregnancy, particularly for vulnerable FGR pregnancies.
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Placenta , Circulación Placentaria , Embarazo , Femenino , Humanos , Placenta/diagnóstico por imagen , Placenta/irrigación sanguínea , Retardo del Crecimiento Fetal/diagnóstico por imagen , Mortinato , Imagen por Resonancia Magnética , OxígenoRESUMEN
Coronavirus disease 2019 (COVID-19), as a pandemic disease and a global concern, is a respiratory tract infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Besides its common symptoms, some other less common symptoms, including genital ulcers, have been reported. Other complications such as autoimmune disease can present with genital ulcers. Differential diagnosis includes Reiter syndrome, Behcet's disease, and psoriasis. We report a 32-year-old married man with genital ulcers due to COVID-19 infection and as a complication of his disease.
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Síndrome de Behçet , COVID-19 , Masculino , Humanos , Adulto , Úlcera/complicaciones , Úlcera/diagnóstico , COVID-19/complicaciones , SARS-CoV-2 , Síndrome de Behçet/complicaciones , Síndrome de Behçet/diagnóstico , GenitalesRESUMEN
Substitution urethroplasty with either a flap or graft is the gold standard for treating long segment urethral strictures. In 1992, Burger and colleagues rediscovered and popularized buccal mucosal graft (BMG). After that El-Kassaby and colleagues, in 1993, used BMG to repair anterior urethral stricture. De la Chapelle syndrome or 46 XX male syndrome is a rare genetic disorder found in 1 in 20,000-25,000 men. This condition described as a presentation of male phenotype along a 46 xx karyotype. In this case report, we report a reconstructive surgery of a 46 XX male syndrome with ambiguous genitalia who presented with the chief complaint of bulbar urethral fistula opened in the perineal space. In this case, we used a buccal mucous graft with the ventral-onlay urethroplasty technique for reconstructing the failed bulbar urethra and closure of the fistula.
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Trastornos Testiculares del Desarrollo Sexual 46, XX , Fístula , Estrechez Uretral , Masculino , Humanos , Uretra/cirugía , Trastornos Testiculares del Desarrollo Sexual 46, XX/complicaciones , Mucosa Bucal/trasplante , Estrechez Uretral/cirugía , Fístula/cirugía , Fístula/complicaciones , Resultado del TratamientoRESUMEN
Objectives: To estimate the proportion of adult diabetics with a skin to deltoid muscle distance (SDMD) of > 25 mm, representing a distance greater than the standard needle length used for intramuscular COVID-19 vaccination, and to assess whether anthropometric measurements predict ultrasound SDMD measurements. Design: Non-interventional cross-sectional study. Setting: Single site, non-clinical setting, Wellington, New Zealand. Participants: One hundred participants (50 females) aged at least 18 years diagnosis with diabetes. All participants completed the study. Main outcome measures: The proportions of participants with a SDMD > 25 mm and a SDMD > 20 mm (indicating that the needle would not have penetrated at least 5 mm into the deltoid, which is considered necessary to ensure deposition of vaccine into muscle); the relationship between anthropometric measurements (body weight, body height, body mass index (BMI), skinfold thickness, arm circumference) and SDMD measured by ultrasound. Results: The proportion (95 %CI) of participants with a SDMD > 25 mm was 6/100; 6 % (2.2 to 12.6), and the proportion with a SDMD > 20 mm was 11 % (5.6 to 18.8), of which 9/11 had a BMI ≥ 30 kg/m2 and 9/11 were female. The strongest relationships between anthropometric measurements and SDMD were with arm circumference (r = 0.76, P < 0.001) and BMI (r = 0.73, P < 0.001). Arm circumference and BMI were the best predictors of SDMD measurements with AUC for ROC curves of 0.99 and 0.94 above the 25 mm cut point, 0.97 and 0.89 above the 20 mm cut point respectively. Conclusions: The standard needle length of 25 mm is likely to be insufficient to ensure deposition of COVID-19 vaccine within the deltoid muscle in a small but important proportion of obese adults with diabetes. Arm circumference and BMI are simple measurements that could identify those that need a long needle to ensure successful intramuscular vaccine administration. Funding: Ruth Maud Ring Spencer Estate; Health Research Council of New Zealand (Independent Research Organisation).
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Accurate acquisition and segmentation of muscles are essential in 3-D freehand ultrasonography (US) to estimate in vivo muscle volume, but the source of segmentation inaccuracy in shape variation has never been the focus. This study was aimed at investigating reliability of 3-D US in the acquisition and segmentation for muscle volume of two muscles of different sizes and in identifying a primary source of measurement difference. The lateral gastrocnemius and flexor pollicis brevis of 12 healthy adults were assessed using freehand 3-D US scans. The motion-tracking data of the probe were synchronized with the B-mode ultrasound scan to reconstruct 3-D muscle volume. Statistical shape modeling was used to provide a spatial segmentation volume difference that further explains the variation around segmentation repeatability. The absolute difference of the flexor pollicis brevis was 3.5 percentage points greater than that for the lateral gastrocnemius. The highest measurement differences were observed when for inter-acquirer analysis. Statistical shape modeling revealed that the primary segmentation volume differences were at the muscle ends and edges, where the muscle interfaces with the surrounding muscles. Three-dimensional US is a reliable tool in the clinical setting, but care must be taken to ensure that acquisition and segmentation are consistent, particularly in a small muscle that interfaces with tendons and other soft tissues.
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Mano , Músculo Esquelético , Adulto , Humanos , Músculo Esquelético/diagnóstico por imagen , Reproducibilidad de los Resultados , Tendones/diagnóstico por imagen , Ultrasonografía/métodosRESUMEN
Urethral foreign bodies are rare with a variety of causes, including psychiatric illness, autoeroticism, intoxication, and perceived contraception, we present a 49-yr-old man who went to the emergency ward 9-month ago inserting an AA battery in his urethra. The battery was removed without surgery after 24 hours. The patient was then referred to the hospital after 5 months with complaints of obstructive urinary symptoms such as decreased urinary caliber, diagnosed with penile urethral stricture, and undergoes dilatation of the urethra. Usually, in the case of urethral stricture, the initial steps including urethral dilation and internal urethrotomy can be used.
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The aim of this review was to report on the imaging modalities used to assess morphological and architectural properties of the m. triceps surae muscle in typically developing children, and the available reliability analyses. Scopus and MEDLINE (Pubmed) were searched systematically for all original articles published up to September 2020 measuring morphological and architectural properties of the m. triceps surae in typically developing children (18 years or under). Thirty eligible studies were included in this analysis, measuring fibre bundle length (FBL) (n = 11), pennation angle (PA) (n = 10), muscle volume (MV) (n = 16) and physiological cross-sectional area (PCSA) (n = 4). Three primary imaging modalities were utilised to assess these architectural parameters in vivo: two-dimensional ultrasound (2DUS; n = 12), three-dimensional ultrasound (3DUS; n = 9) and magnetic resonance imaging (MRI; n = 6). The mean age of participants ranged from 1.4 years to 18 years old. There was an apparent increase in m. gastrocnemius medialis MV and pCSA with age; however, no trend was evident with FBL or PA. Analysis of correlations of muscle variables with age was limited by a lack of longitudinal data and methodological variations between studies affecting outcomes. Only five studies evaluated the reliability of the methods. Imaging methodologies such as MRI and US may provide valuable insight into the development of skeletal muscle from childhood to adulthood; however, variations in methodological approaches can significantly influence outcomes. Researchers wishing to develop a model of typical muscle development should carry out longitudinal architectural assessment of all muscles comprising the m. triceps surae utilising a consistent approach that minimises confounding errors.
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Pierna , Músculo Esquelético , Adolescente , Niño , Humanos , Lactante , Imagen por Resonancia Magnética/métodos , Músculo Esquelético/anatomía & histología , Reproducibilidad de los Resultados , Ultrasonografía/métodos , Adulto JovenRESUMEN
Lumen diameter reduction which is called urethral stricture has several causative agents including iatrogenic, inflammatory, traumatic, and idiopathic factors. The main treatment options are transurethral or open surgical interventions.The UroLume which was introduced around 33 years ago is an intraurethral stent and as a temporary treatment approach has a maximum 9-month durability, but here we present the first ever reported forgotten 16-year remained UroLume without any complication.
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Procedimientos de Cirugía Plástica , Estrechez Uretral , Femenino , Humanos , Masculino , Procedimientos de Cirugía Plástica/efectos adversos , Stents/efectos adversos , Uretra/cirugía , Estrechez Uretral/etiología , Estrechez Uretral/cirugíaRESUMEN
EB is an inherited skin disease that presents with the symptom of skin blisters following trauma, Involvement of the Urogenital system in these people is in the form of meatal stenosis, urinary tract infection and ureteral stricture. We introduce patient with EB and urethral involvement. A 32-year-old man without prenatal problems whose EB skin symptoms started at 6 months of age and urinary symptoms started at 12 years of age. Skin changes on prenatal ultrasound at 18 and 21 weeks of gestation will help in the diagnosis. Follow-up of EB patients with ultrasound will help to identify urogenital complications earlier.
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BACKGROUND: Outer membrane vesicles (OMVs) release from Gram-negative bacteria and are interesting alternatives that can replace those vaccines that contain naturally incorporated bacterial surface antigens, lipopolysaccharides (LPS) and outer membrane proteins (OMPs). Nanoparticles can be used to encapsulate vesicles for slow release and prevent macromolecular degradation. OBJECTIVE: Therefore, encapsulation of OMVs of B. pertussis into sodium alginate nanoparticles was the main aim of the current study. METHODS: The OMVs of B. pertussis extracted and characterized by particle sizer, electron microscopy, SDSPAGE and Western blot assays. The extracted OMVs were encapsulated in sodium alginate nanoparticles (OMV-NP) using unique gelation process and injected into BALB/c mice. Immunogenicity indices such as different classes of antibodies and interleukins related to different T cell lines were evaluated in immunized mice by ELISA. The respiratory challenge was evaluated in the groups of mice. The existence of pertussis toxin (PTX), filamentous haemagglutinin (FHA) and PRN (pertactin) in B. pertussis OMVs was verified using SDSPAGE and Western blot analysis. RESULTS: TEM electron microscopy showed the size of these OMVs to be around 20-80 nm. The OMVs with appropriate quality were encapsulated into sodium alginate nanoparticles (OMV-NP), and the final size was about 500 nm after encapsulation. Immunity indices were significantly higher in the OMV-NP receiving group. In challenge tests, the OMV-NP vaccine was able to show the highest rate of lung clearance compared to the control groups (OMV and wPV) at the lowest injection dose. CONCLUSION: The results indicate the potential of OMV-NP as a novel vaccine delivery system.
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Bordetella pertussis , Nanopartículas , Alginatos , Animales , Humanos , Ratones , Ratones Endogámicos BALB C , Vacuna contra la Tos FerinaRESUMEN
Barefoot running has been associated with lowered joint loading, but it remains unclear whether the biomechanical benefits are evident after mid-distance running. A musculoskeletal model was adopted for estimating lower limb joint loading for barefoot (n = 10) versus shod (n = 10) 5 km running. The barefoot group reduced peak joint reaction force at the hip and knee, and presented muscle force reductions compared to shod controls with significant group effects and interaction effects (p < .05). These changes were primarily group effects as time point effects were not significant. These findings should be considered when designing barefoot running shoes, running programmes, and injury prevention programmes.
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Pie , Carrera , Articulación del Tobillo , Fenómenos Biomecánicos , Marcha , Humanos , Músculos , ZapatosRESUMEN
KEY POINTS: Maternal supine sleep position in late pregnancy is associated with an increased risk of stillbirth. Maternal supine position in late pregnancy reduces maternal cardiac output and uterine blood flow. Using MRI, this study shows that compared to the left lateral position, maternal supine position in late pregnancy is associated with reduced utero-placental blood flow and oxygen transfer across the placenta with an average 6.2% reduction in oxygen delivery to the fetus and an average 11% reduction in fetal umbilical venous blood flow. ABSTRACT: Maternal sleep position in late gestation is associated with an increased risk of stillbirth, though the pathophysiological reasons for this are unclear. Studies using magnetic resonance imaging (MRI) have shown that compared with lateral positions, lying supine causes a reduction in cardiac output, reduced abdominal aortic blood flow and reduced vena caval flow which is only partially compensated for by increased flow in the azygos venous system. Using functional MRI techniques, including an acquisition termed diffusion-relaxation combined imaging of the placenta (DECIDE), which combines diffusion weighted imaging and T2 relaxometry, blood flow and oxygen transfer were estimated in the maternal, fetal and placental compartments when subjects were scanned both supine and in left lateral positions. In late gestation pregnancy, lying supine caused a 23.7% (P < 0.0001) reduction in total internal iliac arterial blood flow to the uterus. In addition, lying in the supine position caused a 6.2% (P = 0.038) reduction in oxygen movement across the placenta. The reductions in oxygen transfer to the fetus, termed delivery flux, of 11.2% (P = 0.0597) and in fetal oxygen saturation of 4.4% (P = 0.0793) did not reach statistical significance. It is concluded that even in healthy late gestation pregnancy, maternal position significantly affects oxygen transfer across the placenta and may in part provide an explanation for late stillbirth in vulnerable fetuses.
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Imagen por Resonancia Magnética , Circulación Placentaria , Femenino , Feto/diagnóstico por imagen , Hemodinámica , Humanos , Placenta/diagnóstico por imagen , Embarazo , Tercer Trimestre del EmbarazoRESUMEN
OBJECTIVES: There remains a lack of consistent evidence surrounding the developmental anatomy of the maxillary sinus (MS), despite its clinical significance in radiological interpretation and surgical intervention. Our aim was to analyse the typical growth trends of the MS throughout development, to better understand its morphological transformation from birth until late adolescence. METHODS: Computed Tomography (CT) scans of individuals aged 0-18 years within the Auckland District Health Board region in New Zealand were examined retrospectively. Sixty-five patients were selected for the study after careful review of criteria. Manual 3D-reconstruction was utilised to replicate the MS from CT images and the mean changes in size, volume and shape with increasing age were analysed. Principal component analysis (PCA) was performed to statistically calculate the mean 3D shapes of the MS in each age group and analyse the independent modes of variation evident in sinus morphology between individuals of each age category. RESULTS: At 0-1 years of age, mean MS size was 1.03 × 1.82 × 1.27 cm (width, length, height) with a mean volume of 0.81 cm3. By 16-18 years, mean size increased to 3.39 × 4.30 × 4.63 cm and mean volume became 21.63 cm3. Growth increased linearly but most rapid expansion occurred in the first 4-5 years of life. No significant differences were observed between males and females or right and left sides. Sinus morphology gradually progressed throughout the course of growth from an ellipsoidal structure at its rudimentary phase to a pyramidal shape when fully matured. PCA revealed that within younger age groups significant variation was observed in MS size, while older age groups had relatively less size variation but increased variability in 3D shape. CONCLUSIONS: The results of this study can be used as a reference for typical MS growth in healthy children and as a basis to study how disease or surgical intervention may influence sinus development in children.
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Seno Maxilar , Tomografía Computarizada por Rayos X , Adolescente , Anciano , Niño , Preescolar , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Seno Maxilar/diagnóstico por imagen , Nueva Zelanda , Radiografía , Estudios RetrospectivosRESUMEN
OBJECTIVE: Since sperm abnormalities are known to be a major reason for recurrent pregnancy loss (RPL), any defects in DNA structure and chromatin condensation can place embryos at risk in the early stage of development and implantation. As antioxidants such as vitamin C may play a protective role against the destruction of protamine genes in sperm chromatin, this study was conducted to evaluate the effects of vitamin C on chromatin and the expression of protamine genes in the male partners of couples with RPL. METHODS: Twenty male partners of couples with RPL were selected as the intervention group and received vitamin C supplementation (250 mg daily for 3 months). Healthy fertile men (n=20) were included as controls. Sperm chromatin, DNA integrity, and the expression levels of protamine genes were evaluated before and after treatment. RESULTS: Significant differences were found in sperm morphology, protamine deficiency, and apoptosis between the two groups and before and after vitamin C administration. A significant change was found in mRNA levels of PRM1, PRM2, and the PRM1/PRM2 ratio after treatment. CONCLUSION: Daily oral administration of vitamin C may improve human sperm parameters and DNA integrity by increasing protamine gene expression levels in the male partners of couples with RPL. The beneficial effects of vitamin C supplementation as an antioxidant for the male partners of couples with RPL could lead to improved pregnancy outcomes in these cases.
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Some serovars of salmonella cause huge global diseases such as enteric fever and invasive non typhoidal Salmonella disease. Flagellin as a key antigenic component of salmonella, can induce humoral and cellular immunity responses. In this research, we performed an opsonophagocytic killing assay (OPKA) as an important mechanism of the host-defense system, for salmonella to study the activity of anti-sera of native FliC, truncated modified recombinant FliC (tmFliC) and full length recombinant FliC proteins (flFliC). Also, the potency of antibodies for inhibiting bacterial movement was evaluated by traditional and newly-designed motility inhibition assay methods. Results showed both recombinant FliC anti-sera and native FliC (nFliC) anti-serum had the ability to opsonize Salmonella typhimurim, which led to bacterial clearance by mice macrophages. Also, inhibition of bacterial motility was observed for all anti-sera. Anti-nFliC and anti-flFliC sera showed higher effects on Salmonella typhimurim motility than that of tmFliC. In traditional method, about 88%, 86% and 80% inhibition were observed by using 5% nFliC, anti-flFliC and anti-tmFliC sera, respectively. In the newly-designed method using SIM (Sulfide indole motility) medium, results confirmed the traditional method for motility inhibition. Our findings suggest that salmonella fliC as a protective antigen may disrupt the flagellum apparatus activity.
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A 3D anatomically-based finite-element foot model was adopted for predicting von Mises stresses within tibiotalar cartilage following 5 km barefoot running. To compare this predicted stress with T2 maps, magnetic resonance scans of the right ankle and plantar pressure were obtained from ten novices and ten marathon-experienced runners before and after running. Following running, tibiotalar cartilage stress was decreased in experienced runners. This corresponded with T2 values that did not change between pre- and post-running suggesting no increase in cartilage fluid levels. In contrast, novices maintained the same level of von Mises stress and this corresponded with a significant T2 increase in tibiotalar cartilage.
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Atletas , Cartílago/fisiopatología , Pie/fisiopatología , Carrera/fisiología , Estrés Mecánico , Astrágalo/fisiopatología , Adulto , Cartílago/diagnóstico por imagen , Femenino , Pie/diagnóstico por imagen , Humanos , Imagen por Resonancia Magnética , Masculino , Presión , Astrágalo/diagnóstico por imagenRESUMEN
BACKGROUND: The protozoan parasite Theileria annulata is the causative agent of tropical theileriosis in cattle. Vaccination is recommended by administration of attenuated schizont-infected cell lines. The expected protective immunity post-vaccination can be demonstrated by challenge test through inoculation of highly virulent infective sporozoites. The aim of this study was to produce Hyalomma anatolicum anatolicum tick infected with T. annulata (local strain) for preparation of tick-derived sporozoite stabilates for molecular characterization and infectivity test assay. METHODS: A local T. annulata strain was used for experimental infection of calves. A field isolate of H. a. anatolicum was isolated, laboratory-reared and infected by blood-feeding on Theileria infected above-mentioned calves. The infectivity of calf, tick and prepared stabilate were confirmed by clinical signs of theileriosis, microscopic inspection, RT-PCR and in vitro cell culture. RESULTS: The tick stabilate was prepared and cryopreserved in liquid nitrogen. The infectivity of the tick stabilate was verified by in vivo bioassay, in vitro cell culture infection, microscopic inspection in salivary glands and RT-PCR assay. The in vitro produced cell line in this study was characterized by T. annulata Cytochrome b gene analyzing. CONCLUSION: The infectivity of a new prepared tick-derived sporozoite stabilate was confirmed in susceptible calves; by microscopically, post mortem, tick microscopic and molecular assays. Moreover, naïve PBMCs were transformed and proliferated by T. annulata infected tick stabilate to immortal T. annulata schizont infected cell line. The potent infective sporozoite tick derived stabilate could be used for vaccine efficacy and challenge test as well as in vaccine development.
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OBJECTIVE: To investigate sperm chromatin/DNA integrity, global DNA methylation, and DNMT mRNA transcription in men with oligoasthenoteratozoospermia (OAT) compared with normozoospermic men. METHODS: Semen samples from 32 OAT patients who comprised the case group and 32 normozoospermic men who comprised the control group were isolated and purified using a standard gradient isolation procedure according to World Health Organization criteria. DNMT1, DNMT3A, and DNMT3B transcripts were then compared between groups using real-time quantitative reverse-transcription polymerase chain reaction. Global DNA methylation in sperm was determined by an enzyme-linked immunosorbent assay. Protamine deficiency and the proportion of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3), aniline blue (AB), and toluidine blue (TB) staining, as well as the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The p-values <0.05 were considered to indicate statistical significance. RESULTS: Significantly higher proportions of AB+, TB+, CMA3+, and TUNEL+ spermatozoa, as well as DNMT3A and DNMT3B transcription, were found in the OAT group. Positive correlations were detected between sperm parameters, DNA/chromatin damage, and DNMT3A and DNMT3B transcripts. Global DNA methylation was significantly higher in the OAT patients and had a significant correlation with abnormal results of all sperm chromatin integrity tests, but was not associated with DNMT1, DNMT3A, or DNMT3B expression. CONCLUSION: Oligoasthenoteratozoospermic men showed abnormal sperm parameters, abnormal chromatin/DNA integrity, and a higher global DNA methylation rate, as well as overexpression of DNMT mRNA.