Subcutaneous and visceral adipose tissue in patients with primary and recurrent incisional hernia

Purpose Visceral obesity rather than body mass index has been reported to be associated with a higher incidence of incisional hernias. The aim of this study was to examine the relationship between CT measured adipose tissue and muscle in primary and recurrent incisional hernia. Methods Patients with a ‘Primary’ or ‘Recurrent incisional hernia’ were obtained from a prospective cohort of patients who were being assessed for incisional hernia repair over a 2-year period. Computerised tomography (CT)-images were analysed using NIH Image-J software to quantify adipose tissue and skeletal muscle cross-sectional areas at the level of lumber vertebra 3/4 using standard Hounsfield units. To test inter-observer ‘absolute agreement’, each parameter was measured independently by two investigators and reliability analysis performed. Results Thirty-six patients were included in the study: 15 had a Primary while 21 had a Recurrent incisional hernia. Both groups had similar baseline characteristics. Reliability analysis for CT-measured areas showed very high interclass correlation coefficient (ICC) between observers. Patients in the recurrent group had significantly greater subcutaneous adipose tissue (SAT) [median = 321.9cm2 vs 230.9cm2, p = 0.04] and visceral adipose tissue (VAT) [median = 221.1cm2 vs 146.8cm2, p = 0.03] than those in the primary group. There was no difference in skeletal muscle areas for right [median = 2.8cm2 vs 2.9cm2] and left [median = 3.7cm2 vs 4.1cm2] rectus muscles between groups. Conclusion Our study shows that patients with a recurrent incisional hernia have significantly more subcutaneous and visceral adipose tissue than those with a primary incisional hernia. Further studies in this area are required if we are to reduce the burden of recurrent hernia following repair of a primary incisional hernia.

Abstract 212: Effects of Therapeutic Hypothermia on Glymphatic Clearance

Objectives: Therapeutic hypothermia is the most potent protective therapy for cerebral ischemia in pre-clinical models, however clinical trials have failed to show a significant benefit in patient outcomes. We sought to reconcile this discrepancy by investigating whether hypothermia impaired glymphatic system mediated waste clearance from brain parenchyma in a rat model. Methods: Sprague -Dawley rats were divided into normothermia and hypothermia groups. Hypothermia was achieved using a perivascular closed-loop cooling circuit implanted one week before the experiments in both the groups. Animals in the normothermia group were kept at 37C while the hypothermia animals were cooled to 33C. Once the targeted temperature was reached, two fluorescent tracers, small molecular weight (Texas Red-Dextran 3KdA) and large molecular weight (FITC -Dextran 2MdA) were injected intracisternally in each animal. Animals in both the groups were monitored for blood pressure and blood gases. Thirty minutes after injection, animals were perfused with 4% paraformaldehyde and 50 micron sections were cut using Leica vibratome. Brain sections were mounted on slides and cover slipped using antifade Fluro gold. Sections were imaged on Carl-Zeiss apotome microscope. For each animal 12 sections were imaged, data was analyzed using NIH Image J software. Results and Discussion: Cerebrospinal fluid tracer efflux in brain was significantly reduced in the hypothermia group compared to the normothermia group. This experiment helps provide insight as to a potential reason clinical trials have failed to show benefit in stroke patients, as hypothermia may have an untoward effect of hindering the brain’s natural clearance system from clearing inflammatory molecules and other waste from brain parenchyma.

Abstract 483: Novel Evaluation Method of Spheroid

Introduction: The 3D culture methods are growing in importance in various fields, such as tissue engineering, cancer research and drug estimation. There are many methods to create 3D cell aggregation, as known as spheroids, e.g. hanging drop method, low attach U-shape plate, microwell low attach plate, spinning flask and magnetic levitation. In either field, researchers require not only high-scale but also high-quality spheroids creation. However, the method for evaluation of spheroids has not yet established. We established an accurate method of quantification of spheroids by measuring cell density and area of them with NIH Image J. Method: Spheroids were created with a hanging drop method (HD) and 96 well low attach U-shape plate (LAP) by using H9C2 cells, NIH 3T3 cells, human cardiac fibroblast and human dermal fibroblast cells using 22000 cells, 33000 cells and 44000 cells per spheroids. Spheroids were created five times using one of these systems.These spheroids were taken pictures in 10 times magnification with a microscope under constant condition using one tenth of spheroids solution created with HD and LAP and then quantified mean gray value and area using NIH image J. We saw relationship between its cell diameter and these data and also measured the cost and time for creating one spheroid and compared them among these methods. We also compared these data with histological analysis. Result: Spheroids created with HD had no relation between cell number and spheroid quality, whereas those with LAP demonstrated that higher cell numbers created larger and higher density of spheroid consistently. The number of spheroids created with the hanging drop method was higher but more inconstant and the success rate was low. However, Spheroids creation with this method took much less cost and time compared with LAP. We could see a positive correlation between area and cell size and negative correlation between gray value and its cell diameter when we create spheroids with HD with the same kinds of cells, i.e. NIH 3T3 cells, human cardiac fibroblast, and human dermal fibroblast cells. Conclusion: Although HD has the potentiality to create a large number of spheroids at once and this method was cost and time effective, the quality of spheroids was not consistent compared to LAP. The cell diameter influenced the quality of spheroids when we created them with HD.

Estudo comparativo da densidade radiográfica de cimentos resinosos

Introdução: A radiopacidade dos cimentos resinosos revela a presença de partículas densas, sendo útil na identificação da adaptação de peças cimentadas. Nosso objetivo é comparar a densidade óptica de diferentes cimentos resinosos pelos métodos radiográficos convencional e digital. Métodos: Para cada cimento escolhido (C&B, BisCem, Enforce) e para o grupo-controle (amálgama) foram confeccionados 5 corpos de prova (cp) idênticos em uma matriz de pvc. Cada um dos cp foi radiografado 3 vezes tanto pelo método convencional (utilizando-se filme periapical e aparelho de raios X intra-oral), quanto pelo digital (utilizando-se placas sensoras periapicais do sistema Digora-Soredex). As radiografias convencionais foram digitalizadas e a análise óptica da densidade dos materiais foi comparada pelo software Image J (NIH Image - Machintoch). Para o tratamento estatístico realizado pela ANOVA com 2 fatores e o teste complementar de Tukey com nível de significância de 5%, os valores considerados foram os obtidos a partir da média dos valores entre as diferentes radiografias e cp. Resultados: os valores de densidade pelo método convencional e digital foram respectivamente para cada material: 232,87 e 255 para o amálgama; 136,73 e 136,24 para o Enforce; 135,44 e 148,51 para o Bis-Cem e 125,24 e 58,97 para o C&B. Conclusões: Não houve diferenças significantes entre os métodos radiográficos (p=0,069%). Foram verificadas diferenças significantes entre os materiais (p=0,00), em ambos os métodos, exceto entre o Enforce e o BisCem, que obtiveram média de tons de cinza semelhantes. O cimento C&B mostrou as menores médias de tons de cinza.

Micromorphological characterization of adhesive interface of sound dentin and total-etch and self-etch adhesives

Introduction. The ultimate goal in restorative dentistry has always been to achieve strong and permanent bond between the dental tissues and filling materials. It is not easy to achieve this task because the bonding process is different for enamel and dentin - dentin is more humid and more organic than enamel. It is moisture and organic nature of dentin that make this hard tissue very complex to achieve adhesive bond. One of the first and most widely used tools for examining the adhesive bond between hard dental tissues and composite restorative materials is scanning electron microscopy. The aim of this study was scanning electron microscopy analyzes the interfacial micro morphology of total-etch and self-etch adhesives. Material and Methods. Micro morphological characteristics of interface between totaletch adhesive (Prime & Bond NT) in combination with the corresponding composite (Ceram X Mono) were compared with those of self-etching adhesive (AdheSE One) in combination with the corresponding composite (Tetric EvoCeram). The specimens were observed under 1000 x magnification of scanning electron microscopy (JEOL, JSM-6460 Low Vacuum). Measurement of the thickness of the hybrid layer of the examined composite systems was performed with the software of the device used (NIH Image ?nalyser). Results. Micromorphological analysis of interface showed that the hybrid layer in sound dentin was well formed, its average thickness being 2.68 ?m, with a large number of resin tags and a large amount of lateral branches for specimens with a composite system Prime & Bond NT - Ceram X Mono. However, the specimens with composite systems Adhese One - Tetric EvoCeram did not show the presence of hybrid layer and the resin tags were poorly represented. Conclusion. The results of this study suggest that total-etch adhesives bond better with sound dentin than self-etch adhesives.

Obesity and menopausal status as determinants of procarcinogenic breast inflammation.

40 Background: Chronic inflammation predisposes to several malignancies. We previously demonstrated an obesity → inflammation → aromatase axis in breast tissue. As obesity is a risk factor for postmenopausal (PoM) but not premenopausal (PreM) breast cancer (BC), we examined whether menopause and body mass index (BMI) independently impact breast white adipose tissue (WAT) inflammation. Methods: WAT was prospectively collected from patients (pts) from 04/10 to 08/13. WAT inflammation, detected by CD68 immunohistochemistry, was defined by the presence of dead or dying adipocytes surrounded by an envelope of macrophages known as crown-like structures of the breast (CLS-B). WAT area was measured with NIH Image J. Adipocyte diameter was measured with Canvas 11 Software. Endpoints were 1) CLS-B (+/-) and 2) CLS-B/cm2. Clinicopathologic associations with CLS-B were analyzed by logistic regression and Fisher’s exact test. Results: WAT (237 mastectomies, 13 abdominal reconstructions) was obtained from 238 pts; median age 48 (range 22 to 90). CLS-B occurrence and number of CLS-B/cm2 were greater in overweight/obese (BMI ≥ 25) and PoM pts compared to lean (BMI < 25) and PreM pts (Table). In multivariable analyses, BMI and PoM state were independently associated with CLS-B presence (p <.01 and p = .04) and greater CLS-B/cm2(p < .01 and p = .01). PoM pts had larger mean adipocyte diameter (105.2 +/- 14.0 μ) than PreM pts (95.7 +/-15.6 μ; p < 0.01). In pts with bilateral breast WAT and abdominal WAT, concordant CLS status (+/-) was found in 49/63 (78%) and 10/13 (77%) pts, respectively. Conclusions: Breast WAT inflammation (both presence and severity), which we have previously linked to increased aromatase activity, is associated with both increased BMI and menopause. These findings can explain the increased risk of estrogen receptor–positive BC with obesity and PoM status and may also provide targets for rational therapies. [Table: see text]

Regulatory effect of Dimethyl Sulfoxide (DMSO) on astrocytic reactivity in a murine model of cerebral infarction by arterial embolization

Introduction: The pathophysiology of cerebral ischemia is essential for early diagnosis, neurologic recovery, the early onset of drug treatment and the prognosis of ischemic events. Experimental models of cerebral ischemiac an be used to evaluate the cellular response phenomena and possible neurological protection by drugs. Objective: To characterize the cellular changes in the neuronal population and astrocytic response by the effect of Dimethyl Sulfoxide (DMSO) on a model of ischemia caused by cerebral embolism. Methods: Twenty Wistar rats were divided into four groups (n= 5). The infarct was induced with α-bovinethrombin (40 NIH/U). The treated group received 90 mg (100 µL) of DMSO in saline (1:1 v/v) intraperitoneally for 5 days; ischemic controls received only NaCl (placebo) and two non-ischemic groups (simulated) received NaCl and DMSO respectively. We evaluated the neuronal (anti-NeuN) and astrocytic immune-reactivity (anti-GFAP). The results were analyzed by densitometry (NIH Image J-Fiji 1.45 software) and analysis of variance (ANOVA) with the Graph pad software (Prism 5). Results: Cerebral embolism induced reproducible and reliable lesions in the cortex and hippocampus (CA1). similar to those of focal models. DMSO did not reverse the loss of post-ischemia neuronal immune-reactivity, but prevented the morphological damage of neurons, and significantly reduced astrocytic hyperactivity in thesomato-sensory cortex and CA1 (P <0.001). Conclusions: The regulatory effect of DMSO on astrocyte hyperreactivity and neuronal-astroglial cytoarchitecture, gives it potential neuroprotective properties for the treatment of thromboembolic cerebral ischemiain the acute phase.