Avaliação dos testículos de ratos submetidos à torção testicular antes, durante e após a puberdade, e o efeito do tratamento com L-arginina

Torção testicular (TT) é uma síndrome urológica comumente encontrada em recém nascidos, crianças e adolescentes. Neste trabalho foi estudada as lesões morfológicas e a função reprodutiva em ratos adultos que sofreram TT, em diferentes idades de maturidade sexual e o efeito protetor da L-arginina, contra os danos causados pela isquemia/reperfusão na torção testicular. Dezoito ratos pré-púberes (4 semanas de vida) dezessete púberes (6 semanas de vida) e dezessete adultos (9 semanas de vida) foram submetidos à TT. Sob anestesia o testículo direito foi rotacionado em 720 e fixado, sendo então destorcido após 4 horas. Vinte e quatro ratos (ARG4, n=8, ARG6, n=8 e ARG9 n=8) foram submetidos ao tratamento com 650mg/kg de L-arginina, por via oral, durante 7 dias. Outros trinta ratos de mesma idade sofreram cirurgia simulada (SH4, n=10, SH6, n=10 e SH9, n=10). Com 12 semanas de idade, foram submetidos ao acasalamento controlado com 3 fêmeas e ao vigésimo dia de gestação o número de fetos, corpos lúteos, absorções e implatações foram contados. Na 14 semana, os ratos foram mortos e os espermatozóides coletados da cauda dos epidídimos, foi anotado o peso corporal, o peso e volume testicular. O soro foi usado para dosagem de testosterona. Foram avaliadas a concentração, motilidade e a viabilidade espermática. Os testículos coletados foram fixados em Bouin, pós-fixados em formalina e processados em parafina. Utilizando o programa de imagem Image J, lâminas coradas com HE, mensuramos a altura do epitélio, densidade volumétrica e diâmetro do túbulo seminífero. Para avaliar a integridade do epitélio seminífero foi utilizado a frequência dos estágios do ciclo do epitélio e o escorre de Johnsen. Também foi avaliado a proliferação do compartimento tubular e intertubular, através da imunomarcação com PCNA. Os dados foram tabulados e as médias dos grupos comparadas pelo teste de ANOVA com pós-teste de Bonferroni ou teste de Kruskal-Wallis com pós teste de Dunns (programa Graphpad Prism, com p < 0,05). Os resultados revelaram grandes danos produzidos pela injuria testicular, no testículo ipsilateral, com diminuição da capacidade reprodutiva, da concentração, viabilidade e mobilidade, do peso e volume testicular. Animais TT9 não apresentaram espermatozoides nas amostras coletas. Houve diminuição do diâmetro dos túbulos seminíferos e da altura do epitélio, aumento do compartimento intertubular, com ênfase dos vasos sanguíneos, aumento da proliferação celular estromal e diminuição da proliferação epitelial. Houve diminuição da concentração sérica de testosterona no grupo TT4, quando comparado como grupo TT9. Em geral, os animais submetidos à torção na fase adulta foram os mais acometidos. Não foram encontradas alterações dignas de nota, nos testículo contralaterais. Animais tratados com L-arginina obtiveram melhora dos índices reprodutivos, com aumento da potência em todas as idades. Houve aumento da concentração espermática no testículo contralateral de ARG4 e ARG6, mostrando que a L-arginina atuou como antioxidante. Não houve proteção para as lesões causadas pela torção na maioria dos grupos, mas os animais tratados ARG9 apresentaram concentração espermática mensuravel, quando comparados aos ratos TT9, que tinham azospermia.

Pathology of mixed infections of saprolegniasis-myxosporidiosis in Indian major carp (Catla catla Ham.)

An outbreak of saprolegniasis in Catla catla in composite carp culture ponds were recorded during winter season. The typical cotton wool growths were observed on whole body surfaces of catla along with sporadic mortality. The fungal invasion was only restricted to skin and no fungal elements were visible in any internal organs after periodic acid schiff staining. On histology, periportal accumulation of mononuclear cells in liver, presence of myxosporidean cysts in antieror kidney, eosinophilic granular cells reaction in submucosa of stomach and intestine, dilated and engorged blood vessels of brain along with sloughing of epidermis and hyperplasia at gill lamellar base were pronounced changes. The possible role of release of Saprolegnia toxin in producing internal organs pathology has been discussed.

Pattern of cutaneous wound healing in a live fish Clarias batrachus (L.)(Clariidae, Pisces.)

Incisional wounds of the same length and depth were made on skin between dorsal fin and the lateral line canal of Clarias batrachus and the pattern of wound closure has been studied histologically. Following infliction, a marked change in the colour of the skin surrounding the wound was observed which lasted for about 30 h and restored thereafter. Mucus and blood cells plugged the wound gap shortly after infliction. The epidermis surrounding the wound was found to be detached from the basement membrane. Mass movement of epidermal cells was observed from both side of the wound gap. The epidermal cells at the margin of the wound became hypertrophied. The epidermis became normal by 32 days. The dealing of sub-epidermal tissue indicated degenerative and regenerative changes of muscle fibres. The mucus and blood cells were accumulated in the wound gap and later fine blood vessels were formed. Gradually granulation tissue was formed and fibroblasts and myoblasts appeared. Myoblast differentiated into muscle bundles. The epidermal repair was completed within 35 days.

The study of maturational progress in the whitespotted rabbitfish (Siganus sutor) under different photoperiod and light intensity conditions

The rule of light on the timing of maturation and spawning in tropical and subtropical regions is not clear well, because the reproductive cycle in these systems is lunar synchronized. In this study, thus, the effects of different light regimes were investigated on maturational progress of whitespotted rabbitfish, Siganus sutor, the commercial species in Persian Gulf and Oman Sea. During prespawning season, 50 adult fish were randomly divided into ten 300-l tanks (n=5). The fish in control tank received indoor light condition and the fish in each other tanks were exposed to nine different combinations of photoperiod (8L: 16D, 12L: 12D, 16L: 8D) and light intensity (1000, 2000, 3000 lux). After 60 days, GSI and HSI values, serum levels of estradiol (E2), testosterone (T), 17-α hydroxyprogestrone (OHP), calcium (Ca2+) and gonad histology were evaluated for females and males. In females the GSI mean values of exposed fish increased in comparison with control except for fish were kept under 8L, 2000 lux (tank 8). These differences were significant only for fishes in tank 7 (8L, 3000 lux). In the cases of HSI, the results were converse, so that, the most of thanks showed significant decreasing than fishes reared under indoor condition. Morphology and histology study of Ovaries showed three developmental phases including 3, 4 and chiefly 5 that were parallel with GSI values. Fortheremore the serum levels of E2 was recorded between 0.54-15.8 ng/ml in different fish and their mean values were lower than control in all treatmants (P> 0.05). In males, the similar results were obtained. The GSI and HSI mean values in experimental regimes compared with control were upper and lower, respectively, except for fishes were reared in tank 1 (16L, 3000 lux). Testes histology of fishes were reared under different regimes showed signs of stage 5, since no blood vessels observed and thick milt exuded on slight pressure. The mean values of testosterone consentration in fishes were kept in tanks 1 and 6 (12L, 1000 lux) were higher and in other ones were lower than control group. It is also noted that the OHP and Ca2+ had diverse results including increasing and decreasing mean values than control. So, these factors contrary to E2 and T were not suitable to evaluation of maturity in both sexes. On the basis of ovarian structure in stage 5, oocyte development pattern in this species was group synchronous. So, increased mean of GSI versus decreased values of HSI, E2 and perhaps Ca2+ were the signs of improved maturation. But in males, reduced levels of T and similarity of testes morphology in all samples caused that GSI mean value was the only indicator for differentiating among treatments. These findings suggest that alternations were used in light regimes have been the reason of improved maturity in all treatments except fishes reared in tank 8. The ١٠٧ rule of light intensity on induction of maturity was cleared by comparision between fishes in tanks 4, 5, 6 and control group. Because day length was the same whereas fishes in tanks 4, 5 and 6 were exposed to increased light intensity compared with control. This fact verified by results was obtained from fishes in tanks 9 and especially 7, since photoperiod in this group was lower than control. So, higher intensity was considered as the reason of alternations. Contrasting with indoor condition, Induced maturity was also cleared for fishes were kept in tanks 1, 2 and 3, where both of light duration and intensity were increased. But, the rule of photoperiod was individually demonstrated when obtained results were compared with similar light intensities in other treatments. In conclusion, with comparison among different light regimes it is declared that siganids were kept under light condition of tank 2 including 16h light duration combined with 2000 lux intensity showed the best signs of sexual maturation in both sexes. On the basis of this study, setting up the spring light condition during prespawning season induces maturation in withspotted rabbitfish. This improvement not only by influence of photoperiod or light intensity, separately, but obtained through interaction between them. Thus, determination of threshold and resistance to light be recommended before exposure, although using proper regime during suitable time are necessary to achieve purposes considerably.

Impact of phosphamidon and its metabolites on histopathology of the liver, gill and intestine of Labeo rohita

Impact of phosphamidon, an organophosphorus pesticide and its metabolites viz. dimethyl phosphoric acid and 2-chloro 2-diethyl carbamoylmethyl vinyl acid on histopathology of a common teleost, Labeo rohita was studied by exposing the fish to sub-lethal concentrations which were taken as 1/3rd of LC50 and were equal to 0.0123 ppm for phosphamidon, 0.0160 ppm for dimethyl phosphoric acid and 0.0167 ppm for 2-chloro 2-diethyl carbamoylmethyl vinyl acid respectively. The results revealed that hepatocytes in the liver were markedly swollen and exhibited hydropic degeneration. Fusion of primary lamellae and moderate congestion of blood vessels were evident in the gill. Intestine showed degeneration of mucosa and cellular infiltration in sub-mucosa. LC50 values and histopathological photomicrographs suggest that phosphamidon is more toxic as compared to dimethyl phosphoric and 2-chloro 2-diethyl carbamoylmethyl vinyl acid.

Ontogeny of IgM-producing cells in the mandarin fish Siniperca chuatsi identified by in situ hybridisation

The ontogeny of IgM-producing cells was studied in juvenile mandarin fish Simperca chuatsi, an important fish in China's aquaculture sector. The IgM-producing cells were localised through in situ hybridisation with a probe complementary to the Ig mu-chain in lymphoid-related tissues, including head kidney, spleen, thymus, intestine and gills. In head kidney, transcripts of Ig mu were first detected at 20 days post-hatching (dph) with a few positive signals. and the number of IgM-producing cells increased obviously from 39 dph onwards. At 136 dph, a large amount of positive cells were observed in the entire organ with clusters of these cells located around the blood vessels. In spleen, IgM-producing cells were found from 26 dph onwards, followed by an increase until 67 dph: clusters of positive cells were also detected around blood vessels at 102 dph. In thymus, IgM-producing cells were first observed at 39 dph; thereafter, no obvious increase was detected until 78 dph. The positive cells in thymus were distributed mainly in the outer zone of thymus. A few IgM-producing cells were still observed in thymus of 1-year-old mandarin fish. IgM-producing cells were not detected in the intestine until 87 dph, with several discrete positively stained cells distributed in the lamina propria. IgM-producing cells, scattered mainly in primary gill filaments around blood vessels, were detected in gills from 90 dph. As in other teleosts, these results indicated that the head kidney appears to be the primary organ for IgM production in mandarin fish, and IgM-producing cells exist in all organs examined in the present study, implying their lymphoid role in fish. In addition, it is suggested that vaccination after 20 dph may be much more effective in mandarin fish. (C) 2009 Elsevier B.V. All rights reserved.

C1q-like inhibits p53-mediated apoptosis and controls normal hernatopoiesis during zebrafish embryogenesis

Except for the complement C1q, the immunological functions of other C1q family members have remained unclear. Here we describe zebrafish C1q-like, whose transcription and translation display a uniform distribution in early embryos, and are restricted to mid-hind brain and eye in later embryos. In vitro studies showed that C1q-like could inhibit the apoptosis induced by ActD and CHX in EPC cells, through repressing caspase 3/9 activities. Moreover, its physiological roles were studied by morpholino-mediated knockdown in zebrafish embryogenesis. In comparison with control embryos, the C1q-like knockdown embryos display obvious defects in the head and cramofacial development mediated through p53-induced apoptosis, which was confirmed by the in vitro transcribed C1q-like mRNA or p53 MO co-injection. TUNEL assays revealed extensive cell death, and caspase 3/9 activity measurement also revealed about two folds increase in C1q-like morphant embryos, which was inhibited by p53 MO co-injection. Real-time quantitative PCR showed the up-regulation expression of several apoptosis regulators such as p53, mdm2, p21, Box and caspase 3, and down-regulation expression of hbae1 in the C1q-like morphant embryos. Knockdown of C1q-like in zebrafish embryos decreased hemoglobin production and impaired the organization of mesencephalic vein and other brain blood vessels. Interestingly, exposure of zebrafish embryos to UV resulted in an increase in mRNA expression of C1q-like, whereas over-expression of C1q-like was not enough resist to the damage. Furthermore, C1q-like transcription was up-regulated in response to pathogen Aeromonas hydrophila, and embryo survival significantly decreased in the C1q-like morphants after exposure to the bacteria. The data suggested that C1q-like might play an antiapoptotic and protective role in inhibiting p53-dependent and caspase 3/9-mediated apoptosis during embryogenesis, especially in the brain development, and C1q-like should be a novel regulator of cell survival during zebrafish embryogenesis. (c) 2008 Elsevier Inc. All rights reserved.

Effects of Ionizing Radiation on Three-Dimensional Human Vessel Models: Differential Effects According to Radiation Quality and Cellular Development

Little is known about the effects of space radiation on the human body. There are a number of potential chronic and acute effects, and one major target for noncarcinogenic effects is the human vasculature. Cellular stress, inflammatory response, and other radiation effects on endothelial cells may affect vascular function. This study was aimed at understanding the effects of space ionizing radiation on the formation and maintenance of capillary-like blood vessels. We used a 3D human vessel model created with human endothelial cells in a gel matrix to assess the effects of low-LET protons and high-LET iron ions. Iron ions were more damaging and caused significant reduction in the length of intact vessels in both developing and mature vessels at a dose of 80 cGy. Protons had no effect on mature vessels up to a dose of 3.2 Gy but did inhibit vessel formation at 80 cGy. Comparison with gamma radiation showed that photons had even less effect, although, as with protons, developing vessels were more sensitive. Apoptosis assays showed that inhibition of vessel development or deterioration of mature vessels was not due to cell death by apoptosis even in the case of iron ions. These are the first data to show the effects of radiation with varying linear energy transfer on a human vessel model. (C) 2011 In Radiation Research Society

On Combining Morphological Component Analysis and Concentric Morphology Model for Mammographic Mass Detection

Mammographic mass detection is an important task for the early diagnosis of breast cancer. However, it is difficult to distinguish masses from normal regions because of their abundant morphological characteristics and ambiguous margins. To improve the mass detection performance, it is essential to effectively preprocess mammogram to preserve both the intensity distribution and morphological characteristics of regions. In this paper, morphological component analysis is first introduced to decompose a mammogram into a piecewise-smooth component and a texture component. The former is utilized in our detection scheme as it effectively suppresses both structural noises and effects of blood vessels. Then, we propose two novel concentric layer criteria to detect different types of suspicious regions in a mammogram. The combination is evaluated based on the Digital Database for Screening Mammography, where 100 malignant cases and 50 benign cases are utilized. The sensitivity of the proposed scheme is 99% in malignant, 88% in benign, and 95.3% in all types of cases. The results show that the proposed detection scheme achieves satisfactory detection performance and preferable compromises between sensitivity and false positive rates.

The experimental research of R-phycoerythrin subunits on cancer treatment: A new photosensitizer in PDT

The mouse tumor cell 5180 and human liver carcinoma cell SMC 7721 cells were first treated with R-PE and its subunits (alpha, beta, gamma subunits), then irradiated with Argon laser (496 nm, 28.8 J/cm(2)). Survival rate was measured by MTT method. In order to compare the phototoxicity in normal cells, the mouse marrow cells were treated with photofrin II and beta-subunit, irradiated with 45 J/cm(2) of light; survival rate was also measured by MTT method. The result showed that R-PE subunits had better PDT effect on s180 cells than R-PE and lower phototoxicity in marrow cells than photofrin II Flow cytometric analysis showed that PDT results in a growth inhibition and a G(0)-G(1) cell cycle arrest in SMC 7721 cells. The tumor cells inhibited by PDT in vivo were morphologically observed by TEM, the tumor cell death was daze to the occlusion of tumor blood vessels and inducement of cell programmed death in nuclei. Therefore, with the advantage in special fluorescence activity, loth molecular weight, good light absorbent character and weak phototoxicity, R-PE subunit is art attractive option for improving the selectivity of PDT.

靶向斑马鱼VEGF的shRNA表达载体的构建及其对斑马鱼胚胎血管发育的影响

血管内皮生长因子(vascular endothelial growth factor, VEGF)是一种多功能的细胞因子，其主要作用是促进血管内皮细胞增殖和增加血管通透性，是肿瘤及正常组织血管生成的中心调控因素，以VEGF为靶点的肿瘤血管靶向性治疗成为近几年肿瘤治疗的新途径。RNAi是近年来新发展的一项反向遗传学技术，是一种研究基因功能的有力工具。斑马鱼作为一种重要的模式生物，被广泛用于胚胎的分子发育机制、疾病模型的构建以及药物筛选等研究中。然而在斑马鱼中运用RNAi技术进行基因功能研究是一个相对较新的领域，研究资料较少，并且目前进行的斑马鱼RNAi实验中，siRNA大都是通过化学方法或体外转录合成的。体外合成的siRNA在进入体内后会被降解而无法达到持久阻抑基因表达的目的。因此本研究旨在探讨VEGF特异性siRNA表达载体对斑马鱼VEGF基因的沉默作用，通过分析表型及相关细胞因子的变化，阐明VEGF对斑马鱼胚胎血管生成的影响及作用机制。 研究通过计算机辅助设计软件，针对斑马鱼VEGF mRNA不同位点设计合成了4段含siRNA特异序列的DNA单链，经退火，克隆入pSilencer 4.1-CMV neo载体CMV启动子下游，构建了重组质粒pS1-VEGF、pS2-VEGF、pS3-VEGF及pS4-VEGF。 通过显微注射的方法将载体导入1-2细胞期斑马鱼体内，于胚胎发育的48 h采用RT-PCR的方法检测VEGF基因的表达量，研究不同干扰序列对VEGF基因表达的干涉作用。结果显示，针对不同位点的表达载体对VEGF基因表达的抑制效率有显著差异。它们对VEGF mRNA的抑制率分别为80.5%，42.8%，12.5%，40.7%。通过筛选我们得到了一条具有高效抑制作用的载体pS1-VEGF，该载体的相应序列靶向斑马鱼两个主要异构体VEGF165和VEGF121的共有外显子序列。 形态学检测结果显示，注射了pS1-VEGF的胚胎出现了心包膜水肿、血流速度减慢、循环红细胞堆积等症状。定量碱性磷酸酶染色显示，注射pS1-VEGF能够抑制斑马鱼胚胎新生血管的形成，当注射剂量为0.4 ng时，血管生成的抑制率为31.8%。NBT/BCIP血管染色显示，注射该载体后72 h，50%的斑马鱼肠下静脉、节间血管以及其它血管的发育受到不同程度的抑制。随着注射剂量的加大，血管发育受抑制的情况也随之加重，当注射剂量为1 ng时，只有心脏、头部及卵黄有血液循环。对干扰效果的特异性进行了研究，结果表明pS1-VEGF对斑马鱼内源基因胸苷酸合成酶（thymidylate synthase, TS）基因的表达没有明显的抑制作用。针对TS基因的shRNA表达载体及与斑马鱼没有同源性的对照载体对VEGF基因表达也没有明显的抑制作用。浓度梯度实验表明在0-1.2 ng的范围内干扰效果具有剂量依赖性。 以胚胎整体原位杂交的方法检测质粒对VEGF基因受体NRP1基因表达的影响，发现VEGF特异性shRNA表达载体能够引起NRP1基因表达的降低，说明斑马鱼中VEGF所介导的血管生成作用至少在部分上是依赖于NRP通路所调节的。 本研究工作为进一步研究斑马鱼基因功能、VEGF调控网络提供了一个快速、有效的手段，为阐明斑马鱼的血管生成机制提供了新的资料，为采用RNAi技术，以VEGF为靶点，以斑马鱼为模型对肿瘤进行基因治疗研究奠定了基础。

玻璃海鞘多肽PCI的分离纯化及其抗血管生成活性研究

血管生成是肿瘤生长、发展的必经之路，并且与实体瘤的发生、转移有着密切的关系，抑制肿瘤新生血管生成具有特异性高、疗效好、不易产生耐药性以及毒副作用低等特点，因此抑制肿瘤血管形成可望成为治疗癌症的一个突破点，以抗血管生成为主的肿瘤生物治疗研究已成为近十年的研究热点。玻璃海鞘（Coina intestinalis）属于内性目、玻璃海鞘科，因其进化上的独特地位常作为研究神经发育、免疫系统进化的材料。在其它种属的海鞘中分离发现了多种具有抗肿瘤活性的多肽，但关于玻璃海鞘抗血管生成活性多肽的分离纯化和活性研究未见报道。本文利用多种分离纯化手段，采用活性追踪的方法首次从玻璃海鞘中分离得到具有抗新生血管生成作用的多肽，并对其抗血管生成活性做了初步研究。 本研究利用冷丙酮分级沉淀，超滤截留分子量小于5kDa的蛋白，再经SephadexG25、Superdex75柱层析，µRPC C2/C18反相柱层析等分离手段，采用活性追踪的方法，由玻璃海鞘（Coina intestinalis）中分离纯化出抗血管生成多肽PCI，据保留时间计算其分子量为1.8 kDa。MTT检测表明其对人脐静脉血管内皮细胞（HUVEC）具有强烈的抑制作用，IC50为7.5 μg/ml，并对多种肿瘤细胞有直接的抑制作用。斑马鱼胚胎体内实验进一步表明PCI在40 μg/ml的浓度下作用12h，斑马鱼胚胎新生血管生成受到显著抑制，肠下静脉血管长度为正常组的30%，斑马鱼胚胎血管生成率为正常组的45%。

冠状动脉旁路移植术后抑郁焦虑状态随访和分析

To explore the presence of depression, anxiety and cardiac events after coronary artery bypass grafting (CABG) prospectively and comparably. To analysis influential factors of depression and anxiety ( peri-operation and 1 year after operation ), and the effection on cardiac events in the year after operation. We followed the patients who underwent scheduled consecutive CABG. We interviewed patients to assess depression, anxiety symptoms using self-rating depression scale( SDS) and self-rating anxiety scale（SAS）before operation , before discharge and 1 year after operation. And cardiac events were also identified. All patients were divided into the group with depression and/or anxiety symptoms or the other group without depression and/or anxiety symptoms 3 times according to depression and anxiety symptoms before operation , before discharge and 1 year after operation. 69 patients completed the follow-up.24 patients (34.8%) had depression and/or anxiety symptoms before CABG, 31 patients (44.9%) had depression and/or anxiety symptoms before discharge, and 10 patients (14.5%) had depression and/or anxiety symptoms 1 year after operation. 6 patients(8.7%) had cardiac events, and 5 patients were re-admitted. Arhythmia before and after operation, the quantity of blood vessel grafted, length of stay , depression and anxiety symptoms before operation, cardiac events may lead to depression and anxiety symptoms 1 year after operation. The use of cardiopulmonary bypass grafting (CPB), Arhythmia after operation, the prolonged length of stay are influential factors of cardiac events. Though the incidence of depression and/or anxiety symptoms 1 year after operation is lower than before operation and before discharge, it is high still. Arhythmia before and after operation, the more blood vessels grafted, the prolonged length of stay , depression and anxiety symptoms before operation and cardiac events could be risk factors of depression and anxiety symptoms 1 year after operation. Much attention must be paid to this phenomenon. Heavy patient's condition, complicated surgery are risk factors of cardiac events, and bad emotion before operation and before discharge are independent to cardiac events. Key words： perspective research; depression；anxiety; coronary artery bypass grafting; follow up

LED (660 nm) and laser (670 nm) use on skin flap viability: angiogenesis and mast cells on transition line

Skin flap procedures are commonly used in plastic surgery. Failures can follow, leading to the necrosis of the flap. Therefore, many studies use LLLT to improve flap viability. Currently, the LED has been introduced as an alternative to LLLT. the objective of this study was to evaluate the effect of LLLT and LED on the viability of random skin flaps in rats. Forty-eight rats were divided into four groups, and a random skin flap (10 x 4 cm) was performed in all animals. Group 1 was the sham group; group 2 was submitted to LLLT 660 nm, 0.14 J; group 3 with LED 630 nm, 2.49 J, and group 4 with LLLT 660 nm, with 2.49 J. Irradiation was applied after surgery and repeated on the four subsequent days. On the 7th postoperative day, the percentage of flap necrosis was calculated and skin samples were collected from the viable area and from the transition line of the flap to evaluate blood vessels and mast cells. the percentage of necrosis was significantly lower in groups 3 and 4 compared to groups 1 and 2. Concerning blood vessels and mast cell numbers, only the animals in group 3 showed significant increase compared to group 1 in the skin sample of the transition line. LED and LLLT with the same total energies were effective in increasing viability of random skin flaps. LED was more effective in increasing the number of mast cells and blood vessels in the transition line of random skin flaps.

HEATING IN VASCULAR TISSUE AND FLOW-THROUGH TISSUE PHANTOMS INDUCED BY FOCUSED ULTRASOUND

High intensity focused ultrasound (HIFU) can be used to control bleeding, both from individual blood vessels as well as from gross damage to the capillary bed. This process, called acoustic hemostasis, is being studied in the hope that such a method would ultimately provide a lifesaving treatment during the so-called "golden hour", a brief grace period after a severe trauma in which prompt therapy can save the life of an injured person. Thermal effects play a major role in occlusion of small vessels and also appear to contribute to the sealing of punctures in major blood vessels. However, aggressive ultrasound-induced tissue heating can also impact healthy tissue and can lead to deleterious mechanical bioeffects. Moreover, the presence of vascularity can limit one’s ability to elevate the temperature of blood vessel walls owing to convective heat transport. In an effort to better understand the heating process in tissues with vascular structure we have developed a numerical simulation that couples models for ultrasound propagation, acoustic streaming, ultrasound heating and blood cooling in Newtonian viscous media. The 3-D simulation allows for the study of complicated biological structures and insonation geometries. We have also undertaken a series of in vitro experiments, in non-uniform flow-through tissue phantoms, designed to provide a ground truth verification of the model predictions. The calculated and measured results were compared over a range of values for insonation pressure, insonation time, and flow rate; we show good agreement between predictions and measurements. We then conducted a series of simulations that address two limiting problems of interest: hemostasis in small and large vessels. We employed realistic human tissue properties and considered more complex geometries. Results show that the heating pattern in and around a blood vessel is different for different vessel sizes, flow rates and for varying beam orientations relative to the flow axis. Complete occlusion and wall- puncture sealing are both possible depending on the exposure conditions. These results concur with prior clinical observations and may prove useful for planning of a more effective procedure in HIFU treatments.