A new organ preservation solution for static cold storage of the liver. Amniotic fluid

Abstract Purpose: To evaluate the effect of amniotic fluid in liver preservation in organ transplantation, and compare it with standard preservation solutions. Methods: The groups consisted of Group 1: Ringer Lactate (RL) group, Group 2: HTK group, Group 3: UW group, Group 4: AF group. The livers of rats from Group 1, 2, 3, and 4 were perfused and placed into falcon tubes containing RL, HTK, UW, and AF solutions at +4‎°C, respectively. The tubes were stored for 12 hours in the refrigerator at +4°C. Tissue samples were taken at the 6th and 12th hours for histopathological examinations of the perfused livers, and storage solutions for biochemical analyzes at 6th and 12th hours. Results: AF was shown to maintain organ viability by reducing the number of cells undergoing apoptosis. Histopathological changes such as sinusoidal dilatation, hydropic degeneration, and focal necrosis were found to be similar to the groups in which the standard organ preservation solutions were used. Additionally, the results of INOS, IL-10, and TNF-α,which were evaluated immunohistochemically, have been shown to be similar to the UW and HTK groups. Conclusions: AF provided conservation similar to UW and HTK in the 12-hour liver SCS process. The fact that apoptosis values are comparable to standard preservation solutions supports the success of AF in the cold storage of the liver.

Practical Implications of Myocardial Viability Studies / Implicações Práticas da Pesquisa de Viabilidade Miocárdica

Abstract Many non-invasive methods, such as imaging tests, have been developed aiming to add a contribution to existing studies in estimating patients' prognosis after myocardial injury. This prognosis is proportional to myocardial viability, which is evaluated in coronary artery disease and left ventricular dysfunction patients only. While myocardial viability represents the likelihood of a dysfunctional muscle (resulting from decreased oxygen supply for coronary artery obstruction), hibernation represents post-interventional functional recovery itself. This article proposes a review of pathophysiological basis of viability, diagnostic methods, prognosis and future perspectives of myocardial viability. An electronic bibliographic search for articles was performed in PubMed, Lilacs, Cochrane and Scielo databases, according to pre-established criteria. The studies showed the ability of many imaging techniques in detecting viable tissues in dysfunctional areas of left ventricle resulting from coronary artery injuries. These techniques can identify patients who may benefit from myocardial revascularization and indicate the most appropriate treatment.

Resumo Diversos métodos não invasivos, como novos exames de imagem, vem sendo aprimorados, a fim de somar esforços com os atuais em estimar o prognóstico de pacientes pós-injúria miocárdica. Este prognóstico é proporcional à viabilidade miocárdica, a qual tem sua avaliação reservada para pacientes portadores de doença arterial coronariana e insuficiência ventricular esquerda. Enquanto a viabilidade miocárdica se mostra como a capacidade de recuperação funcional do músculo com disfunção por redução de oxigênio fornecido por artérias coronárias obstruídas, a hibernação consiste na própria recuperação funcional após intervenções. Este artigo propõe uma revisão sobre as bases fisiopatológicas do processo de viabilidade, métodos diagnósticos disponíveis, prognóstico e perspectivas para o futuro acerca dessa condição. Realizou-se pesquisa de busca bibliográfica informatizada em bases eletrônicas de dados, como PubMed, Lilacs, Cochrane e Scielo, onde foram selecionados os estudos de acordo com critérios pré-determinados. Os estudos demonstram a capacidade de várias técnicas de imagem de identificar tecido viável em regiões disfuncionais do ventrículo esquerdo em decorrência de lesões em artérias coronárias. Estas técnicas podem identificar pacientes com potencial benefício da revascularização miocárdica e orientar o tratamento mais adequado.

Follicle Viability after Vitrification of Bovine Ovarian Tissue / Viabilidade folicular após a vitrificação de tecido ovariano bovino

Abstract Purpose The present study aimed to evaluate the impact of vitrification on the viability of follicles using a three-dimensional (3D) in vitro culture. Methods Bovine ovarian tissue samples (n = 5) obtained from slaughterhouses were utilized. The cortex was cut into small fragments of 2 x 3 x 0.5 mm using a tissue slicer. From these fragments, secondary follicles were first isolated by mechanical and enzymatic methods, then encapsulated in alginate gel and individually cultured for 20 days. Additional fragments of the same ovarian tissue were vitrified in a solution containing 25% glycerol and 25% ethylene glycol. After warming, the follicles underwent the same follicular isolation process that was performed for the fresh follicles. Results A total of 61 follicles were isolated, 51 from fresh ovarian tissue, and 10 from vitrified tissue. After the culture, the vitrified and fresh follicles showed 20% and 43.1% survival rates respectively (p = 0.290),with no significant differences. At the end of the culture, therewere no significant differences in follicular diameter between the vitrified (422.93 ± 85.05 μm) and fresh (412.99 ± 102.55 μm) groups (p = 0.725). Fresh follicles showed higher mean rate of antrum formation when compared with vitrified follicles (47.1% and 20.0% respectively), but without significant difference (p = 0.167). Conclusions The follicles were able to develop, grow and form antrum in the 3D system after vitrification, despite the lower results obtained with the fresh tissue.

Resumo Objetivo O presente estudo teve como objetivo avaliar o impacto da vitrificação na viabilidade dos folículos utilizando a cultura in vitro tridimensional (3D). Métodos Foi utilizado tecido ovariano bovino (n = 5) obtido de abatedouros. O córtex foi cortado em pequenos fragmentos de 2 x 3 x 0,5 mm, utilizando o tissue slicer e a partir destes fragmentos foram isolados folículos secundários por meio de método enzimático e mecânico, encapsulados em gel de alginato e cultivados individualmente durante 20 dias. Outros fragmentos do mesmo tecido ovariano foram vitrificados em solução contendo 25% de glicerol e 25% de etilenoglicol. Após aquecimento, os folículos passaram pelo mesmo processo de isolamento folicular realizado a fresco. Resultados Foram isolados 61 folículos, sendo 51 originários de tecido ovariano a fresco, e 10 de tecido vitrificado. Após a cultura, os folículos vitrificados apresentaram taxa de sobrevida de 20%, e o grupo a fresco apresentou taxa de 43,1% (p = 0,290). O diâmetro folicular ao final da cultura também não apresentou diferença significativa entre o grupo vitrificado (422,93 ± 85,05 μm) e a fresco (412,99 ± 102,55 μm) (p = 0,725). Os folículos a fresco apresentarammaior taxa média de formação de antro do que os folículos vitrificados (47,1% e 20,0%, respectivamente), mas sem diferença significativa (p = 0,167). Conclusões Os folículos foram capazes de se desenvolver, crescer e formar antro em sistema 3D após a vitrificação.

Effects of dimethylsulfoxide and pentoxifylline in the vitality of cutaneous flaps in rats / Efeitos do dimetilsulfóxido e da pentoxifilina na vitalidade de retalhos cutâneos em ratos

ABSTRACT Objectives: to verify the influence of dimethylsulfoxide and pentoxifylline on the vitality of cutaneous flaps in rats and the tissue repair process. Methods: were studied 30 Wistar rats, submitting them to a 2cm wide by 8cm long dorsal cutaneous flap, of caudal base. We distributed the animals in three groups: Control Group (n=10) with application gauze moistened with 0.9% Saline in the flap bed for 30 seconds; Dimethylsulfoxide group (n=10), with administration of 1ml of 5% dimethylsulfoxide divided into five injections of 0.2ml in the transition of the flap segments; Pentoxifylline group (n=10), with administration of pentoxifylline 20mg/kg, diluted to 1ml and divided into five injections of 0.2ml in the transition of the flap segments. Drugs were administered intraoperatively, in a single dose and subcutaneously. We observed the skin flaps for changes in color and texture. On the 10th postoperative day, we checked the dimensions of viable and necrotic tissues, followed by excision of the specimen for histological analysis. Results: the measurements of length of the viable and necrotic tissues between groups showed no differences. Histological analysis showed that the Dimethylsulfoxide group presented neovascularization, inflammatory infiltrate with leukocytes and more structured conjunctival stroma. The Pentoxifylline group showed neovascularization and inflammatory infiltrate, with moderate to intense granulation. The control group evolved with a higher rate of necrosis in the distal segment. Conclusion: dimethylsulfoxide and pentoxifylline influenced the vitality of the flap and the tissue repair process. However, they did not prevent necrosis macroscopically.

RESUMO Objetivos: verificar a influência do dimetilsulfóxido e da pentoxifilina na vitalidade e no processo de reparo tecidual de retalhos cutâneos em ratos. Método: foram estudados 30 ratos Wistar, nos quais foi confeccionado retalho cutâneo dorsal de 2cm de largura por 8cm de comprimento, de base caudal, e distribuídos em três grupos: Grupo Controle (n=10) com aplicação de gaze umedecida com solução salina a 0,9%, no leito do retalho, por 30 segundos; Grupo dimetilsulfóxido (n=10) com injeção de 1ml de dimetilsulfóxido a 5% divididos em cinco injeções de 0,2ml na transição dos segmentos do retalho; Grupo pentoxifilina (n=10) com injeção de 1ml pentoxifilina 20mg/kg, divididos em cinco injeções de 0,2ml na transição dos segmentos do retalho. Os fármacos foram administrados no transoperatório, em dose única e por via subcutânea. Os retalhos cutâneos foram observados quanto às alterações de cor e textura. No décimo dia de pós-operatório aferiu-se a dimensão do tecido viável e de necrose, seguido da exérese da peça para análise histológica. Resultados: a medida da dimensão de tecido viável e de necrose dos grupos não apresentou diferenças. A análise histológica mostrou que o grupo dimetilsulfóxido apresentou neovascularização, infiltrado inflamatório com leucócitos e estroma conjuntivo mais estruturado. O grupo pentoxifilina, mostrou neovascularização e infiltrado inflamatório com granulação moderada e intensa. O grupo controle evoluiu com maior índice de necrose no segmento distal. Conclusão: dimetilsulfóxido e pentoxifilina influenciaram na vitalidade do retalho e no processo de reparo tecidual. Entretanto, não evitaram a necrose macroscopicamente.

Myocardial Viability on Cardiac Magnetic Resonance / Viabilidade Miocárdica pela Ressonância Magnética Cardíaca

Abstract The study of myocardial viability is of great importance in the orientation and management of patients requiring myocardial revascularization or angioplasty. The technique of delayed enhancement (DE) is accurate and has transformed the study of viability into an easy test, not only for the detection of fibrosis but also as a binary test detecting what is viable or not. On DE, fibrosis equal to or greater than 50% of the segmental area is considered as non-viable, whereas that below 50% is considered viable. During the same evaluation, cardiac magnetic resonance (CMR) may also use other techniques for functional and perfusion studies to obtain a global evaluation of ischemic heart disease. This study aims to highlight the current concepts and broadly emphasize the use of CMR as a method that over the last 20 years has become a reference in the detection of infarction and assessment of myocardial viability.

Resumo O estudo de viabilidade miocárdica é de grande importância para a orientação e manejo de pacientes que necessitam de cirurgia de revascularização miocárdica ou angioplastia. A técnica de realce tardio (RT) é precisa e transformou o estudo de viabilidade em um teste fácil, não só para a detecção de fibrose, mas também como um modelo binário para a detecção do que é ou não é viável. Uma fibrose identificada pelo RT é considerada como não viável quando igual ou maior do que 50% da área segmentar e como viável quando menor que 50%. A ressonância magnética cardíaca (RMC) também pode lançar mão de outras técnicas para estudo funcional e de perfusão para uma avaliação global da doença isquêmica do coração no mesmo exame. Este estudo tem como objetivo destacar os conceitos atuais e enfatizar amplamente o uso da RMC como um método que nos últimos 20 anos se tornou referência na detecção de infarto e avaliação de viabilidade miocárdica.

Avaliação das simetrizações imediatas em reconstrução de mama / Assessment of immediate symmetrization in breast reconstruction

Introdução: O tratamento cirúrgico do câncer de mama resulta com frequência em mutilação. A reconstrução de mama em mulheres mastectomizadas objetiva criar nova mama esteticamente aceitável e simétrica à mama contralateral. O objetivo deste estudo é estudar a viabilidade da simetrização da mama contralateral simultaneamente à reconstrução de mama, discutir possíveis complicações e fazer uma breve revisão da literatura. Métodos: Estudo retrospectivo realizado no Serviço de Cirurgia Plástica do Hospital Daher, de outubro de 2013 a fevereiro de 2015. Avaliadas as reconstruções mamárias imediatas pós-mastectomia por câncer de mama e selecionadas para inclusão todas as pacientes submetidas à simetrização da mama oposta no mesmo tempo cirúrgico, com mesma técnica cirúrgica e análise estatística. Resultados: Estudados 42 pacientes dentro dos critérios estabelecidos, totalizando 21 reconstruções com simetrização simultânea (Grupo 1) e 21 simetrizações em dois tempos (Grupo 2). A média de idade foi de 53,86 anos para o Grupo 1 e 52,62 anos para o grupo controle. Os grupos foram comparáveis para todas as variáveis. Os dados referentes às complicações pósoperatórias foram analisados. De todas as pacientes estudadas, algumas não finalizaram todas as etapas da reconstrução. O grupo que realizou simetrização imediata alcançou mais simetria. Pacientes com até 45 anos e com índice de massa corporal menor alcançaram mais simetria no Grupo 1. Conclusão: A execução de procedimentos de simetrização no mesmo tempo de uma reconstrução de mama unilateral está associada a baixa taxa de complicação e de cirurgias de revisão. Em casos selecionados, a simetrização imediata pode ser preferível ao procedimento em outra etapa cirúrgica.

Introduction: The surgical treatment of breast cancer frequently results in mutilation. Breast reconstruction in mastectomized women aims to create a new esthetically acceptable breast symmetrical to the contralateral breast. The objective of this study was to assess the feasibility of symmetrization of the contralateral breast simultaneously with breast reconstruction, discuss possible complications, and perform a brief review of the literature. Methods: A retrospective study was conducted in the Department of Plastic Surgery of Hospital Daher from October 2013 to February 2015. Breast reconstruction outcomes immediately after mastectomy for breast cancer were assessed, and all patients undergoing symmetrization of the contralateral breast in the same surgical stage using the same surgical technique were selected for inclusion and statistical analysis. Results: The study comprised 42 patients within the established criteria, totaling 21 reconstructions with simultaneous symmetrization (Group 1) and 21 symmetrization procedures in two stages (Group 2). The mean age was 53.86 years in Group 1 and 52.62 years in the control group. The groups were comparable in all variables. Data regarding postoperative complications were analyzed. Some of the studied patients did not complete all stages of reconstruction. The group that underwent immediate symmetrization attained more symmetry. The patients aged 45 years and with lower body mass index attained more symmetry in Group 1. Conclusion: The implementation of symmetrization procedures at the same stage of unilateral breast reconstruction is associated with low complication rates and revision surgeries. In selected cases, immediate symmetrization may be preferable to the procedure in another surgical stage.

Salvage of Unilateral Complete Ear Amputation with Continuous Local Hyperbaric Oxygen, Platelet-Rich Plasma and Polydeoxyribonucleotide without Micro-Revascularization

In many cases of complete ear amputation, microvascular surgery is required for tissue perfusion and organ survival. However, microvascular reconstruction is not always feasible in the absence of suitable vessels. Here, we present the case of a 76-year-old man who underwent complete amputation of the left ear after a collapse at home because of cardiogenic syncope. He was treated with primary replantation and underwent a postoperative salvage course including continuous local hyperbaric oxygen therapy (HBOT), platelet-rich plasma (PRP) injections, and polydeoxyribonucleotide (PDRN) injections. The ear was almost completely salvaged, with a tiny eschar at the mid-scapha on both the anterior and posterior aspects. This case demonstrates the efficacy of local HBOT with PRP and PDRN injections.

CT Angiography of Collateral Vessels and Outcomes in Endovascular-Treated Acute Ischemic Stroke Patients

BACKGROUND AND PURPOSE: Measuring the extent of the collateral blood vessels using computed tomography (CT) angiography source images may promote tissue survival and functional gain in acute ischemic stroke patients who are candidates for endovascular recanalization treatment. METHODS: Of 5,558 acute stroke patients registered in a prospective clinical stroke registry, 104 met the selection criteria of endovascular recanalization treatment for internal cerebral artery or middle cerebral artery main-stem (M1) occlusions and presented for treatment ≤4 hours after the event. Using CT angiography source images, two independent and blinded reviewers measured the extent of collateral circulations at four regions, with good interrater reliability. The functional recovery at 3 months after stroke was used as an outcome variable. RESULTS: Cases with a sufficient collateral circulation at the Sylvian fissure showed significantly increased likelihood of having a modified Rankin Scale score of ≤2 at 3 months after stroke (adjusted odds ratio=3.03, 95% confidence interval=1.19–7.73, p=0.02), but the association became nonsignificant after adding the infarct volume to the model (p=0.65). The association between leptomeningeal convexity collaterals and functional recovery was no longer significant after adjusting for the infarct volume (p=0.28). The natural indirect effect of infarct volume on functional recovery was significant for both the Sylvian fissure (p=0.03) and leptomeningeal convexity (p=0.02) collaterals. CONCLUSIONS: The extent of collateral circulation at the Sylvian fissure was significantly associated with functional recovery, which may be mediated via the volume of the final infarction.

Functional investigation of bone implant viability using radiotracers in a new model of osteonecrosis

OBJECTIVES: Conventional imaging methods are excellent for the morphological characterization of the consequences of osteonecrosis; however, only specialized techniques have been considered useful for obtaining functional information. To explore the affinity of radiotracers for severely devascularized bone, a new mouse model of isolated femur implanted in a subcutaneous abdominal pocket was devised. To maintain animal mobility and longevity, the femur was harvested from syngeneic donors. Two technetium-99m-labeled tracers targeting angiogenesis and bone matrix were selected. METHODS: Medronic acid and a homodimer peptide conjugated with RGDfK were radiolabeled with technetium-99m, and biodistribution was evaluated in Swiss mice. The grafted and control femurs were evaluated after 15, 30 and 60 days, including computed tomography (CT) and histological analysis. RESULTS: Radiolabeling achieved high (>95%) radiochemical purity. The biodistribution confirmed good blood clearance 1 hour after administration. For 99mTc-hydrazinonicotinic acid (HYNIC)-E-[c(RGDfK)2, remarkable renal excretion was observed compared to 99mTc-methylene diphosphonate (MDP), but the latter, as expected, revealed higher bone uptake. The results obtained in the control femur were equal at all time points. In the implanted femur, 99mTc-HYNIC-E-[c(RGDfK)2 uptake was highest after 15 days, consistent with early angiogenesis. Regarding 99mTc-MDP in the implant, similar uptake was documented at all time points, consistent with sustained bone viability; however, the uptake was lower than that detected in the control femur, as confirmed by histology. CONCLUSIONS: 1) Graft viability was successfully diagnosed using radiotracers in severely ischemic bone at all time points. 2) Analogously, indirect information about angiogenesis could be gathered using 999mTc-HYNIC-E-[c(RGDfK)2. 3) These techniques appear promising and warrant further studies to determine their potential clinical applications.

Comparison between Slow Freezing and Vitrification in Terms of Ovarian Tissue Viability in a Bovine Model / Comparação da viabilidade do tecido ovariano após congelamento lento e vitrificação em modelo bovino

Abstract Objective To assess the viability of bovine ovarian tissue after cryopreservation through either slow freezing or vitrification, and to compare it to that of control tissue by performing morphological analyses. Methods The study included 20 bovine ovarian cortex fragments that were divided into control, vitrification, and slow freezing groups. Each group consisted of four fragments of the same ovary, two fixed without cultivation, and two fixed with cultivation. Tissues were evaluated based on follicular morphology immediately after heating and after 7 days of culture, and compared with the control group. Results A total of 240 fragments were analyzed, generating a sample of 1,344 follicles without cultivation and 552 with cultivation. When the non-cultivated samples were classified as non-atretic follicles, 572 were found in the control group, 289 in the vitrification group, and 373 in the slow freezing group, showing no significant differences. When classified as atretic, 46 follicles were found in the control group, 23 in the vitrification group, and 41 in the slow freezing group, also showing no statistical difference. In the post-culture sample, an evolution of the follicular stages could be observed. This finding was important to support that the follicles considered non-atretic in the non-cultivated group were actually viable in the morphological evaluation. Conclusion With no differences between the protocols, vitrification was shown to be an advanced and alternative method for patients who will undergo treatments that.

Resumo Objetivo avaliar a viabilidade do tecido ovariano bovino após a criopreservação, utilizando congelamento lento e vitrificação, e comparando com o tecido controle por meio de análises morfológicas. Métodos o estudo incluiufragmentos de córtex de vinte ovários bovinos divididos em grupos controle, vitrificação e congelamento lento. Cada grupo foi composto por quatro fragmentos do mesmo ovário, sendo dois fragmentos fixados sem cultivo e dois fragmentos fixados pós-cultivo. Os tecidos foram avaliados pela morfologia folicular logo após o aquecimento e após sete dias de cultivo, e comparados com o grupo controle. Resultados um total de 240 fragmentos foi analisado, gerando uma amostra de 1.344 folículos sem cultivo e 552 pós-cultivo. Quando a amostra sem cultivo teve seus folículos agrupados em não atrésicos, obtivemos 572 no grupo controle, 289 no vitrificação, e 373 no congelamento lento, não apresentando diferença estatística. Quando agrupados em atrésicos, o grupo controle apresentou 46 folículos, o vitrificação, 23, e o congelamento lento, 41, não apresentando também diferença estatística. Na amostra pós-cultivo, podemos observar uma evolução dos estágios foliculares: esse achado foi importante para sustentar que os folículos considerados não atrésicos na avaliação morfológica sem cultivo estavam realmente viáveis. Conclusão não havendo diferenças entre os protocolos, a vitrificação se mostra um avanço e um método alternativo para pacientes que irão se submeter a tratamentos que podem levar a uma falência ovariana, uma vez que a metodologia é mais barata, mais rápida e mais bem adaptável a uma rotina de um laboratório.

Antioxidant effect of Legalon(r) SIL in ischemia-reperfusion injury of rat skeletal muscle

PURPOSE: To evaluated the potential antioxidant agent Legalon (r) SIL (silibinin-C-2',3-bis(hydrogensuccinat)) in the skeletal muscle of rats. METHODS: IRI was achieved via tourniquet application in Wistar-albino rats. Experimental groups were chosen as (i) sham control, (ii) IRI (3+2 h), (iii) IRI and Legalon (r) SIL-50 (50 mg/kg/i.p.), (iv) IRI and Legalon (r) SIL-100 (100 mg/kg/i.p.), and (v) IRI and Legalon (r) SIL-200 (200 mg/kg/ i.p.). Muscle viability (evaluated by triphenyltetrazolium chloride dye method), malondialdehyde, superoxide dismutase, catalase, and glutathione peroxidase were assessed in muscle samples using a spectrophotometer. RESULTS: Although viability of the injured limb non-significantly declined in the IRI group, administration of Legalon (r) SIL did not prevent injury. However, dramatic increase observed in malondialdehyde levels in the IRI group was prohibited by Legalon (r) SIL in a statistically significant manner. In comparison with the sham-control group, IRI and Legalon (r) SIL administration did not cause any significant alterations in the levels of superoxide dismutase, catalase, and glutathione peroxidase. CONCLUSION: Although Legalon (r) SIL was not sufficient to prevent muscle injury in terms of viability, it is found to be an effective option to reduce reactive oxygen species-induced cell injury.

Intraoperative indocyanine green angiography for the objective measurement of blood flow

PURPOSE: Intraoperative assessment of tissue perfusion is important to predict wound healing or improvement of symptoms in patients with peripheral arterial disease (PAD) or vascular trauma. There is no widely accepted standard for intraoperative measurement of tissue perfusion. Here, we report the use of indocyanine green (ICG)-based angiography to determine the blood flow in patients with PAD and vascular trauma. METHODS: The SPY fluorescent imaging system was utilized. A dose of 3-5 mL of ICG (2.5 mg/mL) was injected intravenously followed by a 10 mL normal saline flush. The SPY imaging system was used to quantitatively assess perfusion. During the study period, the SPY imaging system was applied in 4 patients with PAD and one patient with vascular trauma. RESULTS: In 3 patients with PAD associated with an ischemic wound, complete wound healing was achieved with the indication of viable tissue by the SPY system. In one patient with severe claudication in both lower extremities, the ICG angiography was used to determine the increased blood flow after revascularization. In the case of vascular trauma, this imaging system enabled the delineation of viability of the injured tissue. CONCLUSION: ICG angiography can determine the surface tissue viability in PAD patients. In cases of severe vascular trauma,the SPY system can be used to determine tissue perfusion. Further study is warranted to define the definite utility of this technology to assess perfusion, response to revascularization, and potentially, to predict the likelihood of wound healing.

EFFECTS OF DIFFERENT PERIODS OF GASTRIC ISCHAEMIA IN THE VIABILITY OF THE TISSUE OF BODY, FUNDUS AND ANTRUM REGION OF RABBIT STOMACH / EFEITOS DE DIFERENTES TEMPOS DE ISQUEMIA GÁSTRICA SOBRE A VIABILIDADE DOS TECIDOS DO CORPO, FUNDO E ANTRO DO ESTÔMAGO DE COELHOS

BACKGROUND: Despite the rich vascular arcade of the stomach, gastric ischemia represents an important medical challenge and can be the consequence of obstructive or non-obstructive vascular processes of pathological or iatrogenic origin.AIM: To assess the effects of acute gastric ischaemia on the different regions of the stomach.METHOD: Fifteen New Zeland rabbits were divided into three groups: group 1, animals were observed during 3 h; group 2, during 6 h; group 3, during 12 h. Rabbit stomachs were subjected to devascularization of the greater and lesser curvatures. After predetermined time, the stomachs were removed for macro and microscopic studies.RESULTS: Haemorrhagic necrosis was more marked in the gastric fundus and body. In contrast, the antropylorus remained preserved in 80% of the animals. Necrosis of the gastric body and fundus mucosa were observed in all animals after 6 h and 12 h of ischaemia.CONCLUSION: Acute gastric ischaemia in rabbits produces haemorrhagic necrosis of the gastric fundus and body even in a short period of time. Beside this, the antropyloric region was significantly more resistant to ischaemia.

RACIONAL: Isquemia gástrica representa importante desafio médico e geralmente é decorrente de processos vasculares obstrutivos ou não-obstrutivos. Apesar da rica arcada vascular do estômago, lesões de isquemia gástrica têm sido observadas. Quando a isquemia progride, pode culminar em necrose do órgão.OBJETIVO: Avaliar os efeitos dos diferentes tempos de isquemia gástrica sobre os tecidos da parede do estômago de coelhos mediante desvascularização total de ambas as curvaturas gástricas.MÉTODO: Foram utilizados 15 coelhos machos da raça Nova Zelândia. Os animais foram distribuídos nos seguintes grupos: grupo 1, observados por 3 h; grupo 2, por 6 h; grupo 3, por 12 h. A técnica cirúrgica foi a mesma em todos os animais e consistiu na ligadura e secção de todas as veias e artérias da parede gástrica. Após o período pré-determinado de observação, o estômago foi removido por completo para estudo macro e microscópico.RESULTADOS: As alterações macroscópicas e histológicas tornaram-se mais intensas com o aumento do tempo de isquemia e foram mais evidentes nas regiões do fundo e do corpo. Por outro lado, o antro manteve-se preservado na maioria dos animais operados. Necrose de mucosa do corpo e do fundo foi observada em todos os animais estudados, e foi acompanhada por hemorragia em 60% dos coelhos dos grupos 2 e 3.CONCLUSÃO: O modelo experimental de isquemia gástrica foi eficaz para produzir necrose hemorrágica de fundo e corpo do estômago de coelhos mesmo em curto tempo. Por outro lado, a região do antro pilórico desses animais mostrou-se significativamente mais resistente à isquemia do que as demais regiões.

Analysis of effect isoxsuprine hydrochloride and nicotine in the Transverse Rectus Abdominis Myocutaneous flap (TRAM) in rats

PURPOSE: To evaluate the effects of isoxsuprine and nicotine on TRAM. METHODS: Forty eight 48 Wistar rats distributed into four Groups (n=12). All rats received medication managed daily for 20 days: saline solution (SA), nicotine solution (NI), isoxsuprine solution (IS) and nicotine solution (NI) + isoxsuprine solution (IS). On day 21st the rats were submitted to the caudally based, right unipedicled TRAM flap and after 48 hours, made the macroscopic evaluation of the surface of the flap, photographic documentation and collection of material for histology. Data from macroscopic evaluation were analyzed by ANOVA and microscopic evaluation by Kruskal-Wallis test, with significance level of 5%. RESULTS: In the macroscopic evaluation of isoxsuprine Group retail presented absolute numbers: final area (p=0.001*) and viable area (p=0.006*) with the highest values; necrosis (p=0.001*) had the lowest value. Microscopic examination revealed no significant findings in the study of TRAM under the action of isoxsuprine and nicotine to the percentage of necrosis in the left and right cranial and caudal regions. CONCLUSIONS: There was significant improvement in viability of TRAM using the isoxsuprine solution alone. No influence using nicotine alone and in association with isoxsuprine. .

Predictive role of hematologic parameters in testicular torsion

PURPOSE: To evaluate the predictive role of the neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), mean platelet volume (MPV), and platelet count (PLT) in the diagnosis of testicular torsion (TT) and testicular viability following TT. MATERIALS AND METHODS: We analyzed two study groups in this retrospective study: 75 patients with a diagnosis of TT (group 1) and 56 age-matched healthy subjects (group 2). We performed a complete blood count as a part of the diagnostic procedure, and NLR, PLR, MPV, and PLT values were recorded. We compared the patient and control groups in terms of these parameters. Then, TT patients were divided into two subgroups according to the time elapsed since the onset of symptoms. Subsequently, we evaluated the relationship between the duration of symptoms and these parameters. RESULTS: There were significant differences between groups 1 and 2 in NLR, PLR, and PLT (p<0.001 for all). There was no predictive role of MPV in the diagnosis of TT (p=0.328). We determined significantly high sensitivity and specificity levels for NLR in the prediction of TT diagnosis (84% and 92%, respectively). Furthermore, NLR was significantly related to the duration of symptoms in TT patients (p=0.01). CONCLUSIONS: NLR may be a useful parameter in the diagnosis of TT. Furthermore, NLR may be used as a predictive factor for testicular viability following TT.

Usefulness of Skin Explants for Histologic Analysis after Fractional Photothermolysis

BACKGROUND: Fractional laser resurfacing treatment has been extensively investigated and is widely used. However, the mechanism underlying its effects is poorly understood because of the ethical and cosmetic problems of obtaining skin biopsies required to study the changes after laser treatment. OBJECTIVE: To evaluate the usefulness of human skin explants for the investigation of fractional photothermolysis. METHODS: Full-thickness discarded skin was treated in 4 ways: no treatment (control), fractional carbon dioxide laser, fractional Er:YAG laser, and fractional 1,550-nm erbium-doped fiber laser. Both treated and non-treated skin samples were cultured ex vivo at the air-medium interface for 7 days. Frozen tissue was sectioned and stained with hematoxylin & eosin for histologic examination and nitro blue tetrazolium chloride for viability testing. RESULTS: Skin explants cultured for up to 3 days exhibited histologic changes similar to those observed in in vivo studies, including microscopic treatment zones surrounded by a thermal coagulation zone, re-epithelialization, and formation of microscopic epidermal necrotic debris. However, the explant structure lost its original form within 7 days of culture. The viability of skin explants was maintained for 3 days of culture but was also lost within 7 days. CONCLUSION: The skin explant model may be a useful tool for investigating the immediate or early changes following fractional photothermolysis, but further improvements are required to evaluate the long-term and dermal changes.

Successful parathyroid tissue autograft after 3 years of cryopreservation: a case report / Implante bem-sucedido de paratireoide criopreservada por 3 anos: relato de caso

After a total parathyroidectomy, well-established protocols for the cryopreservation of parathyroid tissue and for the delayed autograft of this tissue exist, especially in cases of secondary hiperparathyroidism (HPT) or familial or sporadic parathyroid hyperplasia. Although delayed autografts are effective, the published success rates vary from 10% to 83%. There are numerous factors that influence the viability, and therefore the success, of an autograft, including cryopreservation time. Certain authors believe that the tissue is only viable for 24 months, but there is no consensus on how long the parathyroid tissue can be preserved. A 63-year-old male who was diagnosed with sporadic multiple endocrine neoplasia type 1 and primary hyperparathyroidism, and was submitted to a total parathyroidectomy and an autograft in the forearm. The implant failed, and the patient developed severe hypoparathyroidism in the months following the surgery. Thirty-six months after the total parathyroidectomy, the cryopreserved autograft was successfully transplanted, and hypoparathyroidism was reversed (most recent systemic parathyroid hormone, PTH, of 36 pg/mL, and total calcium of 9.1 mg/dL; no oral calcium supplementation). The case presented here indicates that cryopreserved parathyroid tissue may remain viable after 24 months in storage, and may retain the capacity to reverse permanent postsurgical hypoparathyroidism. These data provide reasonable evidence that the time limit for cryopreservation remains undetermined and that additional research would be valuable. Arq Bras Endocrinol Metab. 2014;58(3):313-6.

O implante de tecido paratireoideano criopreservado após paratireoidectomia total é um procedimento bem estabelecido e, embora tenha sua eficácia comprovada, as taxas de sucesso variam de 10% a 83% na literatura. O tempo de criopreservação é um dos diversos fatores relacionados ao sucesso do implante. Alguns autores defendem que o tecido permanece viável até 24 meses de criopreservação, no entanto, não há consenso. Homem de 63 anos diagnosticado com neoplasia endócrina múltipla tipo I e hiperparatireoidismo primário foi submetido a paratireoidectomia total e autoimplante em membro superior. O implante falhou e o paciente desenvolveu hipoparatireoidismo. Após 36 meses da paratireoidectomia total, foi realizado o implante de paratireoide criopreservada, com sucesso. O hipoparatireoidismo foi revertido e o paciente permanece sem suplementação de cálcio e PTH sistêmico de 36 pg/mL e cálcio total de 9,1 mg/dL. O caso apresentado mostra que o tecido paratireoideano criopreservado pode permanecer viável após 24 meses e há possibilidade de reverter o hipoparatireoidismo pós-cirúrgico. Isso traz evidência de que o tempo limite de criopreservação permanece incerto e que novas pesquisas seriam de grande valia. Arq Bras Endocrinol Metab. 2014;58(3):313-6.

Pre-ejection mitral annular motion velocity responses to dobutamine infusion: a quantitative approach for assessment of myocardial viability

Dobutamine stress echocardiography [DSE] is widely used for detection of myocardial viability. The main limitation of DSE is its subjective interpretation. Assessment of mitral annular motion velocities with tissue Doppler imaging is a simple and quantitative measurement. To determine the relationship between myocardial viability and regional systolic mitral annular motion tissue Doppler velocities responses to dobutamine stress. Our study group included 42 patients with previous myocardial infarction referred for coronary angiography and revascularization. We did dobutamine stress tissue Doppler echocardiography [DSTDE] measuring velocities of pre-ejection wave [pre-Ej] and peak ejection wave [Ej] at rest and during low-dose dobutamine infusion. We did follow up echocardiography after 1 month. After exclusion of the normokinetic walls, we analyzed 196 walls. Using receiver operator characteristic ROC curves, the optimal cut-off value for viability assessment was an increase of 1.75cm/s in pre-ejection velocity during DSTDE [area under the curve 0.70, p<0.001]. On the other hand, the optimal cut-off value for viability assessment was an increase of 1.75cm/s in ejection velocity during DSTDE [area under the curve 0.613, p=0.01]. The sensitivity, specificity, and total accuracy of the DSTSE [pre-Ej] versus the gold standard for detection of myocardial viability were 66.15%, 67.94%, and 67.35%, respectively. The sensitivity, specificity, and total accuracy of the DTSE [Ej] were 56.92%, 64.12%, and 61.43%, respectively. There was a good correlation between the pre-Ej at 5 ug/kg/min dobutamine infusion and the pre-Ej after revascularization [r=0.64, p=0.01] while the correlation with the Ej was moderate [r=0.50, p=0.01]. Viable left ventricular myocardium could be identified easily and quantitatively with pre-ejection mitral annular velocity during dobutamine infusion. The pre-ejection wave during DSTDE showed greater sensitivity and specificity for the prediction of myocardial viability than the ejection wave

Adipose-derived stem cells (ADSC) in the viability of a random pattern dorsal skin flap in rats

PURPOSE: To evaluate the viability of random pattern dorsal skin flaps in rats after injection of adipose-derived stem cells (ADSC). METHODS: Thirty five adult male Wistar EPM rats (weight 250-300 g) were distributed, at random, in two groups. I- Control (flap elevation with injection of saline solution) with fifteen animals and II- Experimental (flap elevation with injection of ADSC ) with fifteen animal. The ADSC were isolated from others five adult male rats. A dorsal skin flap measuring 10x4 cm was raised and a plastic barrier was placed between the flap and its bed in both groups and the injection (cells or saline solution) were perfomed immediately after the surgery. The percentage of flap necrosis was measured on the seventh postoperative day. RESULTS: The ADSC were able to replicate in our culture conditions. We also induced their adipogenic, osteogenic and chondrogenic differentiation to verify their mesenchymal stem cells potentiality in vitro. The results were statistically significant showing that the ADSC decreased the area of necrosis (p<0.05). CONCLUSIONS: The cells demonstrated adipogenic, osteogenic and chondrogenic differentiation potential in vitro. The administration of adipose-derived stem cells was effective to increase the viability of the random random pattern dorsal skin flaps in rats. .

Adipose-derived stem cells (ADSC) in the viability of random skin flap in rats

PURPOSE: To evaluate the effects of the adipose-derived stem cells (ADSC) in the viability of random skin flap in rats. METHODS: Thirty five adult male Wistar rats (weight 250-300 g) were used. ADSC were isolated from adult male rats (n=5). ADSC were separated, cultured and then analyzed. A dorsal skin flap measuring 10x4 cm was raised and a plastic barrier was placed between the flap and its bed. After the surgical procedure, the animals were randomized into two groups (n=15 each group), group control and group ADSC. In all groups the procedures were performed immediately after the surgery. The percentage of flap necrosis was measured on the seventh postoperative day. RESULTS: The ADSC were able to replicate in our culture conditions. We also induced their adipogenic, osteogenic and chondrogenic differentiation, verifying their mesenchymal stem cells potentiality in vitro. The results were statistically significant showing that the ADSC decreased the area of necrosis (p<0.05). CONCLUSION: The cells demonstrated adipogenic, osteogenic and chondrogenic differentiation potential in vitro. The administration of adipose-derived stem cells was effective to increase the viability of the random skin flaps in rats. .