Ischemic preconditioning and tacrolimus pretreatment as strategies to attenuate intestinal ischemia-reperfusion injury in mice.

The intestine is highly sensitive to ischemia-reperfusion injury (IRI), a phenomenon occurring in different intestinal diseases. Several strategies to mitigate IRI are in experimental stages; unfortunately, no consensus has been reached about the most appropriate one. We report a protocol to study ischemic preconditioning (IPC) evaluation in mice and to combine IPC and tacrolimus (TAC) pretreatment in a warm ischemia model. Mice were divided into treated (IPC, TAC, and IPC + TAC) and untreated groups before intestinal ischemia. IPC, TAC, and IPC + TAC groups were able to decrease postreperfusion nitrites levels (P < .05). IPC-containing groups had a major beneficial effect by preserving the integrity of the intestinal histology (P < .05) and improving animal survival (P < .002) compared with TAC alone or the untreated group. The IPC + TAC group was the only one that showed significant improvement in lung histological analysis (P < .05). The TAC and IPC + TAC groups down-regulated intestinal expression of interleukin (II)-6 and IL1b more than 10-fold compared with the control group. Although IPC and TAC alone reduced intestinal IRI, the used of a combined therapy produced the most significant results in all the local and distant evaluated parameters.

Outstanding survival and regeneration process by the use of intelligent acellular dermal matrices and mesenchymal stem cells in a burn pig model.

A pig model with a deep large burn was used to study the regeneration process induced by mesenchymal stem cells (MSCs) and acellular pig dermal matrices, made intelligent by the combination with biodegradable nanofibers loaded with growth factors (granulocyte-macrophage colony-stimulating factor and epidermal growth factor) and coated with the anti-CD44 monoclonal antibody (intelligent acellular dermal matrices, IADMs). These IADMs are specially designed to integrate in the wound bed as new biological scaffolds as well as to specifically recruit and attach circulating and/or externally applied MSCs through the anti-CD44 antibody while delivering precise amounts of growth factors. In this way, the reparative process as well as the aesthetic and functional results were enhanced in our burn model. The animal survived, the wound was completely closed, and total regeneration of the skin was obtained without much scarring. Surprisingly, hair follicles and other skin appendages developed despite the severity and deepness of the burn. Even burned muscles and ribs seemed to have undergone a regenerative process by the end of the study. Based on these findings, we have proposed the use of IADMs and autologous, allogeneic or xenogeneic MSCs, as a new paradigm for the future treatment of large burns and probably other dermatological and cosmetic human conditions.

The next generation of burns treatment: intelligent films and matrix, controlled enzymatic debridement, and adult stem cells.

We describe a novel technology based on nanoengineered multifunctional acellular biologic scaffolds combined with wound dressings and films of the same kind. This method allows selective delivery and release of shielded biomaterials and bioactive substances to a desired wound or damaged tissue while stimulating the selective anchoring and adhesion of endogenous circulating repairing cells, such as mesenchymal stem cells, to obtain a faster and more physiologic healing process. We also present a new controlled enzymatic debridement process for more effective burned tissue scarolysis. In light of our preliminary in vitro and in vivo data, we are convinced that these approaches can include the use of other kinds of adult stem cells, such as endometrial regenerative cells, to improve the vascularization of the constructs, with great potential in the entire tissue and organ regeneration field but especially for the treatment of severely burned patients, changing the way these lesions may be treated in the future.

Effect of cyclosporine and sirolimus on fatty acid desaturase activities in cultured HEPG2 cells.

The aim of the present work was to evaluate the influence of cyclosporine (CsA) and sirolimus (SRL) on fatty acid (FA) desaturase activities. These enzymes (named Delta9, Delta6, and Delta5 desaturases) catalyze reactions leading to the biosynthesis of n-9, n-6, and n-3 FA families. n-3 FA family, derived from alpha-linolenic acid, is involved in the prevention of vascular events, which appear after successful kidney transplantation. Five groups of HepG(2) cells in culture were treated with either CsA (1 microg/microL and 2 microg/microL) or SRL (10 ng/mL and 20 ng/mL) for 3 days, including a control group without immunosuppressive treatment. We studied the incorporation and metabolic conversion of radioactive [1-(14)C]palmitic, linoleic, and eicosatrienoic acids. We also analyzed fatty acid composition. The distribution of radioactive metabolic products after incubation of these cells with [1-(14)C]palmitic acid revealed a decrease in Delta9 desaturase activity in the presence of each immunosuppressive drug: CsA = 0.61 +/- 0.01; SRL = 0.59 +/- 0.04 versus control = 0.79 +/- 0.05 (P < .01). We observed a significant increase in Delta6 and Delta5 desaturase activities under the influence of the immunosuppressive drugs: radiolabeled linoleic acid (CsA: 0.93 +/- 0.04; SRL: 1.02 +/- 0.03 vs control 0.60 +/- 0.03; P < .01) and eicosatrienoic acid (CsA: 1.12 +/- 0.02; SRL: 1.07 +/- 0.01 vs control 0.75 +/- 0.01; P < .01). In conclusion, CsA and SRL modulated the biosynthesis of polyunsaturated FAs, decreasing Delta9 desaturase and increasing Delta6 and Delta5 desaturase activities.

Use of cryopreserved rat arteries for microsurgical training.

Silastic tubes are used as training material for performing microvascular anastomoses. However, silastic texture differs from that of actual blood vessels. In the present work, we evaluate the use of preserved rat arterial segments for training in microvascular anastomoses. One-centimeter-long rat arterial segments were obtained from femoral, carotid, and abdominal arteries, preserved in cold saline solution, and frozen. Trainees performed microvascular anastomoses using the aforementioned material and answered questions about texture, consistency, and wall resistance to the needle, comparing preserved arterial wall and silastic tubes. They were also asked whether the arterial pedicles had a consistency and texture similar to normal vessels, and if they were a more reliable method for practicing microsurgery techniques than synthetic materials. They preferred frozen arterial pedicles over silastic tubes. We conclude that arterial cadaveric segments are a suitable biologic material for microsurgical training. Since they can be obtained from other experiments, this is an effective way to reduce the number of animals bred and sacrificed for teaching purposes.

Trasplante hepático ortotápico en ratas. Técnica quirúrgica, complicaciones y sus tratamientos / Orthotopic liver transplantation in rats

The orthotopic rat liver transplant model is a widely used technique in transplantation research. It has many advantages over other animal transplant models because of its availability and low cost. However, it must be emphasized that success with the rat model requires thorough training. The aim of this paper is to describe the microsurgical technique involved in 60 rat liver transplants and to discuss the complications and their treatments. Forty-nine liver transplants were performed at the Experimental Laboratory of the University Hospital, Ontario, Canada (ELUH) and 11 were performed at the Laboratorio de Trasplante de Organos de la Facultad de Ciencias Medicas de La Plata, Buenos Aires. Argentina (LTO). Among the transplants performed at the ELUH, the observed complications were haemorrhage (n = 4), pneumothorax (n = 1), anastomotic failure (n = 15), bile leak (n = 3), and bile duct necrosis (n = 9). The remaining 17 rats at the ELUH were healthy at day 7 after surgery. Animal survival immediately postop, at 24 hours postop and at 7 days postop was achieved with the 9th, 20th and 21st transplants respectively. At the LTO, 3 rats died as a result of anaesthetic complications. Seven-day animal survival was achieved with the 11th transplant. We beleive that the description of the orthotopic rat liver transplantation technique, as well as the discussion regarding complications and their management, can be useful for researchers interested in performing liver transplantation in rats

Trasplante hepático ortotápico en ratas. Técnica quirúrgica, complicaciones y sus tratamientos / Orthotopic liver transplantation in rats

The orthotopic rat liver transplant model is a widely used technique in transplantation research. It has many advantages over other animal transplant models because of its availability and low cost. However, it must be emphasized that success with the rat model requires thorough training. The aim of this paper is to describe the microsurgical technique involved in 60 rat liver transplants and to discuss the complications and their treatments. Forty-nine liver transplants were performed at the Experimental Laboratory of the University Hospital, Ontario, Canada (ELUH) and 11 were performed at the Laboratorio de Trasplante de Organos de la Facultad de Ciencias Medicas de La Plata, Buenos Aires. Argentina (LTO). Among the transplants performed at the ELUH, the observed complications were haemorrhage (n = 4), pneumothorax (n = 1), anastomotic failure (n = 15), bile leak (n = 3), and bile duct necrosis (n = 9). The remaining 17 rats at the ELUH were healthy at day 7 after surgery. Animal survival immediately postop, at 24 hours postop and at 7 days postop was achieved with the 9th, 20th and 21st transplants respectively. At the LTO, 3 rats died as a result of anaesthetic complications. Seven-day animal survival was achieved with the 11th transplant. We beleive that the description of the orthotopic rat liver transplantation technique, as well as the discussion regarding complications and their management, can be useful for researchers interested in performing liver transplantation in rats (AU)

[Orthotopic liver transplant in rats. Surgical technique, complications and treatment]. / Trasplante hepàtico ortotòpico en ratas. Técnica quirúrgica, complicaciones y sus tratamientos.

The orthotopic rat liver transplant model is a widely used technique in transplantation research. It has many advantages over other animal transplant models because of its availability and low cost. However, it must be emphasized that success with the rat model requires thorough training. The aim of this paper is to describe the microsurgical technique involved in 60 rat liver transplants and to discuss the complications and their treatments. Forty-nine liver transplants were performed at the Experimental Laboratory of the University Hospital, Ontario, Canada (ELUH) and 11 were performed at the Laboratorio de Trasplante de Organos de la Facultad de Ciencias Médicas de La Plata, Buenos Aires. Argentina (LTO). Among the transplants performed at the ELUH, the observed complications were haemorrhage (n = 4), pneumothorax (n = 1), anastomotic failure (n = 15), bile leak (n = 3), and bile duct necrosis (n = 9). The remaining 17 rats at the ELUH were healthy at day 7 after surgery. Animal survival immediately postop, at 24 hours postop and at 7 days postop was achieved with the 9th, 20th and 21st transplants respectively. At the LTO, 3 rats died as a result of anaesthetic complications. Seven-day animal survival was achieved with the 11th transplant. We beleive that the description of the orthotopic rat liver transplantation technique, as well as the discussion regarding complications and their management, can be useful for researchers interested in performing liver transplantation in rats.

Trasplante hepOtico ortotòpico en ratas. Técnica quirúrgica, complicaciones y sus tratamientos. / [Orthotopic liver transplant in rats. Surgical technique, complications and treatment]

The orthotopic rat liver transplant model is a widely used technique in transplantation research. It has many advantages over other animal transplant models because of its availability and low cost. However, it must be emphasized that success with the rat model requires thorough training. The aim of this paper is to describe the microsurgical technique involved in 60 rat liver transplants and to discuss the complications and their treatments. Forty-nine liver transplants were performed at the Experimental Laboratory of the University Hospital, Ontario, Canada (ELUH) and 11 were performed at the Laboratorio de Trasplante de Organos de la Facultad de Ciencias Médicas de La Plata, Buenos Aires. Argentina (LTO). Among the transplants performed at the ELUH, the observed complications were haemorrhage (n = 4), pneumothorax (n = 1), anastomotic failure (n = 15), bile leak (n = 3), and bile duct necrosis (n = 9). The remaining 17 rats at the ELUH were healthy at day 7 after surgery. Animal survival immediately postop, at 24 hours postop and at 7 days postop was achieved with the 9th, 20th and 21st transplants respectively. At the LTO, 3 rats died as a result of anaesthetic complications. Seven-day animal survival was achieved with the 11th transplant. We beleive that the description of the orthotopic rat liver transplantation technique, as well as the discussion regarding complications and their management, can be useful for researchers interested in performing liver transplantation in rats.

Anticuerpos contra el virus de la hepatitis C - anti HCV. Prevalencia e influencia sobre la morbimortalidad en los pacientes sometidos a trasplante renal, en los dos ultimos anos del programa de trasplante renal de la ciudad de la Plata / Hepatitis C viruses antibodies - anti HCV. Prevalence and their influence on the morbi-mortality in transplantation patients, in the last two years of the kidney transplantation program of the La Plata city

Se estudiaron 34 pacientes con trasplante renal (TxR), 18 varones y 16 mujeres, con el objetivo de conocer la prevalencia de Anti HCV, en este tipo de pacientes y su influencia sobre la morbimortalidad temprana. La media de segmiento fue 8.44 DS 6.7 meses y la de edad 38.32 años DS 13.97. Todos recibieron el mismo esquema inmuno-supresor y los episodios de rechazo se trataron con pulsos de metilprednisolona. Resultaron Anti HCV r (por EIA II) de Abbott e Inmunoblotting de Péptidos Sintéticos LIA TEK Organon Teknika); 7 (20.6 por ciento) pacientes y (NR) 27 (79.4 por ciento). Recibieron injerto de donante cadavérico 4 (57.1 por ciento), Anti HCV R y 10 (37.0 por ciento) Anti HCV NR; de donante vivo relacionado 3 (42.9 por ciento) Anti HCV R y 17 (63.0 por ciento) Anti HCV NR. Tenían antecedentes de haber pedacido hepatitis 6 (85.7 por ciento) de lso 7 Anti HCV R: 2 hepatitis crónicas y 4 agudas (2 HBV y 2 no B (NABV) y 6 (22.2 por ciento) de los 27 Anti HCV NR. El tiempo medio de tratamiento hemodialítico antes del trasplante en el grupo Anti HCV r fue 63.0 DS 27.0 meses y resultó significativamente superior (P<0.05) al del grupo Anti HCV (NR) (27.3 DS 20.7). Episodios de rechazos, hepatopatías post-trasplante y sobrevida del injerto y del paciente no fueron significativamente diferentes entre los pacientes Anti HCV R y los NR.

Anticuerpos contra el virus de la hepatitis C - anti HCV. Prevalencia e influencia sobre la morbimortalidad en los pacientes sometidos a trasplante renal, en los dos ultimos anos del programa de trasplante renal de la ciudad de la Plata / Hepatitis C viruses antibodies - anti HCV. Prevalence and their influence on the morbi-mortality in transplantation patients, in the last two years of the kidney transplantation program of the La Plata city

Se estudiaron 34 pacientes con trasplante renal (TxR), 18 varones y 16 mujeres, con el objetivo de conocer la prevalencia de Anti HCV, en este tipo de pacientes y su influencia sobre la morbimortalidad temprana. La media de segmiento fue 8.44 DS 6.7 meses y la de edad 38.32 años DS 13.97. Todos recibieron el mismo esquema inmuno-supresor y los episodios de rechazo se trataron con pulsos de metilprednisolona. Resultaron Anti HCV r (por EIA II) de Abbott e Inmunoblotting de Péptidos Sintéticos LIA TEK Organon Teknika); 7 (20.6 por ciento) pacientes y (NR) 27 (79.4 por ciento). Recibieron injerto de donante cadavérico 4 (57.1 por ciento), Anti HCV R y 10 (37.0 por ciento) Anti HCV NR; de donante vivo relacionado 3 (42.9 por ciento) Anti HCV R y 17 (63.0 por ciento) Anti HCV NR. Tenían antecedentes de haber pedacido hepatitis 6 (85.7 por ciento) de lso 7 Anti HCV R: 2 hepatitis crónicas y 4 agudas (2 HBV y 2 no B (NABV) y 6 (22.2 por ciento) de los 27 Anti HCV NR. El tiempo medio de tratamiento hemodialítico antes del trasplante en el grupo Anti HCV r fue 63.0 DS 27.0 meses y resultó significativamente superior (P<0.05) al del grupo Anti HCV (NR) (27.3 DS 20.7). Episodios de rechazos, hepatopatías post-trasplante y sobrevida del injerto y del paciente no fueron significativamente diferentes entre los pacientes Anti HCV R y los NR. (AU)

[Hepatitis C viruses antibodies. Prevalence and their influence on morbidity-mortality in renal transplant recipients, in the last two years of the kidney transplantation program in La Plata city]. / Anticuerpos contra el Virus de la Hepatitis C--anti HCV. Prevalencia e influencia sobre la morbimortalidad en los pacientes sometidos a trasplante renal, en los dos ultimos años del programa de trasplante renal de la ciudad de La Plata.

UNLABELLED: A small series of 34 renal transplanted patients (RTx) were studied, 18 males and 16 females in order to know the prevalence of Anti HCV in this type of patients and their influence on early morbi-mortality. The follow-up mean was 8.44 months SD 6.7, and Age 38.32 SD 13.97. All patients were under the same immunosuppressive scheme, and rejection episodes were treated with methilprednisolone pulses. The results were: 7 (20.6%) Anti HCV seroreactives (R) (EIA II Abbott and Immunoblotting of synthetic Peptides LIA TEK Organon Teknika); and 27 (79.4%) non-serorectives (NR), 14 patients received grafts from cadaveric donor; 4 (57.1%) Anti HCV (R), and 10 (37.0%) Anti HCV (NR). 20 patients have received grafts from lived-related donors: 3 (42.9%) Anti HCV (R), and 17 (63.0%) Anti HCV (NR). 6 (85.7%) of the 7 patients Anti HCV (R) had hepatitis history: 2 chronic hepatitis, 4 acute hepatitis (2HBV) and 2 no A no B (NANBV) and 6 (22.2%) of the 27 Anti HCV (NR). The mean time of hemodialysis treatment before transplantation in the Anti HCV (R) group was of 63.0 months SD27.0, and it was significantly superior (P < 0.05) to the Anti HCV (NR) group with 27.3 months SD 20.7. There were no significant differences between the Anti HCV (R) and (NR) patients with regard to rejection episodes, post-transplant hepatopathies, and survival of graft and patient. CONCLUSIONS: 1) Anti HCV prevalence is of 20.6%. 2) Time of hemodialysis prior to transplantation and an hepatitis history during hemodialysis came out to be significantly higher in Anti HCV (R) RTx. 3) Morbi-mortality is no modified by the presence of Anti HCV during a mean follow-up period of 8.44 months.

La patologia toracopleuropulmonar en el trasplante renal

Se presenta la patología toracopleuropulmonar que afectó a 20 pacientes de una serie de 45 casos de trasplante renal y a 3 casos sobre 17 donantes vivos relacionados. Se confirma el predominio del compromiso parenquimatoso 75% sobre el torácico 25%; a su vez el primero corresponde escencialmente a neumopatías infecciosas 88%, mientras que el segundo a osteodistrofia renal 100% con especial participaron del hiperparatirodismo secundario severo en 10%. Las neumopatías infecciosas revelaron un elevado predominio de la etiología bacteriana 80% con respecto a la parasitaria 13,3% y fúngica 6,6%. Así como una marcada prevalencia en el primer semestre 65% sobre el segundo 39% y el segundo año 5% de pos-trasplante. La distribución según origen es semejante 47% hospitalaria y 53% extrahospitalaria. El cuadro clínico mostró a la fiebre 100% tos y expectoración 80% y a un sindrome de condensación inicial (foco de rales creptandes y subcrepitantes ) 82% como los elementos relevantes. El laboratorio aportó al diagnóstico tes aspectos: a) Recuento y fórmula leucocitaria: leucopenia (menos de 5.000/mm3) en el 35% de los casos, aunque sin alcanzar neuropenia absoluta (menos de 500 mm3) en ninguno, leucocitosis 11,8%) correlacionándose con la mortalidad la neutropenia y la neutrofilia. b) Gases en sangre: se consideróa necesario en la mitad de los casos y sólo aportó hipoxemia en ods episodios con evolución opuesta (uno vivo y un fallecido), lo que le restó valor predictivo. c) cultivo de esputo: fue positiv

La patologia toracopleuropulmonar en el trasplante renal

Se presenta la patología toracopleuropulmonar que afectó a 20 pacientes de una serie de 45 casos de trasplante renal y a 3 casos sobre 17 donantes vivos relacionados. Se confirma el predominio del compromiso parenquimatoso 75% sobre el torácico 25%; a su vez el primero corresponde escencialmente a neumopatías infecciosas 88%, mientras que el segundo a osteodistrofia renal 100% con especial participaron del hiperparatirodismo secundario severo en 10%. Las neumopatías infecciosas revelaron un elevado predominio de la etiología bacteriana 80% con respecto a la parasitaria 13,3% y fúngica 6,6%. Así como una marcada prevalencia en el primer semestre 65% sobre el segundo 39% y el segundo año 5% de pos-trasplante. La distribución según origen es semejante 47% hospitalaria y 53% extrahospitalaria. El cuadro clínico mostró a la fiebre 100% tos y expectoración 80% y a un sindrome de condensación inicial (foco de rales creptandes y subcrepitantes ) 82% como los elementos relevantes. El laboratorio aportó al diagnóstico tes aspectos: a) Recuento y fórmula leucocitaria: leucopenia (menos de 5.000/mm3) en el 35% de los casos, aunque sin alcanzar neuropenia absoluta (menos de 500 mm3) en ninguno, leucocitosis 11,8%) correlacionándose con la mortalidad la neutropenia y la neutrofilia. b) Gases en sangre: se consideróa necesario en la mitad de los casos y sólo aportó hipoxemia en ods episodios con evolución opuesta (uno vivo y un fallecido), lo que le restó valor predictivo. c) cultivo de esputo: fue positiv