ABSTRACT
BACKGROUND: Trees within the Calophyllum genus are multi-use trees that produce valuable wood, phytochemicals with a range of biological activities, and seed oil as a source of biodiesel. As a consequence of climate change, there is a need to develop strategies to preserve valuable plant genetic resources. Cryopreservation represents the most suitable option for the long-term storage of germplasm with minimal space and maintenance requirements. OBJECTIVE: To determine appropriate methods to cryopreserve seeds of Calophyllum antillanum and maintain secondary compound production. MATERIALS AND METHODS: Seeds at a moisture content of 6% were used to evaluate two treatments: seeds immersed in liquid nitrogen and control seeds. Biosynthetic pathway efficiency was assessed post-cryo by determining anthraquinone contents in roots, stems and leaves following 30 and 75 d of seedling growth. RESULTS: The results indicated that exposure to liquid nitrogen delayed germination and seedling emergence for a period of up to 45 d after seed sowing. By 60 d of cultivation, no significant differences in plant growth were observed for cryostored and control seeds. The levels of anthraquinones, which were also measured in seeds and seedlings, were lower in plants regenerated from cryostored seeds following 30 d of growth, but there were no differences in roots and stems by 75 d of growth. Furthermore, the difference in leaf anthraquinone levels for cryopreserved and control seeds at 75 d was much smaller than at 30 d. CONCLUSION: The low initial anthraquinone levels in emerging seedlings correlated with the initial slow growth of cryopreserved seeds.
Subject(s)
Calophyllum , Seedlings , Anthraquinones/pharmacology , Cryopreservation/methods , Nitrogen/chemistry , Plant Leaves , SeedsABSTRACT
PURPOSE: To evaluate the vas deferens and testicles of rats submitted to bilateral inguinotomy and polypropylene (PP) mesh placement. METHOD: Sixty Wistar rats were randomized into three groups: Control (inguinotomy only), mesh placement over the vas deferens (Mesh-DD) or under the spermatic funiculus (Mesh-SF). The following analyses were performed: vas deferens morphometry (lumen area and wall thickness), quantification of collagen fibers, spermatogenesis, apoptosis (cleaved caspase-3 and TUNEL) and cellular proliferation (Ki67). Quantitative gene expression (qPCR) for apoptosis and inflammatory cytokines were evaluated by RT-PCR. RESULTS: In the apoptosis pathway, Mesh-DD showed one upregulated gene (Il10) and three downregulated genes (Fadd, Tnfrsf1b and Xiap). In Mesh-SF, 17 genes were downregulated. In the inflammation pathway (Mesh-DD), one gene was upregulated (Il1r1), and one gene was downregulated (Ccl12). In Mesh-SF, three genes were upregulated (Il1r1, Tnfsf13b and Csf1), and two were downregulated (Ccl12 and Csf2). PP mesh placement preserved spermatogenesis and did not alter the vas deferens or the testicle. In the ductus deferens, there was reduced luminal area (30 days), increased wall thickness (90 days), and increased type III collagen and cell proliferation (30 and 90 days) (p < 0.05). In the testicle, cell proliferation was greater in the Mesh-DD (p < 0.05). CONCLUSIONS: PP mesh, whether or not in direct contact with spermatic funicular structures, induces changes that were not sufficient to cause damage to the evaluated organs.
Subject(s)
Genitalia/surgery , Polypropylenes/therapeutic use , Surgical Mesh/standards , Animals , Disease Models, Animal , Male , Rats , Rats, WistarABSTRACT
Itolizumab is a humanized anti-CD6 monoclonal antibody (mAb) that has previously shown encouraging results, in terms of safety and positive clinical effects, in a 6-week monotherapy clinical trial conducted in rheumatoid arthritis (RA) patients. The current Phase I study evaluated the safety and clinical response for a longer treatment of 12 itolizumab intravenous doses in subjects with active RA despite previous disease-modifying anti-rheumatic drug (DMARD) therapy. Twenty-one subjects were enrolled into four dosage groups (0·1, 0·2, 0·4 and 0·8 mg/kg). Efficacy end-points including American College of Rheumatology (ACR)20, ACR50 and ACR70 response rates and disease activity score in 28 joints (DAS28) were monitored at baseline and at specific time-points during a 10-week follow-up period. Itolizumab was well tolerated up to the highest tested dose. No related serious adverse events were reported and most adverse events were mild. Remarkably, itolizumab treatment did not produce lymphopenia and, therefore, was not associated with infections. All patients achieved a clinical response (ACR20) at least once during the study. Eleven subjects (55%) achieved at least a 20% improvement in ACR just 1 week after the first itolizumab administration. The clinical response was observed from the beginning of the treatment and was sustained during 24 weeks. The efficacy profile of this 12-week treatment was similar to that of the previous study (6-week treatment). These results reinforce the safety profile of itolizumab and provide further evidence on the clinical benefit from the use of this anti-CD6 mAb in RA patients.
Subject(s)
Antibodies, Monoclonal, Humanized/therapeutic use , Antirheumatic Agents/therapeutic use , Arthritis, Rheumatoid/drug therapy , Adolescent , Adult , Aged , Antigens, CD/immunology , Antigens, Differentiation, T-Lymphocyte/immunology , Cuba , Disease Progression , Drug-Related Side Effects and Adverse Reactions , Female , Follow-Up Studies , Humans , Lymphopenia , Male , Middle Aged , Young AdultABSTRACT
Intestinal ischemia and reperfusion injury are widely used models, which result into tissue injury and multiple organ failure also observed after trauma and surgery. Ischemic preconditioning (IPC) preceding ischemia and reperfusion (IR) was shown to attenuate this injury and has a potential therapeutic application; however the exact underlying mechanism is not clear. Neutrophils play an important role in the mechanism of injuries caused by ischemia and reperfusion while IPC led to a decrease in neutrophil stimulation and activation. The effect of preconditioning on the neutrophil proteome is unclear. Proteomic analysis has been ratified as an appropriate tool for studying complex systems. In order to evaluate the effect of IPC preceding 45min of ischemia on the proteome of neutrophils we used Wistar rats divided in four experimental groups: Control, sham laparotomy, intestinal ischemia reperfusion and ischemic preconditioning. After neutrophil separation, proteins were extracted, trypsin digested and the resulting peptides were iTRAQ labeled followed by HILIC fractionation and nLC-MS/MS analysis. After database searches, normalization and statistical analysis our proteomic analysis resulted in the identification of 2437 protein groups that were assigned to five different clusters based on the relative abundance profiles among the experimental groups. The clustering followed by statistical analysis led to the identification of significantly up and downregulated proteins in IR and IPC. Cluster based KEGG pathways analysis revealed up- regulation of actin cytoskeleton, metabolism, Fc gamma R mediated phagocytosis, chemokine signaling, focal adhesion and leukocyte transendothelial migration whereas downregulation in ribosome, spliceosome, RNA transport, protein processing in endoplasmic reticulum and proteasome, after intestinal ischemic preconditioning. Furthermore, enzyme prediction analysis revealed the regulation of some important antioxidant enzymes and having their role in reactive oxygen species production. To our knowledge, this work describes the most comprehensive and detailed quantitative proteomic study of the neutrophil showing the beneficial role of ischemic preconditioning and its effects on the neutrophil proteome. This data will be helpful to understand the effect of underlying protective mechanisms modulating the role of PMNs after IPC and provide a trustworthy basis for future studies. BIOLOGICAL SIGNIFICANCE: Preconditioning is a relevant strategy to overcome clinical implications from ischemia and reperfusion. Such implications have the neutrophil as a major player. Although many publications describe specific biochemical and physiological roles of the neutrophil in such conditions, there is no report of a proteomic study providing a broader view of this scenario. Here we describe a group of proteins significantly regulated by ischemia and reperfusion being such regulation prevented by preconditioning. Such finding may provide relevant information for a deeper understanding of the mechanisms involved, as well as serve as basis for future biomarker or drug target assays.
Subject(s)
Ischemic Preconditioning , Neutrophils/chemistry , Proteome/analysis , Proteomics/methods , Animals , Cell Movement , Cluster Analysis , Intestines/pathology , Oxidoreductases , Rats , Rats, Wistar , Reperfusion Injury , Ribosomal Proteins/analysisABSTRACT
La rabdomióisis es una entidad clínica y bioquímica caracterizada por necrosis muscular y liberación del contenido intracelular al torrente circulatorio. Sus causas son múltiples y su presentación clínica muy variable, por 10 que Los síntomas clásicos de dolor muscular, debilidad y orinas oscuras no siempre están presentes. El diagnostico se establece con niveles de creatina-fosfocinasa cinco veces por encima del valor normal y ha de instaurarse un tratamiento precoz para evitar complicaciones graves. Se presenta el caso de un varón joven que refería dolor y debilidad muscular tras practicar ejercicio físico días antes y se revisa la literatura.
Rhabdomyolysis is defined as a clinical and biochemical syndrome resulting from skeletal muscle injury that alters the integrity of the muscle cell membrane sufficiently to allow the release of the muscle cell contents into the plasma. There is a great heterogeneity in the clinical presentation and clasical symptoms as myalgias, weakness and tea-coloured urine, may not always be present. The diagnosis is established with the increase at least five times the normal value of creatine kinase. Treatment aims to prevent complications. We present the case of young male who had a days history of myalgias and weakness after exercise and review the literature.
Subject(s)
Creatine Kinase , Exercise , RhabdomyolysisABSTRACT
The aim of this study was evaluate the beta blocker atenolol (AT) and ischemic preconditioning (IPC) strategies for tissue protection against systemic effects of intestinal ischemia (I) and reperfusion (R) injury. Forty-two rats were pretreated with AT (1.5 mg · kg(-1)), 0.9% saline solution (SS; 0.1 mL), or IPC and then subjected to prolonged occlusion of the superior mesenteric artery for 60 minutes leading to I followed or not by 120 minutes of R, according to the group. For IPC, 5 minutes of I prior to 10 minutes of R were established. After this process of I or I-R, the right lung of each animal was adequately prepared for staining with hematoxylin and eosin and subsequent histologic analysis for quantification of inflammatory infiltrate was done. The left lung was frozen and prepared for assessment of oxidative stress by the quantification of thiobarbituric acid-reactivity substances (TBARS). Histologic analysis showed an important inflammatory infiltrate in the I-R + SS (I-R + SS = 4.5), which was significantly (P < .05) reduced by IPC (I-R + IPC = 3.0) or AT (I-R + AT = 3.0). Likewise, the TBARS levels were decreased by both strategies (I-R + SS = 0.63; I-R + IPC = 0.23; I-R + AT = 0.38; P < .05). Our results showed that AT and IPC attenuate pulmonary lesions caused by intestinal I and R process.
Subject(s)
Atenolol/pharmacology , Intestines/blood supply , Ischemic Preconditioning/methods , Lung Injury/prevention & control , Lung/blood supply , Reperfusion Injury/prevention & control , Adrenergic beta-1 Receptor Antagonists/pharmacology , Animals , Disease Models, Animal , Lung Injury/etiology , Male , Rats , Rats, Wistar , Reperfusion Injury/complicationsABSTRACT
To study the role of heparin and ischemic preconditioning (IPC) in cardiac injury after intestinal ischemia (I) and reperfusion (R), 54 rats underwent 60 minutes of I, which was produced by occlusion of the superior mesenteric artery, and/or 120 minutes of R. The IPC group had the I procedure stimulation for 5 minutes and R for 10 minutes. The control group was subjected to sham surgery only, and the other groups were injected with saline solution (SS; 0.1 mL) or heparin (100 IU/kg) via the inferior cava vein 5 minutes before I and 5 minutes before R and 55 minutes after the R begins in I-R groups. In all animals, cardiac samples were stained with hematoxylin and eosin for optical microscopy analysis, and other sample was processed for lipid peroxidation determination. In I-R groups, both heparin and IPC showed significant protection compared to the SS group; conversely, in animals subjected only to I, no protection was observed. Moreover, when heparin was associated with IPC, I-R protection was compromised and the ischemic injury increased. Data showed that IPC and heparin attenuated cardiac dysfunction caused by intestinal I and I-R, but when used in association did not show beneficial effects.
Subject(s)
Cardiomyopathies/prevention & control , Heparin/therapeutic use , Intestines/blood supply , Ischemic Preconditioning/methods , Reperfusion Injury/prevention & control , Animals , Anticoagulants/therapeutic use , Cardiomyopathies/etiology , Disease Models, Animal , Male , Rats , Rats, Wistar , Reperfusion Injury/complicationsABSTRACT
Surgery to transfer the axillary nerve and the nerve of the long head of the triceps presents two obstacles: 1) the access portals are not standardized and 2) the nerves are for their larger part approached through large incisions. The goal of this study was to explore the feasibility of an endoscopic microsurgical approach. The posterior aspect of a cadaver shoulder was approached through three communicating mini-incisions. The Da Vinci robot camera was installed on a central trocart, and the instrument arms on the adjacent trocarts. A gas insufflation distended the soft tissues up to the lateral axillary space. The branches of the axillary nerve and the nerve to the long head of the triceps brachii muscle were identified. The dissection of the axillary nerve trunk and its branches was easy. The posterior humeral circumflex veins and artery were dissected as well without any difficulty. Finding the nerve to the long head of the triceps brachii was found to be more challenging because of its deeper location. Robots properties allow performing conventional microsurgery: elimination of the physiologic tremor and multiplication of the movements. They also facilitate the endoscopic approach of the peripheral nerves, as seen in our results on the terminal branches of the axillary nerve and the nerve to the long head of the triceps brachii.
Subject(s)
Axilla/innervation , Brachial Plexus/surgery , Microsurgery/methods , Robotics , Axilla/surgery , Cadaver , Feasibility Studies , HumansABSTRACT
To study whether treatment with the beta-blocker atenolol (AT) attenuates intestinal dysfunction caused by ischemia (I) and reperfusion (R), rats were treated with AT (1.5 mg · kg(-1), intravenously) or saline solution (SS) prior to I (60 minutes), which was produced by occlusion of the superior mesenteric artery, and/or R (120 minutes). After I or I/R, 2-cm jejunal segments were mounted in an organ bath to study neurogenic contractions stimulated by electrical pulses or KCl using a digital recording system. Thin jejunal slices were stained with hematoxylin and eosin for optical microscopy analysis. Compared to the sham group, jejunal contractions were similar in the I + AT and the I/R + AT groups, but reduced in the I + SS and the I/R + SS groups. The jejunal enteric nerves were damaged in the I + SS and the I/R + SS groups, but not in the I + AT and the I/R + AT. These results suggest that AT may attenuate intestinal dysfunction caused by I and I/R.
Subject(s)
Adrenergic beta-1 Receptor Antagonists/pharmacology , Atenolol/pharmacology , Gastrointestinal Agents/pharmacology , Jejunum/blood supply , Jejunum/drug effects , Reperfusion Injury/prevention & control , Animals , Cytoprotection , Disease Models, Animal , Electric Stimulation , Enteric Nervous System/drug effects , Enteric Nervous System/physiopathology , Gastrointestinal Motility/drug effects , Jejunum/innervation , Jejunum/pathology , Jejunum/physiopathology , Male , Potassium Chloride/pharmacology , Rats , Rats, Wistar , Reperfusion Injury/pathology , Reperfusion Injury/physiopathologyABSTRACT
BACKGROUND: Dysfunction of the liver after transplantation may be related to the graft size and ischemia/reperfusion (I/R) injury. N-Acetylcysteine (NAC) exerts beneficial effects on livers undergoing ischemia reperfusion. We sought to evaluate NAC modulation on reduced livers associated with I/R injury. METHODS: Male C57BL/6 mice of 8 weeks of age were divided into groups: 50% hepatectomy (G-Hep); NAC (G-Hep + NAC [150 mg/kg]) via vena cava 15 minutes before hepatectomy; ischemia (G-Hep + IR); NAC with hepatectomy (G-IR + Hep + Nac); and IR using 30 minutes selective hepatic occlusion and reperfusion for 24 hours. After 24 hours, the remaining liver was removed, for staining with hematoxylin and eosin or labeling by proliferating cell nuclear antigen. Blood was collected for biochemical evaluations. Significance was considered for P ≤ .05. RESULTS: Aspartate aminotransferase was high in all studied groups reflecting the hepatectomy and intervention. injuries. However, when assessing alanine aminotransferase, which depicts liver function, induction of IR promoted a greater increase than hepatectomy (P = .0003). NAC decreased ALT activity in all groups, even in association with I/R (P < .05), reflecting a modulation of the injury. Necrosis resulting from IR was mitigated by NAC. The experimental model of 50% reduced live promoted regeneration of the hepatic remnant, which was accentuated by NAC, according to the total number of hepatocytes and PCNA values. CONCLUSION: NAC preserved the remnant liver in mice and stimulates regeneration even after IR injury.
Subject(s)
Acetylcysteine/pharmacology , Cell Proliferation/drug effects , Hepatectomy , Liver Regeneration/drug effects , Liver/drug effects , Liver/surgery , Reperfusion Injury/drug therapy , Alanine Transaminase/blood , Animals , Aspartate Aminotransferases/blood , Biomarkers/blood , Biomarkers/metabolism , Cytoprotection , Disease Models, Animal , Immunohistochemistry , Liver/blood supply , Liver/metabolism , Liver/pathology , Male , Mice , Mice, Inbred C57BL , Necrosis , Proliferating Cell Nuclear Antigen/metabolism , Reperfusion Injury/metabolism , Reperfusion Injury/pathology , Time FactorsABSTRACT
Anti-idiotypic monoclonal antibodies (mAb) have been evaluated for actively induced immunotherapy with encouraging results. However, rational combination of cancer vaccines with chemotherapy may improve the therapeutic efficacy of these two approaches used separately. The main objective of this study was to evaluate the antitumor effect of the co-administration of 1E10 (Racotumomab), a monoclonal anti-idiotype tumor vaccine against an IgM mAb, named P3 that reacts specifically with NeuGc-containing gangliosides and low-dose Cyclophosphamide in a mammary carcinoma model. F3II tumor-bearing mice were immunized subcutaneously with 100 microg of 1E10 mAb in Alum or with 150 mg/m(2) of Cyclophosphamide intravenously 7 days after the tumor inoculation. While a limited antitumor effect was induced by a single 1E10 mAb immunization; its co-administration with low-dose Cyclophosphamide reduced significantly the F3II mammary carcinoma growth. That response was comparable with the co-administration of the standard high-dose chemotherapy for breast cancer based on 60 mg/m(2) of Doxorubicin and 600 mg/m(2) of Cyclophosphamide, without toxicity signs. Combinatorial chemo-immunotherapy promoted the CD8(+) lymphocytes tumor infiltration and enhanced tumor apoptosis. Furthermore, 1E10 mAb immunization potentiated the antiangiogenic effect of low-dose Cyclophosphamide. Additionally, splenic myeloid cells Gr1(+)/CD11b(+) associated with a suppressor phenotype were significantly reduced in F3II tumor-bearing mice immunized with 1E10 mAb alone or in combination with low-dose Cyclophosphamide. This data may provide a rational for chemo-immunotherapy combinations with potential medical implications in breast cancer.
Subject(s)
Antibodies, Anti-Idiotypic/therapeutic use , Antibodies, Monoclonal/therapeutic use , Cancer Vaccines/therapeutic use , Gangliosides/immunology , Immunotherapy/methods , Mammary Neoplasms, Experimental/therapy , Animals , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Apoptosis/drug effects , Cancer Vaccines/immunology , Cyclophosphamide/pharmacology , Female , Immunohistochemistry , Mammary Neoplasms, Experimental/immunology , Mammary Neoplasms, Experimental/pathology , Mice , Mice, Inbred BALB C , Neovascularization, Pathologic/drug therapyABSTRACT
PURPOSE: The purpose of this study was to establish morphologically the best time of vascular occlusion to induce ischemic preconditioning (IPC) for rat small bowel undergoing ischemia and reperfusion injury. METHODS: After approval by the Ethics Committee, 36 EPM-1 young adult Wistar rats from 300-350 g were distributed into 6 groups: sham (S); ischemia and reperfusion (IR), with 50 minutes of cranial mesenteric artery occlusion and 30 minutes of reperfusion; IPC with 1 cycle of 2 minutes (IPC-2), 5 minutes (IPC-5), 10 minutes (IPC-10), or 15 minutes (IPC-15), followed by sustained IR. The animals anesthetized with ketamine (60 mg/kg) and xylazine (10 mg/kg) intramuscular (IM), were maintained on mattress heat, hydrated with saline (80 mL/kg), and injected with 100 IU heparin. Samples of jejunum were stained with hematoxylin and eosin (HE) and classified according to Park et al. Statistical analysis of results was performed using Kruskal-Wallis tests (P < .05). RESULTS: The histological evaluation showed no difference between IR and IPC15 rats (5.2 and 5, respectively; P = .84). Greater jejunal injury was observed with IPC15 (5) compared with other groups (IPC2 = 3, P = .03; IPC5 = 3.2, P = .05; IPC10 = 2.8, P = .02, respectively). There was no difference between groups IPC2 x IPC5 x IPC10. CONCLUSION: Morphologically, IPC with short times promoted greater intestinal protection against the IR lesion in rats.
Subject(s)
Intestine, Small/physiopathology , Ischemic Preconditioning/methods , Reperfusion Injury/physiopathology , Animals , Intestinal Mucosa/injuries , Intestinal Mucosa/pathology , Intestine, Small/pathology , Ischemia , Male , Rats , Rats, Wistar , Reperfusion , Reperfusion Injury/pathology , Reperfusion Injury/prevention & controlABSTRACT
PURPOSE: The purpose of this study was to evaluate estradiol serum levels and follicular development in rats subjected to ovarian autologous transplantation with or without remote ischemic preconditioning (R-IPC). METHODS: Seventy-two adult female Wistar EPM-1 rats were distributed into 3 groups: (1) controls; (2) ovarian transplantation; and (3) ovarian transplantation + R-IPC. The groups were divided into subgroups, according to the prefixed date for euthanasia: 24 hours, 48 hours, 72 hours, and 7th postoperative day (PO). R-IPC was performed by clamping the common iliac artery for a 15-minute period of ischemia followed by 15 minutes of reperfusion, before undergoing ovarian transplantation. The graft was fixed to the retroperitoneum with a simple 8-0 prolene thread. Blood samples were collected from the vena cava. For evaluation of follicular development, the ovarian follicles were classified as immature and mature follicles besides corpora lutea. Only the viable follicles and functioning corpora lutea were counted. RESULTS: At 72 hours, the R-IPC group showed higher estradiol values than the other groups, which were similar. After 24 hours the mean values were similar among all groups, and at 48 hours the R-IPC group was similar to the transplanted group without IPC. Animals undergoing R-IPC showed superior morphologic aspects, but 7 days after transplantation the morphology was worse in all groups. R-IPC enhanced the number of immature follicles at 48 hours (P > .05) and number of mature follicles from 24 hours to 48 hours after transplantation (P < .01). Functioning corpora lutea number was increased as well. CONCLUSION: R-IPC increased the estradiol levels in autologous ovarian transplants associated with better graft morphology and more mature follicles.
Subject(s)
Estradiol/blood , Ischemic Preconditioning/methods , Ovarian Follicle/physiology , Ovary/transplantation , Animals , Corpus Luteum/pathology , Corpus Luteum/physiology , Female , Inflammation/pathology , Necrosis , Ovarian Follicle/pathology , Ovariectomy , Ovary/pathology , Rats , Rats, Wistar , Reference Values , Transplantation, Isogeneic/pathologyABSTRACT
As epidermal growth factor receptor (EGFR) has been reported to be a radiation response modulator, HER inhibitors are regarded to act as potential radiosensitisers. Our study examined the role of nimotuzumab and cetuximab both, the two monoclonal antibodies (mAbs) to EGFR, as radiosensitisers in a murine glioma model in vivo. Co-administration of both the antibodies with radiation increased the radiosensitivity of U87MG, resulting in a significant delay of subcutaneous (s.c.) tumour growth. Furthermore, the addition of antibodies to the radiation decreased brain tumour sizes and is inhibited by 40-80% the increased tumour cell invasion provoked by radiotherapy, although promoted tumour cell apoptosis. Whereas nimotuzumab led to a reduction in the size of tumour blood vessels and proliferating cells in s.c. tumours, cetuximab had no significant antiangiogenic nor antiproliferative activity. In contrast, cetuximab induced a more marked inhibition of EGFR downstream signalling compared with nimotuzumab. Moreover, both antibodies reduced the total number of radioresistant CD133+ cancer stem cells (CSCs). These results were encouraging, and showed the superiority of combined treatment of mAbs to EGFR and radiation over each single therapy against glioblastoma multiforme (GBM), confirming the role of these drugs as radiosensitisers in human GBM. In addition, we first showed the ability of mAb specifics against EGFR to target radioresistant glioma CSC, supporting the potential use in patients.
Subject(s)
Antibodies, Monoclonal/therapeutic use , Brain Neoplasms/radiotherapy , ErbB Receptors/antagonists & inhibitors , Radiation-Sensitizing Agents/therapeutic use , AC133 Antigen , Animals , Antibodies, Monoclonal, Humanized , Antigens, CD/analysis , Brain Neoplasms/pathology , Cell Line, Tumor , Cetuximab , ErbB Receptors/physiology , Female , Glycoproteins/analysis , Humans , Mice , Neoplasm Invasiveness , Neoplastic Stem Cells/chemistry , Neoplastic Stem Cells/drug effects , Peptides/analysis , Xenograft Model Antitumor AssaysABSTRACT
Anti-idiotype monoclonal antibody (mAb) 1E10 was generated by immunizing BALB/c mice with an Ab1 mAb which recognizes NeuGc-containing gangliosides, sulfatides and some tumor antigens. 1E10 mAb induces therapeutic effects in a primary breast carcinoma and a melanoma model. However, the tumor immunity mechanisms have not been elucidated. Here we show that aluminum hydroxide-precipitated 1E10 mAb immunization induced anti-metastatic effect in the 3LL-D122 Lewis Lung carcinoma, a poorly immunogenic and highly metastatic model in C57BL/6 mice. The therapeutic effect was associated to the increment of T cells infiltrating metastases, the reduction of new blood vessels formation and the increase of apoptotic tumor cells in lung nodules. Interestingly, active immunization does not induce measurable antibodies to the 1E10 mAb, the NeuGc-GM3 or tumor cells, which may suggest a different mechanism which has to be elucidated. These findings may support the relevance of this target for cancer biotherapy.
Subject(s)
Antibodies, Anti-Idiotypic/immunology , Carcinoma, Lewis Lung/immunology , Gangliosides/immunology , Lung Neoplasms/immunology , Animals , Antibodies, Anti-Idiotypic/metabolism , Apoptosis/immunology , Carcinoma, Lewis Lung/metabolism , Cell Line, Tumor , Female , Gangliosides/metabolism , Lung Neoplasms/metabolism , Mice , Mice, Inbred C57BLABSTRACT
OBJECTIVE: Usually an experimental necrotizing enterocolitis experimental model, we Investigated nitric oxide levels in intestinal tissues of newborn mice with or without l-arginine therapy during sessions of ischemia and reoxygenation. METHODS: Twenty-six newborn mice from the Wistar EPM-1 lineage, weighing from 4.5 to 6.2 g, were randomly assigned to three groups: G-I/R, hypoxia and reoxygenation; G-Arg, l-arginine treatment I/R; and G-CTL, controls. G-I/R and G-Arg mice underwent twice a day during their first 3 days of life exposure to gas chambers with 100% CO(2) for 5 minutes at 22 degrees C before reoxygenation with 100% O(2) for another 5 minutes. After 12 hours, all animals were sedated, laparotomized, and had samples of ileum and colon taken and- either formalin fixed histopathologic examinations or frozen to -80 degrees C for estimation of tissue nitric oxide levels. Intestinal injuries were classified according to the criteria of Chiu et al. RESULTS: The G-I/R and G-Arg groups showed injuries characteristic of necrotizing enterocolitis (NEC) with an improved structural preservation rate in G-Arg. The concentration of nitric oxide in the Ileum was much higher with G-Arg (16.5 +/- 4.9; P = 0.0019) G-I/R (7.3 +/- 2.0). This effect was not observed in the colon: G-I/R = 10.7 +/- 4.6 versus G-Arg = 15.5 +/- 8.7 (P = .2480). CONCLUSION: Supply of L-arginine increased tissue levels of nitricoxide and reduced morphologic intestinal injury among mice undergoing I/R.
Subject(s)
Arginine/therapeutic use , Intestinal Mucosa/metabolism , Intestines/blood supply , Nitric Oxide/metabolism , Reperfusion Injury/metabolism , Animals , Animals, Newborn , Mice , Mice, Inbred Strains , Transplantation, HomologousABSTRACT
PURPOSE: Verify the optimum remote vascular occlusion time to reduce ovarian injury in autologous transplants in rats. METHODS: Twenty-four adult female rats were assigned to four groups: GC (control group): bilateral oophorectomy followed by ovary transplant; GIPC (ischemic preconditioning group): remote ischemic preconditioning at the iliac artery for 5, 10, and 15 minutes (GIPC-5, GIPC-20, and GIPC-15) previous to bilateral oophorectomy and ovarian transplantation. The right ovary was fixed in the retroperitoneum. Euthanasia was performed 4 days after the surgical procedure. The follicles were counted and classified as developing versus atretic. The immunohistochemical assay identified vascular factor of endothelial growth (VEGF) in the ovarian stroma and assessed the proliferation capacity by means of the Ki-67 in the ovarian follicles. RESULTS: Every group showed an inflammatory infiltrate, luteous body, and ovarian follicles in several phases of development. The ischemic preconditioning groups displayed greater amounts of viable ovarian follicles and increased vascularization and vasodilatation than the control group. GIPC-15 showed the highest amount of viable follicles compared to the others (P < .05 GIPC-15 vs GC; GIPC-15 vs GIPC-5). More VEGF-labeled cells were observed in GIPC-10 than the control group (P < .05, GIPC-10 vs GC). The proliferation index assessed by Ki-67 marking showed GC: 80%; GIPC-5: 76%; GIPC-10: 67%; and GIPC-15: 64% (P > .05). CONCLUSIONS: The PCI-15 cohort seem to be the most adequate timing to achieve functional support and preservation of a greater number of viable ovarian follicles.
Subject(s)
Ischemic Preconditioning/methods , Ovarian Follicle/cytology , Ovary/transplantation , Animals , Female , Models, Animal , Ovarian Follicle/blood supply , Ovariectomy , Rats , Rats, Wistar , Transplantation, AutologousABSTRACT
The efficiency of software for a personal computer in the interactive generation of three-dimensional (3D) images from computer tomography was studied in six pig livers after hepatic resection and catheterization of the hepatic and portal veins. After perfusion the livers were submitted to computed tomography angiography, volumetric measurement by water displacement, and production of an acrylic model of the veins by the injection and corrosion method, by which the lengths of the hepatic and portal veins were measured. From the angiogram, the software generated a 3D image that allowed measurement of the vein lengths. The identified branches of the hepatic and portal veins were correlated with the hepatic sectors and segments, respectively. The virtual measures from the 3D images were compared with the real measures. There were no significant differences between the topography and the vessel length. The mean difference between the volumes calculated from software and those measured by water displacement corresponded to 1.2%, and between the vessel lengths, 0.2%. In conclusion, the software for personal computer (named LIVER3D) is efficient, allowing interactive inspection of 3D images. All virtual measurements of liver vessel length and partial/total liver volume were similar to the actual ones.
Subject(s)
Liver/anatomy & histology , Microcomputers , Animals , Image Processing, Computer-Assisted , Liver Circulation , Portal Vein/anatomy & histology , Software , Swine , User-Computer Interface , Vena Cava, Inferior/anatomy & histologyABSTRACT
AIM: To study the effects of N-acetylcysteine and ischemic preconditioning on the portal triad clamping compared to arterial and portal clamping alone. METHODS: Eighty EPM 1-Wistar rats were randomized into two groups, depending on inclusion (Group 1) or not (Group 2) of the bile duct in the hepatic vascular pedicle occlusion. Each group was divided into four subgroups as follows. IR 1: 20 minutes after celiotomy, the pedicle containing vascular elements and bile duct to the left lateral and median liver lobes was occluded for 40 minutes, followed by 30 minutes of reperfusion. IPC 1: after 10 minutes of ischemia and 10 minutes of reperfusion, the ischemic preconditioning period, the rats were submitted to the same procedure described for IR 1 Group. NAC 1: the rats received N-acetylcysteine (150 mg/kg) 15 minutes before 40 minutes of ischemia and 5 minutes before 30 minutes of reperfusion. SHAM 1: The hepatic pedicle for the lateral and median liver lobes was dissected after 20 minutes, the bile duct alone was clamped for 40 minutes, and released for an additional 30 minutes. In the IR 2, IPC 2, and NAC 2 groups, ischemia was achieved with an exclusive vascular occlusion. SHAM 2: dissection and observation for 90 minutes. The blood was sampled for liver enzyme levels. Statistical analysis was done (P = .05). RESULTS: Hepatic IR injury was less severe for animals from the classic portal triad clamping (group 1), with regard to AST (IR 1 Group 766 vs IR 2 Group 1380 U/L) and ALT (IR 1 Group 840 vs IR 2 Group 1576 U/L); IPC, but not NAC administration, was able to protect the liver from IR injury for animals from the classic portal triad clamping group, with regard to AST (IPC 1 Group 421 vs NAC 1 Group 1131 U/L) and ALT (IPC 1 Group 315 vs NAC 1 Group 1085 U/L). CONCLUSIONS: IPC protects the liver from IR injury; classic portal triad clamping results in a less severe hepatic IR injury when compared to bile duct exclusion.
Subject(s)
Acetylcysteine/pharmacology , Bile Ducts/physiology , Ischemic Preconditioning , Liver Circulation/physiology , Reperfusion Injury/prevention & control , Alanine Transaminase/metabolism , Alkaline Phosphatase/metabolism , Animals , Aspartate Aminotransferases/metabolism , Bilirubin/metabolism , Liver Circulation/drug effects , Male , Portal System , Rats , Rats, Wistar , gamma-Glutamyltransferase/metabolismABSTRACT
El ha investigado la utilidad terapéutica de las plantas basado principalmente en el uso folklórico por su aplicación en un amplio rango de enfermedades. En el presente trabajo, se evaluó la toxicidad y la actividad inmunológica in vivo e in vitro del extracto metanílico de la Cordia sebetena. Cultivos de celuals mononucleares de sangre periférica humana, donde se investigó el efecto de la viabilidad (coloración supravital azul tripán), linfoproliferación (Timidina Tritiada) y concentraciones del Factor Estimulador de Colonias Granulocítico-Macrofágico (FEC-GM) en los sobrenadantes, empleando la técnica de ELISA de captura, así como la actividad quimiotáctica de celulas polimorfonucleares humanas a través de la técnica en gel de agarosa. Se estudio la toxicidad in vivo en hígado, vaso y riñón en ratones bajo tratamiento oral e intraperitoneal a través de estudios histológicos, niveles sanguíneos de leucocitos totales y plaquetas/mm3 y el efecto en la ingesta diaria y el peso corporal de dichos ratones. Los resultados indican que el extracto metanolico de Cordia sebestena es inocuo a las celulas ensayadas, estimula la producción del FEC-GM, y por ende a la formación de nuevas células linfoides comportándose como un quimioatractante celular, hallazgos que podrían, en parte, explicar los efectos farmacológicos atribuidos a esta especie vegetal