Changes in the levels of free sialic acid during ex vivo lung perfusion do not correlate with pulmonary function. Experimental model.

BACKGROUND: Ex vivo lung perfusion (EVLP) constitutes a tool with great research potential due to its advantages over in vivo and in vitro models. Despite its important contribution to lung reconditioning, this technique has the disadvantage of incurring high costs and can induce pulmonary endothelial injury through perfusion and ventilation. The pulmonary endothelium is made up of endothelial glycocalyx (EG), a coating of proteoglycans (PG) on the luminal surface. PGs are glycoproteins linked to terminal sialic acids (Sia) that can affect homeostasis with responses leading to edema formation. This study evaluated the effect of two ex vivo perfusion solutions on lung function and endothelial injury. METHODS: We divided ten landrace swine into two groups and subjected them to EVLP for 120 min: Group I (n = 5) was perfused with Steen® solution, and Group II (n = 5) was perfused with low-potassium dextran-albumin solution. Ventilatory mechanics, histology, gravimetry, and sialic acid concentrations were evaluated. RESULTS: Both groups showed changes in pulmonary vascular resistance and ventilatory mechanics (p < 0.05, Student's t-test). In addition, the lung injury severity score was better in Group I than in Group II (p < 0.05, Mann-Whitney U); and both groups exhibited a significant increase in Sia concentrations in the perfusate (p < 0.05 t-Student) and Sia immunohistochemical expression. CONCLUSIONS: Sia, as a product of EG disruption during EVLP, was found in all samples obtained in the system; however, the changes in its concentration showed no apparent correlation with lung function.

Endothelin-Converting Enzyme 1 and Vascular Endothelial Growth Factor as Potential Biomarkers during Ex Vivo Lung Perfusion with Prolonged Hypothermic Lung-Sparing.

Lung transplantation requires optimization of donor's organ use through ex vivo lung perfusion (EVLP) to avoid primary graft dysfunction. Biomarkers can aid in organ selection by providing early evidence of suboptimal lungs during EVLP and thus avoid high-risk transplantations. However, predictive biomarkers of pulmonary graft function such as endothelin-converting enzyme (ECE-1) and vascular endothelial growth factor (VEGF) have not been described under EVLP with standard prolonged hypothermic preservation, which are relevant in situations where lung procurement is difficult or far from the transplantation site. Therefore, this study is aimed at quantifying ECE-1 and VEGF, as well as determining their association with hemodynamic, gasometric, and mechanical ventilatory parameters in a swine model of EVLP with standard prolonged hypothermic preservation. Using a protocol with either immediate (I-) or delayed (D-) initiation of EVLP, ECE-1 levels over time were found to remain constant in both study groups (p > 0.05 RM-ANOVA), while the VEGF protein was higher after prolonged preservation, but it decreased throughout EVLP (p > 0.05 RM-ANOVA). Likewise, hemodynamic, gasometric, mechanical ventilatory, and histological parameters had a tendency to better results after 12 hours of hypothermic preservation in the delayed infusion group.

In vivo performance of decellularized tracheal grafts in the reconstruction of long length tracheal defects: Experimental study.

BACKGROUND: The repair of long-segment tracheal lesions remains an important challenge. Nowdays no predictable and dependable substitute has been found. Decellularized tracheal scaffolds have shown to be a promising graft for tracheal transplantation, since it is non-immunogenic. OBJECTIVE: Evaluate in vivo decellularized tracheal allografts performance to replace long tracheal segment. METHODS: Forty-five swines underwent surgery as follows: Fifteen trachea donors and 30 receptors of decellularized trachea allografts. The receptors were randomly divided in five groups (n = 6). In groups I and II, donor trachea segment was decellularized by 15 cycles with sodium deoxycholate and deoxyribonuclease, in group II, the allograft was reinforced with external surgical steel wire. Groups, III, IV, and V decellularization was reduced to seven cycles, supplemented with cryopreservation in group IV and with glutaraldehyde in group V. A 10 rings segment was excised from the receptor swine and the decellularized trachea graft was implanted to re-establish trachea continuity. RESULTS: Both decellularization cycles caused decreased stiffness. All trachea receptors underwent euthanasia before the third post-implant week due to severe dyspnea and trachea graft stenosis, necrosis, edema, inflammation, hemorrhage, and granulation tissue formation in anastomotic sites. Histologically all showed total loss of epithelium, separation of collagen fibers, and alterations in staining. CONCLUSIONS: Both decellularization techniques severely damaged the structure of the trachea and the extracellular matrix of the cartilage, resulting in a no functional graft, in spite of the use of surgical wire, cryopreservation or glutaraldehyde treatment. An important drawback was the formation of fibrotic stenosis in both anastomosis.

Metabolic syndrome enhances endoplasmic reticulum, oxidative stress and leukocyte-endothelium interactions in PCOS.

OBJECTIVE: Polycystic ovary syndrome (PCOS) is associated with insulin resistance, which can lead to metabolic syndrome (MetS). Oxidative stress and leukocyte-endothelium interactions are related to PCOS. Our aim was to evaluate whether the presence of MetS in PCOS patients can influence endoplasmic reticulum (ER) and oxidative stress and leukocyte-endothelium interactions. MATERIAL AND METHODS: This was a prospective controlled study conducted in an academic medical center. The study population consisted of 148 PCOS women (116 without/32 with MetS) and 112 control subjects (87 without / 25 with MetS). Metabolic parameters, reactive oxygen species (ROS) production, ER stress markers (GRP78, sXBP1, ATF6), leukocyte-endothelium interactions, adhesion molecules (VCAM-1, ICAM-1, E-Selectin), TNF-α and IL-6 were determined. RESULTS: Total ROS, inflammatory parameters and adhesion molecules were enhanced in the presence of MetS (p<0.05), and the PCOS+MetS group showed higher levels of IL-6 and ICAM-1 than controls (p<0.05). Increased adhesion and leukocyte rolling flux were observed in PCOS and PCOS+MetS groups vs their respective controls (p<0.05). GRP78 protein expression was higher in the PCOS groups (p<0.05 vs controls) and sXBP1 was associated with the presence of MetS (p<0.05 vs controls without MetS). Furthermore, PCOS+MetS patients exhibited higher GRP78 and ATF6 levels than controls and PCOS patients without MetS (p<0.05). In PCOS women, HOMA-IR was positively correlated with ICAM-1 (r=0.501; p<0.01), ROS (r=0.604; p<0.01), rolling flux (r=0.455;p<0.05) and GRP78 (r=0.574; p<0.001). CONCLUSION: Our findings support the hypothesis of an association between altered metabolic status, increased ROS production, ER stress and leukocyte-endothelium interactions in PCOS, all of which are related to vascular complications.

Insulin Resistance in PCOS Patients Enhances Oxidative Stress and Leukocyte Adhesion: Role of Myeloperoxidase.

Cardiovascular diseases and oxidative stress are related to polycystic ovary syndrome (PCOS) and insulin resistance (IR). We have evaluated the relationship between myeloperoxidase (MPO) and leukocyte activation in PCOS patients according to homeostatic model assessment of IR (HOMA-IR), and have explored a possible correlation between these factors and endocrine and inflammatory parameters. This was a prospective controlled study conducted in an academic medical center. The study population consisted of 101 PCOS subjects and 105 control subjects. We divided PCOS subjects into PCOS non-IR (HOMA-IR<2.5) and PCOS IR (HOMA-IR>2.5). Metabolic and anthropometric parameters, total and mitochondrial reactive oxygen species (ROS) production, MPO levels, interactions between human umbilical vein endothelial cells and leukocytes, adhesion molecules (E-selectin, ICAM-1 and VCAM-1) and proinflammatory cytokines (IL-6 and TNF-α) were evaluated. Oxidative stress was observed in PCOS patients, in whom there was an increase in total and mitochondrial ROS production and MPO levels. Enhanced rolling flux and adhesion, and a decrease in polymorphonuclear cell rolling velocity were also detected in PCOS subjects. Increases in IL-6 and TNF-α and adhesion molecules (E-selectin, ICAM-1 and VCAM-1) were also observed, particularly in the PCOS IR group, providing evidence that inflammation and oxidative stress are related in PCOS patients. HOMA-IR was positively correlated with hsCRP (p<0.001, r = 0.304), ROS production (p<0.01, r = 0.593), leukocyte rolling flux (p<0.05, r = 0.446), E-selectin (p<0.01, r = 0.436) and IL-6 (p<0.001, r = 0.443). The results show an increase in the rate of ROS and MPO levels in PCOS patients in general, and particularly in those with IR. Inflammation in PCOS induces leukocyte-endothelium interactions and a simultaneous increase in IL-6, TNF-α, E-selectin, ICAM-1 and VCAM-1. These conditions are aggravated by the presence of IR.

Effects of metformin on mitochondrial function of leukocytes from polycystic ovary syndrome patients with insulin resistance.

OBJECTIVE: Oxidative stress and mitochondrial dysfunction are implicated in polycystic ovary syndrome (PCOS). The present study assesses the effect of metformin treatment on mitochondrial function in polymorphonuclear cells from PCOS subjects. Additionally, we evaluate endocrine parameters and levels of interleukin 6 (IL6) and tumour necrosis factor alpha (TNFα). DESIGN AND METHODS: Our study population was comprised of 35 women of reproductive age diagnosed with PCOS and treated with metformin for 12 weeks, and their corresponding controls (n=41), adjusted by age and BMI. We evaluated the alteration of endocrinological and anthropometrical parameters and androgen levels. Mitochondrial O2 consumption (using a Clark-type O2 electrode), membrane potential, mitochondrial mass, and levels of reactive oxygen species (ROS) and glutathione (GSH) (by means of fluorescence microscopy) were assessed in poymorphonuclear cells. H2O2 was evaluated with the Amplex Red(R) H2O2/Peroxidase Assay kit. IL6 and TNFα were measured using the Luminex 200 flow analyser system. RESULTS: Metformin had beneficial effects on patients by increasing mitochondrial O2 consumption, membrane potential, mitochondrial mass and glutathione levels, and by decreasing levels of reactive oxygen species and H2O2. In addition, metformin reduced glucose, follicle-stimulating hormone, IL6 and TNFα levels and increased dehydroepiandrosterone sulfate levels. HOMA-IR and mitochondrial function biomarkers positively correlated with ROS production (r=0.486, P=0.025), GSH content (r=0.710, P=0.049) and H2O2 (r=0.837, P=0.010), and negatively correlated with membrane potential (r=-0.829, P=0.011) at baseline. These differences disappeared after metformin treatment. CONCLUSIONS: Our results demonstrate the beneficial effects of metformin treatment on mitochondrial function in leukocytes of PCOS patients.

Metformin modulates human leukocyte/endothelial cell interactions and proinflammatory cytokines in polycystic ovary syndrome patients.

OBJECTIVE: We aim to assess the effect of metformin treatment on metabolic parameters, endothelial function and inflammatory markers in polycystic ovary syndrome (PCOS) subjects. METHODS: The study population consisted of 40 reproductive-age women with PCOS, who underwent treatment with metformin during a 12-week period, and their corresponding matched controls (n = 44). We evaluated endocrinological parameters, adhesion molecules (vascular cell adhesion molecule 1 (VCAM-1), intercellular cell adhesion molecule 1 (ICAM-1) and E-selectin) and proinflammatory cytokines (interleukin 6 (IL-6) and tumor necrosis factor alpha (TNFα)) in serum. In addition, interactions between human umbilical vein endothelial cells and polymorphonuclear (PMN) cells were assessed by flow chamber microscopy. In addition, a group of type 2 diabetes patients who underwent treatment with metformin during a 12-week period was incorporated into the study. RESULTS: Metformin produced beneficial effects on PCOS patients by decreasing polymorphonuclear (PMN) rolling flux and adhesion. It also decreased levels of ICAM-1, E-selectin, IL-6 and ΤΝFα. In addition, metformin induced an improvement of endocrine and anthropometric parameters in PCOS subjects by reducing glucose, follicle-stimulating hormone (FSH) and androstendione, and by increasing dehydroepiandrosterone-sulfate (DHEA-S). Metformin also had beneficial effects in type 2 diabetic subjects by reducing body weight, waist circumference and PMN adhesion, and by increasing PMN rolling velocity. CONCLUSION: Our results highlight the modulating effect of metformin on leukocyte/endothelium interactions. These findings may explain the potential beneficial effect of metformin in reducing the risk of vascular events in PCOS patients and in insulin resistance conditions.

Mitochondrial impairment and oxidative stress in leukocytes after testosterone administration to female-to-male transsexuals.

INTRODUCTION: Testosterone undecanoate (T) treatment is common in female-to-male transsexuals (FtMs) but can induce impairment of mitochondrial function and oxidative stress. AIM: The effect of T treatment on the mitochondrial function and redox state of leukocytes of FtMs subjects was evaluated. METHODS: This was an observational study conducted in a university hospital. Fifty-seven FtMs were treated with T (1,000 mg) for 12 weeks, after which anthropometric and metabolic parameters and mitochondrial function were evaluated. MAIN OUTCOME MEASURES: Anthropometric and metabolic parameters were evaluated. Mitochondrial function was studied by assessing mitochondrial oxygen (O2) consumption, membrane potential, reactive oxygen species (ROS) production, glutathione levels (GSH), and the reduced glutathione/oxidized glutathione (GSH)/(GSSG) ratio in polymorphonuclear cells. RESULTS: T treatment led to mitochondrial impairment in FtMs as a result of a decrease in mitochondria O2 consumption, the membrane potential, GSH levels, and the (GSH)/(GSSG) ratio and an increase in ROS production. Mitochondrial O2 consumption and membrane potential negatively correlated with T levels, which was further confirmed that the T treatment had induced mitochondrial dysfunction. T also produced a significant increase in total testosterone, free androgenic index, and atherogenic index of plasma, and a decrease in sex hormone-binding globulin and high-density lipoprotein cholesterol. CONCLUSIONS: Treatment of FtMs with T can induce impairment of mitochondrial function and a state of oxidative stress. This effect should be taken into account in order to modulate possible comorbidities in these patients.

Induction of oxidative stress and human leukocyte/endothelial cell interactions in polycystic ovary syndrome patients with insulin resistance.

CONTEXT: Insulin resistance is a feature of polycystic ovary syndrome (PCOS) and is related to mitochondrial and endothelial function. OBJECTIVE: We tested whether hyperandrogenic insulin-resistant women with PCOS, who have an increased risk of vascular disease, display impaired leukocyte-endothelium interactions, and mitochondrial dysfunction. DESIGN AND SETTING: This was a prospective controlled study conducted in an academic medical center. PATIENTS: The study population consisted of 43 lean reproductive-age women with PCOS and 39 controls subjects. MAIN OUTCOME MEASURES: We evaluated anthropometric and metabolic parameters, adhesion molecules, and interactions between leukocytes and human umbilical vein endothelial cells. Mitochondrial function was studied by assessing mitochondrial oxygen consumption, membrane potential, reactive oxygen species production, glutathione levels (GSH), and the oxidized glutathione (GSSG)/GSH ratio in polymorphonuclear cells. RESULTS: Impairment of mitochondrial function was observed in the PCOS patients, evident in a decrease in oxygen consumption, an increase in reactive oxygen species production, a decrease in the GSH/GSSG ratio and GSH levels, and an undermining of the membrane potential. PCOS was related to a decrease in polymorphonuclear cell rolling velocity and an increase in rolling flux and adhesion. Increases in IL-6 and TNFα and adhesion molecules (vascular cell adhesion molecule-1 and E-selectin) were also observed. CONCLUSION: This study supports the hypothesis of an association between insulin resistance and an impaired endothelial and mitochondrial oxidative metabolism. The evidence obtained shows that the inflammatory state related to insulin resistance in PCOS induces a leukocyte-endothelium interaction. These findings may explain the increased risk of vascular disease in women with PCOS.

Mitochondrial complex I impairment in leukocytes from polycystic ovary syndrome patients with insulin resistance.

CONTEXT: Insulin resistance is a feature of polycystic ovary syndrome (PCOS) and is related to mitochondrial function. OBJECTIVE: Our objective was to assess mitochondrial function by evaluating mitochondrial oxygen (O(2)) consumption, reactive oxygen species (ROS) production, levels of glutathione (GSH), the oxidized glutathione/GSH ratio, TNFalpha levels, and membrane potential. Additionally, we have evaluated mitochondrial complex I as a target of the oxidative stress responsible for PCOS in polymorphonuclear cells. DESIGN AND SETTING: This was a prospective controlled study conducted in an academic medical center. PATIENTS: The study population consisted of 20 lean reproductive-age women with PCOS and 20 body composition-matched controls. MAIN OUTCOME MEASURES: We evaluated mitochondrial O(2) consumption using the Clark-type O(2) electrode; levels of ROS, GSH, and membrane potential by means of fluorescence microscopy; TNFalpha levels by ELISA; and complex I activity by spectrophotometric assay. RESULTS: An impairment in mitochondrial function was observed in PCOS patients, evident by a decrease in mitochondrial O(2) consumption; an increase in ROS production, oxidized glutathione/GSH ratio, and TNFalpha levels; a drop in GSH levels; and an undermining of membrane potential. Furthermore, an impairment of mitochondrial complex I was identified. CONCLUSION: This study supports the hypothesis of an association between insulin resistance and an impaired mitochondrial oxidative metabolism. We also propose that the oxidative stress responsible for PCOS takes place at complex I. These abnormalities may contribute to the increased risk of type 2 diabetes among women with PCOS.

Características clínicas, histológicas y evolutivas de una serie de 11 casos de cáncer diferenciado de tiroides con hipertiroidismo / Clinical and histopathological characteristics and outcomes in eleven patients with hyperthyroidism and concurrent differentiated thryoid cancer

Introducción: La coexistencia de cáncer diferenciado de tiroides (CDT) e hipertiroidismo se considera infrecuente. El objetivo de este trabajo es describir las características de los casos detectados en nuestro centro. Pacientes y métodos: Estudio descriptivo con 11 pacientes (3 varones y 8 mujeres), diagnosticados de CDT, en los que coexistía hipertiroidismo. Se les realizó de forma preoperatoria ecografía tiroidea, gammagrafía, analítica con TSH, T4 libre, T3 libre, anticuerpos antitiroperoxidasa (ATPO) y anticuerpos antirreceptor de TSH (TSI). Se realizó punción-aspiración con aguja fina (PAAF) de tiroides a los pacientes con nódulos fríos iguales o mayores de 2 cm (5 pacientes). Se les practicó tiroidectomía total y ablación posterior con radioyodo. La mediana de seguimiento fue de 64 meses (límites 17 a 161). Resultados: Tres pacientes tenían un nódulo tóxico único (NT), 4 un bocio multinodular tóxico (BMN-T) y 4 una enfermedad de Graves-Basedow (EGB). De los 3 pacientes con NT, se intervino a 2 por sospecha de malignidad y el restante por nódulo superior a 3 cm. De los 4 pacientes con BMN-T, se reintervino a 3 por nódulos fríos, y el restante por crecimiento del bocio y mal control del hipertiroidismo. De los 4 pacientes con EGB, se intervino a 3 por presentar nódulos (1 con citología sospechosa de malignidad) y al restante por recidiva de la enfermedad. La anatomía patológica definitiva fue, en todos los casos, de carcinoma papilar, 2 de variedad papilofolicular y uno esclerosante. El estadio según la clasificación TNM fue de riesgo bajo en 6 pacientes y riesgo alto riesgo en 5 (un caso con metástasis pulmonares). La evolución ha sido favorable en todos los casos. Conclusiones: El hipertiroidismo no excluye la posibilidad de CDT, especialmente si hay nódulos fríos en la gammagrafía. En la EGB, la existencia de nódulos en la ecografía nos obliga a plantearnos tratamiento quirúrgico. El estadio al diagnóstico no es siempre de riesgo bajo. La evolución del CDT no parece diferente de la que se presenta en pacientes eutiroideos (AU)

Introduction: The coexistence of differentiated thyroid cancer (DTC) and hyperthyroidism is considered a rare event. The aim of this study was to analyze the characteristics of the patients with both entities attended at our institution. Patients and methods: A descriptive study of 11 patients (3 men and 8 women) with concurrent hyperthyroidism and DTC was performed. Ultrasound scan, thyroid scintigraphy and measurement of thyroid-stimulating hormone (TSH), free triiodothyronine, free thyroxine, autoantibodies to thyroid peroxidase, and TSH receptor autoantibodies were performed preoperatively. Thyroid fine-needle biopsy was performed in patients with cold nodules equal to or larger than 2 cm (5 patients). All patients underwent total thyroidectomy and 131I therapy. The median follow-up was 64 months (range 17 to 161). Results: Toxic uninodular goiter (TUG) was found in 3 patients, toxic multinodular goiter (TMG) in 4 patients, and Graves' disease (GD) in 4 patients. The 3 patients with TUG underwent surgery, 2 for suspected malignancy and the third due to a nodule larger than 3 cm. Of the 4 patients with TMG, 3 underwent surgery due to the presence of cold nodules and the fourth because of goiter growth and uncontrolled hyperthyroidism. Of the 4 patients with GD, 3 underwent surgery due to the presence of nodules (one suspicious for malignancy on cytological analysis), and the fourth because of GD recurrence. Postsurgical histological diagnosis was papillary carcinoma in all patients, a follicular variant of papillary carcinoma in two and sclerosing carcinoma in one. According to TNM classification, 6 patients had a low risk and 5 had a high risk of recurrence (one patient had lung metastases). Outcome was favorable in all patients. Conclusions: Diagnosis of hyperthyroidism does not exclude DTC, especially in the presence of cold nodules on scintigraphy. When nodules are detected on ultrasound scan in GD patients, surgery should be considered. At diagnosis, not all patients were low risk. The behavior of DTC in patients with hyperthyroidism seems to be similar to that in euthyroid patients (AU)

[Epidemiologic and situational panorama of hepatitis C in Mexico]. / Panorama epidemiológico y situacional de la hepatitis C en México.

OBJECTIVE: To determine the epidemiological situation of Chronic Hepatitis C (CHC) in our country. BACKGROUND DATA: Chronic Hepatitis C affects 170 million people worldwide, and about 0.7% of Mexican population. There is no enough epidemiological information about CHC in our country, and it is very probable that some cases are not even detected. METHODS: An investigation poll was performed. Age, gender, birthday, weight, race, residence and birth place, routes of transmission, ALT levels, histological, serological and molecular diagnosis, evidence of complications and previous treatments were recorded. A data recollection sheet was dispatched to different country provinces; they had 6 months to answer it, in order to recollect all information. RESULTS: 831 patients were analized (58.6% female and 41.4% male) with the following distribution in our country provinces: Aguascalientes 15, Chihuahua 12, Distrito Federal 495, Durango 10, Jalisco 89, Guanajuato 78, Yucatán 8, Querétaro 11, Sonora 40, Tabasco 15, Baja California 5, Veracruz 13, Tamaulipas 2 and 38 patients of Nuevo León. The highest incidence of CHC was found at fifth and sixth decade of life (28.5% y 26.7% respectively. The weight distribution was 36.2% < 65kg, 34.6% 65-75 kg and 29.2% > 75 kg. 86.5% had chronic hepatitis and 13.2% cirrhosis. The risk factors for HCV infection analysis showed that the main route of transmission was via contaminated blood (64.2%); when we excluded the patients that were exposed before 1995, the incidence was lowered to 4.5%. The higher incidence was showed between 1970 and 1990 (68%). The intravenous drug users were predominantly male and on those patients in the provinces near the north border line of our country. The predominant genotype was gen- 1 no matter the province (72.2%), in the intravenous drug users genotype 3 was found in 25%. The viral load was similar in all the provinces. 75% of the patients had have treatment and 22.5% had have two cycles, 50% of cirrhotic patients had have treatment whereas only 28% of the patients with late complications had have it. The most common treatment was pegylated alpha-2a interferon plus ribavirine. CONCLUSIONS: 1. The main route of transmission was blood transfusion. There is a marked decrease in the incidence of post-transfusional hepatitis since the introduction of anti-VHC antibody screening of blood donors (4.5%). 2. The time between the infection and diagnosis was 23 years for chronic hepatitis and 26 years for cirrhosis. 3. Intravenous drugs use was an important route of transmission in the north of our country. 4. The predominant genotype was gen-1. 5. Almost all the patients with chronic hepatitis received treatment, the most common used was pegylated interferon alpha-2a and ribavirin. 6.50% of the patients with CHC have late complications.