Endothelial ILK induces cardioprotection by preventing coronary microvascular dysfunction and endothelial-to-mesenchymal transition.

Endothelial dysfunction is an early event in coronary microvascular disease. Integrin-linked kinase (ILK) prevents endothelial nitric oxide synthase (eNOS) uncoupling and, thus, endothelial dysfunction. However, the specific role of endothelial ILK in cardiac function remains to be fully elucidated. We hypothesised that endothelial ILK plays a crucial role in maintaining coronary microvascular function and contractile performance in the heart. We generated an endothelial cell-specific ILK conditional knock-out mouse (ecILK cKO) and investigated cardiovascular function. Coronary endothelial ILK deletion significantly impaired cardiac function: ejection fraction, fractional shortening and cardiac output decreased, whilst left ventricle diastolic internal diameter decreased and E/A and E/E' ratios increased, indicating not only systolic but also diastolic dysfunction. The functional data correlated with extensive extracellular matrix remodelling and perivascular fibrosis, indicative of adverse cardiac remodelling. Mice with endothelial ILK deletion suffered early ischaemic-like events with ST elevation and transient increases in cardiac troponins, which correlated with fibrotic remodelling. In addition, ecILK cKO mice exhibited many features of coronary microvascular disease: reduced cardiac perfusion, impaired coronary flow reserve and arterial remodelling with patent epicardial coronary arteries. Moreover, endothelial ILK deletion induced a moderate increase in blood pressure, but the antihypertensive drug Losartan did not affect microvascular remodelling whilst only partially ameliorated fibrotic remodelling. The plasma miRNA profile reveals endothelial-to-mesenchymal transition (endMT) as an upregulated pathway in endothelial ILK conditional KO mice. Our results show that endothelial cells in the microvasculature in endothelial ILK conditional KO mice underwent endMT. Moreover, endothelial cells isolated from these mice and ILK-silenced human microvascular endothelial cells underwent endMT, indicating that decreased endothelial ILK contributes directly to this endothelial phenotype shift. Our results identify ILK as a crucial regulator of microvascular endothelial homeostasis. Endothelial ILK prevents microvascular dysfunction and cardiac remodelling, contributing to the maintenance of the endothelial cell phenotype.

A novel study, textbook outcome in adrenalectomy: retrospective observational study in an endocrine surgical unit.

Textbook outcome is a multidimensional quality management tool that uses a set of traditional surgical measures to reflect an "ideal" surgical result for a particular pathology. Retrospective study of all patients undergoing scheduled for adrenal tumors surgery at an endocrine surgery unit from January 2010-December 2022. The definition of Textbook Outcome were: R0 resection, no Clavien-Dindo ≥ IIIa complications, no prolonged stay (< P75), no readmissions, and no mortality in the first 30 days. The main objective was to analyze the rate of Textbook Outcome obtained. One hundred and five patients were included in the study. Textbook Outcome was achieved in 71.4%. Surgical approach (p < 0.001), Charlson scale (p = 0.031), American Society of Anesthesiologists Classification (p = 0.047) and surgical time (p < 0.001) were all significantly associated with the achievement of Textbook Outcome. The laparoscopic approach was associated as an independent factor with obtaining Textbook Outcome (OR:5.394; p = 0.016), as was surgical time (OR:0.986; p = 0.004). Textbook Outcome is a novel, useful, easy-to-interpret tool for measuring results in adrenal surgery. The laparoscopic approach is associated with a higher rate of "ideal" surgical results. The study was registered in the public accessible database clinicaltrials.gov with the ClinicalTrials.gov ID: NCT05888753.

Growth modeling approach with the Verhulst coexistence dynamic properties for regulation purposes.

For this research, the properties of the logistic growth model for independent and coexisting species were used to set definitions for the possible regulation of one or two growth variables through their coupling parameters. The present analysis is done for the single-species Verhulst model without coupling, the single-species Verhulst model coupled with an exogenous signal, and the two-species Verhulst coexistence growth model which represents six different ecological regimes of interaction. The models' parameters, such as the intrinsic growth rate and the coupling, are defined. Finally, the control results are expressed as lemmas for regulation, and they are shown using a simulation example of a fish population growing independent of human interaction (no harvesting, no fishing) and the simulation of the regulation of said population when the coupling of fish and humans is involved (harvesting, fishing).

Two unusual cases of canine prostatitis: prostatitis in a castrated dog and preputial oedema in an intact male.

In this study, two unusual presentations of canine prostatitis are described; in the first case a 10-years-old neutered Boxer dog was presented to the Veterinary Teaching Hospital of the University of Extremadura with a complaint of anorexia, apathy and preputial discharge. In the second case, a local veterinarian referred an 8-years-old male Labrador to the Veterinary Teaching Hospital of the University of Extremadura. The dog had a history of pain in the caudal abdomen and preputial oedema. The final diagnosis in both cases was acute prostatitis. It is concluded that although canine prostatitis is a common disease, sometimes can have presentations that may differ from those classically described in the literature.

Bisphenol A induces coronary endothelial cell necroptosis by activating RIP3/CamKII dependent pathway.

Epidemiological studies link long term exposure to xenoestrogen Bisphenol-A to adverse cardiovascular effects. Our previous results show that BPA induces hypertension by a mechanism involving CamKII activation and increased redox stress caused by eNOS uncoupling. Recently, CamKII sustained activation has been recognized as a central mediator of programmed cell death in cardiovascular diseases, including necroptosis. However, the role of necroptosis in cardiac response to BPA had not yet been explored. Mice exposed to BPA for 16 weeks showed altered heart function, electrical conduction, and increased blood pressure. Besides, a stress test showed ST-segment depression, indicative of cardiac ischemia. The hearts exhibited cardiac hypertrophy and reduced vascularization, interstitial edema, and large hemorrhagic foci accompanied by fibrinogen deposits. BPA initiated a cardiac inflammatory response, up-regulation of M1 macrophage polarization, and increased oxidative stress, coinciding with the increased expression of CamKII and the necroptotic effector RIP3. In addition, cell death was especially evident in coronary endothelial cells within hemorrhagic areas, and Evans blue extravasation indicated a vascular leak in response to Bisphenol-A. Consistent with the in vivo findings, BPA increased the necroptosis/apoptosis ratio, the expression of RIP3, and CamKII activation in endothelial cells. Necrostatin-1, an inhibitor of necroptosis, alleviated BPA induced cardiac dysfunction and prevented the inflammatory and hemorrhagic response in mice. Mechanistically, silencing of RIP3 reversed BPA-induced necroptosis and CamKII activation in endothelial cells, while inhibition of CamKII activation by KN-93 had no effect on RIP3 expression but decreased necroptotic cell death suggesting that BPA induced necroptosis is mediated by a RIP 3/CamKII dependent pathway. Our results reveal a novel pathogenic role of BPA on the coronary circulation. BPA induces endothelial cell necroptosis, promotes the weakening of coronary vascular wall, which caused internal ventricular hemorrhages, delaying the reparative process and ultimately leading to cardiac dysfunction.

Nitric oxide inhibition of coxsackievirus replication in vitro.

Nitric oxide is a radical molecule with antibacterial, -parasitic, and -viral properties. We investigated the mechanism of NO inhibition of Coxsackievirus B3 (CVB3) replication in vitro by determining the effect of NO upon a single replicative cycle of CVB3 grown in HeLa cells. Transfection of inducible NO synthase cDNA into HeLa cells reduces the number of viral particles produced during a single cycle of growth. Similarly, a noncytotoxic concentration of the NO donor S-nitroso-amino-penicillamine reduces the number of viral particles in a dose-dependent manner. To explore the mechanisms by which NO exerts its antiviral effect, we assayed the attachment, replication, and translation steps of the CVB3 life cycle. NO does not affect the attachment of CVB3 to HeLa cells. However, NO inhibits CVB3 RNA synthesis, as shown by a [3H]uridine incorporation assay, reverse transcription-PCR, and Northern analysis. In addition, NO inhibits CVB3 protein synthesis, as shown by [35S]methionine protein labeling and Western blot analysis of infected cells. Thus, NO inhibits CVB3 replication in part by inhibiting viral RNA synthesis by an unknown mechanism.

Effects of Oral Prednisone Administration on Serum Cystatin C in Dogs.

BACKGROUND: Oral administration of glucocorticoid alters serum cystatin C (sCysC) concentration in humans. OBJECTIVE: To determine if oral administration of prednisone alters sCysC in dogs without pre-existing renal disease. ANIMALS: Forty six dogs were included: 10 dogs diagnosed with steroid responsive meningitis arteritis (SRMA; group A), 20 dogs diagnosed of pituitary-dependent hyperadrenocorticism (PDH; group B), and 16 healthy control dogs (group C). METHODS: Retrospective observational study. SRMA diagnosed dogs were administered prednisone 4 mg/kg/24 h PO 7 days, reducing the dose to 2 mg/kg/24 h 7 days before medication withdrawal. In group A, sampling was performed at days 0, 7, 14 and a final control at day 21. Blood and urine samples were collected in the 3 groups, and in group A, sampling was performed at all time points (days 1, 7, 14, and 21). RESULTS: In group A, sCysC was significantly higher at day 7 compared to the control group (0.4 ± 0.04 mg/L vs. 0.18 ± 0.03 mg/L mean ± SEM respectively P < 0.01); sCysC values decreased to basal at day 14 when the dose was decreased and after 1 week of withdrawal of prednisone (0.27 ± 0.03 mg/L for group A at day 14 and 0.15 ± 0.02 mg/L at day 21; P > 0.05). Dogs with PDH included in group B did not have significant differences in sCysC (0.22 ± 0.03 mg/L) compared to control (P > 0.05). CONCLUSIONS AND CLINICAL IMPORTANCE: Oral administration of prednisone unlike altered endogenous glucocorticoid production, increases sCysC in dogs in a dose-dependent fashion.

Interaction of interferon regulatory factor-1 and nuclear factor kappaB during activation of inducible nitric oxide synthase transcription.

We investigated the molecular mechanism for the synergistic induction of inducible nitric oxide synthase transcription by TNF-alpha and IFN-gamma. Since TNF-alpha and IFN-gamma stimulate cells in part by activating NF-kappaB and IRF-1, we hypothesized that these two transcription factors interact with each other. IRF-1 and NF-kappaB co-localize in the nucleus of stimulated macrophages. Co-immunoprecipitation experiments show that IRF-1 and NF-kappaB interact in stimulated but not resting cells. Super-shift experiments show that IRF-1 and NF-kappaB interact while binding to their respective DNA binding sites. These results demonstrate the existence of a physical interaction between IRF-1 and NF-kappaB proteins in vivo. We next suggested that this interaction between IRF-1 and NF-kappaB bends the DNA of the iNOS promoter region. Using a cyclization assay, we demonstrate that nuclear extracts from stimulated cells accelerate the rate of conversion of a linear to circular DNA, compared to extracts from resting cells. However, stimulated nuclear extracts cannot affect the rate of cyclization of a promoter with a mutant IRE or kappaB site. Furthermore, stimulated nuclear extracts depleted of IRF-1 and NF-kappaB cannot induce cyclization. We conclude that IRF-1 and NF-kappaB interact in vivo, and that this interaction physically bends the indicible nitric oxide synthase promoter DNA. This interaction may explain the mechanism by which IFN-gamma synergistically augments inducible nitric oxide synthase transcription.

Native and oxidized low density lipoproteins oppositely modulate the effects of insulin-like growth factor I on VSMC.

OBJECTIVE: Changes in the local expression and signaling activity of the insulin-like growth factor-I (IGF-I) axis regulate growth and survival of plaque-derived vascular smooth muscle cells (VSMC) and influence plaque fate. Recent evidence suggests that accumulation of low density lipoproteins (LDL) in VSMC during the progression of atherogenesis is linked to local changes in IGF-I signaling. We investigated the effects of LDL on the biological actions and downstream signaling pathways mediated by this growth factor in A10 VSMC. METHODS AND RESULTS: We first characterized the effects of LDL on the proliferative and anti-apoptotic actions of IGF-I in A10 VSMC. Native LDL were mitogenic and synergistically enhanced DNA synthesis induced by IGF-I from 4-, 9- up to 7.8-fold, while having no effect on its anti-apoptotic actions. In contrast, oxidized LDL, at oxidation levels that did not modify these actions by themselves, significantly reduced the mitogenic and survival effects of IGF-I by 40% and 60%, respectively. These observations correlated with opposite changes exerted by native and oxidized LDL on the insulin receptor substrate-1 (IRS)-associated PI3 kinase/Akt response to IGF-I. The extracellular signal-regulated kinase (ERK) signaling response was not affected. CONCLUSIONS: Our study demonstrates a previously unidentified modulation of the actions of IGF-I on A10 VSMC by LDL, dependent on their extent of oxidative modification. Our findings suggest that the differential modulation of the PI3 kinase/Akt response to IGF-I play a pivotal role.

Hydrogen peroxide regulation of bovine endothelin-converting enzyme-1.

Vascular injury leads to the production of reactive oxygen species (ROS), but the mechanisms by which ROS contribute to vascular pathology are not completely understood. We hypothesized that ROS increase endothelin converting enzyme (ECE-1) expression. We found that glucose oxidase (GO) increases ECE-1 mRNA, protein, and activity in bovine aortic endothelial cells. Catalase abolishes this effect. Glucose oxidase treatment of endothelial cells transactivates the ECE-1 promoter. The ECE-1 promoter element that mediates this response to GO is located between -444 and -216 bp. This region contains a STAT response element, and GO activates STAT-3 binding to this STAT response element. Our data suggest that STAT3 mediates hydrogen peroxide induction of ECE-1 expression.

Cardiovascular morbidity and obesity in adult liver transplant recipients.

There is a direct relationship between the grade of obesity and mortality based on the increased cardiovascular diseases, cancer, etc. However, the results of studies in renal and liver allograft recipients relating obesity to morbidity and mortality are contradictory. A retrospective cohort study of 170 patients transplanted between March 1987 and July 1997 showed obesity to be identified in 77 (45.3%) patients. During the mean follow-up of 5 years posttransplantation, 16 (9.4%) patients experienced cardiovascular complications, including 10 patients with ischemic cardiac syndromes (five acute infarctions and five angina), five patients with acute cerebrovascular accidents, and one patient with intermittent lower limbs claudication. The prevalence of obesity at 1, 3, 5, 7, and 9 years after transplantation was 58.2%, 56.9%, 60.3%, 59.5%, and 66.4%, respectively. Compared with the baseline value, the BMI was increased at 1 year posttransplantation (25.78), a significant difference. No significant differences were found between the mean BMI values of patients with and without cardiovascular diseases, or overweight and morbidly obese patients compared to the normal weight population. Among liver transplant recipients, obesity was a frequent complication after transplantation, but it was not clearly associated with increased morbidity and mortality secondary to cardiovascular disease.

Canine pyometra: a study of the urinary proteins by SDS-PAGE and Western blot.

Canine pyometra often causes glomerulonephritis by immune complex deposition in the glomeruli. Proteinuria, ranging from moderate to severe, may be present secondary to renal damage. To determine urinary protein excretion due to pyometra, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was conducted on urine from 15 bitches with pyometra and 10 healthy bitches. To characterize urinary immunoglobin excretion, Western blot analysis of the urine samples using antibodies to canine IgG and IgA was also performed. Nine bands were detected by electrophoresis in bitches with pyometra, while only four were detected in the healthy animals. The urinary proteins from bitches with pyometra were primarily of glomerular origin; 58% were of medium-high molecular weight (MW), and the remainder were low MW. None of the healthy dogs had IgG or IgA in their urine, whereas three bitches with pyometra had IgG in their urine and another bitch with pyometra had both IgG and IgA. The low proportion of bitches with urinary immunoglobins was probably be due to early diagnosis of the disease. Although only a limited number of dogs was used, this study is apparently the first to characterize the electrophoretic pattern of urinary proteins and to quantify urinary excretion of IgG and IgA in bitches with pyometra.

Characterization of renal damage in canine leptospirosis by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting of the urinary proteins.

Canine leptospirosis is a zoonotic disease that can cause interstitial nephritis. As a consequence of the renal damage, proteinuria may occur. To determine the urine protein pattern in the disease we performed sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) on the urine from 10 dogs with leptospirosis and 20 healthy dogs. Western blotting analysis of the urine samples with antibodies against canine IgG and IgA was also performed to identify these immunoglobulins in the urine. Urine electrophoresis showed three new bands in the dogs suffering from leptospirosis. Only two of the dogs with leptospirosis showed detectable concentrations of IgG and IgA in urine, while a third animal showed IgG alone. The study showed a 36.7% increase in the excretion of low molecular weight proteins in dogs with leptospirosis but almost no change in the high molecular weight protein pattern. These results, together with the low number of animal with detectable concentrations of IgG and IgA in the urine, support the view that canine leptospirosis is characterized by interstitial nephritis.

[The femoral vein route: an exceptional alternative for the implantation of permanent pacemakers]. / La vía venosa femoral: una alternativa de excepción para el implante de marcapasos definitivos.

A case of thrombosis in the superior vena cava after two permanent pacemaker implantations is described. Ten years ago the patient had a permanent pacemaker implanted because of disturbances in his cardiac rhythm. Five months later he was readmitted with pacemaker malfunction due to failure of the electrode and a new lead was introduced. In 1989 the generator presented "end of life" and the depleted battery was changed. In February, 1993 he presented a new pacemaker malfunction and thrombosis in the superior vena cava. We suggest that in similar cases the iliac vein approach should be performed and short-term anticoagulant therapy should be considered.

[A prospective and comparative study of 262 laparoscopic cholecystectomies]. / Estudio prospectivo y comparativo de 262 colecistectomías laparoscópicas.

A prospective, comparative study is made of 262 patients (195 males, 67 females) subjected to laparoscopic cholecystectomy. Two groups were considered: simple cholelithiasis (SC) (187 cases), and complicated cholelithiasis (CC) (75 cases: 63 acute cholecystitis, 8 hydrocholecystitis, 4 pancreatitis). Mean age was 51 among the SC cases, and 57 in the CC patients. Mean operating time was 67 and 96 minutes for the SC and groups, respectively. Preoperative complications were more frequent in the CC group (51.4%) than in the SC patients (24%)--immediate laparotomies being performed in 2 and 25% of the SC and CC patients, respectively. Mortality was zero, with similar morbidity in both groups. Mean hospitalization time was 4.9 and 3.4 days for the CC and SC groups, respectively.

Giant inguino-scrotal bladder hernia. Report of a case.

Bladder hernia is very uncommon. It tends to affect patients over the age of 50 and is predisposed by cervico-urethral obstruction. The condition is often diagnosed during inguinal hernia surgery. The authors report a case of massive inguino-scrotal bladder herniation. The different types of bladder hernia are described, and the clinical-radiological findings and surgical management are discussed.

[Corpus luteum defect. Analysis of 30 cases in clinical pregnancy]. / Cuerpo lúteo deficiente. Análisis de 30 casos con embarazo clínico.

Deficient corpus luteum (DCL) is an ovulatory dysfunction little defined but real. It is said that is frequency is 3 and 10% of sterile couples and 30 to 40% of habitual abortion. Is part of the group of ovulatory aberrations together with in situ luteinization and syndrome of not broken luteinized folicle. The diagnosis is based in endometrial morphology and progesterone determination. The objective of this work is to analyze 30 cases of DCL that achieved pregnancy. The diagnosis was done based on low determination of progesterone in serum, biopsy of irregular or indysphase endometrium and all the factors, remaining normal of esterility. Average age of patients was 36 years and for sterility 3.1. Treatment consisted in clomiphen cytrate (CC) administration 100 mg daily of hCG in 21 cases, CC only in eight cases and associated to bromocriptine in one case (concomitant hyper-prolactinemia); 26 pregnancies went to term, seven of them with support of exogenous progesterone the first weeks. There were three abortions and an ectopic one. It is concluded that with a complete study of sterility that only shows low seric progesterone and/or endometrium in dysphase or irregular, the diagnosis of DCL is probable and should be treated first with CC plus hCG. The evolution of pregnancies is normal and according to these results is little acceptable the use of progesterone as a support in pregnancy.

[Heterotopic pregnancy in intrauterine insemination. Presentation of a case]. / Embarazo heterotópico con inseminación intrauterina. Presentación de un caso.

The heterotopic (ectopic and orthotopic simultaneous) pregnancy shows a frequency of 1 to 15,000 to 1 to 30,000 gestations. The clinical diagnosis is difficult due to the lack of precise indicators, as to diagnose an intrauterine pregnancy eliminates the possibility of ectopic pregnancy. The methods of Assisted Reproduction seem to be factors that have influenced on the increment of this type of gestations. A case of a 32 year patient with primary sterility by pelvic adhesions process, that was surgically treated, as there was no pregnancy after surgery, she was given intrauterine insemination with her husband's semen (IU) pregnancy was obtained, determined at 15 days of menstrual lack by presence of subunit B of HCG in serum and vaginal ultrasound that confirmed gestational sac. One month after she presented at Urgencies with an acute abdominal condition; laparotomy was done and salpingectomy was carried out for ruptured tubal pregnancy confirmed by histopathology. The evolution on intrauterine pregnancy was normal culminating with cesarean section at week 35 by inminence of eclampsia/Mother and child in good conditions.

[Presentation of a mathematics formula of duplicated times of estradiol predictive for ovular harvesting]. / Presentación de una fórmula matemática de tiempos de duplicación de estradiol pronóstica para captura ovular.

30 patients were studied, that were included in the IVF program of the INPer. The estradiol response was studied and was correlated with the number of retrieved oocytes. The previously known E2 values considered as indicative for cancellation did no were observed in this series, and there are proposed new ones, also prognostic criterion for oocyte retrieval are proposed.