Pharmacokinetics/pharmacodynamics and therapeutic drug monitoring of ceftazidime/avibactam administered by continuous infusion in patients with MDR Gram-negative bacterial infections.

BACKGROUND: Therapeutic drug monitoring (TDM) of ß-lactams in critically ill patients has been correlated with better clinical outcomes. Evidence on TDM of newer ß-lactams such as ceftazidime/avibactam administered by continuous infusion (CI) is very limited. OBJECTIVES: To describe our experience with TDM of ceftazidime/avibactam and pharmacokinetic/pharmacodynamic (PK/PD) target attainment in patients with MDR bacterial infections. Clinical outcomes of ceftazidime/avibactam administered by CI were also assessed. METHODS: Patients treated with ceftazidime/avibactam administered by CI and undergoing TDM of ceftazidime plasma concentrations were included. Blood samples were obtained as part of the TDM program. The PK/PD therapeutic target of ceftazidime/avibactam was defined as 100%fT > 4 × MIC of the causative pathogen, and 100%fT > 10 × MIC was considered overexposure. Dose changes were made according to the TDM results. RESULTS: Thirty-one patients were included. Ceftazidime/avibactam total daily doses ranged from 1 g/0.25 g to 6 g/1.5 g. Twenty-six patients (83.9%) achieved a 100%fT > 4 × MIC, 15 (48.4%) of which were overexposed (100%fT > 10 × MIC). Dose reduction was suggested in 16/28 (57.1%) patients and dose maintenance in 12/28 (42.9%). Overall clinical cure was observed in 21 (67.7%) patients, and 18 of these (85.7%) achieved a 100%fT > 4 × MIC. CONCLUSIONS: Administering ceftazidime/avibactam by CI enabled the desired PK/PD target to be achieved in a large proportion of patients, even at lower doses than those recommended for a 2 h extended infusion. We suggest that the use of CI with TDM may be a useful tool for reducing initial doses, which could help to reduce antimicrobial-related adverse effects and treatment costs.

Review & meta-analysis: isopropanolic black cohosh extract iCR for menopausal symptoms - an update on the evidence.

A systematic literature search revealed 35 clinical studies and one meta-analysis comprising 43,759 women, of which 13,096 were treated with isopropanolic Cimicifuga racemosa extract (iCR). Compared to placebo, iCR was significantly superior for treating neurovegetative and psychological menopausal symptoms, with a standardized mean difference of -0.694 in favor of iCR (p < 0.0001). Effect sizes were larger when higher dosages of iCR as monotherapy or in combination with St. John's wort (Hypericum perforatum [HP]) were given (-1.020 and -0.999, respectively), suggesting a dose-dependency. For psychological symptoms, the iCR+HP combination was superior to iCR monotherapy. Efficacy of iCR was comparable to low-dose transdermal estradiol or tibolone. Yet, due to its better tolerability, iCR had a significantly better benefit-risk profile than tibolone. Treatment with iCR/iCR+HP was well tolerated with few minor adverse events, with a frequency comparable to placebo. The clinical data did not reveal any evidence of hepatotoxicity. Hormone levels remained unchanged and estrogen-sensitive tissues (e.g. breast, endometrium) were unaffected by iCR treatment. As benefits clearly outweigh risks, iCR/iCR+HP should be recommended as an evidence-based treatment option for natural climacteric symptoms. With its good safety profile in general and at estrogen-sensitive organs, iCR as a non-hormonal herbal therapy can also be used in patients with hormone-dependent diseases who suffer from iatrogenic climacteric symptoms.

Assessment of Cardiovascular Safety of Anti-Osteoporosis Drugs.

The incidence of osteoporosis and cardiovascular disease increases with age, and there are potentially shared mechanistic associations between the two conditions. It is therefore highly relevant to understand the cardiovascular implications of osteoporosis medications. These are presented in this narrative review. Calcium supplementation could theoretically cause atheroma formation via calcium deposition, and in one study was found to be associated with myocardial infarction, but this has not been replicated. Vitamin D supplementation has been extensively investigated for cardiac benefit, but no consistent effect has been found. Despite findings in the early 21st century that menopausal hormone therapy was associated with coronary artery disease and venous thromboembolism (VTE), this therapy is now thought to be potentially safe (from a cardiac perspective) if started within the first 10 years of the menopause. Selective estrogen receptor modulators (SERMs) are associated with increased risk of VTE and may be related to fatal strokes (a subset of total strokes). Bisphosphonates could theoretically provide protection against atheroma. However, data from randomised trials and observational studies have neither robustly supported this nor consistently demonstrated the potential association with atrial fibrillation. Denosumab does not appear to be associated with cardiovascular disease and, although parathyroid hormone analogues are associated with palpitations and dizziness, no association with a defined cardiovascular pathology has been demonstrated. Finally, romosozumab has been shown to have a possible cardiovascular signal, and therefore post-market surveillance of this therapy will be vital.

[Vitamin D deficiency rickets complicating Dorfman-Chanarin syndrome]. / Rachitisme carentiel compliquant une maladie de Chanarin-Dorfman.

Vitamin D deficiency rickets remains a public health issue in many parts of the world. In France, this diagnosis has almost disappeared since 1992 with routine vitamin D supplementation for children. Therefore, it is more difficult for doctors to identify risk factors and early signs of this disease. In this article, we report a rickets diagnosis acquired by vitamin D deficiency in a child who presented with the onset of a genu valgum and difficulty walking at the age of 9½ years. This patient was a Comorian child followed up from his birth for Dorfman-Chanarin syndrome. Dorfman-Chanarin syndrome is a rare disease, with about 80 cases reported in the literature. It belongs to the group of neutral lipid storage diseases (NLSD) characterized especially on the skin by ichthyosis. This child presented risk factors for vitamin D deficiency (dark skin color, prolonged and exclusive breastfeeding, premature end of supplementation, and particularly severe ichthyosis) that should have alerted us to the risk of vitamin D deficiency and the need for supplementation. This case highlights the importance of vitamin D, especially if there are risk factors such as ichthyosis, and the need to remain watchful in monitoring all chronic diseases.

[Postinfectious family case of acute necrotizing encephalopathy caused by RANBP2 gene mutation]. / Un cas familial d'encéphalopathie nécrosante aiguë post-infectieuse associé à une mutation du gène RANBP2.

UNLABELLED: Acute necrotizing encephalopathy is a rare neurologic disease most often triggered by a febrile viral event affecting an otherwise healthy infant. The clinical course is characterized by rapid deterioration of the neurological condition that often leads to coma and requires intensive care. The diagnosis is usually suggested by MRI, which shows symmetrical and focal necrotic lesions of thalami. Acute necrotizing encephalopathy has been linked in recent studies to an autosomal-dominant mutation of the gene for the protein RAN-binding protein 2. CASE REPORT: We report three cases in siblings of Tunisian origin. Two of them presented with acute necrotizing encephalopathy at the age of 9 months in the immediate aftermath of a viral infection. The molecular study conducted in the family showed that both patients and their mother were carriers of the missense mutation gene RAN-binding protein 2. COMMENTS: Although the role of Ran BP2 protein is incompletely known, mutation of the RANBP2 gene causes rare, reversible central neurologic disorders. Suspected diagnosis is facilitated by MRI, which shows specific lesions of multifocal, symmetric involvement of the thalami, brainstem tegmentum, supratentorial white matter, and cerebellum. Due to the low frequency of the disease and its non-specific clinical presentation, the diagnosis of acute necrotizing encephalopathy is a major challenge, while preventative measures can be proposed in familial mutation.

The impact of coffee on health.

OBJECTIVE: Coffee is a beverage used worldwide. It includes a wide array of components that can have potential implication on health. We have reviewed publications on the impact of coffee on a series of health outcomes. METHODS: Articles published between January 1990 and December 2012 were selected after crossing coffee or caffeine with a list of keywords representative of the most relevant health areas potentially affected by coffee intake. RESULTS: Caffeine, chlorogenic acids and diterpenes are important components of coffee. Tolerance often acts as a modulator of the biological actions of coffee. There is a significant impact of coffee on the cardiovascular system, and on the metabolism of carbohydrates and lipids. Contrary to previous beliefs, the various forms of arterial cardiovascular disease, arrhythmia or heart insufficiency seem unaffected by coffee intake. Coffee is associated with a reduction in the incidence of diabetes and liver disease. Protection seems to exist also for Parkinson's disease among the neurological disorders, while its potential as an osteoporosis risk factor is under debate. Its effect on cancer risk depends on the tissue concerned, although it appears to favor risk reduction. Coffee consumption seems to reduce mortality. CONCLUSION: The information gathered in recent years has generated a new concept of coffee, one which does not match the common belief that coffee is mostly harmful. This view is further supported by the discovery of a series of phyto-components with a beneficial profile. Reasonable optimism needs to be tempered, however, by the insufficiency of the clinical data, which in most cases stem from observational studies.

The impact of chocolate on cardiovascular health.

Cardiovascular disease is the leading determinant of mortality and morbidity in women. Functional foods are attracting interest as potential regulators of the susceptibility to disease. Supported by epidemiological evidence, chocolate has emerged as a possible modulator of cardiovascular risk. Chocolate, or cocoa as the natural source, contains flavanols, a subclass of flavonoids. The latter years have witnessed an increasing number of experimental and clinical studies that suggest a protective effect of chocolate against atherogenesis. Oxidative stress, inflammation, and endothelial function define three biological mechanisms that have shown sensitivity to chocolate. Moreover, the consumption of chocolate has been involved in the protective modulation of blood pressure, the lipid profile, the activation of platelets, and the sensitivity to insulin. Dark chocolate seems more protective than milk or white chocolate. Despite this array of benefits, there is a lack of well designed clinical studies demonstrating cardiovascular benefit of chocolate. The high caloric content of chocolate, particularly of some less pure forms, imposes caution before recommending uncontrolled consumption.

[Nutrition therapy for adult patients with caustic injuries to gastrointestinal tract]. / Terapia nutricional en pacientes adultos con quemaduras del tracto gastrointestinal por cáusticos.

OBJECTIVES: To present here the experience of our Nutrition Therapy Team of the Hospital El Tunal, for the nutritional management of adult patients with caustic injuries to gastrointestinal tract. MATERIALS AND METHODS: This is a retrospective, descriptive study of patients with caustic injuries to gastrointestinal tract managed by our Nutrition Therapy Team between January 2000 and December 2007. We revisited the clinical history of patients with diagnosis of caustic injury. Various nutritional variables, as well as the evolution and outcome were pooled and analyzed. RESULTS: A total of 30 patients, 17 male y 13 female with a mean age of 34.4 +/- 17.2 years old were found. The ingestion of caustics was suicidal intent in 22 (73.3%) and accidental in 8 (26.7%). The global mortality was high (43.3%). Weight loss was found in 46.9% of the patients and a negative nitrogen balance in 62.5%. Sixteen patients (53.12%) were managed with mixed nutrition (enteral and/or parenteral) for a mean time of 24 +/- 22 days. We compared two groups Moderate vs Severe, according to the severity of the caustic injury to gastrointestinal tract and found that mortality, the length of hospital stay and the final albumin value were significantly different among groups. CONCLUSIONS: Caustic injuries to gastrointestinal tract are not frequent, they are found mainly in young patients with suicidal intent and are associated with high mortality, especially in severe injuries. This aggression causes important catabolic state leading to a negative nitrogen balance and weight loss. These patients require early nutritional intervention sometimes extended for months.

Comparative effects of 17beta-estradiol, raloxifene and genistein on bone 3D microarchitecture and volumetric bone mineral density in the ovariectomized mice.

UNLABELLED: This study assessed the effect of estradiol, raloxifene and genistein on the preservation of bone 3D-microarchitecture and volumetric bone mineral density (vBMD) in the ovariectomized mouse model. Our results indicated that raloxifene was more effective in preserving bone ovariectomized-induced changes, the advantage being concentrated in both bone microarchitecture and vBMD. INTRODUCTION: This study assessed the effect of different estrogen receptor (ER) agonists on the preservation of bone 3D-microarchitecture and volumetric bone mineral density (vBMD) in the ovariectomized (OVX) mouse model. METHODS: Twelve-week-old female C57BL/6 mice were randomly assigned to one of five groups: (1) SHAM-operated + vehicle; (2) OVX + vehicle; (3) OVX + 17beta-estradiol (5 microg/kg); (4) OVX + raloxifene (1 mg/kg); (5) OVX + genistein (25 mg/kg), during 4-weeks. Bone microarchitecture and trabecular, cortical and total vBMD of distal femur were imaged by ex vivo microcomputed tomography (micro-CT). RESULTS: Ovariectomy produced a global deterioration involving both trabecular and cortical 3D-microarchitecture and vBMD. Raloxifene maintained both microarchitecture and vBMD, whereas estradiol prevented deterioration of some microstructural parameters, such as trabecular thickness (Tb.Th), trabecular bone pattern factor (Tb.Pf), and cortical periosteal perimeter (Ct.Pe.Pm), but did not completely block the loss in vBMD. Mice treated with genistein exhibited the less favourable profile in both vBMD and microstructural parameters preserving only cross-sectional bone area (B.Ar) and Ct.Pe.Pm in cortical bone. CONCLUSION: Our data indicate that, at the selected doses, raloxifene was more effective in preserving bone OVX-induced changes than either estradiol or genistein, the advantage being concentrated in both bone microarchitecture and vBMD.

[Effect of the administration of cholecystokinin on the cholestasis associated with total parenteral nutrition in experimental short bowel]. / Efecto de la administración de colescistoquinina sobre la colestasis asociada a la nutrición parenteral total en el intestino corto experimental.

INTRODUCTION: Total parenteral nutrition (TPN) is not free of complications. One of the most serious is cholestasis; the cause of this complication is unclear but it may be due to a lack of an enteral stimulus for cholecystokinin (CCK) production. CCK is essential for contraction of the gallbladder and also stimulates intrahepatic bile flow. Its absence may contribute to cholestasis. After any hepatic aggression, the Kupffer cells respond and release proinflammatory cytokines, such as interleukin-1 (IL-1) and tumour necrosis factor alpha (TNF-alpha), which increase the hepatic damage. The objective of this experimental study has been to observe the effect that the exogenous administration of CCK could have on hepatic damage in experimental short bowel with and without TPN, determined using the serum levels of IL-1 and TNF-alpha. MATERIAL AND METHODS: A resection of 80% of the small bowel was performed on 53 Wistar rats and a continuous infusion of saline or TPN was initiated. The rats were divided into the following groups: SHAM (N = 14): normal saline infusion and free access to food and water. TPN (N = 15): Standard TPN. SHAM-CCK (N = 14): same as the SHAM group but with a daily dose of CCK. TPN-CCK (N = 10): same as the TPN group but with a daily dose of CCK. At the end of the experiment, the animals were sacrificed and blood samples were obtained to determine the IL-1 and TNF-alpha values by ELISA. RESULTS: The IL-1 and TNF-alpha levels were higher in the TPN group (7.537 and 5.899 pg/mL, respectively) than in the SHAM group (6.509 and 4.989 pg/mL, respectively) (p > 0.05). The TNF-alpha values were higher in the SHAM group (4.989 pg/mL) than in the SHAM-CCK group (4.583 pg/mL) (p < 0.001). The IL-1 and TNF-alpha levels were higher in the TPN group than in the TPN-CCK group (6.709 and 4.794 pg/mL, respectively) (p < 0.001 for TNF-alpha). CONCLUSIONS: 1. There is a rise in the serum levels of the pro-inflammatory cytokines IL-1 and TNF-alpha in animals with short bowel on TPN or enteral nutrition. 2. The administration of CCK causes a fall in the IL-1 and TNF-alpha levels, and could be used such as a further measure to prevent TPN-associated cholestasis.

Classic and late-onset neurological disease in two siblings with glutaryl-CoA dehydrogenase deficiency.

Late-onset neurological disease has rarely been reported in patients with glutaryl-CoA dehydrogenase (GCDH) deficiency. We present two siblings with GCDH deficiency. One of them presented with the classic neurological disease (patient 1). Routine investigation of family members revealed that her apparently unharmed 13-year-old sister was also affected (patient 2). Patient 2 started to have academic difficulties in the months prior to our assessment. Her clinical examination was normal, with the exception of a cranial circumference of 57 cm (slightly over the 98 th centile). A severe leukoencephalopathy was demonstrated on MRI. Neuropsychological assessment showed an IQ within the normal-low range and a mild impairment of memory and executive function. Previous reports on late-onset neurological disease in GCDH deficiency have revealed that progressive leukoencephalopathy develops over time. Following the recently published guideline for the diagnosis and management of GCDH deficiency, both patients are receiving dietary treatment in combination with L-carnitine supplementation. We emphasize the need to search for chronic neurological changes of late-onset type in apparently unaffected GCDH deficiency cases diagnosed in routine family investigations.

Efecto de la administración de colescistoquinina sobre la colestasis asociada a la nutrición parenteral total en el intestino corto experimental / Effect of the administration of cholecystokinin on the cholestasis associated with total parenteral nutrition in experimental short bowel

Introducción. La nutrición parenteral total (NPT) no está libre de complicaciones. Una de las más serias es la colestasis, cuyo origen no es muy claro y puede deberse a la falta de estimulo enteral para la producción de colescistoquinina (CCQ). La CCQ es fundamental para la contracción de la vesícula biliar y como estimulante del flujo biliar intrahepático. Su falta puede contribuir a la colestasis. Ante toda agresión hepática, las células de Kupffer responden y liberan citoquinas proinflamatorias, como la interleukina-1 (IL-1) y el factor de necrosis tumoral alfa (TNF-α), que aumentan el daño hepático. El objetivo de este estudio experimental ha sido observar el efecto que la administración exógena de CCQ pudiera tener sobre el daño hepático en el intestino corto experimental con y sin NPT, medido por los niveles séricos de IL-1 y TNF-α. Material y métodos. Cincuenta y tres ratas Wistar fueron sometidas a una resección del 80% del intestino delgado y a una infusión continua de suero o NPT y se asignaron a los siguientes grupos: • SHAM (N=14): infusión de suero fisiológico y acceso libre a comida y agua. • NPT (N=15): NPT estándar. • SHAM-CCQ (N=14): como el grupo SHAM y una administración diaria de CCQ. NPT-CCQ (N=10): como el NPT y una administración diaria de CCQ. Al final del experimento, los animales fueron sacrificados y se obtuvieron muestra de sangre para determinar los valores IL-1 y TNF-α por ELISA. Resultados. Los niveles de IL-1 y TNF-α fueron mayores en el grupo NPT (7,537 y 5,899 pg/mL, respectivamente) que en el grupo SHAM (6,509 y 4,989 pg/mL, respectivamente) (p > 0,05). Los valores de TNF-α fueron mayores en el grupo SHAM (4,989 pg/mL) que en el grupo SHAM-CCQ (4,583 pg/mL), (p < 0,001). Los niveles de IL-1 y TNF-α fueron mayores en el grupo NPT que en el grupo NPT-CCQ (6,709 y 4,794 pg/mL, respectivamente) (p< 0,001 para TNF-α). Conclusiones. 1. En los animales con intestino corto bajo NPT o con nutrición enteral, se elevan los niveles séricos de las citoquinas proinflamatorias IL-1 y TNF-α. 2. La administración de CCQ, disminuye los niveles de IL-1 y TNF- α, pudiendo ser utilizada como una medida más para combatir la colestasis asociada a la NPT

Introduction. Total parenteral nutrition (TPN) is not free of complications. One of the most serious is cholestasis; the cause of this complication is unclear but it may be due to a lack of an enteral stimulus for cholecystokinin (CCK) production. CCK is essential for contraction of the gallbladder and also stimulates intrahepatic bile flow. Its absence may contribute to cholestasis. After any hepatic aggression, the Kupffer cells respond and release proinflammatory cytokines, such as interleukin-1 (IL-1) and tumour necrosis factor alpha (TNF-α), which increase the hepatic damage. The objective of this experimental study has been to observe the effect that the exogenous administration of CCK could have on hepatic damage in experimental short bowel with and without TPN, determined using the serum levels of IL-1 and TNF-α. Material and methods. A resection of 80% of the small bowel was performed on 53 Wistar rats and a continuous infusion of saline or TPN was initiated. The rats were divided into the following groups: • SHAM (N=14): normal saline infusion and free access to food and water. • TPN (N=15): Standard TPN. • SHAM-CCK (N=14): same as the SHAM group but with a daily dose of CCK. • TPN-CCK (N=10): same as the TPN group but with a daily dose of CCK. At the end of the experiment, the animals were sacrificed and blood samples were obtained to determine the IL-1 and TNF-α values by ELISA. Results. The IL-1 and TNF-α levels were higher in the TPN group (7.537 and 5.899 pg/mL, respectively) than in the SHAM group (6.509 and 4.989 pg/mL, respectively) (p>0.05). The TNF-α values were higher in the SHAM group (4.989 pg/mL) than in the SHAM-CCK group (4.583 pg/mL) (p<0.001). The IL-1 and TNF-α levels were higher in the TPN group than in the TPNCCK group (6.709 and 4.794 pg/mL, respectively) (p< 0.001 for TNF-α). Conclusions. 1. There is a rise in the serum levels of the pro-inflammatory cytokines IL- 1 and TNF-α in animals with short bowel on TPN or enteral nutrition. 2. The administration of CCK causes a fall in the IL-1 and TNF-α levels, and could be used such as a further measure to prevent TPN-associated cholestasis

Pérdida ósea posmenopáusica: estudio del polimorfismo genético y efecto de la administración de fitoestrógenos sobre los marcadores de metabolismo óseo / Postmenopausal bone loss: Study of genetic polymorphism and effect of phytoestrogen administration upon bone metabolism markers

Objetivo: Estudiar el efecto de la administración de fitoestrógenos (flavonoides) sobre determinados marcadores bioquímicos de metabolismo óseo a corto plazo (3 meses) en mujeres posmenopáusicas y analizar la asociación de algunos polimorfismos genéticos sobre el índice de masa corporal (IMC). Material y método: Se seleccionaron 54 mujeres en los primeros años de la menopausia para el estudio bioquímico y 102 formaron parte del estudio genético. El estudio bioquímico, lo finalizaron 49 mujeres, 22 en el grupo control y 27 tratadas con extracto de isoflavonas de soja (Phyto- Soya®). Se determinó el IMC (Kg/m2), varios marcadores de metabolismo óseo como el fragmento amino-terminal del colágeno tipo I (NTx), fosfatasa alcalina ósea y total, hormona paratiroidea, etc., hormonas como la testosterona y el estradiol y citoquinas como la interleuquina-6, la osteoprotegerina, y RANKL. El ADN se obtuvo de células de sangre periférica y los polimorfismos en los genes de receptor de estrógenos alfa (ERα), CD40 y Runx2 se genotiparon mediante la técnica de PCR-RFLP. Resultados: A los 3 meses de administrar fitoestrógenos, el perfil de metabolismo óseo no se modificó aunque disminuyó el nivel de interleuquina-6. El genotipo de los polimorfismos de restricción del gen del ERα (PvuII y XbaI) y el polimorfismo de restricción MspA1I y el de longitud (deleción de 11 alaninas) en el gen Runx2 están asociados al IMC. Conclusión: Los fitoestrógenos no han modificado el perfil de los marcadores de metabolismo óseo en un grupo de mujeres pequeño y a corto plazo. Sin embargo, el grupo de mujeres tratadas con fitoestrógenos han mostrado menores niveles de IL-6, una citoquina proinflamatoria relacionada con mayores tasas se resorción ósea. En el estudio genético, el genotipo del gen ERα se ha asociado y predice el IMC y el polimorfismo C/T del gen CD40 se relaciona con la excreción de NTx, un marcador de resorción ósea

Objective: To study the short-term (3 months) effect of phytoestrogen (flavonoid) administration upon certain biochemical markers of bone metabolism in postmenopausal women, with an analysis of the association of certain genetic polymorphisms to body mass index (BMI). Material and method: Fifty-four women in the first years of menopause were selected for the biochemical study, while 102 formed part of the genetic study. The biochemical study was completed by 49 women, 22 in the control group and 27 in the group treated with soy isoflavone extract (Phyto-Soya®). Determinations were made of BMI (kg/m2), several bone metabolism markers such as amino-terminal fragment of collagen type I (NTx), total and bone alkaline phosphatase, parathyroid hormone, etc., hormones such as testosterone and estradiol, and cytokines such as interleukin-6 (IL-6), osteoprotegerin, and RANKL. DNA was obtained from peripheral blood cells, and the polymorphisms of the alpha-estrogen receptor (ERα), CD40 and Runx2 genes were genotyped using the PCR-RFLP technique Results: After three months of phytoestrogen administration, the bone metabolic profile showed no changes, though the IL-6 levels decreased. The genotypes of the restriction polymorphisms of the ER· gene (PvuII and XbaI) and the restriction polymorphism MspA1I and length polymorphism (deletion of 11 alanines) in the Runx2 gene are associated to BMI. Conclusion: Phytoestrogens have not modified the bone metabolic marker profile in a small group of women over the short term. However, the women treated with phytoestrogens showed lower levels of IL-6, a proinflammatory cytokine related to increased bone resorption rates. In the genetic analysis, the ERα gene genotype has been associated with, and predicts BMI, while C/T polymorphism of the CD40 gene is related to the excretion of NTx, a bone resorption marker

Immune response to Neospora caninum in naturally infected heifers and heifers vaccinated with inactivated antigen during the second trimester of gestation.

The objective of this study was to compare the immune response to Neospora caninum in naturally infected heifers and heifers inoculated with a killed whole N. caninum tachyzoite preparation during the second trimester of gestation. Nine Holstein heifers were used in this study; three naturally infected heifers were born from seropositive dams, and six seronegative heifers were born from seronegative dams. Four seronegative heifers were subcutaneously vaccinated with a killed whole N. caninum tachyzoite preparation at weeks 13, 15 and 17 of gestation. A killed whole N. caninum tachyzoite preparation containing 45 mg of protein/5 ml dose was formulated with 70% of mineral oil adjuvant (13% consisting of Arlacel C, 85% Marcol 52 and 2% Tween-80). Similarly, two seronegative heifers (negative controls) were inoculated with mock-infected bovine monocytes in oil adjuvant. Humoral immune responses were tested by using an indirect fluorescent antibody test (IFAT) and an indirect enzyme-linked immunosorbent assay (ELISA) for detecting isotype specific antibodies. Cellular immune responses were assessed by lymphocyte proliferation test (LPT) and IFN-gamma production. N. caninum-specific antibody responses increased in immunized cattle by week 15 of gestation (mean reciprocal antibody titers 450+/-252), peaked at week 23 (mean 16,000+/-6400). Maximum antibody response in naturally infected heifers was observed at week 19 of gestation (mean: 3467+/-2810). Mean serum IFAT titers were significantly higher in immunized heifers compared with those in naturally infected heifers from weeks 17 to 25 (P < 0.05). Analysis of isotype specific antibodies in naturally infected heifers revealed a predominant IgG1 response in one heifer and a predominant IgG2 response in the other two. Similar titers of IgG1 and IgG2 occurred in immunized heifers. Control heifers remained seronegative throughout the study by IFAT and ELISA. Significant antigen-specific proliferation responses were only detected in naturally infected heifers in week 19 of gestation. Peripheral mononuclear blood cells (PMBC) from immunized animals produced IFN-gamma in similar concentrations to those of infected animals (P > 0.05). No abortion was seen in any experimental group; however, one calf from a vaccinated heifer died due to dystocia. All calves from vaccinated and control heifers were seronegative by IFAT at 6 months of age; in contrast, calves born from naturally infected heifers remained seropositive with titers > or = 200. Killed vaccine induced similar immune responses to those found in chronically, naturally infected cattle which did not abort; however, different immune pathways may be followed in vaccinated and natural infected heifers with differences in degree of protective immunity.

Utilidad clínica del videoelectroencefalograma en régimen ambulatorio / Clinical utility of outpatient videoelectroencephalogram monitoring

Objetivo. Analizar la utilidad de los registros por video-electroencefalograma (VEEG) en régimen ambulatorio realizados en un servicio de neurología general para la detección de un episodio crítico. Pacientes y métodos. Desde el 1 de junio de 1999 hasta el 1 de junio de 2003 realizamos 105 exploraciones por VEEG, de 30 minutos a 5 horas de duración, en pacientes con crisis de etiología no aclarada, ante la sospecha de pseudocrisis o en presencia de una epilepsia farmacorresistente y crisis muy frecuentes. No modificamos la medicación del paciente para realizar la exploración. Resultados. En 33 pacientes se registraron eventos clínicos patológicos; en 14 se trató de crisis epilépticas, en 12 de pseudos crisis, en 4 de síncopes y en 3 de movimientos anormales no epilépticos. La duración del registro fue de 30 minutos en 12, de entre 30 minutos y 2 horas en 12 y de más de 2 horas en 9 pacientes. En 18 pacientes el VEEG fue la exploración diagnóstica. En un caso cambió el diagnóstico del tipo de crisis epiléptica que sufría el paciente, y en 11 pacientes nos ayudó a caracterizar sus crisis epilépticas. Conclusión. Si bien el porcentaje de registro de eventos patológicos durante un estudio por VEEG ambulatorio de 30 minutos a 5 horas de duración es bajo, su repercusión clínica es muy alta y el coste añadido, escaso

Objective. To analyze the utility of outpatient videoelectroencephalogram (VEEG) in a general neurology department to detect an ictal event. Patients and methods. One hundred and five patients with ictal phenomenology of unknown etiology, suspicion of pseudoseizures, refractory epilepsy with very frequent seizures, underwent outpatient VEEG monitoring from 30 minutes to five hours of duration, between June 1, 1999 and June 30, 2003. Patient medication was not modified to perform the recording. Results. Among the 105 outpatient VEEG monitoring, 33 clinical pathologic events were identified; these comprised 14 epileptic seizures, 12 pseudoseizures, four syncopes, and three non epileptic abnormal movements. Outpatient VEEG monitoring duration was as follows: 30 minutes in 12 patients, between 30 minutes and two hours in another 12, and more than two hours in 9. In 19 patients, the VEEG recording allowed a definitive diagnosis; in one case, it changed the epileptic seizure type, and in 11 patients, it helped to better characterize the epileptic seizure type. Conclusion. Although the percentage of pathologic events during an outpatient VEEG monitoring of 30 minutes to five hours of duration is low, its clinical repercussion is very important and the added cost is low

Oral administration of pharmacological doses of vitamins C and E reduces reproductive fitness and impairs the ovarian and uterine functions of female mice.

This study aims to ascertain whether oral administration of pharmacological doses of Vitamins C and E has any detrimental effect on reproductive fitness of female mice. We fed hybrid female mice from the first day of weaning a standard diet supplemented or not supplemented with pharmacological doses of Vitamins C and E. At the age of 28 weeks, we individually caged females with a male for the rest of their reproductive life. We performed a series of mating experiments to ascertain the number of oocytes ovulated and the potential for embryo development in vitro to the blastocyst stage and in vivo to Day 12 of gestation. The antioxidant diet decreased the frequency of litters, litter size, total number of offspring born and survival of male pups to weaning. This effect was associated with lower number of corpora lutea in the left ovary, decreased percentage of viable fetuses, and higher number of fetal resorptions in the left uterine horn when compared to the control group. The strategy of supplementing the diet with antioxidant vitamins to prevent the age associated decrease in reproductive potential should not be implemented in human beings until a safe and efficient diet is designed.

Effect of dietary supplementation with a mixture of Vitamins C and E on fertilization of tertiary butyl hydroperoxide-treated oocytes and parthenogenetic activation in the mouse.

The present study aims to analyze the effect of dietary supplementation with a mixture of Vitamins C and E on fertilization and later development of tertiary butyl hydroperoxide (tBH)-treated mouse oocytes and on parthenogenetic activation of freshly ovulated mouse oocytes. We fed hybrid mice a standard diet supplemented or not supplemented with Vitamins C and E from the first day of weaning until the age of 12 weeks. We noted no significant effect of diet on fertilization rate, percentage of total and hatching blastocysts, total number of cells, mitotic index and percentage of apoptotic nuclei at 120 h post-insemination of oocytes incubated for 15 min in the presence of 0, 1, 5 and 10 microM tBH. Furthermore, diet did not affect the percentage of activated oocytes after treatment with Ca2+ ionophore, acid Tyrode's solution or ethanol. The percentage of parthenogenetically activated oocytes that progressed to the pronuclear stage was significantly higher in the antioxidant group. Oocytes from antioxidant females exhibited a significantly lower mitogen-activated protein kinase (MAPK) activity than oocytes from control females. We detected no significant differences between groups in M-phase-promoting factor (MPF) activity. These results show that oral administration of antioxidants decreases MAPK activity and increases the probability of reaching the pronuclear stage after parthenogenetic activation.

Superoxide scavenging by polyphenols: effect of conjugation and dimerization.

The superoxide anion scavenging capacity of two flavonols (quercetin and kaempferol) and some of their conjugates (quercetin-3-rhamnoglucoside, quercetin-3-sophoroside, quercetin-3-sulphate, quercetin-3-glucuronide, kaempferol-3-sophoroside, kaempferol-3-glucuronide) and of several hydroxycinnamic acids (caffeic acid, ferulic acid, 5-5 diferulic acid, 8-O-4 diferulic acid and 8-8 diferulic acid) were studied. Superoxide anions were generated non-enzymatically in a phenazine methosulphate-NADH system and assayed by reduction of nitro-blue tetrazolium. Among the flavonols examined, the most effective scavengers of superoxide anions were the sophoroside, glucuronide and rhamnoglucoside conjugates. Conversely, quercetin-3-sulphate and the flavonol aglycones, exhibited some pro-oxidant activity at the range of concentrations tested (0.5-10 microM). These results show that conjugation has a marked effect on the scavenging capacity of flavonols and that the type of conjugate at the 3-position determines the final superoxide scavenging capacity. Caffeic acid and ferulic acid showed no effect on the generation of superoxide anions by phenazine methosulphate-NADH. However, dimerization of ferulic acid enhanced the superoxide scavenging capacity of this hydroxycinnamic acid, but this depended on the type of linkage between the monomers. The order, from highest to lowest, of superoxide radical scavenging capacity for the dimers of ferulic acid was: 5-5-diferulic acid > 8-O-4-diferulic acid > 8-8-diferulic acid.

Estimation of free radical-quenching activity of leaf pigment extracts.

The chromogen 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) radical (ABTS.+) can be directly generated by the enzymatic system formed by hydrogen peroxide and horseradish peroxidase in ethanolic media. The reaction is time- and concentration-dependent, and the ABTS.+ generated shows excellent stability. The method is an adaptation of the decolouration method previously reported (Cano et al. 1998. Phytochem Anal 9:196-202), which was used for estimating anti-oxidant activity in aqueous media. When the assay described here was used to quantify the anti-oxidant activity of two anti-oxidants, Trolox (an analogue of vitamin E) and beta-carotene, the latter had 2.5 times better anti-oxidant capacity than Trolox at the same concentration. The free radical quenching activity (anti-oxidant activity) of leaf pigment extracts was measured and related to the the carotenoid contents in leaf extracts of barley, oat and citrus. The possibilities of using this experimental approach to evaluate the status of photoprotective pigments are discussed.