Quantification of low levels of methionine oxidation in erythropoietin drug substance.

A pharmacopoeial monograph under development for recombinant human erythropoietin (rhEPO) drug substance is likely to contain a specification limit for the proportion of the methionine-oxidised variant. Methionine oxidation has no effect on the folded structure and global thermodynamic stability of rhEPO but can decrease biological activity [1]. We describe here the development of a reference standard, a calibrated mixture of the native and oxidised tryptic peptides which contain methionine-54, and an optimised peptide mapping procedure to support this assay. The approach may be developed for analysis of drug product or generalised for other assays in which product-related impurities are quantified by peptide mapping.

Efecto de un agua rica en fibra y L-carnitina como adyuvante en una terapia de control de peso, sobre medidas antropométricas en pacientes con sobrepeso / Effect of a fiber and L-carnitina-enriched mineral water as adjuvant of a weight-control therapy on overweight patients

El sobrepeso y la obesidad son trastornos metabólicos caracterizados por una excesiva acumulación de energía en forma de grasa en el organismo, que conlleva un aumento del peso corporal con respecto al valor esperado según sexo, talla y edad y que están ligados a un gran número de patologías como diabetes tipo 2, dislipemia, hipertensión arterial, etc. En los últimos años la incidencia de estos trastornos ha aumentado de manera alarmante, llegando a ser, en España, del 50% en el año 2000 según los datos de la SEEDO. La relación directa entre la obesidad y el incremento del riesgo enfermedad hace que el consumidor demande tratamientos y productos, ya sean complementos alimenticios o fármacos, que le permitan superar esa situación y mejorar tanto su aspecto físico como su estado de salud, por lo que resulta de gran interés el desarrollo y evaluación de productos que junto con una modificación en la dieta y estilo de vida ayuden a conseguir una disminución en el IMC y una mejora en parámetros asociados al sobrepeso y obesidad. El objetivo de este estudio fue evaluar el efecto de uno de estos productos, un agua enriquecida con fibra y L-carnitina, como adyuvante en una terapia de control de peso. Se llevó a cabo un estudio aleatorizado, doble ciego, controlado con placebo en el que participaron 40 sujetos, hombres y mujeres, con sobrepeso a los que se les indicó el seguimiento de una dieta hipocalórica durante 56 días, complementada, en 20 de los voluntarios con la ingesta de 1 litro de Vitalis Elegante, mientras que en los otros 20 se complementó con la ingesta de 1 litro de agua. Se llevaron a cabo determinaciones de peso, IMC y medidas antropométricas. Ambos tratamientos resultaron eficaces en la reducción de diversas medidas antropométricas relacionadas con el sobrepeso, no encontrándose diferencias estadísticamente significativas entre ambos tratamientos en la evolución de ninguno de los parámetros estudiados, aunque sí se encontraron algunos resultados reseñables. Sólo el grupo que tomó Vitalis Elegante como adyuvante de su terapia de control de peso fue capaz de reducir de manera estadísticamente significativa el perímetro de cintura y el pliegue tricipital respecto a los valores iniciales, lo que no ocurrió en el grupo que al que se le administró placebo como adyuvante. Teniendo en cuenta estos datos, Vitalis Elegante podría contribuir a la mejora de las estrategias de control de peso que se utilizan en la actualidad (AU)

Overweight and obesity are metabolic disorders characterized by an excessive energy accumulation in the organism in the form of fat, which leads to an increase in body weight according to sex, age and height. These disorders are usually linked to different pathologies, such as diabetes, dyslipemia, hypertension, etc. In last years, the incidence of these disorders has reached 50 % of adult population in Spain, according to SEEDO data. This relationship between obesity and the increase of pathology risk has caused an increase in consumer’s demand of products or drugs that allow them to overcome this situation and improve not only their appearance but their health. In that context is interesting the development of products that, together with a modification in diet and life style, allow getting a decrease in BMI and an improvement in parameters associated to overweight and obesity. The aim of this study was to evaluate the effect of a fiber and L-carnitine enriched mineral water as adjuvant of a weight control therapy in overweight patients. A double-blind, placebo-controlled, randomized clinical trial was carried out, in which 40 overweight persons were enrolled. They were indicated to follow a hypocaloric diet during 56 days, supplemented, in one group, with 1liter per day of Vitalis Elegante, while the other group was supplemented with 1 liter per day of placebo. During the intervention weight, BMI and anthropometric measures were determined. Both, Vitalis Elegante and placebo were effective in the reduction of several measures related to overweight, not having found statistically significant differences between them as regards to evolution of parameters during the treatment. However, we found some interesting results. Only the group supplemented with Vitalis Elegante was able to decrease the waist perimeter and the tricipital fold in comparison with basal values, what did not happen in the group supplemented with placebo. Taken together, these data show that Vitalis Elegante could contribute to the improvement of weight control used at the moment (AU)

Targets of cytoprotection in acute ischemic stroke: present and future.

Although the management of stroke has improved remarkably over the last decade due mainly to the advent of thrombolysis, most neuroprotective agents, although successful in animal studies, have failed in humans. Our increasing knowledge concerning the ischemic cascade is leading to a considerable development of pharmacological tools suggesting that each step of this cascade might be a target for cytoprotection. Glutamate has long been recognized to play key roles in the pathophysiology of ischemia. However, although some trials are still ongoing, the results from several completed trials with drugs interfering with the glutamatergic pathway have been disappointing. Regarding the inhibition of glutamate release as a possible target for cytoprotection, it might be afforded either by decreasing glutamate efflux or by increasing glutamate uptake. In this context, it has been shown that glutamate transport is the primary and only mechanism for maintaining extracellular glutamate concentrations below excitotoxic levels. This transport is executed by the five high-affinity, sodium-dependent plasma membrane glutamate transporters. Among them, the transporter EAAT2 is responsible for up to 90% of all glutamate transport. We will discuss the effect of different neuroprotective tools (membrane stabilizers or endogenous neuroprotection) affecting glutamate efflux and/or expression of EAAT2. We will also describe the finding of a novel polymorphism in the EAAT2 promoter region which could be responsible for differences in both gene function and regulation under pathological conditions such as cerebral ischemia, and which might well account for the failure of glutamate antagonists in the clinical practice. These results may possess important therapeutic implications in the management of patients at risk of ischemic events, since it has been demonstrated that those patients with progressing stroke have higher plasma concentrations of glutamate which remain elevated up to 24 h when compared to the levels in patients without neurological deterioration.

TNFR1 mediates increased neuronal membrane EAAT3 expression after in vivo cerebral ischemic preconditioning.

A short ischemic event (ischemic preconditioning) can result in subsequent resistance to severe ischemic injury (ischemic tolerance). Glutamate is released after ischemia and produces cell death. It has been described that after ischemic preconditioning, the release of glutamate is reduced. We have shown that an in vitro model of ischemic preconditioning produces upregulation of glutamate transporters which mediates brain tolerance. We have now decided to investigate whether ischemic preconditioning-induced glutamate transporter upregulation takes also place in vivo, its cellular localization and the mechanisms by which this upregulation is controlled. A period of 10 min of temporary middle cerebral artery occlusion was used as a model of ischemic preconditioning in rat. EAAT1, EAAT2 and EAAT3 glutamate transporters were found in brain from control animals. Ischemic preconditioning produced an up-regulation of EAAT2 and EAAT3 but not of EAAT1 expression. Ischemic preconditioning-induced increase in EAAT3 expression was reduced by the TNF-alpha converting enzyme inhibitor BB1101. Intracerebral administration of either anti-TNF-alpha antibody or of a TNFR1 antisense oligodeoxynucleotide also inhibited ischemic preconditioning-induced EAAT3 up-regulation. Immunohistochemical studies suggest that, whereas the expression of EAAT3 is located in both neuronal cytoplasm and plasma membrane, ischemic preconditioning-induced up-regulation of EAAT3 is mainly localized at the plasma membrane level. In summary, these results demonstrate that in vivo ischemic preconditioning increases the expression of EAAT2 and EAAT3 glutamate transporters the upregulation of the latter being at least partly mediated by TNF-alpha converting enzyme/TNF-alpha/TNFR1 pathway.

Importancia de los índices periodontales en la evaluación de los implantes osteointegrados / Importance of periodontal indices in the evaluation of osseointegrated implants

El objetivo del estudio es conocer la relación existente entre la tasa de fracaso de una muestra de implantes recubiertos de hidroxiapatita (H.A.) y los parámetros periodontales del paciente; así como el grado de asociación entre estos últimos. La muestra está formada por 52 pacientes, a los que hemos colocado 158 implantes recubiertos de H.A. (Integral de Calcitek), que han sido revisados a los tres años de su colocación. El fracaso de los implantes se ha establecido según los criterios de SCHNITMANY SHULMAM de 1979.El estudio realizado es observacional analítico retrospectivo de seguimiento, a través del cual se evalúa la evolución clínica y radiológica de los implantes; resultando que la recesión periimplante medida en vestibular, lingual, mesial y distal de cada fijación es el único parámetro periodontal asociado al fracaso del implante (P= 0.008). La reabsorción ósea y la recesión periimplante se asocian de forma significativa (P < 0.05); así como la profundidad de sondaje lo hace con el índice de gingivitis en todas sus posiciones, y con el índice de placa en posición lingual. Hemos hallado una correlación positiva entre estos dos índices r=0.519, P<0.01 de forma que al aumentar uno lo hace el otro y viceversa. Podemos concluir que la aparición de recesión alrededor de un implante aumenta las probabilidades de que éste fracase. Existe además una correlación positiva entre el índice de gingivitis, de placa y la profundidad de bolsa, lo que indica la salud del surco gingival pero no las probabilidades de éxito del implante (AU)

The aim of this study was to determine, firstly,the relation between the failure of a sample of hydroxyapatite-coated implants (HA) and the periodontal parameters of the patient, and secondly, the degree of association between such parameters. The sample was comprised of 52 patients and 158 H.A. coated implants (Integral® by Calcitek), which were revised three years after their insertion. Implant failure was determined according to the criteria of Schnitman and Shulman (1979). A retrospective analytical observational follow-up study was carried out, by means of which the clinical and radiological evolution of the implants was determined. It was found that the peri-implant recession measured facial, lingual, mesial and distal from each implant is the only periodontal parameter associated with the failure of the implant (p=0.008). Bone resorption and periimplant recession were significantly associated (p<0.05),as was probe depth with the gingivitis index in all positions and with the plaque index in the lingual position. A positive correlation was found between the latter two indices (r=0.519; p<0.01). It may be concluded that the appearance of recession around an implant increases the probability of its failure. There was also a positive correlation between the gingivitis index, the plaque index and pocket depth, which is indica tive of the state of the gingival sulcus but not of the success rate of the implant (AU

[Evaluation of the DTP vaccination technique in infants]. / Evaluación de la técnica de vacunación con DTP en el lactante.

OBJECTIVE: To evaluate the techniques of DPT vaccination in the nursing child. DESIGN: An observation study of a crossover type. Evaluation by means of an anonymous survey of those responsible for administering the vaccinations. Statistical analysis using the precise Fisher test. PARTICIPANTS: The thirteen official vaccination centres in Health Areas 11 and 12 in the Community of Valencia. MEASUREMENTS AND MAIN RESULTS: 12 centres (91%) answered the questionnaire. Four of them (41%) used different needles to aspirate the contents of the vial and give the injection to the nursing child. The DPT was always administered in the gluteal region. 33% used needles which were 16 mm long. Prophylactic paracetamol was used in two of the Centres as a matter of course. The Centre's size or length of time in use did not affect the techniques used. CONCLUSIONS: There is no uniformity in the technique of administering the DPT vaccine to the nursing child in the different Vaccination Centres of Areas 11 and 12 in the Community of Valencia. The techniques used for vaccinations often differ from those recommended by groups of experts.