Pharmacokinetics of genistein distribution in blood and retinas of diabetic and non-diabetic rats.

Genistein, a natural tyrosine kinase inhibitor, may act as an intraocular antiangiogenic agent. Its therapeutical use, however, is limited by its nonlinear pharmacokinetics. We aimed to determine genistein's kinetics and retinal tissue distributions in normal and diabetic rats. We developed an isocratic, reverse-phase C18 HPLC system to measure genistein concentration in blood and retinas of streptozotocin (65 mg/kg IV)-diabetic and non-diabetic rats receiving two types of genistein-rich diet (150 and 300 mg/kg) for ten days. Genistein's decay exhibited a two-compartmental open model. Half-lives of distribution and elimination were 2.09 and 71.79 min, with no difference between groups. Genistein steady-state concentration in blood for 150 and 300 mg/kg diet did not differ between diabetic (0.259 ± 0.07 and 0.26 ± 0.06 µg/ml) and non-diabetic rats (0.192 ± 0.05 and 0.183 ± 0.09 µg/ml). In retina, genistein concentration was significantly higher in diabetic rats (1.05 ± 0.47 and 0.997 ± 0.47 µg/gm wt. vs. 0.087 ± 0.11 and 0.314 ± 0.18 µg/gm wt., p < 0.05). The study determined that increasing genistein dose did not change its bioavailability, perhaps due to the poor aqueous solubility. The retina's increased genistein could be due to increased permeability of blood-retinal barrier that occurs early in diabetes.

Role of observational studies in supporting extrapolation of efficacy data from adults to children with epilepsy - A systematic review of the literature using lacosamide as an example.

Extrapolation of efficacy data from adults to children is accepted for focal epilepsy - the antiepileptic drug, lacosamide, has been approved for the treatment of children ≥4 years of age on this basis. Since many small-scale, open-label studies are reported in the literature before approval, a systematic review was conducted to ascertain whether results of these could be used to support extrapolation in epilepsy in the future. In the absence of randomised trials, a second analysis was conducted for reports on lacosamide use in adults with generalized epilepsies. Twenty-seven articles were included in the paediatric qualitative synthesis, and 14 in the adult. Paediatric studies were analysed separately based on seizure type: focal, generalised and mixed. In focal epilepsy, safety and seizure-related findings mirrored those observed in the adult Phase II/III trials, supporting the feasibility of data extrapolation. Few studies reported outcomes in children with epilepsies associated with generalised seizures, and those that included children with different seizure types, mostly did not provide results separately. Lacosamide treatment appeared beneficial for children and adults experiencing tonic-clonic and myoclonic seizures. Reports of seizure aggravation were inconsistent and, in many cases, could not be clearly attributed to lacosamide. Given the absence of sufficient data, evidence for the feasibility of extrapolation was not as clear-cut as it was in focal epilepsy. These results highlight the complexities of conducting trials in the generalised epilepsy setting, and the importance of studies in the real-life setting and of analysing efficacy data per generalized seizure type and syndrome.

SURF-COPD: the recording of cardiovascular risk in patients with chronic lung disease.

BACKGROUND: Cardiovascular disease (CVD) is the leading cause of death in patients with chronic obstructive pulmonary disease (COPD) and such patients tend to carry a heavy burden of risk factors for CVD. There is little information on the documentation and management of CVD risk factors in COPD patients. AIM: To audit the recording and management of CVD risk factors in 200 COPD patients attending the respiratory out-patient clinics at two University teaching hospitals. DESIGN: Retrospective analysis of hospital records. RESULTS: While there was reasonable recording of a history of CVD risk factors such as hypertension and hyperlipidaemia, the actual recording of risk factor levels was poor-blood pressure and heart rate were recorded in less than half of subjects; lipids, waist circumference, HbA1c and height were essentially not recorded at all. CONCLUSIONS: Best practice guidelines and standard operating procedures for CVD risk factor recording and control require to be developed if the major cause of death in COPD subjects, CVD, is to be addressed. An outline for a standard operating procedure is presented in the Appendix. Serial audits will be required to assess the efficacy of such measures.

Harmonisation of ethics committees' practice in 10 European countries.

BACKGROUND: The Directive 2001/20/EC was an important first step towards consistency in the requirements and processes for clinical trials across Europe. However, by applying the same rules to all types of drug trials and transposing the Directive's principles into pre-existing national legislations, the Directive somewhat failed to meet its facilitation and harmonization targets. In the field of ethics, the Directive 2001/20/EC conditioned the way of understanding and transposing the "single opinion" process in each country. This led to a situation in which two models of research ethics committees organisation systems exist, being the model in which the "single opinion" is considered to be the decision made by a single ethics committee more effective and simpler in terms of administrative and logistic workload. METHOD: A survey was conducted in 10 European countries. Members of the European Clinical Research Infrastructures Network working party number 1, with expertise in the field of ethics, responded. RESULTS: There is a major heterogeneity in the composition of ethics committees among the surveyed countries based on the number of members, proportion of experts versus lay members and expertise of the scientific members. A harmonized education of the ethics committees' membership based in common curricula is recommended by the majority of countries. CONCLUSIONS: Despite the efforts for harmonization of the European Clinical Trial Directive, from an ethical point of view, there remains a plurality of ethics committees' systems in Europe. It is important to comprehend the individual national systems to understand the problems they are facing.

Dyslipidemia therapy update: the importance of full lipid profile assessment.

Lipid guidelines typically focus on total cholesterol +/- low-density lipoprotein cholesterol levels with less emphasis on high-density lipoprotein cholesterol (HDL-C) or triglyceride assessment, thus potentially underestimating cardiovascular (CV) risk and the need for lifestyle or treatment optimization. In this article, we highlight how reliance on isolated total cholesterol assessment may miss prognostically relevant lipid abnormalities; we describe from the European Systematic COronary Risk Evaluation (SCORE) data set how incorporation of HDL-C may improve estimation of CV risk; and, finally, we critically evaluate the evidence base surrounding triglycerides and CV risk.

EUROASPIRE (European Action on Secondary Prevention through Intervention to Reduce Events) III--a comparison of Irish and European results.

The EUROASPIRE III audit was a Europe-wide study which took place in 2006/2007. The objective was to examine the control of risk factors in subjects with established cardiovascular disease. Here, we compare the Irish results to those of the other 21 European countries which participated. Control of blood cholesterol was significantly better in Irish participants, with 73% below the target of 4.5 mmol/l. Blood pressure control was less satisfactory in both Irish and European individuals, with an average of 52% of Irish participants not achieving blood pressure targets. Medication usage was high throughout, particularly anti-platelet agents, beta-blockers and, especially in Ireland, statins. Obesity figures were particularly high in Ireland and throughout Europe, with 82% Irish men and women either overweight or obese. Smoking figures in Irish women were also of concern, with 24% continuing to smoke. Cardiac rehabilitation attendance was particularly high in Ireland, with 68% attending; substantially higher than the European figure of 34%. In common with the rest of Europe, current control of body weight and blood pressure in Ireland is unsatisfactory and in need of increased consideration on the part of both patients and healthcare professionals.

HDL cholesterol protects against cardiovascular disease in both genders, at all ages and at all levels of risk.

OBJECTIVE: We aimed to clarify some previous inconsistencies regarding the role of high density lipoprotein cholesterol (HDL-C) as a CVD protective factor. METHODS: The SCORE dataset contained data on HDL-C for 104,961 individuals (45% women) without pre-existing coronary heart disease (CHD). These were from 7 pooled European prospective studies. The effect of HDL-C, both in quintiles and as a continuous variable, on risk of CVD and CHD mortality was examined, using Cox proportional hazards model, adjusted for age, total cholesterol, systolic blood pressure, smoking, diabetes and body mass index and stratified by gender, age group, country and category of SCORE CVD risk. RESULTS: A strong, graded, independent, inverse relationship between HDL-C and both CVD and CHD mortality was demonstrated. Adjusted hazard ratios per 0.5mmol/l increase in HDL-C were 0.60 (0.51, 0.69) and 0.76 (0.70, 0.83) in women and men, respectively for the CVD mortality endpoint. The corresponding hazard ratios were 0.53 (0.42, 0.68) and 0.79 (0.64, 0.98) in elderly women and men, respectively. The relationship was significant in all SCORE CVD risk strata and age groups. CONCLUSIONS: This multivariable analysis, the largest of its kind to date, has confirmed the inverse, independent, strong and graded relationship between HDL-C and both CVD and CHD mortality. We have clarified previous suggestions that the relationship is stronger in women and that it applies in all age groups. This is the first prospective study to demonstrate the independent relationship specifically in healthy elderly women and to show that the relationship holds at all levels of total CVD risk.

Analysis of the expression of putatively imprinted genes in bovine peri-implantation embryos.

The application of assisted reproductive technologies (ART) has been shown to induce changes in the methylation of the embryonic genome, leading to aberrant gene expression, including that of imprinted genes. Aberrant methylation and gene expression has been linked to the large offspring syndrome (LOS) in bovine embryos resulting in increased embryonic morbidity and mortality. In the bovine, limited numbers of imprinted genes have been studied and studies have primarily been restricted to pre-implantation stages. This study reports original data on the expression pattern of 8 putatively imprinted genes (Ata3, Dlk1, Gnas, Grb10, Magel2, Mest-1, Ndn and Sgce) in bovine peri-implantation embryos. Two embryonic developmental stages were examined, Day 14 and Day 21. The gene expression pattern of single embryos was recorded for in vivo, in vitro produced (IVP) and parthenogenetic embryos. The IVP embryos allow us to estimate the effect of in vitro procedures and the analysis of parthenogenetic embryos provides provisional information on maternal genomic imprinting. Among the 8 genes investigated, only Mest-1 showed differential expression in Day 21 parthenogenetic embryos compared to in vivo and IVP counterparts, indicating maternal imprinting of this gene. In addition, our expression analysis of single embryos revealed a more heterogeneous gene expression in IVP than in in vivo developed embryos, adding further to the hypothesis of transcriptional dysregulation induced by in vitro procedures, either by in vitro maturation, fertilization or culture. In conclusion, effects of genomic imprinting and of in vitro procedures for embryo production may influence the success of bovine embryo implantation.

Audit of the effectiveness of command and control arrangements for medical evacuation of seriously ill or injured casualties in southern Afghanistan 2007.

AIMS: The effectiveness of the command and control of medical evacuation by helicopter (MEDEVAC) of casualties sustained in southern Afghanistan each month from 1 May to 31 July 2007 was audited. In this period 762 casualties of all categories were evacuated to International Security Assistance Force (ISAF) field hospital facilities under the direction of Operations and medical staff of NATO Regional Command (South) (RC-S). The criterion for the audit was the time taken from notification in the RC-S Combined Joint Operations Centre (CJOC) until the helicopter landed ("Wheels Down") at the destination field hospital's helicopter landing site. The standard to be met was 90 minutes for all "9-liner" Category A (URGENT) and Category B (URGENT - surgical) cases (in hospital within 2 hours of wounding) allowing for time from injury to first notification in the CJOC, and time from landing to transfer to the Emergency Department (30 minutes together) at the designated destination hospital. Those that did not meet this target were assessed in order to review their outcome and to identify means for improving performance. RESULTS: Analysis of evacuation times for all missions each month from May to July revealed that three quarters of A and B category missions met the 90 minute target. No adverse outcome resulted from those which did not meet this target, reasons for which included distance (more than 30 minutes flying time each way), delay in securing a hostile landing site, delay in obtaining sufficient information, incorrect categorization of the casualty's priority, and on one occasion, an overmatch of assets available at that time. No casualties died who were recoverable. Comparison with data from the two previous RC-S rotations (prior to 1 May 07) showed an improvement in mean response time, but little change in median response on the rotation of RC-S staff on 1 May 07. The major change that had occurred on this rotation was to move the medical operations staff into the CJOC. The convergence of median and mean at this time indicates a reduction in "outliers", providing evidence that collocation of medical and operations staff improves incident response and should be the "default setting" in deployed tactical formation headquarters. CONCLUSION: Regular audit of MEDEVAC response should be routine for Medical Operations staff, in order to ensure the optimal casualty care pathway from point of wounding to field hospital.

An evaluation of the taxi service available to wheelchair users.

Adverse accounts of wheelchair user's experience of the taxi service have been reported. It is not clear whether these experiences are "once off" or reflect those of the user group. Wheelchair users living in cities throughout Ireland were surveyed. Of 306 questionnaires a response rate of 40.3% was achieved. Nearly 50% felt they had been refused a taxi because of their condition. Fifty one percent felt inadequately secured in a taxi with 35% rating driver's knowledge of handling as poor or very poor. Eight people (7%) had been injured as a result of unsafe handling by the taxi driver. Thirty percent said the modifications to the vehicles were inadequate and 45% said further modifications were needed. The results suggest that wheelchair users do not have access to a reliable, safe taxi service. The most important finding of the study was that that taxi driver's lack knowledge of how to handle and secure wheelchairs and people are being injured as a result. This study illustrates two things; the need to educate taxi drivers and the need to revisit the specifications for wheelchair accessible taxis to ensure modifications are adequate for users needs.

Time to pregnancy and multiple births.

BACKGROUND: Mothers of multiples are alleged to be more fecund than mothers of singletons. Some authors have suggested monitoring twinning rates for assessing temporal changes in a population's reproductive health. METHODS: Using a nested case-control design, we estimated the odds of a multiple birth in relation to fecundity in the US Collaborative Perinatal Project inclusive of 8546 pregnant women who reported a known time-to-pregnancy (TTP) upon enrolment in the cohort, 1959-1966. Case mothers comprised 81 women giving birth to twins/triplets; control mothers comprised 243 women giving birth to singletons matched to case mothers on maternal age at a ratio of 3:1. The odds ratio (OR) for a multiple birth within 6 months of trying adjusting for maternal age and prior pregnancies was estimated using logistic regression. Discrete time Cox regression analysis was also utilized to estimate the fecundability OR. RESULTS: Women with a TTP of 6 months [OR=1.95; 95% confidence interval (95% CI)=1.09-3.51]. Excluding pregnancies after 13+ months resulted in a loss of precision (OR=2.14; 95% CI=0.90-5.04). CONCLUSIONS: These data support higher fecundity among mothers of multiples than mothers of singletons.

Birth of a cloned calf derived from a vitrified hand-made cloned embryo.

The hand-made cloning (HMC) technique describes a simplified nuclear transfer process without the need for micromanipulators. The technique involves manual bisection of zona-free oocytes, selection of cytoplasts by Hoechst staining and fusion of a single somatic cell and two cytoplasts. In this proof-of-principle experiment, the objective was to examine the developmental competence of HMC embryos following embryo transfer. Modifications to the original method include not selecting of matured oocytes and simultaneous fusion of cytoplasts and karyoplast. Blastocyst rates for embryos cultured in the glass oviduct system as singles (10.5%; 24/228) or in pairs (16.1%; 36/224) did not differ significantly. Fresh and vitrified-thawed blastocysts were transferred to 16 synchronised recipients (three to four embryos per recipient). Ultrasound examination on Days 35-45 showed an initial pregnancy rate of 43.8% (7/16) and a pregnancy rate >8 months of 12.5% (2/16). A male cloned calf (42 kg) derived from a vitrified HMC blastocyst was delivered by Caesarean section on Day 271. The birth and ongoing survival (15 months, 243 kg) of a healthy and apparently normal calf, combining both HMC and vitrification technologies, provides a 'proof of principle' of the technology and a promising alternative to traditional nuclear-transfer techniques.

Industrial control of recombinant E. coli fed-batch culture: new perspectives on traditional controlled variables.

Three industrially relevant fed-batch algorithms were applied to the production of recombinant proteins in Escherichia coli BL21 DE3. Starvation dissolved-oxygen (DO)-transient control sustained growth rates greater than those in pH-stat (0.16 h(-1) versus 0.11-0.13 h(-1)) while feed-up DO-transient control better tracked the measured threshold for acetate production (mu approximately 0.2 h(-1)). All controllers supported growth without acetate production, resulting in end concentrations of recombinant protein up to 20 times greater than in batch culture. Both DO-transient control systems were judged superior to pH-stat for their ability to detect and track the acetate threshold. Results also showed that although high cell density at the stationary phase is desirable, this parameter may be dictated by the choice of media and reactor design as opposed to controller type. Controller selection, however, has a great impact on the capacity to track the acetate threshold and therefore to enhance productivity without concomitant acetate production.

Normalization of retinal vascular permeability in experimental diabetes with genistein.

PURPOSE: To study the effects of genistein, a tyrosine kinase inhibitor, on retinal vascular permeability in an experimental diabetic rat model. METHODS: Seventy-two rats were equally divided into four groups: (1) nondiabetic control group, (2) diabetic control group, (3) diabetic rats receiving 150 mg genistein/kg food, and (4) diabetic rats receiving 300 mg genistein/kg food. Diabetes was induced by streptozotocin injection in the three diabetic groups. Rats were fed diets with or without genistein and followed for 6 months. Retinal vascular permeability was assessed by measuring radiolabeled sucrose leakage into the retina and by Western blot analysis for total retinal albumin. Retinal phosphotyrosine levels and proliferating cell nuclear antigen (PCNA) were also evaluated by Western blot analysis. RESULTS: Diabetic control rats had markedly increased retinal vascular leakage of radiolabeled sucrose compared with nondiabetic control rats. Diabetic rats receiving oral genistein had significantly less retinal vascular leakage of radiolabeled sucrose than diabetic control rats in a dose-response fashion. Diabetic control rats had increased levels of phosphotyrosine, retinal albumin, and PCNA by Western blot analysis compared with nondiabetic control rats. Rats receiving 300 mg of genistein had decreased retinal albumin by Western blot analysis. Western blot analysis demonstrated a dose-response decrease in retinal phosphotyrosine levels and PCNA in genistein-treated diabetic rats compared with diabetic control rats. CONCLUSIONS: Long-term oral administration of genistein significantly inhibits retinal vascular leakage in experimentally induced diabetic rats. Tyrosine kinase inhibition may be a useful pharmacological approach for the treatment of diabetic-induced retinal vascular leakage.

Characterisation and phylogenetic analysis of the VP7 proteins of serotype G6 and G8 human rotaviruses.

Serotype G6 and G8 rotaviruses are rarely found in man and may have originated in animals. Human serotype G6 and G8 rotaviruses, isolated from hospitalised children at various locations in Australia, were characterised. Deduced amino acid sequences of the major neutralising antigen, V7, showed significant identity to the cognate proteins of prototype human and bovine G6 and G8 viruses, respectively, and the strains reacted with G6 and G8 serotype-specific neutralising monoclonal antibodies, respectively, in an enzyme immunoassay. The VP4 type was determined as P[14] for all strains tested. Phylogenetic analysis of these and other human and bovine VP7 sequences suggested that a single inter-species transmission event, possibly from cattle, may have led to the emergence of G6 viruses in man. In contrast, the exchange of genes between human and bovine G8 viruses may have occurred onmore than one occasion, or these genes may have originated in a different host.

Dual-test monitoring of hyperglycemia using daily glucose and weekly fructosamine values.

The purpose of this study was to assess the impact of using a dual-test blood glucose/fructosamine home monitoring system to assist individuals identified as having the potential for poor glycemic control to achieve values closer to normal. Forty-eight subjects found to have a fasting blood glucose value of > or = 126 mg/dL, casual blood glucose value of > or = 140 mg/dL, and/or blood fructosamine value of > or = 310 micromol/L, agreed to perform daily self testing for 90 days and were provided a dual-test blood glucose/fructosamine home monitoring system and testing supplies at no charge to them. Medication changes/compliance along with dietary and exercise habits were compared to testing results by the principle investigator at approximate 30-day intervals. The desired goal of this project was to achieve and/or maintain a fasting blood glucose value of < or = 110 mg/dL, a casual blood glucose value of < or = 140 mg/dL and a blood fructosamine value of < or = 310 micromol/L by encouraging each individual to realize the effect of dietary intake and exercise habits, and understand the importance of medication compliance, if appropriate, in achieving better overall glycemic control. Four subjects withdrew from the study prior to completion, 11 of the remaining 44 completed 60 days of testing and 33 of 44 completed 90 days of testing. Regular monitoring and counseling achieved an average reduction in blood glucose of 27.5% and a 16.6% reduction in average blood fructosamine when compared to original screening results of these 44 individuals. This study indicates that the addition of weekly fructosamine values to daily blood glucose values provides both the patient and clinician valuable information to evaluate the impact of dietary, exercise, and medication therapy changes on glycemic control by bridging the existing gap between daily blood glucose values and quarterly HbA1c confirmation of intervention results.

A study of the knowledge that school rugby coaches have in the management and prevention of serious neck injury.

The following study is an analysis of the knowledge school rugby coaches responsible for Senior Cup Team in Leinster, have in the area of neck injury prevention and management. When serious neck injuries affect schoolboys it is particularly tragic and deserves special attention. The study targeted all the coaches of Senior Cup Teams in Leinster. Results showed that coaches lack vital knowledge in the area of neck injury prevention, recognition and management. Only 50% (n = 18) of coaches have a first aid qualification and only 47% (n = 17) carry first aid equipment to deal with neck injuries to matches. More than half of the schools in the study sample do not have neck collars available and a staggering 83% (n = 30) of schools do not have a stretcher available to SCT players. The study highlights the need to provide better first aid facilities in schools and demonstrates the need for further education of school rugby coaches.

Flash vacuum pyrolysis of methoxy-substituted lignin model compounds.

The flash vacuum pyrolysis (FVP) of methoxy-substituted beta-O-4 lignin model compounds has been studied at 500 degrees C to provide mechanistic insight into the primary reaction pathways that occur under conditions of fast pyrolysis. FVP of PhCH(2)CH(2)OPh (PPE), a model of the dominant beta-O-4 linkage in lignin, proceeds by C-O and C-C cleavage, in a 37:1 ratio, to produce styrene plus phenol as the dominant products and minor amounts of toluene, bibenzyl, and benzaldehyde. From the deuterium isotope effect in the FVP of PhCD(2)CH(2)OPh, it was shown that C-O cleavage occurs by homolysis and by 1,2-elimination in a ratio of 1.4:1, respectively. Methoxy substituents enhance the homolysis of the beta-O-4 linkage, relative to PPE, in o-CH(3)O-C(6)H(4)OCH(2)CH(2)Ph (o-CH(3)O-PPE) and (o-CH(3)O)(2)-C(6)H(3)OCH(2)CH(2)Ph ((o-CH(3)O)(2)-PPE) by a factor of 7.4 and 21, respectively. The methoxy-substituted phenoxy radicals undergo a complex series of reactions, which are dominated by 1,5-, 1,6-, and 1,4-intramolecular hydrogen abstraction, rearrangement, and beta-scission reactions. In the FVP of o-CH(3)O-PPE, the dominant product, salicylaldehyde, forms from the methoxyphenoxy radical by a 1,5-hydrogen shift to form 2-hydroxyphenoxymethyl radical, 1,2-phenyl shift, and beta-scission of a hydrogen atom. The 2-hydroxyphenoxymethyl radical can also cleave to form formaldehyde and phenol in which the ratio of 1, 2-phenyl shift to beta-scission is ca. 4:1. In the FVP of o-CH(3)O-PPE and (o-CH(3)O)(2)-PPE, products (ca. 20 mol %) are also formed by C-O homolysis of the methoxy group. The resulting phenoxy radicals undergo 1,5- and 1,6-hydrogen shifts in a ratio of ca. 2:1 to the aliphatic or benzylic carbon, respectively, of the phenethyl chain. In the FVP of (o-CH(3)O)(2)-PPE, o-cresol was the dominant product. It was formed by decomposition of 2-hydroxy-3-hydroxymethylbenzaldehyde and 2-hydroxybenzyl alcohol, which are formed from a complex series of reactions from the 2, 6-dimethoxyphenoxy radical. The key step in this reaction sequence was the rapid 1,5-hydrogen shift from 2-hydroxy-3-methoxybenzyloxy radical to 2-hydroxymethyl-6-methoxyphenoxy radical before beta-scission of a hydrogen atom to give the substituted benzaldehyde. The 2-hydroxybenzyl alcohols rapidly decompose under the reaction conditions to o-benzoquinone methide and pick up hydrogen from the reactor walls to form o-cresol.