Trimethyllysine,vascular risk factors and outcome in acute ischemic stroke (MARK–STROKE)

Trimethyllysine (TML) is involved in the generation of the pro-atherogenic metabolite trimethylamine-N-oxide (TMAO) by gut microbiota. In clinical studies, elevated TML levels predicted major adverse cardiovascular events (MACE) in patients with acute or stable coronary artery disease (CAD). In contrast to cardiovascular patients, the role of TML in patients with acute cerebral ischemia is unknown. Here, we evaluated circulating TML levels in 374 stroke patients from the prospective biomarkers in stroke (MARK–STROKE) study. Compared with 167 matched healthy controls, acute ischemic stroke patients had lower median TML plasma concentrations, i.e. 0.71 vs. 0.47 µmol/L (p < 0.001) and this difference persisted after adjusting for age and sex. TML plasma concentrations were associated with age, serum creatinine, glucose, cholesterol and lysine. Patients with prevalent arterial hypertension, atrial fibrillation or a history of myocardial infarction had increased TML levels, but this observation was not independent of age, sex and GFR. In 274 patients, follow-up data were available. During a median follow-up of 284 [25th–75th percentile: 198, 431] days, TML was not associated with incident MACE (stroke, myocardial infarction, death). In summary, our data suggests a different role of TML in acute ischemic stroke compared with CAD patients.

     

Blood biomarkers role in acute ischemic stroke patients: higher is worse or better?

Background

Thrombolytic therapy (TT) for acute ischemic stroke (AIS) can provoke bleeding’s complication depending on the ischemic lesion (IL) dimension. Inflammation involved in the setting of acute ischaemic stroke, is associated with infarct size. We aimed to study the independent correlation and association between clinical panel of routinely identified biomarkers, including inflammatory parameters, and cerebral IL dimension and site.

Results

We evaluated eleven biomarkers in 105 unrelated patients during their hospitalization after acute stroke event. Our data indicate a significant association of: a) confluent IL size with 4th quartile of Erythrocyte Sedimentation Rate (ESR) (OR = 5.250; 95% CI, 1.002 to 27.514) and an independent correlation with sex; b) confluent IL size with 3rd quartile of fibrinogen (OR = 5.5; 95% CI, 1.027 to 29.451); c) confluent IL size with 3rd quartile of platelets (OR= 0.059; 95% CI, 0.003 to 1.175) and independent correlation with sex; d) smaller IL size (OR = 5.25; 95% CI, 1.351 to 20.396) with 3rd quartile of albumin levels and nodular and parenchimal IL size with 2nd (OR = 0.227; 95% CI, 0.053 to 0.981), 3rd (OR = 0.164; 95% CI, 0.038 to 0.711) and 4th (OR = 0.205; 95% CI, 0.048 to 0.870) quartiles albumin levels; e) smaller IL size with 3rd quartile triglycerides (TG) levels (OR = 9; 95% CI, 2.487 to 32.567) and an independent correlation with anterior location. Smaller IL size, anterior AIS turned out to be independently correlated with high serum albumin levels. Finally, high INR and PTT values were associated with worse NIHSS clinical outcomes in contrast to that observed with higher albumin level.

Conclusions

We provide evidence of routine biomarkers levels correlation with acute IL size, independently of age and sex. In addition, we highlight the importance of differentiation of biomarkers normal interval levels for further improvement not only of the clinical decision making but also in post-acute clinical outcome management.

     

Iodine in early pregnancy--is there enough?

Aims: To evaluate the iodine status of patients in early pregnancy and its dependence on level of thyroid-stimulating hormone (TSH). Methods: Between June 2005 and December 2006, 168 patients with a confirmed vital pregnancy (up to 10(th) week of pregnancy) were included in the study. The entry criteria were no prior thyroid disease, did not take any other medication, had not undergone radio-iodine therapy and did not take multivitamins containing iodine. The iodine status was measured as the amount of iodine in urine over 24 hours. The TSH level was determined from the blood using chemiluminescence. Results: The average ioduria value in patients was found to be 3.04 micromol/24 hr, with the norm 0.6-2.4 micromol/ 24 hr, median 2.9, SD 1.5. None of the patients had a value lower than 0.9 micromol/24 hr. The average TSH value was 1.98 mIU/l, median was 1.31, SD 0.98. The laboratory limits were set to 0.25-3 mIU/l for pregnant women in the first trimester. Three pregnancies ended in miscarriage by week 12, 1 miscarriage occurred in week 22 and the other pregnancies concluded in delivery between weeks 38-41. Fourteen patients had TSH levels above 3 mIU/l with normal levels od free thyroxine (T4) : 10.3-25 pmol/l. Conclusions: The results of this study did not reveal any iodine deficit in any of the patients. However 14 patients had elevated TSH levels signalling subclinical or incipiently clinical hypothyroidism. These pacients underwent levothyroxine therapy after endocrinologist's consultations.     

Admission C – reactive protein after acute ischemic stroke is associated with stroke severity and mortality: The 'Bergen stroke study'

Background

There is growing evidence that inflammation plays an important role in atherogenesis. Previous studies show that C-reactive protein (CRP), an inflammatory marker, is associated with stroke outcomes and future vascular events. It is not clear whether this is due a direct dose-response effect or rather an epiphenomenon. We studied the effect of CRP measured within 24 hours after stroke onset on functional outcome, mortality and future vascular events.

Methods

We prospectively studied 498 patients with ischemic stroke who were admitted within 24 hours after the onset of symptoms. CRP and NIH stroke scale (NIHSS) were measured at the time of admission. Short-term functional outcome was measured by modified Rankin scale (mRS) and Barthel ADL index (BI) 7 days after admission. Patients were followed for up to 2.5 years for long-term mortality and future vascular events data.

Results

The median CRP at admission was 3 mg/L. High CRP was associated with high NIHSS (p = 0.01) and high long-term mortality (p < 0.0001). After adjusting for confounding variables, high CRP remained to be associated with high NIHSS (p = 0.02) and high long-term mortality (p = 0.002). High CRP was associated with poor short-term functional outcomes (mRS > 3; BI < 95) (p = 0.01; p = 0.03). However, the association was not significant after adjusting for confounding variables including stroke severity (p = 0.98; p = 0.88). High CRP was not associated with future vascular events (p = 0.98).

Conclusion

Admission CRP is associated with stroke severity and long-term mortality when measured at least 24 hours after onset. There is a crude association between high CRP and short-term functional outcome which is likely secondary to stroke severity. CRP is an independent predictor of long-term mortality after ischemic stroke.     

Microemboli-monitoring during the acute phase of ischemic stroke: Is it worth the time?

Background  

The prevalence of microembolic signals (MES) during the acute phase of ischemic stroke and its influence on outcome is not well studied. The aim of our study was to determine the prevalence of MES, the different factors that are associated with the presence of MES and the association between MES and outcomes in stroke patients investigated within 6 hours after the onset of ischemic stroke.     

Is there an effect of ischemic conditioning on myocardial contractile function following acute myocardial ischemia/reperfusion injury?

Ischemic conditioning induces cardioprotection; the final infarct size following a myocardial ischemic event is reduced. However, whether ischemic conditioning has long-term beneficial effects on myocardial contractile function following such an ischemic event needs further elucidation. To date, ex vivo studies have shown that ischemic conditioning improves the contractile recovery of isolated ventricular papillary muscle or atrial trabeculae following simulated ischemia. However, in vivo animal studies and studies in patients undergoing elective cardiac surgery show conflicting results. At the subcellular level, it is known that ischemic conditioning improved energy metabolism, preserved mitochondrial respiration, ATP production, and Ca²⁺ homeostasis in isolated mitochondria from the myocardium. Ischemic conditioning also presents with post-translational modifications of proteins in the contractile machinery of the myocardium. The beneficial effects on myocardial contractile function need further elucidation. This article is part of a Special Issue entitled: The power of metabolism: Linking energy supply and demand to contractile function edited by Torsten Doenst, Michael Schwarzer and Christine Des Rosiers.     

Do nasogastric tubes worsen dysphagia in patients with acute stroke?

Background  

Early feeding via a nasogastric tube (NGT) is recommended as safe way of supplying nutrition in patients with acute dysphagic stroke. However, preliminary evidence suggests that NGTs themselves may interfere with swallowing physiology. In the present study we therefore investigated the impact of NGTs on swallowing function in acute stroke patients.     

Why is there persistent disease despite biologic therapy? Importance of early intervention

This short article hypothesizes that the major reason for persistent disease despite biologic therapy is the inappropriately late timing of therapy with biologic agents. There is clear evidence to support this hypothesis. This short review will indicate that patients treated at an earlier phase of disease can achieve a clinical remission rate of 70% and a response rate of above 95%.The majority of studies of rheumatoid arthritis (RA) have shown that when patients with RA are treated with biologics, they achieve a remission rate of only 30% or less. There are many potential reasons why disease activity is incompletely suppressed by biologics. The most commonly used biologics are anti-cytokines, in particular those that block TNF alpha. Therefore, a logical reason for persistent disease activity is either incomplete blockade of the individual cytokine or the fact that multiple cytokines/alternative mechanisms are implicated in the pathogenesis. A pragmatic second reason is that the outcome measures used to measure response include elements other than disease activity/inflammation. Even when inflammation is completely switched off, these outcome measures may not normalize. An example of this is the reduced impact of inflammation-suppressive therapy on outcome measures in later disease, at a time when there is already extensive damage.This short article, representing a personal view, will examine the hypothesis that a major reason there is persistent disease activity is that anti-cytokine biologics are used inappropriately late in the disease. In patients with late disease, the disease activity score (DAS) reflects damage as well as inflammation and therapy has a large irreversible component. More controversially, the pathogenesis of the disease by this time may have evolved, so that the disease process is less reversible with the blockade of a single cytokine. For both of these reasons, TNF blockade will produce full benefits only when given early in the course of disease.     

Is there a role for auxin in early embryogenesis?

The very young embryo of a flowering plant is not an idealsystem in which to study the effects of auxin. Conversely, auxin isusually not considered as a major component of developmental processesin early embryogenesis. However, recent findings from both experimentalstudies in Brassica and analyses of developmental mutants inArabidopsis make it worthwhile to examine critically thepossibility that auxin may have a role in early embryogenesis. In thisreview, we will focus on specific processes, such as formation of anapical-basal axis of polarity and the initiation of the primary rootmeristem. To provide a conceptual framework in which to discuss possibleeffects of auxin, we will first briefly summarise essential features ofearly embryogenesis in Arabidopsis. This will be followed by anevaluation of relevant data suggesting a role for auxin in axisformation and root meristem initiation. Finally, we will discuss a fewexperimental approaches that we believe are necessary to examine whetheror not auxin plays a role in fundamental processes of earlyembryogenesis.     

Does circadian and seasonal variation in occurrence of acute aortic dissection influence in-hospital outcomes?

The risk of acute aortic dissection (AAD) exhibits chronobiological variations with peak onset in the morning and in winter. However, it is not known whether the time of day or season of the year of the AAD affects clinical outcomes. We studied 1,032 patients enrolled in the International Registry of Acute Aortic Dissection from January 1997 to December 2001. For circadian and seasonal analysis, the time and date of symptom onset were available for 741 and 1,007 patients, respectively, and were grouped into four 6 h periods (morning, afternoon, evening, and night) and four seasons (winter, spring, summer, and autumn). The χ² test for goodness of fit was used to evaluate non-uniformity of the time of day and time of year for critical in-hospital clinical events, including death. While highest incidence of AAD occurred in the morning and winter, clinical events (including mortality) were similar during the four different periods of the 24 h (χ²=1.9, p=0.60) and seasonal (χ²=1.2, p=0.75) periods.     

Can environmental conditions experienced in early life influence future generations?

The consequences of early developmental conditions for performance in later life are now subjected to convergent interest from many different biological sub-disciplines. However, striking data, largely from the biomedical literature, show that environmental effects experienced even before conception can be transmissible to subsequent generations. Here, we review the growing evidence from natural systems for these cross-generational effects of early life conditions, showing that they can be generated by diverse environmental stressors, affect offspring in many ways and can be transmitted directly or indirectly by both parental lines for several generations. In doing so, we emphasize why early life might be so sensitive to the transmission of environmentally induced effects across generations. We also summarize recent theoretical advancements within the field of developmental plasticity, and discuss how parents might assemble different ‘internal’ and ‘external’ cues, even from the earliest stages of life, to instruct their investment decisions in offspring. In doing so, we provide a preliminary framework within the context of adaptive plasticity for understanding inter-generational phenomena that arise from early life conditions.     

Does storage time in LN2 influence survival and pregnancy outcome of vitrified rabbit embryos?

Vitrification is one of the most widely used techniques for embryo cryopreservation. The aim of this work was to study the effect of storage time in liquid nitrogen on vitrified rabbit embryos. A total of 1467 vitrified rabbit embryos were transferred into 174 females. The embryos had been maintained in liquid nitrogen during 3 different periods, A) < 1 year (98 transfers, 827 embryos); B) 2-5 years (44 transfers, 360 embryos) and C) > 15 years (32 transfers, 280 embryos). A generalized linear model was used to determine the effect of period on pregnancy and birth rates. A Bayesian approach was applied to analyze the survival rate of the vitrified embryos. In all analyses the number of transferred embryos was included as covariate. It was observed that neither the period of storage nor the number of transferred embryos affected pregnancy rate, and all periods presented similar pregnancy rates (0.85 ± 0.04; 0.86 ± 0.05; 0.78 ± 0.07 for A, B and C). Fertility at birth was affected by the number of transferred embryos, but non-significant differences between periods were detected (0.77 ± 0.04, 0.75 ± 0.07; 0.69 ± 0.08 for A, B and C). Also, the posterior means (highest posterior density intervals at 95%) of embryo survival at birth from pregnant females were similar between the different periods (47 [41 53]; 47 [38 56]; 42 [31 54]; for A, B and C). Results obtained in the present experiment point out that vitrified embryos could be stored in liquid nitrogen during at least fifteen years, achieving good pregnancy rate, fertility and survival at birth.     

Is there an influence of enzyme polymorphisms on ochratoxin A genotoxicity?

Primary cultured human urothelial cells derived from ureter specimens of urological patients were used to evaluate induction of DNA-damage by OTA in the alkaline single-cell gel electrophoresis (comet) assay. With the cultured cells from each donor a separate comet assay was performed and tail length of the damaged DNA was measured. A broad spectrum of effects was detected between the individual cell cultures with effects reaching from tail lengths on control level up to strongly enhanced tail lengths.All donors of urothelial tissue were additionally genotyped for several xenobiotic metabolising enzymes (cytochrome P450 1A2, glutathione S-transferases T1, M1, and P1, N-acetyltransferase 2) in lymphocyte DNA. The genotype was then correlated with the genotoxic effects obtained in the comet assay.No correlation was found with CYP1A2, GSTT1, and GSTM1 genotypes whereas for GSTP1 stronger genotoxic effects were found in cells from donors with hetero-and homozygously mutated (w/m, m/m) genotypes compared to homozygous wildtypes. The strongest hint for a correlation was found for NAT2, as cells from donors with homozygous mutated alleles (m/m), known as slow acetylators, displayed a higher susceptibility to OTA in the comet assay than cells from donors with the heterozygously mutated or wildtype alleles (rapid acetylators).     

Proteomic signatures of neuroinflammation in Alzheimer’s disease,multiple sclerosis and ischemic stroke

Introduction: Inflammation is integral in the neuropathology of both chronic and acute neurological disorders. Knowing the inflammatory profile is important for clarification of disease mechanisms, diagnostic purposes, and ultimately treatment options.

Areas covered: A systematic review was performed on literature from PubMed using the search terms ‘Alzheimer’s disease’ (AD) or ”multiple sclerosis” (MS) or ”ischemic stroke” and ‘proteomics’. Inflammatory proteins were assessed in blood, cerebrospinal fluid (CSF), and post-mortem brain tissue. Regulated inflammatory proteins across compartments and disorders mainly consisted of innate immune proteins, acute phase proteins and oxidative stress response proteins. In addition, immunoglobulin chains were signature proteins of MS, reflecting additional involvement of adaptive immunity. The Chitinase 3-like protein 1 was increased in ten original articles on MS and in three on AD supporting its implication in these diseases. Furthermore, CNS/CSF AD inflammatory proteins were matched to a CNS myeloid cell proteome implicating Alpha-2-Macroglobulin and Annexin A1 in AD pathogenesis.

Expert opinion: Proteomics is an excellent technique for profiling inflammatory proteins in tissues and cells, but still targeted approaches are required for profiling of very low abundance proteins and peptides. Knowing the inflammatory signature of brain tissue, CSF, blood, and CNS myeloid cells holds the potential to point to novel mechanistic aspects of neurological diseases.     

Is there an influence of body mass on digesta mean retention time in herbivores? A comparative study on ungulates

The relation between body mass (BM) and digesta mean retention time (MRT) in herbivores was the focus of several studies in recent years. It was assumed that MRT scaled with BM(0.25) based on the isometric scaling of gut capacity (BM(1.0)) and allometric scaling of energy intake (BM(0.75)). Literature studies that tested this hypothesis produced conflicting results, arriving sometimes at higher or lower exponents than the postulated 0.25. This study was conducted with 8 ruminants (n=2-6 per species) and 6 hindgut fermenting species/breeds (n=2-6, warthog n=1) with a BM range of 60-4000 kg. All animals received a ration of 100% grass hay with ad libitum access. Dry matter intake was measured and the MRT was estimated by the use of a solute and a particle (1-2 mm) marker. No significant scaling of MRT(particle) with BM was observed for all herbivores (32 BM(0.04), p=0.518) and hindgut fermenters (32 BM(0.00), p=1.00). The scaling exponent for ruminants only showed a tendency towards significance (29 BM(0.12), p=0.071). Ruminants on average had an MRT(particle) 1.61-fold longer than hindgut fermenters. Whereas an exponent of 0.25 is reasonable from theoretical considerations, much lower exponents were found in this and other studies. The energetic benefit of increasing MRT is by no means continuous, since the energy released from a given food unit via digestion decreases over time. The low and non-significant scaling factors for both digestion types suggest that in ungulates, MRT is less influenced by BM (maximal allometric exponent ≤0.1) than often reported.     

Protective effects of salusin-α and salusin-β on renal ischemia/reperfusion damage and their levels in ischemic acute renal failure

Salusin-α and salusin-β are expressed in many tissues including the central nervous system, vessels and kidneys; they have been shown to decrease endoplasmic reticulum stress during heart ischemia/reperfusion (I/R) and to decrease apoptosis. We investigated the relation of salusin-α and salusin-β levels to acute ischemic renal failure. We also investigated whether these peptides are protective against renal I/R damage. Fifty-three rats were divided into six groups: control, I/R, I/R + salusin-α1, I/R + salusin-α10, I/R + salusin-β1 and I/R + salusin-β10. After removing the right kidney, the left kidney was subjected to ischemia for 1 h and reperfusion for 23 h. The treatment groups were injected subcutaneously at the beginning of ischemia with 1 or 10 μg/kg salusin-α, and 1 or 10 μg/kg salusin-β. Histopathology was assessed at the end of the experiment. Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) activity and malondialdehyde (MDA) levels were measured in the kidney tissue. Serum levels of blood urea nitrogen (BUN), creatinine (Cre), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1 beta (IL-1β) also were measured. Levels of salusin-α and salusin-β were measured in the serum and kidney tissues of the control and I/R groups. SOD, CAT and GSH-PX activities were decreased and the levels of MDA, TNF-α, IL-6, IL-1β, BUN and Cre were increased in the I/R group compared to controls. Severe glomerular and tubular damage was apparent in the I/R group compared to controls. The level of salusin-β was decreased in the serum and kidney tissue of the I/R group compared to controls, whereas the level of salusin-α was decreased in the serum and increased in the kidney tissue. Salusin-α and salusin-β administration increased SOD and GSH-PX enzyme activation and decreased the levels of MDA, TNF-α, IL-6 and IL-1β compared to the I/R group. BUN and Cre levels were decreased in the I/R + salusin-α1 group and the level of Cre was decreased in I/R + salusin-β10 group compared to the I/R group. We demonstrated a protective effect of salusin-α and salusin-β against renal I/R damage. Changes in the levels of salusin-α and salusin-β in the I/R group suggest that these peptides may be associated with acute renal failure.