The possibility of using citrulline as a new biochemical marker to evaluate the state of kidney under cardiovascular pathologies

The aim of our work was to study the level of citrulline in serum as a biochemical marker of the functional state of the kidneys in patients with cardiovascular pathologies. Methods. The study included 134 patients aged 41-68 years, of which 40 patients with stage II arterial hypertension (AG-II) and 62 patients with chronic heart failure (AG in combination with chronic ischemic heart disease) in the IIa and IIb stages. The control group included 32 practically healthy persons of the corresponding age. Along with standard diagnostic methods, the content of citrulline in serum was determined. Statistical analysis was performed using SPSS 10.0 for Windows. Results. There is a direct relationship between the increase in the content of citrulline and the severity of changes in the parameters of the functional state of the kidneys in patients with AG-II and CHF. At the same time, the fact that the content of citrulline in serum is much higher compared to the control values in patients with normal values of GFR, creatinine and microalbuminuria. As the increase in the content of citrulline directly correlates with the degree of deterioration of the functional state of the kidneys, and the kidneys are the only organ that regulates and controls the exchange of citrulline, the increase in its level in serum can be primarily associated with changes in kidney function in the examined patients. Conclusions. The content of citrulline in serum can be used as an additional marker for the presence of functional renal impairment in patients with cardiovascular and other diseases of the internal organs, especially in the early stages of development, as well as to assess the efficacy and safety of the use of drugs. This indicator needs further study involving more patients with different pathologies of the cardiovascular system and simultaneous monitoring of the functional status of the kidneys by standardized methods

Roles for Drosophila melanogaster myosin IB in maintenance of enterocyte brush-border structure and resistance to the bacterial pathogen Pseudomonas entomophila

Author Posting. © American Society for Cell Biology, 2007. This article is posted here by permission of American Society for Cell Biology for personal use, not for redistribution. The definitive version was published in Molecular Biology of the Cell 18 (2007): 4625-4636, doi:10.1091/mbc.E07-02-0191.Drosophila myosin IB (Myo1B) is one of two class I myosins in the Drosophila genome. In the larval and adult midgut enterocyte, Myo1B is present within the microvillus (MV) of the apical brush border (BB) where it forms lateral tethers between the MV membrane and underlying actin filament core. Expression of green fluorescent protein-Myo1B tail domain in the larval gut showed that the tail domain is sufficient for localization of Myo1B to the BB. A Myo1B deletion mutation exhibited normal larval gut physiology with respect to food uptake, clearance, and pH regulation. However, there is a threefold increase in terminal deoxynucleotidyl transferase dUTP nick-end labeling-positive enterocyte nuclei in the Myo1B mutant. Ultrastructural analysis of mutant midgut revealed many perturbations in the BB, including membrane tethering defects, MV vesiculation, and membrane shedding. The apical localization of both singed (fascin) and Dmoesin is impaired. BBs isolated from mutant and control midgut revealed that the loss of Myo1B causes the BB membrane and underlying cytoskeleton to become destabilized. Myo1B mutant larvae also exhibit enhanced sensitivity to oral infection by the bacterial pathogen Pseudomonas entomophila, and severe cytoskeletal defects are observed in the BB of proximal midgut epithelial cells soon after infection. Resistance to P. entomophila infection is restored in Myo1B mutant larvae expressing a Myo1B transgene. These results indicate that Myo1B may play a role in the local midgut response pathway of the Imd innate immune response to Gram-negative bacterial infection.This work was supported by National Institutes of Health grants DK-25387 (to M.S.M.), DK-55389 (to Jon Morrow, Yale School of Medicine), and GM-52857 (to L.G.T.) and a research grant from the Crohns and Colitis Foundation of America (to M.S.M.)

Why Are Outcomes Different for Registry Patients Enrolled Prospectively and Retrospectively? Insights from the Global Anticoagulant Registry in the FIELD-Atrial Fibrillation (GARFIELD-AF).

Background: Retrospective and prospective observational studies are designed to reflect real-world evidence on clinical practice, but can yield conflicting results. The GARFIELD-AF Registry includes both methods of enrolment and allows analysis of differences in patient characteristics and outcomes that may result. Methods and Results: Patients with atrial fibrillation (AF) and ≥1 risk factor for stroke at diagnosis of AF were recruited either retrospectively (n = 5069) or prospectively (n = 5501) from 19 countries and then followed prospectively. The retrospectively enrolled cohort comprised patients with established AF (for a least 6, and up to 24 months before enrolment), who were identified retrospectively (and baseline and partial follow-up data were collected from the emedical records) and then followed prospectively between 0-18 months (such that the total time of follow-up was 24 months; data collection Dec-2009 and Oct-2010). In the prospectively enrolled cohort, patients with newly diagnosed AF (≤6 weeks after diagnosis) were recruited between Mar-2010 and Oct-2011 and were followed for 24 months after enrolment. Differences between the cohorts were observed in clinical characteristics, including type of AF, stroke prevention strategies, and event rates. More patients in the retrospectively identified cohort received vitamin K antagonists (62.1% vs. 53.2%) and fewer received non-vitamin K oral anticoagulants (1.8% vs . 4.2%). All-cause mortality rates per 100 person-years during the prospective follow-up (starting the first study visit up to 1 year) were significantly lower in the retrospective than prospectively identified cohort (3.04 [95% CI 2.51 to 3.67] vs . 4.05 [95% CI 3.53 to 4.63]; p = 0.016). Conclusions: Interpretations of data from registries that aim to evaluate the characteristics and outcomes of patients with AF must take account of differences in registry design and the impact of recall bias and survivorship bias that is incurred with retrospective enrolment. Clinical Trial Registration: - URL: http://www.clinicaltrials.gov . Unique identifier for GARFIELD-AF (NCT01090362)

Improved risk stratification of patients with atrial fibrillation: an integrated GARFIELD-AF tool for the prediction of mortality, stroke and bleed in patients with and without anticoagulation.

OBJECTIVES: To provide an accurate, web-based tool for stratifying patients with atrial fibrillation to facilitate decisions on the potential benefits/risks of anticoagulation, based on mortality, stroke and bleeding risks. DESIGN: The new tool was developed, using stepwise regression, for all and then applied to lower risk patients. C-statistics were compared with CHA2DS2-VASc using 30-fold cross-validation to control for overfitting. External validation was undertaken in an independent dataset, Outcome Registry for Better Informed Treatment of Atrial Fibrillation (ORBIT-AF). PARTICIPANTS: Data from 39 898 patients enrolled in the prospective GARFIELD-AF registry provided the basis for deriving and validating an integrated risk tool to predict stroke risk, mortality and bleeding risk. RESULTS: The discriminatory value of the GARFIELD-AF risk model was superior to CHA2DS2-VASc for patients with or without anticoagulation. C-statistics (95% CI) for all-cause mortality, ischaemic stroke/systemic embolism and haemorrhagic stroke/major bleeding (treated patients) were: 0.77 (0.76 to 0.78), 0.69 (0.67 to 0.71) and 0.66 (0.62 to 0.69), respectively, for the GARFIELD-AF risk models, and 0.66 (0.64-0.67), 0.64 (0.61-0.66) and 0.64 (0.61-0.68), respectively, for CHA2DS2-VASc (or HAS-BLED for bleeding). In very low to low risk patients (CHA2DS2-VASc 0 or 1 (men) and 1 or 2 (women)), the CHA2DS2-VASc and HAS-BLED (for bleeding) scores offered weak discriminatory value for mortality, stroke/systemic embolism and major bleeding. C-statistics for the GARFIELD-AF risk tool were 0.69 (0.64 to 0.75), 0.65 (0.56 to 0.73) and 0.60 (0.47 to 0.73) for each end point, respectively, versus 0.50 (0.45 to 0.55), 0.59 (0.50 to 0.67) and 0.55 (0.53 to 0.56) for CHA2DS2-VASc (or HAS-BLED for bleeding). Upon validation in the ORBIT-AF population, C-statistics showed that the GARFIELD-AF risk tool was effective for predicting 1-year all-cause mortality using the full and simplified model for all-cause mortality: C-statistics 0.75 (0.73 to 0.77) and 0.75 (0.73 to 0.77), respectively, and for predicting for any stroke or systemic embolism over 1 year, C-statistics 0.68 (0.62 to 0.74). CONCLUSIONS: Performance of the GARFIELD-AF risk tool was superior to CHA2DS2-VASc in predicting stroke and mortality and superior to HAS-BLED for bleeding, overall and in lower risk patients. The GARFIELD-AF tool has the potential for incorporation in routine electronic systems, and for the first time, permits simultaneous evaluation of ischaemic stroke, mortality and bleeding risks. CLINICAL TRIAL REGISTRATION: URL: http://www.clinicaltrials.gov. Unique identifier for GARFIELD-AF (NCT01090362) and for ORBIT-AF (NCT01165710)

Two-year outcomes of patients with newly diagnosed atrial fibrillation: results from GARFIELD-AF.

AIMS: The relationship between outcomes and time after diagnosis for patients with non-valvular atrial fibrillation (NVAF) is poorly defined, especially beyond the first year. METHODS AND RESULTS: GARFIELD-AF is an ongoing, global observational study of adults with newly diagnosed NVAF. Two-year outcomes of 17 162 patients prospectively enrolled in GARFIELD-AF were analysed in light of baseline characteristics, risk profiles for stroke/systemic embolism (SE), and antithrombotic therapy. The mean (standard deviation) age was 69.8 (11.4) years, 43.8% were women, and the mean CHA2DS2-VASc score was 3.3 (1.6); 60.8% of patients were prescribed anticoagulant therapy with/without antiplatelet (AP) therapy, 27.4% AP monotherapy, and 11.8% no antithrombotic therapy. At 2-year follow-up, all-cause mortality, stroke/SE, and major bleeding had occurred at a rate (95% confidence interval) of 3.83 (3.62; 4.05), 1.25 (1.13; 1.38), and 0.70 (0.62; 0.81) per 100 person-years, respectively. Rates for all three major events were highest during the first 4 months. Congestive heart failure, acute coronary syndromes, sudden/unwitnessed death, malignancy, respiratory failure, and infection/sepsis accounted for 65% of all known causes of death and strokes for <10%. Anticoagulant treatment was associated with a 35% lower risk of death. CONCLUSION: The most frequent of the three major outcome measures was death, whose most common causes are not known to be significantly influenced by anticoagulation. This suggests that a more comprehensive approach to the management of NVAF may be needed to improve outcome. This could include, in addition to anticoagulation, interventions targeting modifiable, cause-specific risk factors for death. CLINICAL TRIAL REGISTRATION: http://www.clinicaltrials.gov. Unique identifier: NCT01090362

Risk profiles and one-year outcomes of patients with newly diagnosed atrial fibrillation in India: Insights from the GARFIELD-AF Registry.

BACKGROUND: The Global Anticoagulant Registry in the FIELD-Atrial Fibrillation (GARFIELD-AF) is an ongoing prospective noninterventional registry, which is providing important information on the baseline characteristics, treatment patterns, and 1-year outcomes in patients with newly diagnosed non-valvular atrial fibrillation (NVAF). This report describes data from Indian patients recruited in this registry. METHODS AND RESULTS: A total of 52,014 patients with newly diagnosed AF were enrolled globally; of these, 1388 patients were recruited from 26 sites within India (2012-2016). In India, the mean age was 65.8 years at diagnosis of NVAF. Hypertension was the most prevalent risk factor for AF, present in 68.5% of patients from India and in 76.3% of patients globally (P < 0.001). Diabetes and coronary artery disease (CAD) were prevalent in 36.2% and 28.1% of patients as compared with global prevalence of 22.2% and 21.6%, respectively (P < 0.001 for both). Antiplatelet therapy was the most common antithrombotic treatment in India. With increasing stroke risk, however, patients were more likely to receive oral anticoagulant therapy [mainly vitamin K antagonist (VKA)], but average international normalized ratio (INR) was lower among Indian patients [median INR value 1.6 (interquartile range {IQR}: 1.3-2.3) versus 2.3 (IQR 1.8-2.8) (P < 0.001)]. Compared with other countries, patients from India had markedly higher rates of all-cause mortality [7.68 per 100 person-years (95% confidence interval 6.32-9.35) vs 4.34 (4.16-4.53), P < 0.0001], while rates of stroke/systemic embolism and major bleeding were lower after 1 year of follow-up. CONCLUSION: Compared to previously published registries from India, the GARFIELD-AF registry describes clinical profiles and outcomes in Indian patients with AF of a different etiology. The registry data show that compared to the rest of the world, Indian AF patients are younger in age and have more diabetes and CAD. Patients with a higher stroke risk are more likely to receive anticoagulation therapy with VKA but are underdosed compared with the global average in the GARFIELD-AF. CLINICAL TRIAL REGISTRATION-URL: http://www.clinicaltrials.gov. Unique identifier: NCT01090362

Трирічна динаміка структурних показників лівого шлуночка у хворих на резистентну артеріальну гіпертензію на тлі чотирикомпонентної терапії

 Aim. To study the three-year dynamics of left ventricular (LV) structural parameters with the four-component antihypertensive therapy in patients with resistant arterial hypertension (RAH).Material and methods. A total of 102 patients with true RAH were included. The duration of the treatment was 3.2 ± 0.1 years. Patients received triple single-pill combination of antihypertensive drugs (TSPC), which was alternately added by spironolactone, eplerenone, moxonidine, torasemide, or nebivolol for three-months treatment. Than patients received the TSPC with an addition of the most effective of the four medications listed. Office and ambulatory blood pressure (BP) measurements and echocardiography were performed, clinical characteristics; specifics of neurohumoral and proinflammatory status were assessed.Results. The patients were divided into 2 groups according to the changes in LV mass index (LVMI) within three years. The 1st group included 68 patients who demonstrated regression of LV hypertrophy (LVH). The 2nd group included 28 patients who had LVMI unchanged or increased. LVMI was normalized in 38.2 % of patients, and a degree of LVH was reduced from severe to moderate in 61.8 % of patients in achieving the target BP level in 44 % of patients and restoring the physiological 24-hour BP rhythm in 39 % of patients. In the 2nd group, 28.6 % of patients achieved BP targets; a large proportion of them were patients with concomitant coronary heart disease (42.3 %) and chronic kidney disease (64.3 %).Conclusions. Effective antihypertensive therapy providing 24-hour BP control and restoring the physiological 24-hour BP rhythm contributes to LVH regression in 66.7 % of patients with RAH. Higher baseline LVMI (β = 0.655; P &lt; 0.0001) and plasma active renin (β = 0.442; P = 0.005), lower 24-hour urinary albumin excretion rate (β = -0.475; P &lt; 0.0001) are independent predictors of LVH regression in RAH patients. Цель работы – изучить динамику структурных показателей левого желудочка (ЛЖ) под влиянием трехлетней четырехкомпонентной антигипертензивной терапии у пациентов с резистентной артериальной гипертензией (РАГ).Материалы и методы. В исследование включили 102 пациентов с истинной РАГ. Длительность наблюдения – 3,2 ± 0,1 года. Пациенты принимали тройную фиксированную комбинацию антигипертензивных препаратов (ТФК), к которой поочередно на 3 месяца добавляли спиронолактон, эплеренон, моксонидин, торасемид или небиволол; далее пациенты получали ТФК с добавлением наиболее эффективного из перечисленных четвертого препарата. Выполняли офисное и амбулаторное измерения артериального давления (АД), ЭхоКГ, оценивали клинические характеристики, особенности нейрогуморального, провоспалительного статуса.Результаты. Пациентов сгруппировали в зависимости от изменений индекса массы миокарда ЛЖ (ИММЛЖ) в течение трех лет. Первую группу составили 68 больных, у которых произошел регресс гипертрофии ЛЖ (ГЛЖ), вторую группу – 28 пациентов, у которых ИММЛЖ не изменился или увеличился. На фоне достижения целевого уровня АД у 44 % больных, восстановления физиологического суточного ритма АД у 39 % пациентов нормализацию ИММЛЖ отметили у 38,2 %, снижение степени ГЛЖ от тяжелой к средней – у 61,8 % больных. В группе без регресса ГЛЖ нормализации АД достигли 28,6 % больных, в основном пациенты с сопутствующей ишемической болезнью сердца (42,3 %) и хронической болезнью почек (64,3 %).Выводы. Эффективная антигипертензивная терапия, обеспечивающая контроль АД в течение суток, восстановление физиологического суточного профиля АД, способствует регрессу ГЛЖ у 66,7 % больных с РАГ. Независимые предикторы регресса ГЛЖ: большие исходные значения ИММЛЖ (β = 0,655; p &lt; 0,0001) и уровня активного ренина плазмы (β = 0,442; p = 0,005), меньшая суточная экскреция альбумина с мочой (β = -0,475; p &lt; 0,0001). Мета роботи – вивчити динаміку структурних показників лівого шлуночка (ЛШ) під впливом трирічної чотирикомпонентної антигіпертензивної терапії в пацієнтів з резистентною артеріальною гіпертензією (РАГ).Матеріали та методи. У дослідженні взяли участь 102 пацієнти з істиною РАГ. Тривалість спостереження – 3,2 ± 0,1 року. Пацієнти отримували потрійну фіксовану комбінацію антигіпертензивних препаратів (ПФК), до якої почергово на 3 місяці додавали спіронолактон, еплеренон, моксонідин, торасемід або небіволол; надалі пацієнти отримували ПФК із додаванням найефективнішого з названих четвертого препарату. Виконували офісне й амбулаторне вимірювання артеріального тиску (АТ), ЕхоКГ, оцінювали клінічні характеристики, особливості нейрогуморального, прозапального статусу.Результати. Пацієнтів згрупували залежно від змін індексу маси міокарда ЛШ (ІММЛШ) протягом 3 років. Перша група – 68 хворих, у яких відбувся регрес гіпертрофії ЛШ (ГЛШ), друга група – 28 пацієнтів, у яких ІММЛШ не змінився або збільшився. На тлі досягнення цільового рівня АТ у 44 % хворих та відновлення фізіологічного добового ритму АТ у 39 % пацієнтів нормалізацію ІММЛШ спостерігали у 38,2 % пацієнтів, зниження ступеня ГЛШ від тяжкого до помірного – у 61,8 % хворих. У групі без регресу ГЛШ нормалізації АТ досягли тільки 28,6 % хворих, здебільшого пацієнти з супутньою ішемічною хворобою серця (42,3 %) та хронічною хворобою нирок (64,3 %).Висновки. Ефективна антигіпертензивна терапія, що забезпечує контроль АТ протягом доби та відновлює фізіологічний добовий ритм АТ, сприяє регресу ГЛШ у 66,7 % хворих на РАГ. Незалежними предикторами регресу ГЛШ є вищі вихідні значення ІММЛШ (β = 0,655; p &lt; 0,0001) і рівня активного реніну плазми (β = 0,442; p = 0,005), менша добова екскреція альбуміну з сечею (β = -0,475; p &lt; 0,0001)

Clot formation and lysis in platelet rich plasma of healthy donors and patients with resistant hypertension

Hemostatic balance in blood is affected by numerous factors, including coagulation and fibrinolytic proteins, the wide spectrum of their inhibitors, and blood cells. Since platelets can participate in contradictory processes, they significantly complicate the whole picture. Therefore, nowadays the development of global assays of hemostasis, which can reflect the physiological process of hemostasis and can be used for point-of-care diagnosis of thrombosis, is crucial. This paper outlines a new approach we used to analyze the capabilities of clot waveform analysis tools to distinguish the response of platelet-rich plasma from healthy donors and patients with arterial hypertension caused by stimulation of coagulation and lysis (with exogenous thrombin and recombinant tissue-type plasminogen activator, respectively). In donor plasma, when the clot degradation was accompanied by 40 IU/ml of recombinant tissue-type plasminogen activator, platelets potentiated fibrinolysis more than coagulation, which ultimately shifts the overall balance to a profibrinolytic state. At the same time, for patients with hypertension, platelets, embedded in clot obtained from platelet-rich plasma, showed a weaker ability to stimulate fibrinolysis. The obtained data gives the evidence that platelets can act not only as procoagulants but also as profibrinolytics. By simultaneously amplifying coagulation and fibrinolysis, making their rates comparable, platelets would control plasma procoagulant activity, thereby regulating local hemostatic balance, the size and lifetime of the clot. Moreover, clot waveform analysis may be used to distinguish the effects of platelet-rich plasma on clotting or lysis of fibrin clots in healthy donors and patients with essential hypertension

Determination of plasminogen/plasmin system components and indicators of lipoproteins oxidative modification under arterial hypertension

The present study was investigated of levels of oxidative modification of lipoproteins and content of plasminogen/plasmin system components – tissue-type plasminogen activator (t-PA) and plasminogen activators inhibitor-1 (PAI-I), in patients with stage II arterial hypertension (AHT) and resistant form. It was established that t-PA level in blood plasma of the patients is 2 times lower under stage II hypertension than normal and 2.5 times lower under resistant AHT. The inhibitor activity is 1.5 and 2 times higher consequently. It is concluded that patients with AHT have a decreased fibrinolytic potential, which can cause thrombotic states. Our evaluation showed a significant accumulation of products of lipid and protein oxidation, decrease of activity of antioxidant enzymes and changes of the activity of high density-lipoproteins-associated enzymes (decrease of paraoxonase-1 activity, increase of myeloperoxidase activity). Oxidized lipoproteins, t-PA and PAI-1 can be used as prognostic markers of development of complications and for evaluating the efficacy of therapy in patients with arterial hypertension