Prognostic value of serum nicotinamide phosphoribosyltransferase in patients with bladder cancer

Aim

To analyze the serum nicotinamide phosphoribosyltransferase (Nampt) level and its prognostic value in bladder cancer (BC).

Methods

The study included 131 patients with transitional cell BC and 109 healthy controls from the West China Hospital of Sichuan University in the period between 2007 and 2013. Nampt concentration in serum was measured by commercial ELISA kits for human Nampt.

Results

The serum Nampt protein level in patients with BC (mean ± standard deviation, 16.02 ± 7.95 ng/mL) was significantly higher than in the control group (6.46 ± 2.08 ng/mL) (P < 0.001). Serum Nampt level was an independent prognostic marker of non-muscle-invasive BC, with a higher serum Nampt level (>14.74 ng/mL) indicating shorter recurrence-free survival rate (hazard ratio = 2.85, 95% confidence interval, 1.01-8.06; P = 0.048).

Conclusion

Our results suggest that serum Nampt level may serve as a biomarker of BC and an independent prognostic marker of non-muscle-invasive BC.Bladder cancer (BC) is the ninth most common cancer diagnosis worldwide (1) and the most expensive cancer to treat (2). Among men it is the fourth most common cancer, with incidence four times higher than in women (3). In China, BC caused 17 365 deaths in 2005, with a steady increase in mortality between 1991 and 2005 (4). Of newly diagnosed BC cases, 70%-80% will present with non-muscle-invasive disease, 50%-70% will recur despite endoscopic and intravesical treatments, and 10%-30% will progress to muscle-invasive disease (5,6). Most recurrences occur within 5 years (7). Therefore, to develop improved, more effective prevention and treatments there is a need to find new biomarkers of tumorigenesis and prognosis of BC.Nicotinamide phosphoribosyltransferase (Nampt) is a rate-limiting enzyme in the mammalian NAD⁺ biosynthesis of a salvage pathway (8). Previous studies have shown that it is significantly increased in primary colorectal cancer (9-11), lung cancer (12), breast cancer (13), prostate cancer (14) and gastric cancer (15). Thus, Nampt may be a good biomarker of malignant potential and stage progression (12,16). Our previous study revealed that genetic variants in NAMPT may predict BC risk and prognosis (17). In the present study, we analyzed the serum Nampt level and its prognostic value in BC.     

Ginkgolide B increases hydrogen sulfide and protects against endothelial dysfunction in diabetic rats

AimTo evaluate the effect of ginkgolide B treatment on vascular endothelial function in diabetic rats.MethodsThe study included four groups with 15 male Sprague-Dawley rats: control group; control group treated with ginkgolide B; diabetic group; and diabetic treated with ginkgolide B. The activity of superoxide dismutase (SOD), malondialdehyde content, and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunits, and glutathione peroxidase 1 (GPX1) protein expression were determined in aortic tissues. Vasoconstriction to phenylephrine (PHE) and vasorelaxation to acetylcholine (Ach) and sodium nitroprusside (SNP) were assessed in aortic rings. Nitric oxide (NO) and hydrogen sulfide (H₂S) were measured, as well as cystathionine γ lyase (CSE) and cystathionine β synthetase (CBS) protein expression, and endothelial nitric oxide synthase (eNOS) activity.ResultsDiabetes significantly impaired PHE-induced vasoconstriction and Ach-induced vasorelaxation (P < 0.001), reduced NO bioavailability and H₂S production (P < 0.001), SOD activity, and GPX1 protein expression (P < 0.001), and increased malondialdehyde content and NADPH oxidase subunits, and CSE and CBS protein expression (P < 0.001). Ginkgolide B treatment improved PHE vasoconstriction and Ach vasorelaxation (P < 0.001), restored SOD (P = 0.005) and eNOS (P < 0.001) activities, H₂S production (P = 0.044) and decreased malondialdehyde content (P = 0.014). Vasorelaxation to SNP was not significantly different in control and diabetic rats with or without ginkgolide B treatment. Besides, ginkgolide B increased GPX1 protein expression and reduced NADPH oxidase subunits, CBS and CSE protein expression.ConclusionGinkgolide B alleviates endothelial dysfunction by reducing oxidative stress and elevating NO bioavailability and H₂S production in diabetic rats.Diabetes mellitus is an endocrine disease caused by decreased insulin secretion or action, leading to impaired glucose and lipid metabolism (1). The most dangerous complication of diabetes mellitus is cardiovascular disease, which is the primary factor leading to high mortality and morbidity in diabetic patients (2). A critical role in diabetic cardiovascular complications is played by endothelial dysfunction. Mechanisms responsible for endothelial dysfunction are still poorly understood, but hyperglycemia-induced oxidative stress is hypothesized to be one of them (3). Increased blood glucose increases reactive oxygen species (ROS) production via glucose auto-oxidation (4) and variation in activity of oxidoreductases, such as NADPH oxidase (5). ROS can impair vascular function by damaging endothelial cells, thus playing an important role in diabetes and its cardiovascular complications (6). NADPH oxidase is important because it generates ROS (7,8). Glutathione peroxidase 1 (GPX1) is one of the pivotal antioxidant enzymes in vascular endothelium, which protects against the presence of coronary artery disease (9). Its overexpression reduces ROS formation and enhances phosphorylation of endothelial nitric oxide synthase (eNOS), which improves endothelial function (10).Hydrogen sulfide (H₂S) was previously considered only as a toxic gas, but recent studies have suggested that it plays a variety of important physiological and physiopathological roles (11,12). H₂S is generated from L-cysteine by several enzymes including cystathionine γ lyase (CSE) and cystathionine β synthetase (CBS). Some studies have shown that H₂S takes part in modulation of cardiovascular system (13,14). It has also been shown that H₂S biosynthesis is impaired in diabetes, and that it may be effective to administer different H₂S donors to diabetic animals (15,16).The risk of endothelial dysfunction is increased by sustained progression of hyperglycemia and hyperlipidemia. Endothelial dysfunction is characterized by alterations of endothelium-dependent vascular response to vasoconstrictors and vasodilators in diabetic animals (17,18). Therefore, cardiovascular complications including endothelial dysfunction in patients with diabetes have been treated by decreasing blood glucose and lipid content, and reducing activation of angiotensin. However, these treatments have not prevented the development of complications (19), emphasizing the need for novel approaches.Ginkgolide B, a plant-derived terpenoid, is one of natural bioactive components from the extract of ginkgo biloba leaves. Many studies have demonstrated that ginkgolide B can inhibit platelet-activating factor (PAF)-induced platelet activation via binding with PAF receptor (20,21). Therefore, ginkgolide B is widely used as a natural antagonist of PAF and inhibitor of PAF-induced inflammatory reaction (22). It has been shown that ginkgolide B regulates many physiologic functions, including the antioxidant function, improving the cognitive functions of central nervous system (23,24), repressing atherosclerosis (25), and abating liver cirrhosis (26). In this study, we investigated the effects of ginkgolide B on endothelial function and mediators such as hydrogen sulfide (H₂S), biomarkers of oxidative stress, and oxidoreductase in the aorta of rats with streptozotocin-induced diabetes.     

Urine fibroblast growth factor 23 levels in hypertensive children and adolescents

AimTo determine the correlation of urinary fibroblast growth factor 23 (FGF23) excretion with blood pressure and calcium-phosphorus metabolism.MethodsThe study included 42 hypertensive (17 girls) and 46 healthy children and adolescents (17 girls) aged 6-18 years admitted to the Department of Pediatrics and Nephrology, Medical University of Białystok between January 2013 and December 2013. FGF23 in urine was measured using Human Intact FGF-23 ELISA Kit.ResultsHypertensive participants had significantly higher urine FGF23/creatinine values than the reference group (8.65 vs 5.59 RU/mg creatinine, P = 0.007). Urine FGF23/creatinine positively correlated with systolic blood pressure in all participants. In hypertensive patients, urine FGF23/creatinine positively correlated with serum calcium and negatively with serum 25(OH)D, urinary calcium, phosphorus, and magnesium.ConclusionThis study found that FGF23 may play an important role in the pathogenesis of hypertension in children and adolescents, but our results should be confirmed by further studies.Hypertension is a chronic medical condition and a major risk factor for cardiovascular disease, heart failure, and chronic kidney disease (CKD). Hypertension was found to be associated with several factors, among them calcium-phosphorus imbalance, lack of vitamin D, and serum parathyroid hormone (PTH) (1-5). However, far too little attention has been paid to phosphates and hormonal mechanisms responsible for their regulation, especially since the consumption of phosphorus has considerably increased in recent years. Some studies have shown that serum phosphorus increases BP (11,12). However, recent studies have found that high phosphorus intake reduces BP, when the diet is rich in calcium (6-8), while other have shown that BP was reduced by low phosphorus and high calcium diet (9,10).Phosphate concentration is primarily regulated by PTH and fibroblast growth factor 23 (FGF 23) – phosphatonin, produced by osteoblasts/osteocytes in the bone, which, similarly to PTH, stimulates phosphaturia. FGF23 decreases renal calcitriol production and inhibits PTH secretion. Its main function is to maintain phosphate homeostasis by increasing urinary phosphate excretion and decreasing serum 1,25(OH)₂D (13,14) In patients with CKD, it positively correlated with PTH secretion (15,16). The increase in FGF23 in those patients led to an early development of secondary hypertension by suppression of 1,25(OH)₂D production (17), and low phosphate intake of phosphorus binders caused 35% decrease in plasma FGF23 level (18). However in healthy individuals no changes in FGF23 levels were observed after both phosphate deprivation and loading (19,20).FGF 23 is also involved in renal sodium handling (21) and, what is even more interesting, it suppresses the expression of angiotensin-converting enzyme-2 (ACE2) in CKD-mice and thereby activates renin-angiotensin-aldosterone system (RAAS) (22). FGF23 can also influence the RAAS indirectly through vitamin D (23), which probably reduces renin gene expression and secretory activity of the juxtaglomerular apparatus, the main place of production of renin (24).The investigation of the effect of FGF23 on hypertension is not confined to in vitro models. Hypertensive people were found to have significantly higher plasma FGF23 level than normotensive people (25). FGF23 was shown to have an association with markers of inflammation in individuals with CKD stages 2-4 (26), and with impaired endothelium-dependent vasodilatation in healthy individuals and early CKD patients (27). This effect of FGF23 might also result indirectly from a decrease in 1,25 (OH)₂D (28). FGF23 also correlated with asymmetrical dimethylarginin (ADMA), which is an endogenous inhibitor of NO synthase and a biomarker of endothelial dysfunction (29).So far, however, the relevance of FGF23 in primary arterial hypertension has been under-investigated. What is more, available data focus on adult hypertensive patients and possible relation of phosphorus intake and increased FGF23 concentration to elevated BP (25). There is a paucity of similar data in children and adolescents. The aims of this research were to determine whether urinary excretion of FGF23 in hypertensive children and adolescents was higher than in healthy controls and whether its urinary level correlated with serum calcium, phosphorus, vitamin D, and PTH concentrations. Reference group data were obtained from the OLAF study, which established the reference blood pressure range for Polish children and adolescents. A strong correlation between serum and urine FGF23 was previously confirmed (r = 0.92, P < 0.001) (30).     

Psychiatric heredity and posttraumatic stress disorder: survey study of war veterans

Aim

To explore the prevalence of psychiatric heredity (family history of psychiatric illness, alcohol dependence disorder, and suicidality) and its association with the diagnosis of stress-related disorders in Croatian war veterans established during psychiatric examination.

Methods

The study included 415 war veterans who were psychiatrically assessed and diagnosed by the same psychiatrist during an expert examination conducted for the purposes of compensation seeking. Data were collected by a structured diagnostic procedure.

Results

There was no significant correlation between psychiatric heredity of psychiatric illness, alcohol dependence, or suicidality and diagnosis of posttraumatic stress disorder (PTSD) or PTSD with psychiatric comorbidity. Diagnoses of psychosis or psychosis with comorbidity significantly correlated with psychiatric heredity (φ = 0.111; P = 0.023). There was a statistically significant correlation between maternal psychiatric illness and the patients’ diagnoses of partial PTSD or partial PTSD with comorbidity (φ = 0.104; P = 0.035) and psychosis or psychosis with comorbidity (φ = 0.113; P = 0.022); paternal psychiatric illness and the patients’ diagnoses of psychosis or psychosis with comorbidity (φ = 0.130; P = 0.008), alcohol dependence or alcohol dependence with comorbidity (φ = 0.166; P = 0.001); psychiatric illness in the primary family with the patients’ psychosis or psychosis with comorbidity (φ = 0.115; P = 0.019); alcohol dependence in the primary family with the patients’ personality disorder or personality disorder with comorbidity (φ = 0.099; P = 0.044); and suicidality in the primary family and a diagnosis of personality disorder or personality disorder with comorbidity (φ = 0.128; P = 0.009).

Conclusion

The study confirmed that parental and familial positive history of psychiatric disorders puts the individual at higher risk for developing psychiatric illness or alcohol or drug dependence disorder. Psychiatric heredity might not be necessary for the individual who was exposed to severe combat-related events to develop symptoms of PTSD.There are several risk factors associated with the development of posttraumatic stress disorder (PTSD), such as factors related to cognitive and biological systems and genetic and familial risk (1), environmental and demographic factors (2), and personality and psychiatric anamnesis (3).They are usually grouped into three categories: factors that preceded the exposure to trauma or pre-trauma factors; factors associated with trauma exposure itself; and post-trauma factors that are associated with the recovery environment (2,4).There are many studies which support the hypothesis that pre-trauma factors, such as ongoing life stress, psychiatric history, female sex (3), childhood abuse, low economic status, lack of education, low intelligence, lack of social support (5), belonging to racial and ethnic minority, previous traumatic events, psychiatric heredity, and a history of perceived life threat, influence the development of stress related disorders (6). Many findings suggest that ongoing life stress or prior trauma history sensitizes a person to a new stressor (2,7-9). The same is true for the lack of social support, particularly the loss of support from significant others (2,9-11), as well as from friends and community (12-14). If the community does not have an elaborated plan for providing socioeconomic support to the victims, then the low socioeconomic status can also be an important predictor of a psychological outcome such as PTSD (2,10,15). Unemployment was recognized as a risk factor for developing PTSD in a survey of 374 trauma survivors (16). It is known that PTSD commonly occurs in patients with a previous psychiatric history of mental disorders, such as affective disorders, other anxiety disorders, somatization, substance abuse, or dissociative disorders (17-21). Epidemiological studies showed that pre-existing psychiatric problems are one of the three factors that can predict the development of PTSD (2,22). Pre-existing anxiety disorders, somatoform disorders, and depressive disorders can significantly increase the risk of PTSD (23). Women have a higher vulnerability for PTSD than men if they experienced sexually motivated violence or had pre-existing anxiety disorders (23,24). A number of studies have examined the effects of gender differences on the predisposition for developing PTSD, with the explanation that women generally have higher rates of depression and anxiety disorders (3,25,26). War-zone stressors were described as more important for PTSD in men, whereas post-trauma resilience-recovery factors as more important for women (27).Lower levels of education and poorer cognitive abilities also appear to be risk factors (25). Golier et al (25) reported that low levels of education and low IQ were associated with poorer recall on words memorization tasks. In addition, this study found that the PTSD group with lower Wechsler Adult Intelligence Scale-Revised (WAIS-R) scores had fewer years of education (25). Nevertheless, some experts provided evidence for poorer cognitive ability in PTSD patients as a result or consequence rather than the cause of stress-related symptoms (28-31). Studies of war veterans showed that belonging to racial and ethnic minority could influence higher rates of developing PTSD even after the adjustment for combat exposure (32,33). Many findings suggest that early trauma in childhood, such as physical or sexual abuse or even neglect, can be associated with adult psychopathology and lead to the development of PTSD (2,5,26,34,35). Surveys on animal models confirm the findings of lifelong influences of early experience on stress hormone reactivity (36).Along with the reports on the effects of childhood adversity as a risk factor for the later development of PTSD, there is also evidence for the influence of previous exposure to trauma related events on PTSD (9,26,28). Breslau et al (36) reported that previous trauma experience substantially increased the risk for chronic PTSD.Perceived life threats and coping strategies carry a high risk for developing PTSD (9,26). For instance, Ozer et al (9) reported that dissociation during trauma exposure has high predictive value for later development of PTSD. Along with that, the way in which people process and interpret perceived threats has a great impact on the development or maintenance of PTSD (37,38).Brewin et al (2) reported that individual and family psychiatric history had more uniform predictive effects than other risk factors. Still, this kind of influence has not been examined yet.Keeping in mind the lack of investigation of parental psychiatric heredity on the development of stress-related disorders, the aim of our study was to explore the prevalence and correlation between the heredity of psychiatric illness, alcohol dependence, suicidality, and the established diagnosis of stress-related disorders in Croatian 1991-1995 war veterans.     

Shear-wave sonoelastographic features of invasive lobular breast cancers

AimTo evaluate shear-wave elastographic (SWE) and related gray-scale features of pure invasive lobular breast carcinoma (ILC) and compare them with invasive ductal breast cancers (IDC).MethodsQuantitative SWE features of mean (El-mean), maximum (El-max), minimum (El-min) elasticity values of the stiffest portion of the mass, and lesion-to-fat elasticity ratio (E-ratio) were measured in 40 patients with pure ILC and compared with 75 patients with IDC. Qualitative gray-scale features of lesion size, echogenicity, orientation, and presence of distal shadowing were determined and compared between the groups.ResultsILC were significantly larger than IDC (P = 0.008) and exhibited significantly higher El-max (P = 0.015) and higher El-mean (P = 0.008) than IDC. ILC were significantly more often horizontally oriented, while IDC were significantly more often vertically oriented (P < 0.001); ILC were significantly more often hyperechoic than IDC (P < 0.001). Differences in stiffness between ILC and IDC determined by quantitative SWE parameters were present only in small tumors (≤1.5 cm in size), ie, small ILC had significantly higher El-max (P = 0.030), El-mean (P = 0.014), and El-min (P = 0.045) than small IDC, while tumors larger than 1.5 cm had almost equal stiffness, without significant differences between the groups.ConclusionSpecific histopathologic features of ILC are translated into their qualitative sonographic and quantitative sonoelastographic appearance, with higher stiffness of small ILC compared to small IDC. Gray-scale and sonoelastographic features may help in diagnosing ILC.Invasive ductal cancer (IDC) is the most common breast cancer, while invasive lobular cancer (ILC) is the second most common and accounts for 6%-12% of breast cancers (1-3). ILC differs considerably from IDC by having a unique pathological growth pattern, the so called Indian-file pattern, with sheets of single-cell layers growing along the Cooper ligaments, ductuli, and other breast structures, resembling a spiderweb that diffusely spreads in the breast, producing minor desmoplastic reaction (4,5). This spiderweb-like growth is reflected in imaging features of ILC, as well as in its clinical presentation (6). IDC usually clinically manifests as a firm lump, while ILC usually manifests as a palpable thickening and skin or nipple retraction (3,5). ILC has increased tendency for multifocality and multicentricity, a higher risk of bilateral breast cancer (20%-29%), and older age at onset (7,8). Lymph node metastases are less common in ILC than in IDC of equal size, because ILC tumor cells lack cellular atypia and often have low mitotic rate (9). ILC has the propensity to metastasize to the chest, peritoneum, retroperitoneum, and pelvis (10).Because of its growth pattern of mass infiltrating surrounding tissues, IDC is much more easily detected than ILC also on mammography. ILC has higher false-negative mammographic rates than IDC, since ILC may be invisible or may have quite low mammographic density, and microcalcifications are uncommon (6,11). Due to the higher propensity for multicentric and bilateral lesions, it is generally considered that patients with ILC should be referred to preoperative breast MRI, the best imaging modality to evaluate the tumor extent, while the benefit for preoperative MRI in IDC has not yet been proven (12,13). Fine-needle aspiration is not as sensitive for the diagnosis of ILC as it is for IDC, and core-biopsy should be performed when ILC is suspected, even in cases of palpable lesions (14,15). ILC is associated more often than IDC with positive margins on surgical excision and is more often treated with mastectomy, because of the large size at diagnosis and underestimation of tumor extent with conventional imaging (16).Ultrasound of the breast is widely used in the diagnosis of breast cancer, usually after mammography, and most image-guided core biopsies of breast lesions are routinely performed under the sonographic guidance (17,18). Ultrasound is highly operator-dependent, much more than mammography or MRI. The quality of ultrasonic equipment and transducers is variable, suboptimal examinations are common, and interobserver variability is high; sensitivity of ultrasound in detection of ILC is reported in the range of 68%-88% (6,12,19).Sonoelastography is a relatively new ultrasonographic method, which may help in the detection and differentiation of benign and malignant breast lesions (18,20). Strain elastography allows qualitative estimation of the breast lesion stiffness, while shear-wave elastography (SWE) allows quantification of lesion stiffness in kilopascals (kPa) (18). Multicentric studies found that SWE features can help discriminate breast cancers and benign breast lesions, and breast cancers among themselves (20-22). It was also shown that some IDC, like triple negative breast cancers, differ in their stiffness compared to other IDC (23). Studies evaluating some SWE features of invasive cancers were done in a small number of patients with ILC, but to the best of our knowledge none so far has provided values specific for a larger, homogeneous group of patients with pure ILC (24,25).The aim of this single-center study was to evaluate and establish SWE and related conventional sonographic features of pure ILC of the breast in a group of 40 patients, and to compare these features with the most common invasive breast cancer, IDC. SWE features within ILC group were also correlated with tumor size, extent, histologic grade, and the presence of nodal metastases.     

Performance of several simple,noninvasive models for assessing significant liver fibrosis in patients with chronic hepatitis B

Aim

To compare the performance of several simple, noninvasive models comprising various serum markers in diagnosing significant liver fibrosis in the same sample of patients with chronic hepatitis B (CHB) with the same judgment standard.

Methods

A total of 308 patients with CHB who had undergone liver biopsy, laboratory tests, and liver stiffness measurement (LSM) at the Southwest Hospital, Chongqing, China between March 2010 and April 2014 were retrospectively studied. Receiver operating characteristic (ROC) curves and area under ROC curves (AUROCs) were used to analyze the results of the models, which incorporated age-platelet (PLT) index (API model), aspartate transaminase (AST) to alanine aminotransferase (ALT) ratio (AAR model), AST to PLT ratio index (APRI model), γ-glutamyl transpeptidase (GGT) to PLT ratio index (GPRI model), GGT-PLT-albumin index (S index model), age-AST-PLT-ALT index (FIB-4 model), and age-AST-PLT-ALT-international normalized ratio index (Fibro-Q model).

Results

The AUROCs of the S index, GPRI, FIB-4, APRI, API, Fibro-Q, AAR, and LSM for predicting significant liver fibrosis were 0.726 (P < 0.001), 0.726 (P < 0.001), 0.621 (P = 0.001), 0.619 (P = 0.001), 0.580 (P = 0.033), 0.569 (P = 0.066), 0.495 (P = 0.886), and 0.757 (P < 0.001), respectively. The S index and GPRI had the highest correlation with histopathological scores (r = 0.373, P < 0.001; r = 0.372, P < 0.001, respectively) and LSM values (r = 0.516, P < 0.001; r = 0.513, P < 0.001, respectively). When LSM was combined with S index and GPRI, the AUROCs were 0.753 (P < 0.001) and 0.746 (P < 0.001), respectively.

Conclusion

S index and GPRI had the best diagnostic performance for significant liver fibrosis and were robust predictors of significant liver fibrosis in patients with CHB for whom transient elastography was unavailable.Liver fibrosis is a common pathological process in various chronic liver diseases, including chronic hepatitis B (CHB). In patients with CHB, early detection of liver fibrosis is crucial for therapy planning and prognosis estimation. In particular, the presence of significant liver fibrosis is a strong indication for initiating antiviral therapy (1,2). However, liver biopsies, the gold standard for staging fibrosis, are not performed in all hospitals (especially in primary care) because of their invasiveness, sampling errors, and complications. In addition, biopsies are not appropriate for monitoring disease progression. Transient elastography (FibroScan; Echosens, Paris, France), which measures liver stiffness, is increasingly being recognized as an excellent tool for assessing the degree of fibrosis (3,4). FibroScan’s noninvasive nature, reproducibility, and diagnostic performance have also made it increasingly popular. However, not all hospitals have the means to purchase such expensive equipment.Accordingly, in recent years combinations of serum biomarkers of liver fibrosis have been a hot research topic. Several serological models for liver fibrosis (5-7) that incorporate direct or indirect biomarkers have been developed as alternatives to biopsy. These models reportedly vary considerably in their ability to diagnose fibrosis, and their results are conflicting (8,9). The present study assessed the effectiveness of the following seven fibrosis models, all of which comprise routine serum biomarkers and were found to have predictive value for significant liver fibrosis: age-platelet (PLT) index (API) (10), aspartate transaminase (AST) to alanine aminotransferase (ALT) ratio (AAR) (10,11), AST to PLT ratio index (APRI) (10,11), γ-glutamyl transpeptidase (GGT) to PLT ratio index (GPRI) (11), GGT-PLT-albumin (ALB) index (S index) (12), age-AST-PLT-ALT index (FIB-4) (13), and age-AST-PLT-ALT-international normalized ratio (INR) index (Fibro-Q) (14).     

Single acute stress-induced progesterone and ovariectomy alter cardiomyocyte contractile function in female rats

Aim

To assess how ovarian-derived sex hormones (in particular progesterone) modify the effects of single acute stress on the mechanical and biochemical properties of left ventricular cardiomyocytes in the rat.

Methods

Non-ovariectomized (control, n = 8) and ovariectomized (OVX, n = 8) female rats were kept under normal conditions or were exposed to stress (control-S, n = 8 and OVX-S, n = 8). Serum progesterone levels were measured using a chemiluminescent immunoassay. Left ventricular myocardial samples were used for isometric force measurements and protein analysis. Ca²⁺-dependent active force (F_active), Ca²⁺-independent passive force (F_passive), and Ca²⁺-sensitivity of force production were determined in single, mechanically isolated, permeabilized cardiomyocytes. Stress- and ovariectomy-induced alterations in myofilament proteins (myosin-binding protein C [MyBP-C], troponin I [TnI], and titin) were analyzed by sodium dodecyl sulfate gel electrophoresis using protein and phosphoprotein stainings.

Results

Serum progesterone levels were significantly increased in stressed rats (control-S, 35.6 ± 4.8 ng/mL and OVX-S, 21.9 ± 4.0 ng/mL) compared to control (10 ± 2.9 ng/mL) and OVX (2.8 ± 0.5 ng/mL) groups. F_active was higher in the OVX groups (OVX, 25.9 ± 3.4 kN/m² and OVX-S, 26.3 ± 3.0 kN/m²) than in control groups (control, 16.4 ± 1.2 kN/m² and control-S, 14.4 ± 0.9 kN/m²). Regarding the potential molecular mechanisms, F_active correlated with MyBP-C phosphorylation, while myofilament Ca²⁺-sensitivity inversely correlated with serum progesterone levels when the mean values were plotted for all animal groups. F_passive was unaffected by any treatment.Conclusion Stress increases ovary-independent synthesis and release of progesterone, which may regulate Ca²⁺-sensitivity of force production in left ventricular cardiomyocytes. Stress and female hormones differently alter Ca²⁺-dependent cardiomyocyte contractile force production, which may have pathophysiological importance during stress conditions affecting postmenopausal women.The relation between stress, gender, and cardiovascular diseases is well established (1-4). Some of the known risk factors for cardiovascular disease such as smoking, unhealthy diet, and behavioral and psychosocial stress have deleterious effects on the cardiovascular system via activation of the sympathetic nervous system and the hypothalamic-pituitary-adrenal (HPA) axis (5-8). Acute restraint stress is a preferred and widely used method to induce physical stress in animal models (9). Moreover, restraint and immobilization are important as models for psychological stress, which was shown to adversely affect ovarian function (10) and to play a pivotal role in the pathomechmanism of Takotsubo (stress) cardiomyopathy in postmenopausal women (11).Gender is a very important factor in the development of cardiovascular diseases. Premenopausal women have better lipid profile, endothelial function (12), and a lower risk to develop coronary artery disease and myocardial infarction (MI) than men. These advantages of female gender, however, are abolished after menopause, which is associated with increased prevalence of left ventricular (LV) hypertrophy, decreased LV ejection fraction, and LV contractility (13). One of the explanations for the distinct myocardial responses is the cardioprotective effect of female sex hormones (eg, estrogens) (14,15).Progesterone performs several actions on the heart: it exerts an antiarrhythmic effect by accelerating cardiac repolarization (16) and has a preventive role in ischemia-reperfusion injury via reducing inflammatory response (17). It has been shown to inhibit cardiomyocyte apoptosis (18), induce vasodilation, and reduce blood pressure via increasing nitric oxide (NO) levels in normotensive and hypertensive patients (19). Importantly, progesterone is produced by the both ovaries and the adrenal gland: Moreover, the adrenal progesterone content is similar or even larger than that in the ovaries (20). Adrenal progesterone production and secretion increase along with corticosterone regardless of gender and estradiol under stress conditions (21). Progesterone, being an indirect precursor of cortisol (22), increases in response to adrenocorticotrophic hormone (ACTH) stimulation (23).In the heart, there are multiple estrogen hormone receptor types (24). The expression of aromatase in the heart suggests that estrogen may be synthesized also within the cardiomyocyte to exert autocrine/paracrine actions (25). Myocyte contractility seems to be modulated by systemic estrogen levels and altered in cardiomyocytes derived from ovariectomized (OVX) rats (26). In particular, myofilament Ca²⁺-sensitivity is increased in isolated myofibrillar preparations from OVX rats, and restored to the basal levels with estrogen supplementation (27,28).Activation of the sympathetic nervous system plays a central role in the regulation of cardiomyocyte contractile function and myofilament Ca²⁺-sensitivity through beta-adrenergic receptor stimulation, activating the protein kinase A (PKA). PKA-mediated phosphorylation of Ca²⁺-handling and myofilament proteins (myosin binding protein-C [MyBP-C], troponin I [TnI], titin) were shown to alter cardiomyocyte contractile function (29,30). It has been suggested that female cardiomyocytes operate at lower levels of intracellular Ca²⁺ than those of males, particularly under inotropic conditions (31). This difference in Ca²⁺ homeostasis may be related to the fact that estrogen suppresses the L-type Ca²⁺ current (32,33) and may reduce the amount of Ca²⁺ released from the sarcoplasmic reticulum (SR) (34), which was shown to be larger in myocytes from OVX rats (35). Not only cardiomyocyte contraction, but relaxation may also be affected by estrogen via altered Ca²⁺ re-uptake into the SR and modified Ca²⁺ efflux via increased sarcolemmal Na⁺/Ca²⁺ exchange (36). Interestingly, despite similar SR Ca²⁺ content in males and females (37), studies using OVX models report conflicting results concerning changes in the expression and activity of the SR Ca²⁺-ATPase and its regulator protein phospholamban (38-41). Much less is known about the possible effect of progesterone on cardiomyocyte contractile function. We hypothesized that progesterone affected force production of single isolated cardiomyocytes. Therefore, in the present study we aimed to investigate how sex hormones (particularly progesterone) and single acute restraint stress altered cardiomyocyte contractile function and to identify the consequent posttranslational myofilament protein modifications in OVX rats.     

Association between social capital,health-related quality of life,and mental health: a structural-equation modeling approach

AimTo explore the association(s) between demographic factors, socioeconomic status (SES), social capital, health-related quality of life (HRQoL), and mental health among residents of Tehran, Iran.MethodsThe pooled data (n = 31 519) were extracted from a population-based survey Urban Health Equity Assessment and Response Tool-2 (Urban HEART-2) conducted in Tehran in 2011. Mental health, social capital, and HRQoL were assessed using the 28-item General Health Questionnaire (GHQ-28), social capital questionnaire, and Short-Form Health Survey (SF-12), respectively. The study used a multistage sampling method. Social capital, HRQoL, and SES were considered as latent variables. The association between these latent variables, demographic factors, and mental health was determined by structural-equation modeling (SEM).ResultsThe mean age and mental health score were 44.48 ± 15.87 years and 23.33 ± 11.10 (range, 0-84), respectively. The prevalence of mental disorders was 41.76% (95% confidence interval 41.21-42.30). The SEM model showed that age was directly associated with social capital (P = 0.016) and mental health (P = 0.001). Sex was indirectly related to mental health through social capital (P = 0.018). SES, HRQoL, and social capital were associated both directly and indirectly with mental health status.ConclusionThis study suggests that changes in social capital and SES can lead to positive changes in mental health status and that individual and contextual determinants influence HRQoL and mental health.Mental health is defined by World Health Organization (WHO) as “a state of well-being in which every individual realizes his/her own potential, can cope with the normal pressures of life, can work productively, and is able to make a contribution to his/her community” (1,2). Mental health and associated disorders have received increasing attention worldwide, largely due to their impact on socio-economic and overall health status of patients (3). Mental health problems remain a global concern, and account for a large fraction of diseases (4,5).The overall prevalence of mental disorders in Iran between 2000 and 2008 ranged from 12.5% to 38.9% and was similar in urban (20.9%) and rural areas (21.3%) (6). Anxiety and depression were more prevalent than somatization and social dysfunction (7). The provinces with the highest prevalence of mental problems were Chaharmahal with 38.3% and Golestan with 37.3% (8).Mental health is usually determined by a complex interaction of sociocultural, psychological, environmental, and demographic factors (9). The prevalence of mental health disorders is significantly associated with age, marital status, educational level, employment, and health-related to quality of life (HRQoL) (10). HRQoL incorporates physical and socio-emotional functioning and is used to measure individual''s perception of health status, welfare, and well-being in a society (11). A frequently used psychometrical tool for the assessment of HRQoL is Short-Form Health Survey (SF-12). Its two main components are physical component summary (PCS) and mental component summary (MCS), both of which are associated with mental health (12). Previous studies have confirmed a bidirectional association between physical health and depression (as one of the main dimensions of mental health) (13). However, it is not clear whether there is a causal relationship between them (13,14).The suggested mechanisms by which depression could lead to physical disability and decreased HRQoL are poor health behaviors, increased risk of physical disease, and characteristics of depression (eg, decreased pain threshold) (15). On the other hand, physical disability can lead to depression and deterioration of mental health due to restriction of social activities and loss of social capital (15). Ultimately, this bilateral association between depression and poor physical health can lead to increasing health risks (14).Mental disorders such as depression and anxiety are also influenced by socioeconomic status (SES) (16). SES is commonly conceptualized as an individual or group’s relative social standing or class (16,17). The main predictors of SES are education level, income, and occupation (15,17,18). The correlations between SES and mental health have been explained by various mechanisms. It has been found that negative impact of low SES on mental health (19) can be reduced by the mediating effect of social capital and physical health (4,18).Social capital has been defined as individual’s social networks and social interactions, shared norms, values, and understandings that facilitate collective action within or among groups. It can act as a protective factor, promoting mental health status by reducing socioeconomic inequalities (4,20) and play an important role in reducing the prevalence of mental disorders (4). Previous studies have found that social ties and support significantly improve mental health (9). Nonetheless, the association between social capital, mental health, quality of life, and SES is not consistently reported (21,22). This population-based study aims to explore the association between demographic factors, SES, social capital, HRQoL, and mental health among Tehran residents using structural-equation modeling (SEM).     

Predictors of Increased Left Ventricular Filling Pressure in Dialysis Patients with Preserved Left Ventricular Ejection Fraction

Aim

To study the left and right ventricular function and to assess the predictors of increased left ventricular (LV) filling pressure in dialysis patients with preserved LV ejection fraction.

Methods

This study included 63 consecutive patients (age 57 ± 14 years, 57% women) with end-stage renal failure. Echocardiography, including tissue Doppler measurements, was performed in all patients. Based on the median value of the ratio of transmitral early diastolic velocity to early myocardial velocity (E/E’ ratio), patients were divided into 2 groups: the group with high filling pressure (E/E’>10.16) and the group with low filling pressure (E/E’≤10.16).

Results

Compared with patients with low filling pressure, the group of patients with high filling pressure included a higher proportion of diabetic patients (41% vs 13%, P = 0.022) and had greater LV mass index (211 ± 77 vs 172 ± 71 g/m³, P = 0.04), lower LV lateral long axis amplitude (1.4 ± 0.3 vs 1.6 ± 0.3 cm, P = 0.01), higher E wave (84 ± 19 vs 64 ± 18cm/s, P < 0.001), lower systolic myocardial velocity (S’:8.6 ± 1.5 vs 7.0 ± 1.3 cm/s, P < 0.001), and lower diastolic myocardial velocities (E’: 6.3 ± 1.9 vs 9.5 ± 2.9 cm/s, P < 0.001; A’: 8.4 ± 1.9 vs 9.7 ± 2.5 cm/s, P = 0.018). Multivariate analysis identified LV systolic myocardial velocity – S’ wave (adjusted odds ratio, 1.909; 95% confidence interval, 1.060-3.439; P = 0.031) and age (1.053; 1.001-1.108; P = 0.048) as the only independent predictors of high LV filling pressure in dialysis patients.

Conclusions

In dialysis patients with preserved left ventricular ejection fraction, reduced systolic myocardial velocity and elderly age are independent predictors of increased left ventricular filling pressure.Cardiovascular disorders are the main cause of mortality and morbidity in patients with end-stage renal failure who are in regular hemodialysis programs (1,2). The left ventricular (LV) hypertrophy is a common finding in these patients. It reflects a physiological response to pressure and volume overload (3) and positively correlates with cardiovascular mortality (4). LV hypertrophy is frequently associated with LV dilatation and reduced systolic function (5). An increased incidence of atherosclerotic cardiovascular events in these patients has also been reported (6). Systolic dysfunction and LV hypertrophy have been identified as the best predictors of outcome in dialysis patients (4,7,8). However, the conventional systolic dysfunction appears in the late stages of the chronic renal failure (9).In contrast to conventional echocardiography, tissue Doppler imaging of the myocardial velocities overcomes the load dependence of diastolic parameters (10). The ratio of transmitral early diastolic velocity (E) to early myocardial velocity (E’) (E/E’ ratio) has been shown to be an accurate method of the LV filling pressure estimation (8) and the best predictor of LV diastolic filling in various cardiac pathologies (11,12), thereby serving as one of the best predictors of outcome in heart failure patients (13-15) and patients with end-stage renal disease (16).The aims of this study were to investigate the left and right ventricular function in patients with end-stage renal disease and preserved LV ejection fraction and to assess the predictors of increased LV filling pressure in these patients.     

Human Umbilical Cord Blood-derived Stromal Cells Suppress Xenogeneic Immune Cell Response In Vitro

Aim

To explore immunological properties of human umbilical cord blood-derived stromal cells (hUCBDSC) and their effect on xenogeneic immune cells in vitro.

Methods

Immunological phenotype of freshly isolated and cryopreserved hUCBDSCs was evaluated by flow cytometry. Xenogeneic splenic T-cells were stimulated by phytohemaglutinin A (PHA) or dendritic cells in the absence or presence of hUCBDSCs. T-cell proliferation was measured by cell counting kit-8 after 7-day incubation. The proportion of apoptotic cells and CD4⁺CD25⁺Foxp3⁺ regulatory T-cells (Tregs) was determined in T-cells activated by PHA in the absence or presence of hUCBDSCs by flow cytometry. Phenotype of dendritic cells, cultured alone or with hUCBDSCs, was analyzed by flow cytometry.

Results

Levels of immune molecule expression on freshly isolated hUCBDSCs were as follows: human leukocyte antigen-I (HLA-I) (84.1 ± 2.9%), HLA-II (1.6 ± 0.3%), CD80 (0.8 ± 0.1%), CD86 (0.8 ± 0.1%), CD40 (0.6 ± 0.1%), and CD40L (0.5 ± 0.1%), which was not influenced by cryopreservation. T-cell proliferation in the presence of hUCBDSCs was significantly lower than that of positive control. The coculture led to a 10-fold increase (from 1.2 ± 0.3% to 12.1 ± 1.4%, P < 0.001) in the proportion of CD4⁺CD25⁺Foxp3⁺ regulatory T-cells (Tregs) and a reversion of mature dendritic cells, as indicated by the down-regulation of major histocompatibility complex (MHC)-II molecule (49.3% vs 25.9%, P = 0.001), CD80 (47.2% vs 23.3%, P = 0.001), and CD86 (40.6% vs 25.1%, P = 0.002). When subjected to annexin V binding and propidium iodide uptake assay, the hUCBDSCs did not show the ability to induce apoptosis of xenogeneic T-cells.

Conclusion

These results demonstrate low immunogenicity and immunomodulation effect of the hUCBDSCs. Reversion of mature dendritic cells and increase in Treg proportion, but not cell apoptosis, can possibly contribute to the suppression of xenogeneic T-cell proliferation by the hUCBDSCs.Adherent spindle-shaped, fibroblastoid cells were isolated from the bone marrow by Friedenstein (1) and others (2-4), and are also referred to as multipotent mesenchymal stromal cells (MSC) (5) because of their self-renewal and multilineage differentiation capacity at the population level. One of the main biological functions of bone marrow MSCs is to regulate proliferation, differentiation, and maturation of hematopoietic cells through cell-to-cell interactions and secretion of cytokines and growth factors. Besides, MSCs also probably contribute to regulation of maturation, proliferation, and function of lymphocytes and have been shown to participate in the positive selection of T lymphocytes in the thymus in murine models (6-10).In fact, a variety of studies with human, baboon, and murine MSCs have demonstrated the inability of MSCs to elicit a proliferative response of allogeneic lymphocytes, but instead they showed the capability of MSCs to suppress T lymphocyte activation and proliferation in vitro when stimulated by alloantigens, mitogens, and CD3 and CD28 antibodies (11-17). Different mechanisms are proposed to explain the suppressive effect of MSCs depending on the stimuli. Some animal models, related to alloreactive immunity, autoimmunity, or tumor immunity, have been established to examine the immunomodulatory function of MSCs in vivo. Graft vs host disease (GVHD), induced by donor T-cells, constitutes the most frequent complication after allogeneic hematopoietic cell transplantation (HCT) (18,19). Early and repeated systemic administration of ex-vivo expanded MSCs reduced the incidence and severity of GVHD in mice (20). It was shown clinically that the infusion of MSCs was safe and effective in the treatment of GVHD (21,22). MSCs have been an attractive candidate for cell therapy for GVHD.Although MSCs have also been isolated from other tissues such as adipose tissue (23), placenta (24), amniotic fluid (25), and umbilical cord blood (26,27), most studies on immunomodulatory function focused on bone marrow MSCs. However, the bone marrow collection is an invasive procedure and the number and expansion capacity of bone marrow MSCs decline with age (28,29). Due to easy collection and younger age of donor cells, umbilical cord blood is one of the most attractive alternative sources of MSCs. Our laboratory has previously isolated a novel population of adherent fibroblast-like cells from human umbilical cord blood (hUCB) CD34⁺cells, called hUCB-derived stromal cells (hUCBDSCs) and confirmed their supportive effect on hematopoiesis in vitro (30).Previous studies have usually investigated human MSCs-allogeneic immune cell reaction in vitro (11,14,31-33). To test the feasibility of replacing human immune cells with xenogeneic counterparts in vitro is of significance in establishing an animal model for further in vivo study of immunological properties of human MSCs, such as their inhibitory effect on GVHD. In the present study, we focused on immunological properties of hUCBDSCs and their effect on xenogeneic immune cells and demonstrated that hUCBDSCs suppressed xenogeneic T-cell activation induced by mitogen PHA and dendritic cells, in addition to its low immunogenicity per se, such as the lack of human leukocyte antigen-II (HLA-II) and some costimulator expression.     

Red blood cell distribution width as a simple negative prognostic factor in patients with diffuse large B-cell lymphoma: a retrospective study

AimTo determine the prognostic value of baseline red blood cell distribution width (RDW) in diffuse large B cell lymphoma (DLBCL) patients.MethodsData from 81 DLBCL patients diagnosed from 2006 to 2013 at the University Hospital Center Osijek, Osijek, Croatia, were reviewed. We evaluated disease outcome, overall survival (OS) and event-free survival (EFS), and demographic, clinical and laboratory factors affecting outcome. Univariate analysis and Cox regression analysis were used.ResultsMedian age of patients was 64 years, 29 were men (35.8%). Higher RDW levels (%) were found in patients with advanced Ann Arbor clinical stage (14.94 ± 1.82 vs 13.55 ± 1.54, P = 0.001) and in those with poor response to therapy (14.94 ± 1.82 vs 13.55 ± 1.54, P = 0.001). Patients with RDW>15% (cut-off was calculated by receiver operating characteristics) had significantly worse OS (median [range], 33 months [20-46] vs 74 months [65-82], P < 0.001) and EFS (27 months [15-40] vs 68 months [59-77], P < 0.001). Cox regression analysis showed that RDW>15% was an independent prognostic factor for OS (HR 3.654, 95% CI 1.128-11.836) and EFS (HR 2.611, 95% CI 1.012-6-739).ConclusionHigh baseline RDW is an independent prognostic marker of poor outcome in patients with DLBCL. RDW could be an easily available and inexpensive marker for the risk stratification in patients with DLBCL.Red cell distribution width (RDW) is analyzed routinely as part of the complete blood count (CBC). It is a measure of heterogeneity of the red blood cell (RBC) size and traditionally has played a role in the differential diagnosis of anemia (1). High RDW values are associated with increased mortality in general population and in patients with cardiovascular disease, sepsis, acute kidney injury, chronic obstructive pulmonary disease, hepatitis B, and those on chronic dialysis (2-9). There is also evidence of its prognostic value in various malignancies (10-14). A recent study found a strong relation between high RDW and poor survival in patients with lung cancer (15). RDW was also found to be a significant predictor of poor prognosis in patients with malignant mesothelioma (16). In patients with symptomatic multiple myeloma, elevated RDW values were associated with a higher stage disease according to International Staging System and poor prognosis (17). The mechanism that could explain the relation between RDW and survival or disease activity is not clear, but it is considered that high RDW is caused by chronic inflammation, poor nutritional status, oxidative stress, and age-related diseases that lead to changes in erythropoiesis (2,18-20).Diffuse large B-cell lymphoma (DLBCL) is the most common group of lymphomas, amounting to 25% of all non-Hodgkin’s lymphomas (NHL) (21). It is a type of aggressive lymphoma that usually affects middle-aged and elderly patients. The distribution of NHL subtypes in Croatia corresponds to the European average (22). The most commonly used prognostic index in aggressive NHL is the international prognostic index (IPI) and its variants used in elderly patients (age-adjusted IPI) and in patients treated with rituximab (R-IPI) (23,24).So far, there have been no reports on the prognostic value of RDW in patients with DLBCL. The aim of our study was to determine whether RDW measured at diagnosis was an independent prognostic factor of disease outcome, overall survival (OS), and event-free survival (EFS) in patients with DLBCL.     

Sex-specific chronic stress response at the level of adrenal gland modifies sexual hormone and leptin receptors

Aim

To compare cardiometabolic risk-related biochemical markers and sexual hormone and leptin receptors in the adrenal gland of rat males, non-ovariectomized females (NON-OVX), and ovariectomized females (OVX) under chronic stress.

Methods

Forty six 16-week-old Sprague-Dawley rats were divided into male, NON-OVX, and OVX group and exposed to chronic stress or kept as controls. Weight, glucose tolerance test (GTT), serum concentration of glucose, and cholesterol were measured. Adrenal glands were collected at the age of 28 weeks and immunohistochemical staining against estrogen beta (ERβ), progesterone (PR), testosterone (AR), and leptin (Ob-R) receptors was performed.

Results

Body weight, GTT, serum cholesterol, and glucose changed in response to stress as expected and validated the applied stress protocol. Stressed males had significantly higher number of ERβ receptors in comparison to control group (P = 0.028). Stressed NON-OVX group had significantly decreased AR in comparison to control group (P = 0.007). The levels of PR did not change in any consistent pattern. The levels of Ob-R increased upon stress in all groups, but the significant difference was reached only in the case of stressed OVX group compared to control (P = 0.033).

Conclusion

Chronic stress response was sex specific. OVX females had similar biochemical parameters as males. Changes upon chronic stress in adrenal gland were related to a decrease in testosterone receptor in females and increase in estrogen receptor in males.Maintaining homeostasis is often challenged by different types of stressors (1). Homeostasis is regulated by a complex endocrine processes engaging the hypothalamic-pituitary-adrenal axis (HPA) and sympathetic autonomic system (2-4). Stress can occur either in acute or chronic form with different consequences – the acute stress mostly induces the ˝fight or flight˝ response, while chronic stress promotes long term changes, which can lead to a variety of diseases (5,6). If stress is of sufficient magnitude and duration, the action of HPA is unsuppressed and results in prolonged elevation of cortisol (7), induced production of energy, vasoconstriction, lipolysis, proteolysis, immunosuppression, and suppression of reproductive function to save energy and retain overall homeostasis (8). Women are generally less susceptible to chronic stress up to the period of menopause, when the loss of protective hormones, estrogen and progesterone, occurs and thus they become prone to development of depression, anxiety, or schizophrenia (9). In contrast, men are generally more susceptible and sensitive to chronic stress, showing changes in feeding habits and decreased body weight (10,11).Chronic stress can cause the development of cardiovascular disorder, obesity, and diabetes, which can be reflected in serum cholesterol, glucose, and decreased glucose tolerance (12-14). There is a strong correlation between stress and sexual hormones, but the mechanisms by which estrogen, testosterone, and progesterone exert their possible protective role under stress conditions are not fully explored. Sexual hormones affect stress outcome and stress hormones affect the levels of sexual hormones (15-17). Testosterone is activated during stress response in rats and humans (18,19) and tends to increase more in men than women (20). Estrogen lowers the stress-induced response in women and men (9,21). Estrogens and progesterone are produced even after ovariectomy by adrenal glands (22) but it is not known if such compensation can withstand additional challenge like stress. Another possible player in stress response is leptin (Ob), hormone responsible for maintaining body weight, which is synthesized and secreted by adipose tissue (23), exerting its effects through the leptin receptor (Ob-R) (24). Chronic stress models imply a direct link between stress response and leptin (25,26). Receptors for leptin are present in the adrenal gland (27). The aim of this study was to investigate cardiovascular risk parameters and changes in leptin and sexual hormone receptors in adrenal gland during chronic stress. There is a clinically relevant change in the onset of cardiometabolic risk between healthy women and women with premature ovarian failure (28) and because of that ovariectomized female rats were included in the study.     

Ultrasound measurements of carotid intima-media thickness and plaque in HIV-infected patients on the Mediterranean diet

Aim

To evaluate the influence of food habits, specifically adherence to the Mediterranean diet, on carotid intima-media thickness (CIMT) and the presence of plaques in HIV-infected patients taking antiretroviral therapy (ART) and non-HIV-infected participants and to determine if HIV infection contributes independently to subclinical atherosclerosis.

Methods

We conducted a cross-sectional study of 110 HIV-infected patients on ART and 131 non-HIV-infected participants at the University Hospital for Infectious Diseases in Zagreb, Croatia, from 2009-2011. CIMT measurement and determination of carotid plaque presence was detected by ultrasound. Adherence to the Mediterranean diet was assessed by a 14-point food-item questionnaire. Subclinical atherosclerosis was defined by CIMT≥0.9 mm or ≥1 carotid plaque.

Results

In HIV-infected patients, subclinical atherosclerosis was associated with older age (P < 0.001; Mann-Whitney test), higher body mass index (P = 0.051; Mann-Whitney test), hypertension (P < 0.001; χ² test), and a lower Mediterranean diet score (P = 0.035; Mann-Whitney test), and in non-HIV-infected participants with older age (P < 0.001; Mann-Whitney test) and hypertension (P = 0.006; χ² test). Multivariate analysis showed that decreased adherence to the Mediterranean diet was associated with higher odds of subclinical atherosclerosis (odds ratio [OR] 2.28, 95% confidence interval [CI] 1.10-4.72, P = 0.027) as was current smoking (OR 2.86, 95% CI 1.28-6.40), hypertension (OR 3.04, 95% CI 1.41-6.57), and male sex (OR 2.35, 95% CI 0.97-5.70). There was a significant interaction of age and HIV status, suggesting that older HIV-infected patients had higher odds of subclinical atherosclerosis than controls (OR 3.28, 95% CI 1.24-8.71, P = 0.017 at the age of 60 years).

Conclusion

We confirmed the association between lower adherence to the Mediterranean diet and increased risk of subclinical atherosclerosis and found that treated HIV infection was a risk factor for subclinical atherosclerosis in older individuals.The advent of antiretroviral therapy (ART) for treatment of human immunodeficiency virus (HIV) infection has improved the quality of life and life expectancy of HIV-infected patients (1,2). However, a number of comorbidities have emerged, including atherosclerosis (3,4).The impact of HIV infection and exposure to ART on development of subclinical atherosclerosis is incompletely understood (1,5). Several factors may contribute to an increased risk of coronary and cerebrovascular artery disease in HIV-infected patients: chronic endothelial or myocardial inflammation due to HIV infection per se, dyslipidemia associated with ART, and the interaction of treatment with traditional risk factors for cardiovascular disease (smoking, hypertension, age, etc) (1). Also, several antiretroviral drugs are believed to be partially related to an increase in cholesterol levels (6). The risk and problems associated with ART have led to the study of treatment-sparing strategies, which might provide the benefits of ART while minimizing the risk of adverse advents and other risks (7). The Strategies for Management of Antiretroviral Therapy (SMART) trial was conducted to compare the episodic use of ART according to the CD4+ count with the current practice of continuous ART (7).An early marker of atherosclerosis is an increase in carotid intima-media thickness (CIMT), which enables the risk assessment of coronary and cerebral-artery disease (8). Carotid plaque area is another marker, which is more strongly associated with traditional risk factors and is more predictive of myocardial infarction than of stroke (9,10). There are many studies on the association of HIV infection, HIV disease parameters, and ART use with subclinical carotid artery atherosclerosis, yet there is still controversy over whether chronic HIV infection and ART (mainly protease inhibitors, PI) contributes to subclinical atherosclerosis. Although there are cross-sectional studies reporting an association between HIV infection, ART, and subclinical atherosclerosis (11-16), a number of studies did not find such an association (1,17-19).The Mediterranean diet (20) has been shown to protect against cardiovascular disease and other chronic conditions (21-23). However, only a few studies have investigated the relationship of dietary intake with body fat changes and metabolic abnormalities in HIV-infected patients (24-26). To the best of our knowledge, the association between the adherence to Mediterranean diet and CIMT and the presence of plaques in HIV-infected patients taking ART has never been explored.The aims of the present study are to evaluate the influence of food habits, specifically adherence to the Mediterranean diet, on CIMT and the presence of plaques in HIV-infected patients taking ART and non-HIV-infected participants and to assess if HIV-infection contributes independently to subclinical atherosclerosis.     

Oxidative stress impact on growth hormone secretion in the eye

AimTo evaluate the influence of oxidative stress on extrapituitary growth hormone (GH) secretion in the eye and to analyze the interdependence between eye and serum GH levels under normal and hypoxic conditions.MethodsPars plana vitrectomy (PPV) was performed in 32 patients with developed proliferative diabetic retinopathy (PDR) and 49 non-diabetic controls, both of whom required this procedure as part of their regular treatment in the period from April 2013 to December 2014. During PPV, vitreous samples were taken and blood was simultaneously collected from the cubital vein. GH levels in serum and vitreous samples were measured by electrochemical luminescence assay. Oxidative stress was measured by enzyme-linked immunosorbent assay of advanced oxidation protein products (AOPP) and lipid hydroperoxide (LPO) in serum and vitreous.ResultsSerum AOPP levels were significantly higher than vitreous levels in both groups (P < 0.001 for each group) and LPO levels were significantly higher only in PDR group (P < 0.001). There was a significant positive correlation between serum and vitreous LPO levels in PDR group (r = 0.909; P < 0.001). Serum GH levels were significantly higher than vitreous levels in both groups (P < 0.001 for each group). Serum GH levels were significantly higher in PDR group than in controls (P = 0.012). Vitreous GH values were slightly higher in PDR group, but the difference was not significant.ConclusionOur study confirms that GH production in the eye is autonomous and independent of oxidative stress or pituitary GH influence.The latest research has indicated that the secretion of growth hormone (GH) in human retinal ganglion cells (RGC) is maintained throughout the postembryonic life, and has a neuroprotective role (1-8). Serum human GH levels are elevated in diabetes and their possible correlation with etiopathogenesis of diabetic retinopathy (DR) has been investigated, but not confirmed (9-12). Diabetes is considered to be a primarily metabolic disorder that, besides leading to other complications, affects small and large blood vessels, thus causing hypoxia and oxidative stress in different tissues and organs. Therefore, DR represents an ideal model for exploring extrapituitary GH release, as well as various aspects of serum GH production (13). However, we still do not know the exact mechanism of how serum human GH interferes with the development of DR. The key assumption is that reduced amounts of insulin in the portal circulation reduce insulin growth factor 1 (IGF1) production in the liver with the final lack of feedback inhibition of increased GH secretion (12,13). All this takes place in conditions of hyperglycemia that is being suppressed by introduction of additional amounts of insulin, which provides the necessary condition for increased portal IGF1 production. In this stage of the disease, circulating values of both GH and IGF1 are elevated in serum (11,13,14) and this stage is usually associated with the appearance of DR. It has been proven that IGF1 exerts a promotional effect on neovascularization and proliferative changes in the eye, but it is unclear whether the increased levels of IGF1 measured in diabetic vitreous are derived from the passage of serum IGF1 through the blood retinal barrier (BRB) or it is autonomous growth factor created within the eye (15,16).There are controversial findings on the effects of iatrogenic GH medical suppression in serum on DR. Some earlier studies, primarily on animal models, have shown that suppression of serum GH decreases the synthesis of IGF1 and thus slows down DR progression (17,18). Recent studies on a large number of human participants have shown that suppression of serum GH, despite the accompanying reduction in serum IGF1 levels, does not affect the stage of DR, which implies the existence of an autonomous IGF1 production in the eye (16,19).It should be noted that insulin is significant IGF 1 secretion promoter just like GH, and that patients with type I and some with type II diabetes receive insulin therapy as their standard treatment procedure. Nevertheless, in such conditions insulin increasingly penetrates the BRB and additionally stimulates autonomous creation of IGF1 in the eye (20-23). In normal conditions, this barrier is impermeable to GH and IGF1, but animal model studies showed that in conditions of hypoxia and oxidative stress it became permeable to both molecules (13,21,24). Human model, however, showed that the GH concentration in vitreous was lower in diabetic than in nondiabetic patients. This fact rules out the possibility of excessive serum GH passing through the damaged BRB, but indicates autonomous production of GH within the eye (13,25). It is believed that somatostatin (SST), also known as inhibitor of GH secretion, is an important regulator of GH homeostasis in the eye and its production is confirmed in the retinal ganglion cells and retinal pigment epithelial cells (RPE) cells (26,27).The aim of this study was to explore GH secretion and its dependence on oxidative stress in the eye and also its correlation with GH in the systemic circulation. We assumed that the eye production of GH in oxidative stress was reduced due to the simultaneous apoptosis of neural cells that produce it.     

Significance of Off-hours in Centralized Primary Percutaneous Coronary Intervention Network

Aim

To analyze the efficacy of a regionally organized primary percutaneous coronary intervention (PCI) network at the Heart Center, Semmelweis University Budapest, part of the “Budapest model,” and the factors that influence it.

Methods

In order to investigate the differences between regular and off-hours patient care in a 24-hour myocardial infarction primary care system, we included 1890 consecutive, unselected patients with ST-segment elevation myocardial infarction and followed them until at least one year. The follow-up was complete for all participants.

Results

The difference between regular hours and off-hours mortality was not significant after 30 days (8.6% vs 8.8%, respectively) or after 1 year (15.3% vs 14.7%, respectively). The rate of patients with re-infarction, frequency of re-intervention, and major adverse cardiac events, including death, re-infarction, re-intervention, and coronary artery bypass graft surgery, were similar in both patient groups. The time delay between the onset of chest pain and arrival to the clinic was 5.9 ± 5.8 hours (mean ± standard deviation)during regular hours and 5.2 ± 4.6 hours during off-hours (P = 0.235). Direct transport caused significant decrease in the 30-day and 1-year mortality independent of duty time (7.2% vs 9.9%, P = 0.027; 12.6% vs 16.7%, P = 0.028; respectively).

Conclusion

Centralized primary PCI network of the “Budapest model” achieved the same level of patient care during both off-hours and regular hours.The generally accepted treatment of acute ST elevation myocardial infarction (STEMI) within 12 hours is primary percutaneous coronary intervention (PCI). However, the outcome of PCI may be influenced by several factors, like annual number of procedures (1-4), experience of the operating physician, time delay to treatment (5-8), and organization level of myocardial infarction care (9-16). In case of organized primary PCI network, the most important factor that influences the clinical outcome is the time of the arrival to the PCI center. Patients treated during off-hours can have a higher incidence of failed operation procedure and consequently a worse prognosis than patients treated during regular hours (17-22).Assali et al (18) reported that the unadjusted mortality at 1 month was significantly higher in patients treated during the night than in those treated during the day (9.7% vs 3.1%) (18). Henriques et al (19) demonstrated that the admittance of patients between 8:00 am and 6:00 pm was associated with an angioplasty failure rate of 3.8%, compared with 6.9% between 6:00 pm and 08:00 am Thirty-day mortality was 1.9% in patients with hospital admission between 8:00 am and 6:00 pm, compared with 4.2% in patients with hospital admission between 6:00 pm and 8:00 am (19). A related study of 231 164 STEMI patients (20) showed that the 30-day mortality was significantly higher for patients admitted on weekends (12.9% vs 12.0%). A cohort study of 68 439 patients with STEMI (21) showed that these patients had substantially longer door-to-balloon times during off-hours (116.1 minutes) than during regular hours (94.8 minutes; difference, 21.3 minutes; 95% confidence interval, 20.5-22.2).There are only limited data about organized primary PCI networks in large European cities (11-13,23,24). Despite the fact that it has been shown that high-quality organization of the primary PCI network may improve the outcome (25,26), a well organized, 24-hour available “service” for every citizen is rarely available (11).Five years ago we established a centralized myocardial infarction primary care model for the Budapest region, called the “Budapest model” (16), in which all the patients from the region requiring reperfusion therapy are primarily transported for PCI. The model comprises 5 PCI centers that perform primary PCI care on weekdays during regular hours, while during off-hours 1 of the 5 centers is on duty and admits all the patients.The aim of our study was to analyze the efficacy of the primary PCI network in Budapest and central Hungary, and to study the differences between regular and off-hours patient care in a 24-hour myocardial infarction primary care system.