Green Synthesized Gold and Silver Nanoparticles Increased Oxidative Stress and Induced Cell Death in Colorectal Adenocarcinoma Cells.

The research investigated the effect of gold (Au-CM) and silver nanoparticles (Ag-CM) phytoreduced with Cornus mas fruit extract (CM) on a human colorectal adenocarcinoma (DLD-1) cell line. The impact of nanoparticles on the viability of DLD-1 tumor cells and normal cells was evaluated. Oxidative stress and cell death mechanisms (annexin/propidium iodide analysis, caspase-3 and caspase-8 levels, p53, BCL-2, BAX, NFkB expressions) as well as proliferation markers (Ki-67, PCNA and MAPK) were evaluated in tumor cells. The nanoparticles were characterized using UV-Vis spectroscopy and transmission electron microscopy (TEM) and by measuring zeta potential, hydrodynamic diameter and polydispersity index (PDI). Energy dispersive X-ray (EDX) and X-ray powder diffraction (XRD) analyses were also performed. The nanoparticles induced apoptosis and necrosis of DLD-1 cells and reduced cell proliferation, especially Ag-CM, while on normal cells, both nanoparticles maintained their viability up to 80%. Ag-CM and Au-CM increased the expressions of p53 and NFkB in parallel with the downregulation of BCL-2 protein and induced the activation of caspase-8, suggesting the involvement of apoptosis in cell death. Lipid peroxidation triggered by Ag-CM was correlated with tumor cell necrosis rate. Both nanoparticles obtained with phytocompounds from the CM extract protected normal cells and induced the death of DLD-1 tumor cells, especially by apoptosis.

Arterial stiffness in aortic stenosis - complex clinical and prognostic implications.

Arterial stiffness and degenerative aortic stenosis (AoS) are frequently associated leading to a combined valvular and vascular load imposed on the left ventricle (LV). Vascular load consists of a pulsatile load represented by arterial stiffness and a steady load corresponding to vascular resistance. Increased vascular load in AoS has been associated with LV dysfunction and poor prognosis in pre-intervention state, as well as after aortic valve replacement (AVR), suggesting that the evaluation of arterial load in AoS may have clinical benefits. Nevertheless, studies that investigated arterial stiffness in AoS either before or after AVR used various methods of measurement and their results are conflicting. The aim of the present review was to summarize the main pathophysiological mechanisms which may explain the complex valvulo-arterial interplay in AoS and their consequences on LV structure and function on the patients' outcome. Future larger studies are needed to clarify the complex hemodynamic modifications produced by increased vascular load in AoS and its changes after AVR. Prospective evaluation is needed to confirm the prognostic value of arterial stiffness in patients with AoS. Simple, non-invasive, reliable methods which must be validated in AoS still remain to be established before implementing arterial stiffness measurement in patients with AoS in clinical practice.

Relationship between 25 Hydroxyvitamin D, Overweight/Obesity Status, Pro-Inflammatory and Oxidative Stress Markers in Patients with Type 2 Diabetes: A Simplified Empirical Path Model.

Vitamin D deficiency is highly prevalent in patients with overweight/obesity and type 2 diabetes (T2DM). Herein, we investigated the relationship between vitamin D status and overweight/obesity status, insulin resistance (IR), systemic inflammation as well as oxidative stress (OS). Anthropometric and laboratory assessments of 25-hydroxyvitamin D (25(OH)D) and glycemic, pro-inflammatory and OS biomarkers were performed in a sample of 47 patients with T2DM who were divided into categories based on overweight and degree of obesity. The main findings were: the overweight/obesity status correlated negatively with the degree of serum 25(OH)D deficiency (ρ = -0.27) with a trend towards statistical significance (p = 0.069); the homeostasis model assessment of insulin resistance (HOMA-IR) was significantly different (p = 0.024) in patients with 25(OH)D deficiency, as was total oxidant status (TOS) and oxidative stress index (OSI) in patients with severe serum 25(OH)D deficiency as compared to those with 25(OH)D over 20 ng/mL (TOS: p = 0.007, OSI: p = 0.008); and 25(OH)D had a negative indirect effect on TOS by body mass index (BMI), but BMI was not a significant mediator of the studied relationship. In a setting of overweight and increasing degree of obesity, patients with T2DM did not display decreasing values of 25(OH)D. Subjects with the lowest values of 25(OH)D presented the highest values of BMI. Patients with 25(OH)D deficiency were more insulin resistant and showed increased OS but no elevated systemic inflammation. The negative effect of 25(OH)D on TOS did not seem to involve BMI as a mediator.

Neuronal apoptosis can be prevented by the combined therapy with melatonin and hypothermia in a neonatal rat model of hypoxic-ischemic encephalopathy.

INTRODUCTION: Birth hypoxia is a leading cause of perinatal mortality and neurological morbidity, resulting in central nervous system injury. Cerebral hypoxia and ischemia can produce a severe brain damage following a typical pattern, defined by selective vulnerability of the brain regions. The neonates are most prone to hypoxic-ischemic injuries due to the lack of efficient antioxidant defense. Neonatal hypoxia-ischemia (HI) in a 7-day-old rat HI model can produce cell death by apoptotic or necrotic mechanisms. The degree of apoptotic or necrotic mechanisms responsible for cell death in neonatal hypoxia-ischemia are not very clear as yet. The form of neuronal death may also depend on the severity of ischemic injury. Necrosis predominates in more severe cases, whereas apoptosis occurs in areas with milder ischemic injury. A human study demonstrated apoptotic and necrotic forms of cell death after hypoxic injury, whereas in some brains from stillbirths, only apoptotic figures were observed. The expression of activated caspase-3 reflects the role of apoptosis in neonatal hypoxic ischemic brain injury. OBJECTIVES: The aim of this study was to evaluate the possible neuroprotective effect of melatonin and hypothermia in hypoxic-ischemic encephalopathy in newborn rats. Local damages induced by hypoxia and ischemia were assessed by evaluating the changes in terms of histology and apoptosis. METHODS: The experiment was conducted on 20 newborn Wistar rats premedicated for seven days with melatonin in a dose of 20 mg/kg/day. On the 7th postnatal day (P7), the newborn rats were exposed to ischemia (by clamping the right carotid artery) and hypobaric hypoxia (8% O2 for 90 minutes) and some groups to hypothermia. RESULTS: In this experimental model of neonatal encephalopathy, melatonin, in a dose of 20 mg/kg/day has neuroprotective effect by reducing the number of cells expressing apoptosis in Cornu Ammonis (CA) (Ammon's Horn) CA1, CA2, CA3 and dentate gyrus of the hippocampus when combined with hypothermia. CONCLUSION: The results of this study prove that melatonin is protective in ischemic-hypoxic brain injuries, but the protection is conditioned in most of the brain regions (excepting cerebral cortex) by conjugation with post-injury hypothermia treatment.

The impact of silver nanoparticles phytosynthesized with Viburnum opulus L. extract on the ultrastrastructure and cell death in the testis of offspring rats.

AIM: To investigate the effects of prenatal exposure to AgNPs obtained by green synthesis with Viburnum opulus L. extract on the testis in male offspring rats. MATERIAL AND METHODS: Two different doses of AgNPs (0.8 and 1.5 mg/kg b.w.) and vehicle (PBS) were administered to Wistar female rats on days 3-14 of gestation. At 6 weeks after birth, the ultrastructural changes in correlation with the amount of silver as well as the parameters of oxidative stress, inflammation and cell death mechanisms in the testis of male offspring were evaluated. RESULTS: AgNPs administered during pregnancy crossed the placental and testicular barriers and induced oxidative stress, DNA damage and autophagy as mechanism of cell toxicity. The markers of inflammation and apoptosis decreased after AgNPs exposure while the NFkB activation increased. TEM examination revealed important ultrastructural changes of Sertoli cells, numerous vacuoles and cytoplasmic changes suggestive of the cell's evolution towards necrosis. CONCLUSION: Phytoreduced silver nanoparticles with polyphenols from Viburnum opulus L. fruit extract, administered during the embryological development of the male gonad, have testicular toxic effects in offspring even at 6 weeks after birth.

Quercetin and curcumin effects in experimental pleural inflammation.

BACKGROUND: The inflammatory mechanisms occur with the highest prevalence in pulmonary pathology in addition to oxidative stress and activation of intracellular signaling pathways. The oxidative stress represents the imbalance between pro-oxidants and antioxidants which can lead to the activation of the oxidative mechanisms with noxious potential to the body. Therefore, finding a therapy that would counteract the injurious effects of free radicals and inflammation is highly attractive. Quercetin is the most active flavonoid, with important anti-inflammatory and antioxidant effects, while curcumin has antioxidant effects that are similar to the standard antioxidants and exerts direct anti-inflammatory activity. AIMS: The aim of this study is to evaluate the antioxidant effects of quercetin and curcumin on an experimental model, pleural inflammation induced by carrageenan. METHODS: Eight groups of adult male rats were used: Ia and Ib - control groups, IIa and IIb - with carrageenan administration, IIIa and IIIb - received curcumin and carrageenan, IVa and IVb - quercetin and carrageenan administration. Blood and lung samples were taken at 4 hours (Ia, IIa, IIIa, IVa groups) and at 24 hours (Ib, IIb, IIIb, IVb groups) after carrageenan injection. RESULTS: At 4 and at 24 hours, curcumin and quercetin have shown protective systemic effects, decreasing significantly the oxidative stress (malondialdehyde level) and stimulating significantly the antioxidant protection (ceruloplasmin and glutathione levels) compared to the group that received only carrageenan. In the lungs, at 4 hours, the redox misbalance was significantly reduced only in animals that were treated with quercetin, modifications that were not observed at 24 hours. CONCLUSIONS: In serum, curcumin presented higher antioxidant effects, compared to quercetin. In lungs, quercetin administration showed superior beneficial effects, but only temporarily.

Weight Loss and Late Complications after Silastic Ring Vertical Gastroplasty. A 10 Year Follow-up.

BACKGROUND AND AIM: Severe obesity is a public health care system challenge that requires bariatric surgery. Among the plethora of bariatric surgery techniques silastic ring vertical gastroplasty (SRVG) is a safe and efficient restrictive method that has been successfully used previously. However, it's performance by open approach has been abandoned and replaced by other methods using the laparoscopic method. The aim of the present study was to investigate patients with severe obesity submitted to open SRVG and to report our results over a period of 10 years in terms of weight loss, late complications and surgical re-interventions. MATERIAL AND METHODS: 112 severely obese patients submitted to open SRVG between years 2008- 2009 were investigated retrospectively for body mass index (BMI), percent excess BMI loss (%EBMIL), late surgical complications and reoperations. 41.96% of the patients were followed up 10 years after SRVG. Results: The initial mean BMI was 47.38 +- 7.59 kg/m2 and dropped statistically significant (p 0.001) to 31.05 +- 6.54 kg/m2 by the first year after SRVG. The mean BMI was rather stable along the first 5 years after SRVG when it started to increase gradually, reaching 35.93 +- 7.20 kg/m2 by the 10th year of follow-up when it remained still significantly lower (p 0.001) as compared to the mean baseline value. The %EBMIL was 79% at one year after surgery and reached 51% by the 10th year of follow-up. The most frequent late complications after SRVG were stoma stenosis (8.92%), enlargement of the stoma (8.03%) and incisional hernia (3.36%). As a consequence of stoma stenosis the ring has been removed in all cases. In 2 cases, after the ring removal, the patients underwent gastric bypass. CONCLUSION: SRVG is a safe and efficient restrictive technique of bariatric surgery open to many options to be revised, leading to a successful sustained long term weight loss and maintenance. Stoma stenosis, enlargement of the stoma and incisional hernia are the most frequent late complications after SRVG requesting reoperations.

Effects of sleeve gastrectomy on insulin resistance.

BACKGROUND AND AIM: Obesity is a major risk factor for the onset of insulin resistance (IR), hyperinsulinemia and type 2 diabetes mellitus (T2DM) Evidence data has proven that beyond important weight loss bariatric surgery especially Roux-en-Y gastric bypass (RYGB) and bilio-pancreatic diversion (BPD) leads to significant early reduction of insulinemia and of IR calculated through the homeostatic model assessment (HOMA-IR), independently of fat mass decrease. Sleeve gastrectomy (SG) is now used as a sole weight loss operation with good results. Therefore, the aim of the present study was to investigate the early changes of fasting blood glucose, insulin and HOMA-IR in a group of morbidly obese (MO) patients i.e. at 7, 30 and 90 days after SG. METHODS: The study included 20 MO patients (7 male and 13 female) submitted to SG. Anthropometrical (weight, body mass index -BMI, percent excess BMI loss -%EBMIL) and biochemical (plasma glucose, insulin and calculated HOMA-IR ) evaluation were performed before and at 7, 30 and 90 days after SG. In addition, a second group of 10 normal weight healthy subjects with a BMI ranging form 19 kg/m(2) to 23.14 kg/m(2), matched for age and gender was investigated. RESULTS: Plasma glucose (p=0.018), insulin (p=0.004) and HOMA-IR (p=0.006) values were statistically different between the studied groups. After surgery, at every follow-up point, there were statistically different weight and BMI mean values relative to the operation day (p<0.003). BMI, decreased at 7 days (estimated reduction=2.79; 95% CI:[2.12;3.45]), at 30 days (estimated reduction=5.65; 95% CI:[3.57;7.73]) and at 90 days (estimated reduction=10.88; 95% CI:[7.35;14.41]) respectively after SG. We noted a tendency toward statistical significant change of mean insulin values at 7 days after surgery (corrected p=0.075), no statistical change at 30 days (corrected p=0.327) and a significant change at 90 days (corrected p=0.027) after SG as compared to baseline. There was a significant change in mean values of HOMA-IR at 30 days (corrected p=0.009) and at 90 days (corrected p=0.021) after the operation day. CONCLUSIONS: The present study showed important early changes consisting in reductions of mean values of plasma insulin and HOMA-IR after SG.

Morpho-pathological and physiological changes of the brain and liver after ozone exposure.

This paper presents the consequences of long exposure to ozone in order to draw attention to this matter as far as the brain and liver are concerned. The material used was represented by two batches of 10 rats each that were daily exposed to ozone for 10 minutes at 0.5 ppm O3. From the first group blood was collected after two weeks to determine the indicators of oxidative stress and samples of brain and liver were drawn for histological studies. Tissue changes were highlighted using Hematoxylin-Eosin and argentic impregnation. In addition, the brain and liver samples taken from study subjects were turned into homogeneous preparations in order to determine the intensity of oxidative stress occurred in these organs compared with the witness group. The second batch was exposed for a further two weeks, after which the same sampling techniques and determining methods as for the first group were applied. The results show a correlation between the values of malondialdehyde (MDA) and glutathione (GSH), obtained both in blood and in the homogeneous preparations, and the microscopic changes that implicate a pathological state. Therefore, cerebral edema was discovered in the brain hemispheres and the cerebellum indicating necrotic signs accompanied by a reduction in the molecular layer and Purkinje cells with pale core. The liver presented hepatocellular necrosis, extended from the port area to the centrolobular vein.