Correlation between oxidative stress and G6PD activity in neonatal jaundice.

Fetal distress represents a pathophysiological condition in which oxygen is not available to the fetus in sufficient quantities. In cases of glucose 6-phosphate dehydrogenase (G6PD) deficiency, under conditions of oxidative stress, the residual G6PD and complimentary antioxidant mechanisms may become insufficient to neutralize the large amounts of ROS and to prevent severe hemolysis. Alteration in the oxidant-antioxidant profile is also known to occur in neonatal jaundice. The study group included 22 neonates presented with fetal distress during labor and 24 neonates with no evidence of fetal distress (control group). Umbilical cord blood samples were taken immediately after delivery, and the following blood tests were carried out after birth and at discharge from the hospital: erythrocyte count, total bilirubin, G6PD activity, and parameters presenting oxidative status [thiobarbituric acid reactive substances (TBARS), NO, O2 (-), H2O2, SOD, CAT, O2 (-)/SOD, and H2O2/CAT]. There were no significant differences in TBARS and NO values among neonates with or without fetal distress. However, the values of O2 (-), H2O2, SOD, O2 (-)/SOD, and H2O2/CAT among neonates born after fetal distress were significantly higher than in neonates without fetal distress (p < 0.01). In neonates with fetal distress, the total number of RBCs at delivery was significantly lower, accompanied with higher bilirubin content. Also neonates with fetal distress had lower activity of G6PD and lower CAT activity. Higher values of oxidative stress parameters in newborns delivered after fetal distress do not indicate strictly what occurred first-oxidative stress or basic lower G6PD activity.

A novel semi-quantitative method for measuring tissue bleeding.

In this study, we describe a new semi-quantitative method for measuring the extent of bleeding in pathohistological tissue samples. To test our novel method, we recruited 120 female patients in their first trimester of pregnancy and divided them into three groups of 40. Group I was the control group, in which no dilation was applied. Group II was an experimental group, in which dilation was performed using classical mechanical dilators. Group III was also an experimental group, in which dilation was performed using a hydraulic dilator. Tissue samples were taken from the patients' cervical canals using a Novak's probe via energetic single-step curettage prior to any dilation in Group I and after dilation in Groups II and III. After the tissue samples were prepared, light microscopy was used to obtain microphotographs at 100x magnification. The surfaces affected by bleeding were measured in the microphotographs using the Autodesk AutoCAD 2009 program and its "polylines" function. The lines were used to mark the area around the entire sample (marked A) and to create "polyline" areas around each bleeding area on the sample (marked B). The percentage of the total area affected by bleeding was calculated using the formula: N = Bt x 100 / At where N is the percentage (%) of the tissue sample surface affected by bleeding, At (A total) is the sum of the surfaces of all of the tissue samples and Bt (B total) is the sum of all the surfaces affected by bleeding in all of the tissue samples. This novel semi-quantitative method utilizes the Autodesk AutoCAD 2009 program, which is simple to use and widely available, thereby offering a new, objective and precise approach to estimate the extent of bleeding in tissue samples.

Authors' response to comments on "Factors influencing the removal of divalent cations by hydroxyapatite".

Recently, it has come to our attention, that a polemical note by Milonjic [S.K. Milonjic, Comments on "factors influencing the removal of divalent cations by hydroxyapatite", J. Hazard. Mater. 162 (2009) 1588-1589], has been published raising several critical comments on some experimental procedures, as well as the presentation and interpretation of some data found in the cited paper by Smiciklas et al. [I. Smiciklas, A. Onjia, S. Raicevic, Dj. Janackovic, M. Mitric, Factors influencing the removal of divalent cations by hydroxyapatite, J. Hazard. Mater. 152 (2008) 876-884]. Since we have not received any prior copy of this discussion through the process involved in the acceptance of these comments, we now take the opportunity of responding. We have reviewed some scientific papers dealing with cation sorption, in order to demonstrate variety of experimental approaches used for batch sorption tests, regarding solution pH. Finally, we justified experimental methods and data interpretation methods we chose to perform in the mentioned manuscript.

Theoretical assessment of phosphate amendments for stabilization of (Pb+Zn) in polluted soil.

Contamination of the environment with toxic metals, such as lead (Pb), represents a serious concern for human health. Most of the studies on Pb stabilization were performed using various phosphorus-containing amendments that can reduce Pb mobility and bioavailability by the sorption and precipitation of new, stable pyromorphite-type minerals, presenting very low solubility and bioaccessibility. However, the presence of competing ions, such as zinc (Zn), can reduce stabilization efficacy. The role of chemical composition on the stability of immobilization products of Pb and Zn by the addition of hydroxyapatite (HAP) or fluoroapatite (FAP) has been examined in this paper. In this analysis we used a theoretical criterion which is based on calculation of the ion-ion interaction potential, representing the main term of the cohesive energy of the matrix/pollutant system. It has been demonstrated that the stability of the HAP matrix decreases and that the stability of the FAP matrix increases with the Pb immobilization in the presence of Zn. The results of this analysis point out FAP as an advantageous amendment for the immobilization of Pb in the presence of Zn.

Factors influencing the removal of divalent cations by hydroxyapatite.

The effect of pH, contact time, initial metal concentration and presence of common competing cations, on hydroxyapatite (HAP) sorption properties towards Pb(2+), Cd(2+), Zn(2+), and Sr(2+) ions was studied and compared using a batch technique. The results strongly indicated the difference between the sorption mechanism of Pb(2+) and other investigated cations: the removal of Pb(2+) was pH-independent and almost complete in the entire pH range (3-12), while the sorption of Cd(2+), Zn(2+) and Sr(2+) generally increased with an increase of pH; the contact time required for attaining equilibrium was 30 min for Pb(2+) versus 24h needed for other cations; maximum sorption capacity of HAP sample was found to be an order of magnitude higher for Pb(2+) (3.263 mmol/g), than for Cd(2+) (0.601 mmol/g), Zn(2+) (0.574 mmol/g) and Sr(2+) (0.257 mmol/g); the selectivity of HAP was found to decrease in the order Pb(2+)>Cd(2+)>Zn(2+)>Sr(2+) while a decrease of pH(PZC), in respect to the value obtained in inert electrolyte, followed the order Cd(2+)>Zn(2+)>Pb(2+)>Sr(2+); neither of investigated competing cations (Ca(2+), Mg(2+), Na(+) and K(+)) influenced Pb(2+) immobilization whereas the sorption of other cations was reduced in the presence of Ca(2+), in the order Sr(2+)>Cd(2+)>or=Zn(2+). The pseudo-second order kinetic model and Langmuir isotherm have been proposed for modeling kinetic and equilibrium data, respectively. The sorption of all examined metals was followed by Ca(2+) release from the HAP crystal lattice and pH decrease. The ion exchange and specific cation sorption mechanisms were anticipated for Cd(2+), Zn(2+) and Sr(2+), while dissolution of HAP followed by precipitation of hydroxypyromorphite (Pb(10)(PO(4))(6)(OH)(2)) was found to be the main operating mechanism for Pb(2+) immobilization by HAP, with the contribution of specific cation sorption.

Theoretical stability assessment of uranyl phosphates and apatites: selection of amendments for in situ remediation of uranium.

Addition of an amendment or reagent to soil/sediment is a technique that can decrease mobility and reduce bioavailability of uranium (U) and other heavy metals in the contaminated site. According to data from literature and results obtained in field studies, the general mineral class of apatites was selected as a most promising amendment for in situ immobilization/remediation of U. In this work we presented theoretical assessment of stability of U(VI) in four apatite systems (hydroxyapatite (HAP), North Carolina Apatite (NCA), Lisina Apatite (LA), and Apatite II) in order to determine an optimal apatite soil amendment which could be used for in situ remediation of uranium. In this analysis we used a theoretical criterion which is based on calculation of the ion-ion interaction potential, representing the main term of the cohesive energy of the matrix/pollutant system. The presented results of this analysis indicate (i) that the mechanism of immobilization of U by natural apatites depends on their chemical composition and (ii) that all analyzed apatites represent, from the point of view of stability, promising materials which could be used in field remediation of U-contaminated sites.

In situ stabilization of toxic metals in polluted soils using phosphates: theoretical prediction and experimental verification.

The in situ (in place) immobilization of toxic metals, using inexpensive "reactive" amendments, is considered as a simple and cost-effective approach for the treatment of soils, contaminated by the presence of heavy metals, when these soils are difficult or costly to be removed and treated ex situ. Several application studies have demonstrated that the stabilization of contaminated soils and groundwaters by the addition of apatite minerals has the potential to be a successful and widely applicable remediation strategy for the case of Pb, Cd, as well as for other toxic metals, existing in polluted soils. On the other hand, the specific immobilization mechanism(s) of these toxic metals remains rather elusive. The present work involves an interdisciplinary theoretical and experimental approach, designed to gain at the fundamental (molecular) level the understanding of respective mechanisms, considering the immobilization of Pb and Cd by the addition of apatites. The theoretical analysis of stability, regarding the apatite/Pb or apatite/Cd systems and the relevant results of sorption experiments, pointed out two different mechanisms for the immobilization of Pb or Cd by the use of apatites. The possible practical consequences of these findings for the selection/application of natural apatites for the remediation of contaminated soils by the presence of heavy metals have been also discussed.

Serum leptin levels in patients with acromegaly before and after correction of hypersomatotropism by trans-sphenoidal surgery.

It has been shown that GH excess is associated with decreased leptin levels and decreased body fat mass. Reports regarding the effect of GH on serum leptin levels are inconsistent. We studied leptin secretion in 20 acromegalics before and 2 months after trans-sphenoidal surgery and in 20 gender-, age-, and body mass index (BMI)-matched control subjects. The mean 8-h leptin concentration for each subject was measured from a pool formed of samples collected hourly beginning at 2200 h until 0600 h the next morning. In a subgroup of 10 acromegalics, leptin pulsatility was assessed for the same period of time in 10-min sampling intervals. Basal GH, insulin-like growth factor-I (IGF-I), insulin, glucose, and lipids levels were measured. Area under the curve for insulin (AUCins) during oral glucose tolerance test was calculated. Control subjects and acromegalics had similar BMI, but patients with active acromegaly had significantly lower mean leptin level (mean +/- SEM; in men, 2.6+/-0.4 vs. 7.1+/-1.1 microg/L, P = 0.003; in women, 16.0+/-3.4 vs. 23.5+/-3.1 microg/L; P = 0.036). Mean 8-h leptin correlated with BMI (r = 0.57, P = 0.007, in controls; r = 0.70, P = 0.001, in patients). In stepwise regression analysis with mean 8-h leptin as a dependent variable, BMI (P<0.001) and gender (P = 0.01) in acromegalics entered the equation, whereas in control subjects gender, free fatty acids, insulin, and age accounted for 99.3% in leptin variability. After surgery, BMI did not change significantly; and glucose (P = 0.014), GH (P<0.001), and IGF-I (P<0.001) levels together with AUCins (P = 0.002) decreased, whereas mean leptin concentration rose significantly and attained normal levels (4.1+/-0.8 microg/L, P = 0.028) in acromegalic men and (23.6+/-4.7 microg/L, P = 0.003) in acromegalic women. Correlation between leptin level and BMI was preserved after surgery (r = 0.62, P = 0.005). In stepwise regression analysis, free fatty acids (P = 0.04) contributed to 26.8% of the variance in corrected-leptin (for BMI and gender). Leptin concentration peak height and interpeak nadir level rose significantly (P = 0.033 and P = 0.037) after surgery by Cluster analysis, without significant changes in leptin pulse frequency and incremental peak amplitude. Nocturnal rise of leptin (mathematically described by a cubic curve) was characterized by an acrophase just after midnight, before and after surgery. The amplitude and the average leptin concentration of the cubic fit increased significantly after surgery (P = 0.028 and P< 0.001). In conclusion in acromegalic patients: 1) leptin secretion maintains the pulsatility and nocturnal rise; 2) the gender-based leptin differences are preserved; 3) GH-IGF-I normalization leads to a rise in leptin that is not related to changes in BMI; and 4) the possible role of rise in leptin levels when assessing clinical and metabolic outcome of therapy in acromegalic patients deserves additional studies.

Glutathione adduct of hemoglobin (Hb ASSG) in hemolysates of patients on long-term antiepileptic therapy.

1. A novel, hitherto unrecognized, hemoglobin fraction, HbA1x, has been detected in amounts up to 7% of the total Hb, in hemolysates from epileptic patients with long-term therapy containing phenobarbital (PB) and carbamazepine (Carb). 2. HbA1x was shown to represent glutathione adduct of Hb (Hb ASSG). 3. To our knowledge, this is the first demonstration of the occurrence of Hb ASSG in vivo.

Renal function, protein excretion, and pathology of Balkan endemic nephropathy. II. Protein excretion.

The urinary proteins of 40 patients with Balkan endemic nephropathy from the Tuzla region were examined using ultrathin-layer SDS pore-gradient polyacrylamide gel electrophoresis in combination with silver staining. The typical urinary protein spectrum contained immunoglobulin G, Tamm-Horsfall protein, transferrin, albumin, beta 2-microglobulin (beta 2m), immunoglobulin light chains, retinol-binding protein, and alpha 1-microglobulin (alpha 1m). Densitometric measurements were used to derive glomerular tubular protein ratios (GTPR) and to characterize protein excretion patterns in the 28 patients who excreted more than 150 mg/liter of protein. Results showed that proteinuria of Balkan nephropathy is predominantly tubular, consisting of low-molecular-weight species. The most commonly identified proteins were alpha 1m, light chains, retinol binding protein, and beta 2m. The pattern of proteinuria based on GTPR did not correlate with the underlying histology or the degree of renal failure. These findings, using the ultrathin-layer SDS pore-gradient method of protein separation, more accurately demonstrates the low-molecular-weight proteinuria characteristic for the early stages of BEN.