Do serum and urinary concentrations of kidney injury molecule-1 in healthy newborns depend on birth weight, gestational age or gender?

OBJECTIVE: The aim of work was to establish the normal levels of serum and urinary kidney injury molecule-1 (sKIM-1 and uKIM-1) in healthy full-term newborns. STUDY DESIGN: The study included 88 healthy full-term neonates from normal, uncomplicated pregnancies. The serum and urinary concentrations of KIM-1 in the material obtained in the first or second day of life were determined with a commercially available enzyme-linked immunosorbent assay kits. In addition, uKIM-1 was normalized for urinary creatinine concentration. RESULTS: Male and female newborns, as well as children in whom the samples were obtained in the first or second day of life, did not differ significantly in terms of their sKIM-1 and uKIM-1 levels. Gestational age correlated inversely with sKIM-1 and positively with uKIM-1, but not with uKIM-1/cr. No correlation was found with birth weight and gender. CONCLUSION: This is the first report of sKIM-1 and uKIM-1 levels in healthy full-term newborns during the first postnatal days. The data from healthy newborns may serve as the reference values for future studies in the youngest children.

Effects of fluorides on apoptosis and activation of human umbilical vein endothelial cells.

OBJECTIVE: To determine the effects of fluorides on endothelial functioning. MATERIALS AND METHODS: We analyzed expressions of adhesion molecules, ICAM-1 and ICAM-3, and annexin V, on the surface of human umbilical vein endothelial cells (HUVECs) exposed to various concentrations of NaF and SnF(2) . We compared the effects of fluoride-induced changes with those obtained when stimulating HUVECs with TNF-α and verified whether N-acetyl cysteine (NAC), well-known antioxidant, can prevent both fluoride- and TNF-α-induced alterations. RESULTS: The expressions of annexin V and ICAM-1 increased significantly after adding NaF (5.0 or 7.5mM) or Sn(2) F (0.5 or 0.75mM) to the culture medium. Pre-incubating HUVECs with NAC prevented the effects induced by 5.0 mM of NaF and 0.5 mM of Sn(2) F. Only the highest concentration of NaF (7.5mM) triggered the expression of ICAM-3. The expressions of all three molecules increased significantly upon stimulating the cultures with TNF-α (20ng ml(-1) ); these changes were not reversed by pre-incubation with NAC. CONCLUSIONS: Fluorides induce oxidative stress, resulting in apoptosis and activation of HUVECs, manifested by an elevated expression of ICAM-1. The oxidative stress resulting from a stimulation by the highest NaF concentration triggers ICAM-3 expression on the HUVECs' surface.

TLR4 signaling induced by lipopolysaccharide or paclitaxel regulates tumor survival and chemoresistance in ovarian cancer.

Toll-like receptors (TLRs) expressed on immune cells trigger inflammatory responses. TLRs are also expressed on ovarian cancer (OvCa) cells, but the consequences of signaling by the TLR4/MyD88 pathway in these cells are unclear. Here, TLR4 and MyD88 expression in OvCa tissues (n=20) and cell lines (OVCAR3, SKOV3, AD10, A2780 and CP70) was evaluated by reverse transcriptase-PCR, western blots and immunohistochemistry. Cell growth, apoptosis, nuclear factor-kappaB (NF-kappaB) translocation, IRAK4 and TRIF expression and cJun phosphorylation were measured following tumor cell exposure to the TLR4 ligands, lipopolysaccharide (LPS) or paclitaxel (PTX). Culture supernatants were tested for cytokine levels. TLR4 was expressed in all tumors, tumor cell lines and normal epithelium. MyD88 was detectable in tumor tissues and in 3/5 OvCa lines but not in normal cells. In MyD88(+) SCOV3 cells, LPS or PTX binding to TLR4 induced IRAK4 activation and cJun phosphorylation, activated the NF-kappaB pathway and promoted interleukin (IL)-8, IL-6, vascular endothelial growth factor and monocyte chemotactic protein-1 production and resistance to drug-induced apoptosis. Silencing of TLR4 in SCOV3 cells with small interference RNA resulted in phosphorylated-cJun (p-cJun) downregulation and a loss of PTX resistance. In PTX-sensitive, MyD88(neg) A2780 cells, TLR4 stimulation upregulated TRIF, and TLR4 silencing eliminated this effect. Thus, TLR4/MyD88 signaling supports OvCa progression and chemoresistance, promoting immune escape.

Does prematurity affect platelet indices?

PURPOSE: The current study objective was to compare blood platelet indices in preterm newborns (PTN) and full term newborns (FTN). MATERIALS AND METHODS: We introduced to our study 51 PTN (25 females, 26 males) and 55 FTN (25 females, 30 males). Platelet indices were estimated in blood samples collected from the umbilical artery. RESULTS: PTN demonstrated a decreased count of blood platelets (197 x 103/microL) as compared to FTN (287 x 103/microL), p=0.0001. Platelet hematocrit (PCT) also showed substantial differences in both groups (PTN=0.16% vs. FTN=0.22%; p=0.001). Mean platelet volume (MPV) was found to be nearly the same (PTN=8.02 fl, FTN=7.79 fl). Platelet distribution width (PDW) was higher in PTN (50.64%) than in FTN (46.54%), p=0.021. Large platelet count (LPLT) was diminished in PTN (5.23%) in comparison with FTN (6.12 %). CONCLUSIONS: A decreased count of blood platelets, platelet hematocrit and increased platelet distribution width may result from a low gestational age or a dysfunction of megakaryocytes and the placenta. Blood platelet indices may be vital in the diagnosis of haemostatic disorders.

Expression of FasR, Fas-L and Bcl-2 in CD4+ and CD8+ subpopulations of T lymphocytes in the cord blood of healthy full-term newborns, is gender of influence?

PURPOSE: The expression of FasR, Fas-L and Bcl-2 on CD4+ and CD8+ T lymphocytes subpopulations from the cord blood were assayed. The results in blood from boys and girls were analyses separately and compared. MATERIAL AND METHODS: Twenty four full-term newborns: 13 females and 11 males were included into the study. Blood from the umbilical vein was collected immediately after cutting the umbilical cord. The staining with monoclonal antibodies against CD4, CD8, FasR, Fas-L and Bcl-2 was performed within 2 hours after collection and followed with flow cytometry acquisition and analysis. RESULTS: The percentage of CD4+ and CD8+ T lymphocytes and CD4+:CD8+ ratio was within normal range. The expression of FasR, Fas-L was higher on CD4+ T lymphocytes than on CD8+ T lymphocytes (10,36% vs 6,79% and 6,66% vs 5,63% respectively). The expression of Bcl-2 was comparable (91,9% and 93,75% respectively). The comparison between males and females showed higher percentage of CD4+ lymphocytes on lymphocytes from girls' blood (56% vs 38,69%, p=0.0003). The expression of FasR and Fas-L on CD4+ T lymphocytes was higher on CD4+ T lymphocytes from girls' blood (13,8% vs 7,53% and 6,8% vs 6,52% respectively) but without statistical significance. Bcl-2 expression was higher on CD4+ T lymphocytes from boys' blood (99,65% vs 89,7%) but without statistical significance. Similar pattern of FasR, Fas-L and Bcl-2 expression was noted on CD8+ T lymphocytes analysed separately for girls' and boys' blood origin cells. The difference in Bcl-2 expression was more prominent than on CD4+ T lymphocytes and reached statistical significance. CONCLUSIONS: The lymphocytes from cord blood of boys showed the more immature immunophenotype than T lymphocytes from cord blood of girls'. Impaired apoptosis (as a consequence of low expression of FasR, Fas-L) in neonatal cells may contribute to prolonged inflammation in newborns after oxidative stress or infection.

IRX-2, a novel immunotherapeutic, protects human T cells from tumor-induced cell death.

IRX-2 is a cytokine-based biologic agent that has the potential to enhance antitumor immune responses. We investigated whether IRX-2 can protect T cells from tumor-induced apoptosis. Tumor-derived microvesicles (MV) expressing FasL were purified from supernatants of tumor cells and incubated with activated CD8(+) T cells. MV induced significant CD8(+) T-cell apoptosis, as evidenced by Annexin binding (64.4+/-6.4%), caspase activation (58.1+/-7.6%), a loss of mitochondrial membrane potential (82.9+/-3.9%) and DNA fragmentation. T-cell pretreatment with IRX-2 prevented apoptosis. IRX-2-mediated cytoprotection was dose and time dependent and was comparable to effects of IL-2, IL-7 or IL-15. IRX-2 prevented MV-induced downregulation of JAK3 and TCRzeta chain and induced STAT5 activation in T cells. IRX-2 prevented MV-induced Bax and Bim upregulation (P<0.005-0.05), prevented cytochrome c release and Bid cleavage, and concurrently restored the expression of Bcl-2, Bcl-xL, FLIP and Mcl-1 (P<0.005-0.01) in T cells. In addition, IRX-2 reversed MV-induced inhibition of the PI3K/Akt pathway. An Akt inhibitor (Akti-1/2) abrogated protective effects of IRX-2, suggesting that Akt is a downstream target of IRX-2 signaling. Thus, ex vivo pretreatment of CD8(+) T cells with IRX-2 provided potent protection from tumor-induced apoptosis. IRX-2 application to future cancer biotherapies could improve their effectiveness by bolstering T-cell resistance to tumor-induced immunosuppression.

Hyperthermia-induced reorganization of microtubules and microfilaments and cell killing in CHO AA8 cell line.

The aim of this study was to elucidate the effects of hyperthermic treatment on cell morphology and the cytoskeleton in CHO AA8 cell line. The effects of exposure to elevated temperature were analyzed in CHO AA8 cell line by fluorescence microscopy and flow cytometry. The 30 min, at 44.5 degrees C heat shock treatment resulted in the collapse of microtubules (MTs) and microfilaments (MFs) around the nucleus followed by their recovery 24 h after heating. The initial collapse of these cytoskeletal systems, observed 15 min after treatment, was accompanied by the appearance of cells with reduction of volume, shrunken cytoplasm and condensed chromatin. 24 h afterwards, there was the increase in the number of cells with restored and extended MT and MF cytoskeletons. Most of them were larger in size compared to the control cells and had multiple nuclei. 48 h after heat shock the highest number of the giant cells with alternation in nuclear morphology was seen. Flow cytometry analysis revealed the increase in the number of cells with externalized phosphatidylserine 24 h and 48 h after hyperthermic treatment. These results suggest that following heat shock, CHO AA8 cells undergo mitotic catastrophe that presumably represents one of the events resulting in apoptosis.

Quality of life self-assessment of children living in a children's home, based on own research conducted in the Podlaskie Province.

PURPOSE: The purpose of the study was to diagnose the quality of life (QoL) of children brought up in children's homes, to compare findings with results for peers living in complete families. MATERIAL AND METHODS: In the Stage I was to determine the usefulness of tools for QoL evaluation in a group of 120 children from children's homes in the Podlaskie Province and in a group of 120 children belonging to a control group, brought up in their own families, in the same places where the children's homes were located. Selected research tools were used in Stage II, and the study was carried out in a group of 180 children in the same children's homes and a control group. We used the following survey questionnaires: the standardised CHQ-CF87 survey, standardised KINDL survey and Children's Survey based on WHOQOL-BREF. RESULTS: Significant relationship between the quality of life self-assessment and the place of being brought up for all categories of quality of life was found. A relationship was indicated between the QoL self-assessment and the place of living, age, gender, and physical condition. The charges of a children's home assessed their QoL as significantly lower compared to children living in normal families, mostly in the following categories: health, physical domain and psychological domain, social relations and the ability to function in everyday life. In KINDL survey, strong relationships were found between assessments of QoL categories. CONCLUSIONS: Significant relation between QoL self-assessment and where children were brought up was found. Positive relationship between QoL self-assessment and the place of living, age, gender, and children's physical condition was found.

Relations occurring between health-related behaviour categories and quality of life made by children brought up in a children's home, in the Podlaskie Province.

PURPOSE: The purpose of the study was to determine relations between health-related behaviour categories and quality of life (QoL) categories made by children brought up in a children's home and to compare the results obtained with the results for a group of peers brought up by their own families. MATERIAL AND METHODS: The study was performed on a group of 180 children living in children's homes located in Bialystok, Krasne, Suprasl, Lomza, Nowa Pawlówka; and on a control group of children living with their own families in the same places where children's homes were located. The diagnostic survey method with the Health Behaviour Scale questionnaire, composed of 40 statements defining various behaviours connected with health, and the Children's Questionnaire, based on The World Health Organization Quality of Life (WHOQOL-BREF) was used. RESULTS: Strong correlations between assessments of the Health Behaviour Scale categories and assessments of quality of life categories were found in the group of children living in children's homes, mostly in respect to the relation between health self-assessment and physical activity r = 0.77, mental activity r = 0.74 and environment r = 0.72, and between the physical domain and eating habits r = 0.70, and physical activity and the physical domain r = 0.69. The determination coefficient R2 for the study group had high values for three QoL categories: physical domain 71.5%, mental domain 69.7% and environment 70.1%. CONCLUSIONS: Correlations between Health Behaviour Scale categories and QoL categories were found in the group of children living in children's homes compared to children living with their own families. The relationships for health self-assessment and the physical and mental domains and the environment, and for the physical domain and eating habits and physical activity were found.

Health-related behaviour self-assessment of children living in a children's home; study based on own research realised in the Podlaskie Province.

PURPOSE: The purpose of the study was to diagnose the health-related behaviour of children brought up in children's homes, to compare the obtained results with those obtained from a group of peers brought up in their own families. MATERIAL AND METHODS: The study group included 180 children living in children's homes in the Podlaskie Province and in a control group composed of children brought up in their own families and living in the same places where the children's homes are located. A questionnaire of the Health Behaviour Scale, composed of 40 statements determining health-related issues was used. RESULTS: Self-assessment of health-related behaviour in the studied youth depended on age, for which a statistical significance was shown for: health self-assessment (p = 0.011), categories of stressful situations (p = 0.047), physical activity (p = 0.028) and social support (p = 0.001); gender, for which a statistical significance was shown for the categories of usage of stimulants (p = 0.000) and place of living, in which the factor "place" was significant (p = 0.000) for all categories; and education, where p = 0.000 for the following categories: stressful situations, using stimulants, physical activity, social support and health self-assessment. Relationships between the categories of health-related behaviour were much stronger in the assessments of the children brought up in children's homes were found. CONCLUSIONS: The self-assessment of health-related behaviour of the studied youth depended on age, gender, place of living and education. Relationships between the categories of health-related behaviour were much stronger in assessments of the children brought up in children's homes as compared to controls.

Mid-term effectiveness of behavioral and psychodynamic inpatient treatments of severe obesity based on a randomized study.

BACKGROUND: The objective of this study was to compare the effectiveness of psychodynamic and behavioral inpatient treatments of severely obese patients regarding weight and distress. METHODS: In a longitudinal study obese patients (body mass index, BMI >or=35) were randomly assigned to behavioral or to psychodynamic inpatient treatment. Mostly female (n = 267; 85%) obese patients with psychiatric and somatic comorbidity (age 20-64 years, BMI 35-74) were examined with standardized self-report scales on distress (SCL-90R), interpersonal problems (Inventory of Interpersonal Problems), eating behavior (Fragebogen zum Essverhalten) and body image (Fragebogen zum Korperbild). RESULTS: During 49 days (mean) of inpatient treatment, patients lost an average of 5.6 kg (4.4%) in the behavioral (n = 130) and 5.7 kg (4.4%) in the psychodynamic setting (n = 137). In both settings, eating behavior, well-being and body image also improved significantly. One year after discharge, return rate was 73%. Forty percent had further reduced their weight (by more than 5% compared to intake), 36% had regained weight, but were still below intake level, and another 24% had increased weight above intake. CONCLUSION: Behavioral and psychodynamic treatments were equally effective reducing weight and distress over 1 year.

Tissue localization of Toll-like receptors in biopsy specimens of liver from children infected with hepatitis C virus.

Toll-like receptors (TLR) are important tools of innate immunity, localized mainly on cells of the immune system, but also have been shown on cells of other origin. In the current study, they have been searched in biopsy specimens of liver from children bearing chronic viral hepatitis of C type (HCV). TLR2, TLR3 and TLR4 were traced by means of polyclonal antibodies and avidin-biotin complex (ABC) immunohistochemistry. Besides, mRNA for TLR was looked for using specific primers and polymerase chain reaction. Several controls, including neutralization of primary antibody with respective blocking peptide, confirmed the specificity of the immunohistochemical reaction. All TLR tested could be visualized in a focal distribution in single hepatocytes and some cells of inflammatory infiltrates. There was no reaction whatsoever in liver samples not infected with hepatotropic virus. In molecular studies, mRNA for TLR2 and TLR4 was detected in both noninfected and hepatitis B virus-infected established cell lines of human hepatoma as well as in HCV(+) biopsy samples. These data indicate that TLR can be traced in liver cells, both at the protein and at the mRNA level. Their irregular and focal distribution in HCV(+), but not in HCV(-), liver suggests some role of TLR in the pathogenesis of chronic viral hepatitis, at least in children.

The lack of effect of a prophylactic dose of enoxaparin on thrombin generation in patients subjected to nephrectomy because of kidney cancer.

It is assumed that major surgery, connected with extensive tissue dissection, brings about the release of tissue factor into circulation and subsequent activation of coagulation system. This activation results in the thrombin generation, which is supposed to be suppressed by the low-molecular-mass heparins (LMMH), administered to the surgical patients as the prophylaxis against postoperative venous thromboembolism. We have estimated the concentration of circulating thrombin-antithrombin (TAT) complex in patients subjected to transperitoneal nephrectomy and randomized into controls and who received 40-mg enoxaparin 12 h before and 12 h after the operation and then once daily for 7 days. We have observed a sharp rise of TAT concentration at the end of surgery and it corresponded to the simultaneous drop of antithrombin (AT) activity. TAT concentration gradually decreased and AT activity increased up to the end of observations on the seventh postoperative day, but there were no differences observed between the groups of patients. We have also observed a biphasic increase of plasmin-plasmin inhibitor (PPI) complex concentration in our patients. Again, there were no differences in PPI between the groups of patients. It is our conclusion that under the conditions of this study, the well-known prophylactic effect of enoxaparin against the venous thromboembolic complications was not mediated by the inhibition of intraoperative thrombin generation. The anti-inflammatory or biophysical influence of LMMH may be rather taken into account in surgical patients receiving prophylactic doses of these heparins.

The fibre dimensions of uterine smooth muscle of the rabbit following treatment by female sex steroids.

The effects of female sex hormones on the dimensions of myometrial smooth muscle fibres were studied by using ovariectomized rabbits. After one month of treatment, the fiber dimensions of the outer myometrial layer were measured, using cryostat sections. Calculated smooth muscle fiber volume was found to be in the sequence: control < medroxyprogesterone < estradiol < estradiol + medroxyprogesterone < estradiol alone. The measurements show that medroxyprogesterone-treated uteri contain the narrowest and the longest smooth muscle fibres, while estradiol treatment have the largest cells. This study complements previous observations in showing that medroxyprogesterone alone, or in combination with other modulators, contributes to sustain pregnancy by increasing internal resistance of estradiol-primed myometrial smooth muscle fiber fascicles. Our discussion, based on recent literature, shows that this resistance is ultimately controlled by changes in the myometrium innervation, in the repression of some controlling myofibrillar components, in the expression of specific membrane receptors and ionic channels, and in favoring the switching of molecular connexins in gap junctions, making P paramount in maintaining pregnancy. Moreover, other recent observations have also shown that probably an hcG-like hormone actually control P receptors expression in myometrial smooth muscles.

Postoperative injection of phenylbutazone does not influence fibrinolytic shutdown.

Twenty-nine patients undergoing cholecystectomy because of chronic calculous cholecystitis were randomised to receive phenylbutazone 10 mg/kg intramuscularly or a control injection (vehicle-containing local anaesthetic) immediately after completion of surgery. Fibrinogen and plasminogen concentrations in plasma, plasminogen activator inhibitor activity in plasma and fibrinolytic activity in concentrated euglobulins were determined before surgery and on the first, third and seventh postoperative days. Phenylbutazone delayed the postoperative rise of fibrinogen concentration and reduced the plasminogen level on the first day after surgery. Fibrinolytic activity in euglobulins was decreased after the operation in both groups. The great dispersion of the results of plasminogen activator inhibitor activity was the plausible cause of the lack of any significant difference in this variable. The postoperative fibrinolytic shutdown, reflected by the decrease of fibrinolytic activity, was unaffected by phenylbutazone. It can be speculated, however, that the decline of plasminogen concentration after surgery in patients receiving this drug was the result of its stimulatory influence on the conversion of plasminogen into plasmin.

Postoperative plasma interleukin-6 in patients with renal cancer correlates with C-reactive protein but not with total fibrinogen or with high molecular weight fibrinogen fraction.

The concentration of fibrinogen (Fb) and its fractions, the levels of interleukin-6 (I1-6), C-reactive protein (CRP), and immunoglobulin G (IgG) were determined in 38 patients operated on because of renal cancer. The increased Fb and I1-6 concentrations were found in approximately one-half of the patients with malignancy. The relations among the high molecular weight (HMW) and two low molecular weight (LMW and LMW') fibrinogen fractions in these patients before surgery did not differ from the corresponding relations in normal subjects. The levels of all (except IgG) compounds studied increased after surgery and the peak of I1-6 was observed on the first postoperative day but that of CRP on the third day. The concentrations of total Fb and of its HMW fraction were the highest also on the third postoperative day and this was in contrast with the decline of low molecular weight fractions at the same time. These variations of estimated variables can be regarded as being relevant to the acute phase response. We have noted a correlation between the maximal concentrations of I1-6 and CRP, but not between the corresponding concentrations of Il-6 and total Fb or HMW Fb; this may suggest that the concentration of Fb is also under the control of a factor other than I1-6.

Antiphospholipid antibodies and lipoprotein (a) in women with recurrent fetal loss.

OBJECTIVE: To estimate the occurrence of lupus anticoagulant, both isotypes of anticardiolipins, IgG and IgM, and lipoprotein (a) in women with recurrent fetal loss. METHOD: 43 women with two or more fetal losses were included in this open, prospective study. They were divided into a group of 32 nullipara who have had only miscarriages and a group of 11 women who besides the fetal losses were able to give birth to at least one live newborn. Two control groups were also added: 35 women who were never pregnant and 15 parous who did not have any miscarriage. The presence of lupus anticoagulant was assessed by the Staclot LA clotting test. IgG and IgM isotypes of anticardiolipins and lipoprotein (a) were determined using the immunoenzymatic methods. RESULTS: Lupus anticoagulant was discovered in only one of 23 examined patients. Abnormally high values of IgG isotype of anticardiolipins were found in two subjects of each control group and in two nullipara with miscarriages only. The increased level of IgM isotype was noted in approx. 50% of patients with recurrent fetal loss, but at least one successful delivery and in control subjects who were never pregnant. In contrast, the frequency of the increased level of this antibody was diminished by half in the remaining two groups: women who delivered a live child (children) without any miscarriage and women who had only miscarriages. Elevated levels of lipoprotein (a) were found in approx. 30% of non-pregnant control subjects and in the patients of both groups; the abnormally high concentration of this lipoprotein was encountered in only 13% of women who gave birth to live newborn(s) and did not have any miscarriage. CONCLUSION: Of all four determined variables, only abnormally high values of lipoprotein (a) may have some prognostic significance in women predisposed to the recurrent fetal loss.

Prevalence of high IgM anticardiolipins in patients with ischemic stroke.

Lupus anticoagulant (LA), IgG and IgM isotypes of anticardiolipins (aCL), lipoprotein (a), and the resistance to activated protein C were determined in patients with ischemic stroke. The raised concentration of the aCL-IgM isotype was noted in 42% of patients with this type of stroke, and it was in contrast with an 8% frequency of an increased level of aCL-IgG isotype in these cases. The high level of lipoprotein (a) was found with similar frequency in stroke patients and in age-matched control subjects. It is concluded that the elevated concentration of IgM isotype of anticardiolipin antibodies can be regarded as significant in the ethiological work-up in elderly stroke patients.Copyright Lippincott-Raven Publishers