Uteroplacental insufficiency in rats induces renal apoptosis and delays nephrogenesis completion.

AIM: Uteroplacental insufficiency in rats reduces nephron endowment, leptin concentrations and programmes cardiorenal disease in offspring. Cross-fostering growth-restricted (Restricted) offspring onto a mother with normal lactation restores leptin concentrations and nephron endowment. This study aimed to determine whether the reduced nephron endowment in Restricted offspring is due to delayed glomerular formation and dysregulation of renal genes regulating branching morphogenesis, apoptosis or leptin signalling. Furthermore, we aimed to investigate whether cross-fostering Restricted offspring onto Control mothers could improve glomerular maturation and restore renal gene abundance. METHODS: Uteroplacental insufficiency was induced by bilateral uterine vessel ligation (Restricted) or sham (Control) surgery on gestation day 18 (E18). Kidneys were collected at E20, postnatal day 1 (PN1) and PN7. An additional cohort was cross-fostered onto separate mothers at birth and kidneys collected at PN7. RESULTS: Kidneys were lighter in the Restricted group, but weight was restored with cross-fostering. At E20, abundance of Bax, Flt1 and Vegfa was increased in Restricted offspring, while Ret and Bcl2 transcripts were increased only in Restricted females. At PN7, abundance of Gdnf and Ret was higher in Restricted offspring, as was Casp3. Restricted offspring had a wider nephrogenic zone with more immature glomeruli suggesting a delayed or extended nephrogenic period. Cross-fostering had subtle effects on gene abundance and glomerular maturity. CONCLUSION: Uteroplacental insufficiency induced apoptosis in the developing kidney and delayed and extended nephrogenesis. Cross-fostering Restricted offspring onto Control mothers had beneficial effects on kidney growth and renal maturity, which may contribute to the restoration of nephron endowment.

Synthesis and characterization of a trinuclear iridium(III) based catalyst for the photocatalytic reduction of CO2.

A trimetallic Ir(iii) based complex () was synthesized and fully characterized by spectroscopic and electrochemical methods. A detailed comparison to its mono- () and bimetallic () analogue regarding the photocatalytic reduction of CO2 is outlined. In particular, the effect of intramolecular quenching, provided by ethyl tethers, was investigated. Moreover, the relationship between the photophysical properties, the lifetime of the excited state, the quenching efficiency and the catalytic performance is presented. Notably, the covalent linkage of the Ir(iii) moieties within the three-armed ligand structure (complex ) leads to a twofold increase of the turn over number (TON) compared to its monometallic analogue . Taking in account the quantum efficiency of 10% and the TONCO = 60 (per Ir(iii) center), complex is a highly active Ir(iii) based photocatalyst.

Growth restriction alters adult spatial memory and sensorimotor gating in a sex-specific manner.

In Western society, impaired uteroplacental blood flow is the major cause of human intrauterine growth restriction. Infants born small and who experience late childhood accelerated growth have an increased risk of developing adult diseases. Recent studies also suggest a link between birth weight and altered adult behavior, particularly relating to motor function, learning and memory, depression and schizophrenia. The aim of this study was to determine the relative influence of prenatal and postnatal growth restriction on adult behavioral outcomes in male and female rats. Uteroplacental insufficiency was induced in Wistar Kyoto rats by bilateral uterine vessel ligation on day 18 of gestation producing growth-restricted offspring (Restricted group). The Control group had sham surgery. Another group underwent sham surgery, with a reduction in litter size to five at birth equivalent to the Restricted litter size (Reduced Litter group). At 6 months of age, a series of behavioral tests were conducted in male and female offspring. Growth restriction did not impair motor function. In fact, Restricted and Reduced Litter males showed enhanced motor performance compared with Controls (P < 0.05). Spatial memory was greater in Restricted females only (P < 0.05). The Porsolts test was unremarkable, however, males exhibited more depressive-like behavior than females (P < 0.05). A reduction in sensorimotor gating function was identified in Reduced Litter males and females (P < 0.05). We have demonstrated that growth restriction and/or a poor lactational environment can affect adult rat behavior, particularly balance and coordination, memory and learning, and sensorimotor gating function, in a sex-specific manner.

In vitro effects of antiepileptic drugs on acetylcholinesterase and ectonucleotidase activities in zebrafish (Danio rerio) brain.

Carbamazepine (CBZ), phenytoin (PHT), and gabapentine (GBP) are classical antiepileptic drugs (AEDs) that act through a variety of mechanisms. We have tested the in vitro effects of CBZ, PHT, and GBP at different concentrations on ectonucleotidase and acetylcholinesterase activities in zebrafish brain. CBZ inhibited ATP hydrolysis at 1000 microM (32%) whereas acetylcholine hydrolysis decreased at 500 microM (25.2%) and 1000 microM (38.7%). PHT increased AMP hydrolysis both at 500 microM (65%) and 1000 microM (64.8%). GBP did not promote any significant changes on ectonucleotidase and acetylcholinesterase activities. These results have shown that CBZ can reduce NTPDase (nucleoside triphosphate diphosphohydrolase) and PHT enhance ecto 5'-nucleotidase activities. Therefore, it is possible to suggest that the AEDs induced-effects on ectonucleotidases are related to enzyme anchorage form. Our findings have also shown that high CBZ concentrations inhibit acetylcholinesterase activity, which can induce an increase of acetylcholine levels. Taken together, these results showed a complex interaction among AEDs, purinergic, and cholinergic systems, providing a better understanding of the AEDs pharmacodynamics.

Evaluation of Sinos River water genotoxicity using the comet assay in fish.

The Sinos River, in southern Brazil, is polluted by industrial discharges and untreated urban wastes. Fish genotoxicity biomarkers are valuable parameters for environmental risk assessment. In this study, we used the comet assay to detect genotoxicity due to multiple sources of pollution in the peripheral blood of a native fish species (Hyphessobrycon luetkenii). In addition, we analysed possible DNA damage from aluminum, lead, chromium, copper, nickel, iron and zinc contamination. Water samples were collected seasonally from three sampling sites and the fish were assessed under laboratory conditions. Water chemical analysis showed an increased level of aluminum and iron in most of the samples at sites 2 and 3, located in the middle and lower river course, respectively. The index of DNA damage assessed by the comet assay demonstrated no significant differences in different seasons or at the different sampling sites, while the frequency of cells with DNA damage was higher in water samples collected at sites 1 and 2 during the spring season. None of the metals studied seems to be associated with the increase in the frequency of cells with DNA damage observed during the spring season. The results of this study indicate that the Sinos River is contaminated with substances that are genotoxic to fish, including the waters near the river spring.

A multibiomarker approach in rats to assess the impact of pollution on Sinos River, Southern Brazil.

The aim of this study was to determine the feasibility of combining water quality analysis with different biomarkers to characterise the relationship between anthropogenic contamination and biotic response in the Sinos River, southern Brazil. Wistar rats were studied using three biomarkers combined with physical, chemical and microbiological analysis to assess the effects of pollution at four sampling sites. The induction of oxidative stress was quantified by MDA levels in peripheral blood, lymphocyte DNA damage was determined using the comet assay, and histopathological changes were analysed in the liver. After sampling, animals were allowed to drink the river water during a 48 hours period. No increase in oxidative stress and DNA damage was observed. However, liver damage was observed in the animals exposed to water samples, indicating that the Sinos River is contaminated with hepatotoxic substances. Water analyses confirmed that water quality decreased downriver.

Cross-fostering and improved lactation ameliorates deficits in endocrine pancreatic morphology in growth-restricted adult male rat offspring.

Uteroplacental insufficiency and poor postnatal nutrition impair adult glucose tolerance and insulin secretion in male rat offspring, which can be partially ameliorated by improving postnatal nutrition. Uteroplacental insufficiency was induced in the WKY rat on day 18 of pregnancy (Restricted) compared to sham-operated Controls. Pups were then cross-fostered onto Control or Restricted mothers one day after birth resulting in: (Pup-on-Mother) Control-on-Control, Control-on-Restricted, Restricted-on-Control and Restricted-on-Restricted. Endocrine pancreatic morphology and markers of intrinsic ß-cell function and glucose homeostasis were assessed in male offspring at 6 months. Pancreatic and hepatic gene expression was quantified at postnatal day 7 and 6 months. Restricted pups were born 10-15% lighter than Controls and remained lighter at 6 months. Relative islet and ß-cell mass were 51-65% lower in Restricted-on-Restricted compared to Controls at 6 months. Non-fasting plasma C-reactive protein levels were also increased, suggestive of an inflammatory response. Overall, the average number of islets, small islets and proportion of ß-cells per islet correlated positively with birth weight. Intrinsic ß-cell function, estimated by insulin secretion relative to ß-cell mass, was unaffected by Restriction, suggesting that the in vivo functional deficit was attributable to reduced mass, not function. Importantly, these deficits were ameliorated when lactational nutrition was normalized in Restricted-on-Control offspring, who also showed increased pancreatic Igf1r, Pdx1 and Vegf mRNA expression at 7 days compared to Control-on-Control and Restricted-on-Restricted. This highlights lactation as a critical period for intervention following prenatal restraint, whereby deficits in endocrine pancreatic mass and associated impaired in vivo insulin secretion can be ameliorated.

Steroid-independent regulation of uterine oxytocin receptors.

The oxytocin receptor is an important contractile-associated protein, up-regulated at term in the myometrium in many mammalian species. We conducted studies in a novel animal model to challenge the general view that gonadal steroids are a major regulatory factor of uterine oxytocin receptors. Female marsupials have separate uteri and, in monovular species such as the tammar wallaby, the conceptus is present in one uterus whereas the contralateral uterus is empty. A marked increase in myometrial oxytocin receptors occurs only in the gravid uterus. Fetectomy experiments demonstrated that local embryo-derived factors stimulate this gravid uterus-specific increase in oxytocin receptors, and that uterine distension is probably not a key component in this regulatory pathway. Unilateral ovariectomy has no significant effect on uterine oxytocin receptors, emphasizing the impact of the conceptus on oxytocin receptor regulation and the minimal influence of gonadal steroids on parturition in this species. Our data highlight that regulation of uterine oxytocin receptor expression is multifactorial, and does not necessarily rely on gonadal steroids.

Pulmonary graft function after long-term preservation of non-heart-beating donor lungs.

BACKGROUND: Critical organ shortage in lung transplantation could be attenuated by the use of non-heart-beating donor (NHBD) lungs. In addition, prolonged ischemic tolerance of the organs would contribute to the alleviation of organ shortage. The aim of this study was to investigate pulmonary graft function of NHBD lungs after long-term hypothermic storage. METHODS: Twelve native-bred pigs (bodyweight 20 to 30 kg) underwent left lung allotransplantation. In the heart-beating donor (HBD) group, lungs were harvested immediately after cardiac arrest. In the NHBD group, lungs were subjected to a warm ischemic period of 90 minutes before harvesting. After a total ischemic time of 19 hours, pulmonary grafts in both groups were reperfused and pulmonary graft function was assessed. All values were compared with a sham-operated control group. RESULTS: Pulmonary graft function in the HBD group was excellent. In the NHBD group, pulmonary gas exchange was impaired, but still provided good graft function compared with the excellent graft function in the HBD group. Pulmonary vascular resistance was even lower in the NHBD group. In the NHBD group, calculated intrapulmonary shunt fraction (Qs/Qt) was significantly increased compared with the sham-group. Histologic alteration and wet-to-dry ratio did not differ significantly between the HBD and NHBD group. CONCLUSIONS: We conclude that NHBD lungs (90 minutes of warm ischemic time) have the potential to alleviate organ shortage in lung transplantation even after an extended total ischemic time.

Hip deformities in walking patients with cerebral palsy.

Hip deformities in walking patients with cerebral palsy are rare. Nineteen diplegic and four hemiplegic patients with unilateral hip subluxation were studied to determine whether or not characteristic gait patterns could be identified. All were examined clinically and radiologically as well as undergoing observational and instrumented three dimensional gait analysis. Twenty one of the patients compensated for the subluxation with an ipsilateral trunk lean and contralateral pelvic drop. These patterns were quantified in the kinematic and kinetic variables measured. We conclude that hip abductor weakness should be considered as the cause of the deformity.

Quantitative assessment of vascular surface density in renal cell carcinomas.

OBJECTIVE: To evaluate the extent of vascularization by assessing vascular surface density in renal cell carcinomas (RCCs) of different nuclear grades, and in normal renal cortex and medulla. MATERIALS AND METHODS: Specimens of 79 RCCs of different nuclear grades (16 of G1, 42 of G2 and 21 of G3) were immunostained with the lectin Ulex europaeus agglutinin-I (UEA I). The vascular surface density of tumour tissue was assessed stereologically using a test grid at x400 magnification and compared to the values obtained in normal renal tissue. RESULTS: G3 tumours had a lower vascular surface density than had G1 and G2 RCCs and normal renal tissue of the cortex and medulla (P < 0.001, respectively). G1 tumours had a significantly higher vessel density than had normal medullary parenchyma and G2 carcinomas (P < 0.001). Vessel density was not significantly different among G1 tumours and cortical parenchyma in controls and among normal medullary tissue and G2 tumours. Statistical analysis showed that the vascular surface density was independent of tumour stage and size and the age and sex of the patients. CONCLUSION: The degree of vascularization in RCCs decreased with their grade of differentiation, suggesting that the extent of neovascularization in tumour tissue reflects the relationship between tumour cell proliferation and vascular growth. The values of vascular surface density in normal renal tissue of the cortex and medulla partially overlapped with those obtained in tumour tissue.

Nuclear morphometry of renal cell carcinomas.

In this study, the correlation between different morphometric nuclear parameters and grading of renal cell carcinoma was investigated. Nuclear area, the standard deviation of the nuclear area and the form factor (FF) were assessed using a computer assisted image analyzer system. A statistically significant difference between G2 and G3 carcinomas could be confirmed for the nuclear area and the standard deviation of the nuclear area, with the significance level being lower for the latter parameter. Normal tubular epithelia and G1 carcinomas showed no significant difference in the nuclear area and the standard deviation of the nuclear area. In contrast, the form factor discriminated between normal tubular epithelium and G1 carcinomas, whereas no difference of the FF between G2 and G3 carcinomas was found. Reclassification of the subjectively assessed tumor grading was performed by means of linear discriminant analysis using all possible combinations of the different nuclear parameters. This procedure disclosed an increasing rate of correctly reclassified grading with rising number of parameters applied. We conclude that the isolated assessment of nuclear area does not suffice for the correct morphometric tumor grading, and that further parameters describing the nuclear shape have to be applied additionally in studies concerning morphometric nuclear grading.

HTLV-1-negative pleomorphic T-cell lymphoma of the skin: the clinicopathological correlations and natural history of 15 patients.

The peripheral T-cell lymphomas represent a heterogeneous group and include, besides mycosis fungoides and Sézary syndrome, large-cell anaplastic lymphoma. We report 15 cases from our files that fulfil the histological criteria of pleomorphic T-cell lymphoma with primary skin involvement. Most of the cases were elderly with a male-to-female ratio of 1.5:1. The HTLV-1 serology was negative. The clinical features of these patients differed from those with mycosis fungoides and Sézary syndrome, in that eczematous and precursor lesions such as parapsoriasis en plaque were lacking apart from one exception. All the patients with small-cell pleomorphic T-cell lymphomas were alive, although three of the nine patients with medium-to-large tumour cells have died. Pleomorphic T-cell lymphoma should be regarded as a distinct entity among the lymphoproliferative disorders of the skin.

[Analysis of antibiotic therapy in 500-bed hospital: indications, administration and possibilities of cost reduction]. / Analyse der Antibiotikatherapie in einem 500-Betten-Krankenhaus: Indikation, Durchführung und Möglichkeiten zur Kostenreduktion.

The introduction of a justification form in a 500-bed acute hospital before starting therapy with antibiotics resulted in a more than 30% decrease of current antibiotics costs. The cost reduction was mainly caused by reduced consumption of very expensive antibiotics. Moreover, analysis of the justification form revealed that more than 20% of diseases such as pneumonia, bronchitis, postsurgical complications, or skin and bone infections were diagnosed and treated without sufficient clinical and bacteriological documentation. The use of a justification form for antibiotics is therefore not only useful for cutting costs, but also for educating physicians in how to administer rational prophylaxis and therapy with antibiotics.