Complement genes strongly predict recurrence and graft outcome in adult renal transplant recipients with atypical hemolytic and uremic syndrome.

Atypical hemolytic and uremic syndrome (aHUS) is a severe disease strongly associated with genetic abnormalities in the complement alternative pathway. In renal posttransplantation, few data are available on recurrence risk and graft outcome according to genetic background in aHUS patients. The aim of this study was to identify risk factors for recurrence and transplant outcome and, in particular, the role of complement gene abnormalities. We retrospectively studied 57 aHUS patients who had received 71 renal transplants. A mutation in complement gene was identified in 39 (68%), in factor H (CFH), factor I (CFI), membrane cofactor-protein (MCP), C3 and factor B (CFB). At 5 years, death-censored graft survival was 51%. Disease recurrence was associated with graft loss (p = 0.001). Mutations in complement genes were associated with higher risk of recurrence (p = 0.009). Patients with CFH or gain of function (C3, CFB) mutations had a highest risk of recurrence. M-TOR inhibitor was associated with significant risk of recurrence (p = 0.043) but not calcineurin inhibitor immunosuppressive treatment (p = 0.29). Preemptive plasmatherapy was associated with a trend to decrease recurrence (p = 0.07). Our study highlights that characterization of complement genetic abnormalities predicts the risk of recurrence-related graft loss and paves the way for future genetically based individualized prophylactic therapeutic strategies.

Randomized trial of polychromatic blue-enriched light for circadian phase shifting, melatonin suppression, and alerting responses.

Wavelength comparisons have indicated that circadian phase-shifting and enhancement of subjective and EEG-correlates of alertness have a higher sensitivity to short wavelength visible light. The aim of the current study was to test whether polychromatic light enriched in the blue portion of the spectrum (17,000 K) has increased efficacy for melatonin suppression, circadian phase-shifting, and alertness as compared to an equal photon density exposure to a standard white polychromatic light (4000 K). Twenty healthy participants were studied in a time-free environment for 7 days. The protocol included two baseline days followed by a 26-h constant routine (CR1) to assess initial circadian phase. Following CR1, participants were exposed to a full-field fluorescent light (1 × 1014 photons/cm2/s, 4000 K or 17,000 K, n = 10/condition) for 6.5 h during the biological night. Following an 8 h recovery sleep, a second 30-h CR was performed. Melatonin suppression was assessed from the difference during the light exposure and the corresponding clock time 24 h earlier during CR1. Phase-shifts were calculated from the clock time difference in dim light melatonin onset time (DLMO) between CR1 and CR2. Blue-enriched light caused significantly greater suppression of melatonin than standard light ((mean ±â€¯SD) 70.9 ±â€¯19.6% and 42.8 ±â€¯29.1%, respectively, p < 0.05). There was no significant difference in the magnitude of phase delay shifts. Blue-enriched light significantly improved subjective alertness (p < 0.05) but no differences were found for objective alertness. These data contribute to the optimization of the short wavelength-enriched spectra and intensities needed for circadian, neuroendocrine and neurobehavioral regulation.

Comparison of effectiveness and safety between conbercept and ranibizumab for treatment of neovascular age-related macular degeneration. A retrospective case-controlled non-inferiority multiple center study.

PurposeTo compare the efficacy and safety of conbercept and ranibizumab when administered according to a treat-and-extend (TREX) protocol for the treatment of neovascular age-related macular degeneration (AMD) in China.Patients and methodsBetween May 2014 and May 2015, 180 patients were treated in a 1 : 1 ratio using conbercept or ranibizumab from four hospitals. Patients received either conbercept 0.5 mg or ranibizumab 0.5 mg intravitreal injections. Follow-up time was 1 year and treated based on a TREX approach. Main outcomes and measures include best-corrected visual acuity (BCVA), using Early Treatment Diabetic Retinopathy Study (ETDRS); number of injections; central retinal thickness (CRT); and leakage of choroidal neovascularization before and after the treatment was analyzed by fluorescein fundus angiography and indocyanine green angiography.ResultsThe 1-year visit was completed by 168 (93.3%) of patients. Mean BCVA was equivalent between two cohorts, and were improved by 12.7±7.770 and 12.3±7.269 letters in the conbercept and ranibizumab cohorts, respectively (P=0.624). There was no significant difference in measured CRT, with a mean decrease of 191.5 µm for conbercept and 187.8 µm for ranibizumab (P=0.773). There was a statistically significant difference (P=0.001) between the drugs regarding the number of treatments: 7.4 for conbercept and 8.7 for ranibizumab. The difference in the distribution of injection intervals was statistically significant between two groups (P=0.011). During the study, there were no cases of endophthalmitis or intraocular inflammation.ConclusionBoth drugs had equivalent effects in visual and anatomic gains at 1 year when administered. In the conbercept group, longer treatment intervals were achieved with more patients.

[Progressive multifocal leukoencephalopathy in a non-AIDS patient: high efficiency of combined cytarabine and cidofovir]. / Leucoencéphalite multifocale progressive en dehors du sida: efficacité de l'association cytarabine-cidofovir.

INTRODUCTION: Progressive multifocal leukoencephalopathy (PML) is an opportunistic infection of the central nervous system, occurring in immunocompromised patients. Treatment, not codified to date, is more often inefficient with a rapid and fatal deterioration. CASE RECORD: A 48-year-old woman, treated with immunosuppressant agents for systemic lupus, presented with PML mimicking neurolupus flare. A complete remission was obtained with cytarabine and cidofovir. CONCLUSION: Combined cytarabine and cidofovir appears a promising therapeutic option in PML associated with autoimmune systemic disorders.

Weakly and strongly associated nonfreezable water bound in bones.

Water bound in bone of rat tail vertebrae was investigated by 1H NMR spectroscopy at 210-300 K and by the thermally stimulated depolarization current (TSDC) method at 190-265 K. The 1H NMR spectra of water clusters were calculated by the GIAO method with the B3LYP/6-31G(d,p) basis set, and the solvent effects were analyzed by the HF/SM5.45/6-31G(d) method. The 1H NMR spectra of water in bone tissue include two signals that can be assigned to typical water (chemical shift of proton resonance deltaH=4-5 ppm) and unusual water (deltaH=1.2-1.7 ppm). According to the quantum chemical calculations, the latter can be attributed to water molecules without the hydrogen bonds through the hydrogen atoms, e.g., interacting with hydrophobic environment. An increase in the amount of water in bone leads to an increase in the amount of typical water, which is characterized by higher associativity (i.e., a larger average number of hydrogen bonds per molecule) and fills larger pores, cavities and pockets in bone tissue.

Persistent expression of cyclin D1 disrupts normal photoreceptor differentiation and retina development.

The differentiation of neuronal cells in the developing mammalian retina is closely coupled to cell cycle arrest and proceeds in a highly organized manner. Cyclin D1, which regulates cell proliferation in many cells, also drives the proliferation of photoreceptor progenitors. In the mouse retina, cyclin D1 protein normally decreases as photoreceptors mature. To study the importance of the down-regulation of cyclin D1 during photoreceptor development, we generated a transgenic mouse in which cyclin D1 was persistently expressed in developing photoreceptor cells. We observed numerous abnormalities in both photoreceptors and other nonphotoreceptor cells in the retina of these transgenic mice. In particular, we observed delayed opsin expression in developing photoreceptors and alterations in their number and morphology in the mature retina. These alterations were accompanied by disorganization of the inner nuclear and plexiform layers. The expression of cyclin D1 caused excess photoreceptor cell proliferation and apoptosis. Loss of the p53 tumor suppressor gene decreased cyclin D1-induced apoptosis and led to microscopic hyperplasia in the retina. These findings are distinct from other mouse models in which the retinoblastoma gene pathway is disrupted and suggest that the IRBP-cyclin D1 mouse model may recapitulate an early step in the development of retinoblastoma.

Pigment epithelium-derived factor supports normal development of photoreceptor neurons and opsin expression after retinal pigment epithelium removal.

Dysfunction of the retinal pigment epithelium (RPE), its loss, or separation from the underlying neural retina results in severe photoreceptor degeneration. Pigment epithelium-derived factor (PEDF) is a glycoprotein with reported neuroprotective and differentiation properties that is secreted in abundance by RPE cells. The "pooling" of PEDF within the interphotoreceptor matrix places this molecule in a prime physical location to affect the underlying neural retina. The purpose of this study was to analyze the morphogenetic activity of PEDF in a model of photoreceptor dysmorphogenesis induced by removal of the RPE. Eyes were dissected from embryonic Xenopus laevis, and the RPE was removed before culturing in medium containing PEDF, PEDF plus anti-PEDF antibodies, or medium alone. Control retinas were maintained with an adherent RPE. Light and electron microscopic analysis was used to examine retinal ultrastructure. Opsin was localized immunocytochemically and quantified as an index of outer segment membranous material and photoreceptor protein expression. Removal of the RPE resulted in an aberrant assembly of photoreceptor outer segments, loss of fine subcellular ultrastructure in photoreceptors, and a reduction in opsin protein levels when compared with control retinas. The addition of PEDF prevented the dysmorphic photoreceptor changes induced by RPE removal. In particular, photoreceptor ultrastructure, outer segment membrane assembly, and steady-state levels of opsin were equivalent to control conditions. Anti-PEDF antibodies completely blocked the morphogenetic activity of PEDF. These results indicate that PEDF is able to mimic the supportive role of the RPE on photoreceptors during the final stages of retinal morphogenesis.

Lactose supports Muller cell protein expression patterns in the absence of the retinal pigment epithelium.

PURPOSE: We have previously shown that lactose, but not mannose, promotes the assembly of nascent photoreceptor outer segments in the absence of the retinal pigment epithelium (RPE). The purpose of the present study was to determine if, in addition to the improved outer segment assembly observed in the presence of lactose, biosynthetic changes in Muller cells could also be detected. METHODS: The RPE was removed from intact isolated Xenopus embryonic eyes that were allowed to complete differentiation in Niu-Twitty medium, Niu-Twitty with mannose, or Niu-Twitty with lactose. Control retinas matured in vitro with an adherent RPE. Retinal morphology was evaluated for organized folding of outer segment membranes and cell loss. In addition, the expression of three Muller cell proteins, glial fibrillary acidic protein (GFAP), cellular retinaldehyde-binding protein (CRALBP), and glutamine synthetase, was examined. RESULTS: In control retinas, GFAP is undetectable, CRALBP heavily immunolabels Muller cells, and radial patterns of glutamine synthetase immunoreactivity are present. In the absence of the RPE, Muller cells upregulate GFAP expression, CRALBP labeling is present at a slightly reduced level, and glutamine synthetase immunolabeling is negligible. Neither mannose nor lactose modify significantly the expression of CRALBP. Similarly, both compounds completely prevent the upregulation of GFAP. However, normal glutamine synthetase expression was observed only in the presence of lactose, but not in the presence of mannose. Statistical analyses of slot blot-based protein quantification confirmed our immunochemical results. CONCLUSIONS: In RPE-deprived retinas supplemented with lactose, Muller cells were morphologically normal. The proper photoreceptor outer segment morphogenesis observed under these conditions was uniquely associated with normal levels of glutamine synthetase expression. The exact significance of this finding with respect to photoreceptor outer segment morphogenesis is unknown. We suggest that glutamine synthetase may be a marker of Muller cell metabolic or structural integrity that may reflect the enhanced ability of these cells, in the presence of lactose, to support photoreceptor outer segment morphogenesis.

Lactose promotes organized photoreceptor outer segment assembly and preserves expression of photoreceptor proteins in retinal degeneration.

PURPOSE: We have previously shown that lactose promotes the proper assembly of photoreceptor outer segments in the absence of the retinal pigment epithelium (RPE). The purpose of this study was to determine if the difference between organized and disorganized membranes was a variation in the amounts of two structural proteins, opsin and rds/peripherin. METHODS: Eye rudiments were dissected from Xenopus laevis embryos and the RPE was removed prior to culturing in the following media: Niu-Twitty medium; Niu-Twitty with mannose; Niu-Twitty with lactose. Controls included retinas that matured in vitro with an adherent RPE. Photoreceptor ultrastructure was evaluated with emphasis on outer segment membrane organization. The relative amounts of opsin and rds/ peripherin, two outer segment-specific proteins, were determined, as were their immunolabeling patterns. RESULTS: In control retinas, outer segments were composed of stacked, flattened membranous saccules. Opsin labeling of rod outer segments was very dense, indicative of normally organized disc membranes, and rds/peripherin labeling was heavy at the outer segment disc periphery and incisures. In the absence of the RPE, a whorl-like profile of outer segments is present in what would be the sub-retinal space. Opsin immunolabeling was patchy and disorganized. Immunolabeling of rds/peripherin was present, but in a disorderly array. Mannose showed no protective effect. In contrast, lactose promoted the formation of organized outer segments and allowed for near normal expression of both photoreceptor markers. In retinas with disorganized outer segments, the expression of opsin is downregulated while the expression of rds/peripherin is maintained or upregulated. CONCLUSIONS: Lactose protects against the retinal degeneration induced by RPE removal by preserving the outer segment structure and the photoreceptor immunolabeling patterns. It also maintains constant the relative amounts of opsin and rds/peripherin. It is possible that in degenerating retinas, photoreceptors upregulate rds/peripherin expression in attempt to provide additional support for the proper folding of nascent membranes, however this is insufficient to permit organization of the photoreceptor outer segments. Our results suggest that rescue-effect of lactose is mediated by a non-rds/peripherin related mechanism.

Comparison of conjunctival and corneal surface areas in rabbit and human.

This comparative study shows the surface area ratio of conjunctiva to cornea to be two times larger in humans than in rabbits. This large heretofore unrecognized interspecies difference may affect the applicability of drug pharmacokinetic data obtained using rabbit models and should be taken into consideration in topical drug development and future comparative drug penetration studies between rabbit and man.

The concentrations of osteocalcin and degradation products of type I collagen in pregnant women with pre-eclampsia.

OBJECTIVES: To investigate the concentrations of osteocalcin and collagen type I C-terminal telopeptides in pregnant women with pre-eclampsia. STUDY DESIGN: 26 patients with severe pre-eclampsia and 24 healthy pregnant women were included in the study. Serum concentrations of osteocalcin and C-telopeptides--degradation products of type I collagen were determined using the ELISA method. Statistical analysis was performed using Mann-Whitney U-test. RESULTS: In pre-eclamptic patients, the concentrations of osteocalcin and degradation products of collagen type I were significantly higher (P<0.005) when compared to healthy pregnant women. CONCLUSION: These results could suggest that there are alterations in bone metabolism in pregnant women with pre-eclampsia.

Hypoplasia of G cells in long term steroid-treated rats after ultra-high dose of salmon calcitonin.

Male young adult Wistar rats were treated parenterally with cortisol and cortisol combined with salmon calcitonin for 28 and 56 days and the stomach was investigated histologically as well as the G cells using immunohistochemistry. After 28 days of cortisol administration desquamation of the superficial layers of gastric mucosa was found and after 56 days ulcer-like changes developed and increased gastrin immunoreactivity was observed. Administration of high doses of salmon calcitonin together with cortisol resulted in a significant hypoplasia of G cells and prevented the pathological changes in the gastric mucosa.

Ukrain (NSC-631570) influences on bone status: a review.

Ukrain, a thiophosphoric acid alkaloid derivative of Chelidonium majus L., was shown to affect bone tissue metabolism as assessed in densitometric and biomechanical studies in rats. Its action could be slightly osteopenic at the highest doses administered to intact animals for a prolonged period of time. This phenomenon is possibly related to Ukrain's inhibitory effect on spontaneous locomotor activity of treated animals and/or to the stimulatory effect of the drug on the osteoclastic activity via the macrophage system. By far, the most important finding seems to be the anabolic effect of Ukrain on bone in ovariectomized rats, which is most probably related to induced increase in the production of gonadal hormones, predominantly estrogens. In this regard, the postmenopausal population of female patients treated for malignancies with Ukrain (and obviously the most numerous one) meritis clinical attention as far as the antiosteoporotic effects of this drug are concerned.

Intermittent three-month treatment with Ukrain in intact and ovariectomized rats. Part I: Effect on selected biomechanical parameters of the femur.

Ukrain, an acid alkaloid derivative from Chelidonium majus L., was administered intraperitoneally to intact and ovariectomized sexually mature female rats at doses of 7, 14 and 28 mg/kg once daily for 10 days, followed by 10-day break. This procedure was repeated five times. At the end of this Ukrain treatment (24 h after the last dose of the drug), the right femora of the rats were harvested and a three-point bending test until shaft failure was performed under standard conditions. Assembled data, including ultimate strength, ultimate energy at failure and ultimate displacement, were assessed. Strength and energy were adjusted to accommodate differences between groups in the end body weight. These results showed a decrease in adjusted bone strength of ovariectomized rats as compared to sham-operated animals. None of the three long-term doses of Ukrain (i.e., 7 mg/kg, 14 mg/kg and 28 mg/kg) significantly altered the bone strength when it was adjusted to the end body weight of the ovariectomized animals. Intact rats that received a 28 mg/kg dose of Ukrain demonstrated decrease of bone strength of 13.5% when it was adjusted to the end body weight.

Intermittent three-month treatment with Ukrain in intact and ovariectomized rats. Part II: Effect on bone mineral density of the femur.

Ukrain, an acid alkaloid derivative of Chelidonium majus L., was administered intraperitoneally to ovariectomized and control sexually mature female rats at doses of 7, 14 and 28 mg/kg once daily for 10 days, followed by 10-day break. This procedure was repeated five times. At the end of the Ukrain treatment (24 h after the last dose of the drug) the right femora of the rats were harvested and the bone densitometric parameters of the whole bone and distal metaphysial and intertrochanteric subregions were assessed using the dual energy x-ray absorptiometry densitometric method. The results showed no apparent decrease in bone mineral density in groups of rats studied. A nearly significant (p = 0.08) decrease of bone mineral content was observed in ovariectomized rats treated with 14 mg/kg of Ukrain.

Intermittent three-month treatment with Ukrain in intact and ovariectomized rats. Part III: Effect on the native electron paramagnetic resonance signal intensity of the femur.

Ukrain, an acid alkaloid derivative of Chelidonium majus L., was administered intraperitoneally to ovariectomized and sexually mature female control rats at doses of 7, 14 and 28 mg/kg once daily for 10 days, followed by a 10-day break. This procedure was repeated five times. At the end of the long-term treatment with Ukrain (24 h after the last dose of the drug) the left femora of the rats were harvested. Dried bones were studied by electron paramagnetic resonance (EPR) method. Significant changes in the intensity of the native signal were observed within the groups (ANOVA, p = 0.0001); the lowest value was observed in the ovariectomized rats treated with 28 mg/kg of Ukrain (decrease of 40.8% compared to the ovariectomized group, p = 0.005). A significant increase in the intensity of the signal was observed in the intact 7 mg/kg Ukrain-treated group (34.7% compared to controls, p = 0.004).

Effect of intermittent three-month treatment with different doses of Ukrain on subregional femoral bone mineral density of sexually mature female rats.

Sexually mature but still growing female Wistar rats received i.p. injections of Ukrain (7, 14 or 28 mg/kg in a volume of 0.5 ml/100g) every other day for 10 days, followed by a 10-day break, and this procedure was performed five times. The control animals received the same volume of injected water. At the end of the experiment the rat right femora were harvested and the bone densitometric parameters of the entire bone, distal metaphyseal and basicervical subregions were assessed using the dual-energy X-ray absorptiometry (DXA) densitometric method. No significant changes were observed in the bone mineral density in experimental groups in comparison with control animals that received the vehicle. A slight decrease in the bone mineral content value was observed in the distal metaphyseal region in animals that were treated with the highest dose of Ukrain.

Effect of intermittent three-month treatment with different doses of Ukrain on subregional bone mineral density of the femur of ovariectomized rats.

Ukrain, thiophosphoric acid alkaloid derivative from Chelidonium majus L., was administered i.p. to ovariectomized rats in doses of 7, 14 and 28 mg/kg every other day for 10 days, followed by a 10-day break, and the procedure was performed five times. At the end of long-term treatment with Ukrain (24 h after the last dose of the drug) the rats' right femora were harvested and the bone densitometric parameters of the whole bone and distal metaphyseal and basicervical subregions were assessed using the dual-energy X-ray absorptiometry (DXA) densitometric method. The present results show a decrease in bone mineral density in groups of ovariectomized rats that received 7 mg/kg and 14 mg/kg of Ukrain versus untreated ovariectomized animals. Administration of Ukrain at a dose of 28 mg/kg did not significantly alter bone parameters of ovariectomized rats.

Effect of six-month treatment with Ukrain on early osteoporosis induced by ovariectomy in rats. Part I: Preliminary studies of bone parameters.

Ukrain, thiophosphoric acid alkaloid derivatives from Chelidonium majus L. was administered intraperitoneally in a dose of 28 mg/kg (equivalent to 0.1 LD50) every other day for six months to female rats with ovariectomy-induced early osteoporosis. Administration of Ukrain was started on the second day after the surgical operation. At the end of the long-term treatment with Ukrain each rat was tested for the strength of both humeri and some parameters of rat femur were measured. The body weight of ovariectomized rats was also examined. The present results show that the decrease in the mechanical strength of the humeral bones and some changes in the femur caused by ovariectomy were prevented by the six-month treatment with Ukrain. However, in both ovariectomized groups and in ovariectomized rats pretreated with Ukrain an increase of body weight was observed.

Effect of six-month treatment with Ukrain on early osteoporosis induced by ovariectomy in rats. Part II: Preliminary studies of peripheral blood parameters.

Studies on Wistar rats have demonstrated that six months after ovariectomy no changes were observed in peripheral blood morphology other than a decreased leukocyte count. A similar effect was noted in the sham-operated group. The present results show that Ukrain treatment started on the second postoperative day and continued every other day for six months significantly increased leukocyte count in comparison with the untreated ovariectomized and sham-operated groups. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also estimated in this study. In none of the groups studied was any change of ALT activity noted. The activity of AST was significantly increased in the sham-operated and untreated ovariectomized rats. After six-month treatment with Ukrain, the activity of this enzyme was significantly decreased in the serum of untreated ovariectomized rats. In rats pretreated with Ukrain some of the studied parameters of peripheral blood were similar to those of the control group.