The biologics anakinra and etanercept prevent cytokine-induced growth retardation in cultured fetal rat metatarsal bones

Background/Aims: Chronic inflammation during childhood often leads to impaired bone growth and reduced adult height. Proinflammatory cytokines interleukin (IL)-1β and tumor necrosis factor (TNF)-α synergistically impair bone growth in vitro. We hypothesized that biologic agents may rescue bones from cytokine-induced growth impairment and that insulin growth factor (IGF)-I may potentiate such an effect. Methodology: Metatarsal bones from fetal Sprague-Dawley rats (19-20 days p.c.) were treated with IL-1β plus TNF-α, or the combination of these cytokines with anakinra (IL-1 receptor antagonist), etanercept (TNF-inhibitor) and/or IGF-I. The bones were measured and growth expressed as percent increase in bone length over the 7-day culture period. Results: When exposed to IL-1β plus TNF-α (10 + 10 ng/ml), bone growth was markedly suppressed (6.6 ± 1.4 vs. 50.6 ± 2.5% in control bones; p < 0.001). The growth of cytokine exposed bones (IL-1β plus TNF-α) was dose-dependently rescued by anakinra (0.05-500 μg/ml) or etanercept (0.5-500 μg/ml); at the highest concentrations, growth was similar as in control bones never exposed to cytokines. Also when combining IGF-I (100 ng/ml) and relatively low concentrations of anakinra (0.05 μg/ml) or etanercept (5 μg/ml), growth was rescued in an additive way. Conclusion: Etanercept and anakinra efficiently and dose-dependently prevent cytokine-induced bone growth impairment, and combination with IGF-I further improves bone growth.     

Amnesia induced by morphine in spatial memory retrieval inhibited in morphine-sensitized rats

The present study investigated the effect of morphine sensitization on the impairment of spatial memory retrieval induced by acute morphine in adult male rats. Spatial memory was assessed by 2-day Morris water maze task which included training and test day. On the training day, rats were trained by a single training session of 8 trials. On the test day, a probe trial consisting of 60s free swim period without a platform and the visible test were administered. Morphine sensitization was induced by subcutaneous (s.c.) injection of morphine, once daily for 3 days followed by 5 days without drug treatment before training. The results indicated that acute administration of morphine (7.5mg/kg, s.c.) before testing impaired spatial memory on the test day. Pre-test morphine-induced amnesia decreased in morphine-sensitized (15 and 20mg/kg, s.c.) rats. Improvement in spatial memory retrieval in morphine-sensitized rats was inhibited by once daily administration of naloxone (1 and 2mg/kg, s.c.) 30 min prior to the injection of morphine for three days. The results suggest that morphine sensitization reverses the impairment of spatial memory retrieval induced by acute morphine and it is implied that mu-opioid receptors may play an important role in this effect.     

Effect of fragment removal on blastocyst formation and quality of human embryos

Blastomere fragmentation is one of the most significant defects in cleaving embryos. Scientists believed that removing the fragments was a possible way to reduce their unwanted effects. This hypothesis has been tested in some studies in which the development of human fragmented embryos was followed in vivo after all fragments were removed, but little is known about the potential for in-vitro development of such embryos, which is the subject of the present study. For this purpose, 4-6 cell surplus human embryos were scored according to the degree and pattern of fragmentation into four grades, allocated into two groups of control and fragmentation removal (experimental) and cultured sequentially. At the end of day 6 of culture, in the experimental group especially in grade IV blastocyst rate, size and number of blastomeres in each blastocyst were all improved compared with those of the control group (42.3 versus 20.0%; 19,205.7 +/- 1060.3 versus 15,825.9 +/- 448.7 microm(2) and 100.14 +/- 13.48 versus 63.75 +/- 19.79 respectively, P < 0.05). In the grade IV embryos, apoptotic index was also significantly reduced after embryo fragmentation removal (3.40 +/- 0.88 versus 22.99 +/- 4.45, P < 0.05). In conclusion, fragmentation removal had a positive effect on human fragmented embryos and produced the best quality blastocysts.     

Air-assisted Descemet-stripping automated endothelial keratoplasty with posterior chamber iris-fixation of aphakic iris-claw intraocular lens

We report a combination of surgical techniques during Descemet-stripping automated endoithelial keratoplasty and intraocular lens (IOL) exchange in patients with pseudophakic bullous keratopathy and angle-supported anterior chamber IOLs. During this procedure, the anterior chamber IOL is exchanged for a posterior chamber iris-claw IOL enclavated to the posterior iris; the anterior chamber is kept filled with air using an air-fluid exchange machine during descemetorhexis and insertion of the donor endothelial disk.     

Mutation screening of TGFBI in two Iranian Avellino corneal dystrophy pedigrees

Purpose: Genetic analysis and phenotypic features of Avellino corneal dystrophy patients from Japan and some European countries have been published. We report for the first time the genetic analysis and phenotypic features of two Avellino corneal dystrophy pedigrees from the Middle East. Methods: Slit‐lamp biomicroscope photographs of cornea were obtained, and corneal tissue sections were stained with masson‐trichrome and Congo red. DNA was isolated from peripheral blood leucocytes and exons 4 and 12 of TGFBI were screened for mutations by direct sequencing. Results: The probands of the pedigrees had phenotypic features consistent with diagnosis of Avellino corneal dystrophy. They were homozygous for the same R124H mutation in TGFBI as previously reported in Avellino patients from Japan and European countries. Heterozygous carriers of the mutation were identified in the pedigree and shown to have symptoms of disease milder than those of the probands. Conclusion: The finding of R124H in the Middle Eastern (Iranian) population supports the proposal that perhaps only substitution of histidine for arginine at position 124 of tumour growth factor beta induced protein results in the Avellino corneal dystrophy phenotype. As both probands were originally diagnosed with granular corneal dystrophy, and as heterozygous carriers of R124H were unaware of their disease status prior to genetic analysis, the importance of genetic analysis is emphasized.     

Health and ecological risk assessment and simulation of heavy metal-contaminated soil of Tehran landfill

The toxic effects of heavy metals in landfill soils have become a significant concern for human health. The present study aimed to estimate the health and ecological risk associated with soil heavy metal in Tehran landfill. A total of 48 soil samples were taken from the landfill and residential area and were analyzed using inductively coupled plasma-optical emission spectroscopy. The results showed the following order for heavy metal levels in landfill soil: Al > Fe > Mn > Zn > Cr > Cu > Pb > Ni > Co > As > Cd. The investigated ecological indices showed moderate to high heavy metal pollution. The principal component analysis revealed that the concentration of Pb, Cu, Zn, Cr, and Ni in the investigated soil was mainly affected by anthropogenic activities. Although the hazard index (HI) value in children was 6.5 times greater than that of adults, this value for both landfill workers and residents of the target area was at a safe level (HI ≤ 1). In the residential area, the Incremental Lifetime Cancer Risk (ILCR) value of adults (1.4 × 10⁻⁴) was greater than children ILCR value (1.2 × 10⁻⁴). Monte Carlo simulation and sensitivity analysis showed input variables such as exposure duration, exposure frequency, Ni concentration, soil ingestion rate, and As concentration have a positive effect on ILCR of 41.3, 24.3, 9.4, 9.0, and 2.9% in children, respectively. These results indicate that the landfill soil and the adjacent residential area are affected by heavy metal contamination and that the current solid waste management policies need to be revised.

The toxic effects of heavy metals in landfill soils have become a significant concern for human health.     

Tamoxifen Impairs Both Longitudinal and Cortical Bone Growth in Young Male Rats

Tamoxifen (Tam) has been used experimentally to treat boys with gynecomastia and girls with McCune‐Albright syndrome. This drug was recently shown to inhibit the growth of cultured fetal rat metatarsal bones and thus might also affect bone growth in vivo. Four‐week‐old Sprague‐Dawley rats were gavaged daily with vehicle alone (peanut oil), Tam (40 mg/kg/d; 1 or 4 wk), or estradiol (40 μg/kg/d; 4 wk). Five of the 10 rats in each group were killed after 4 wk and the other five after 14 wk of recovery. Bone growth was followed by repeat DXA scans, whereas other bone parameters and spine length were evaluated by pQCT and X‐ray at the time of death. Four‐week Tam treatment significantly decreased body weight, nose‐anus distance, spinal and tibial bone lengths, trabecular BMD, cortical periosteal circumference, and bone strength and also reduced serum IGF‐I levels (424 ± 54 versus 606 ± 53 ng/ml in control; p < 0.05). Analysis of the tibial growth plate of treated rats showed elevated chondrocyte proliferation (BrdU) and apoptosis (TUNEL), as well as decreases in the number of hypertrophic chondrocytes and in the size of terminal hypertrophic chondrocytes. Despite a complete catch‐up of body weight after 14 wk of recovery, the tibia was still shorter (p < 0.001) and its cortical region was smaller. We conclude that, when administered at a clinically relevant dose, Tam causes persistent retardation of longitudinal and cortical radial bone growth in young male rats. Our findings suggest that this inhibition results from local effects on the growth plate cartilage and systemic suppression of IGF‐I production. Based on these rat data, we believe that Tam, if given to growing individuals, might compromise cortical bone growth, bone strength, and adult height.