The necdin gene is deleted in Prader-Willi syndrome and is imprinted in human and mouse

Human chromosome 15q11-q13 contains genes that are imprinted and expressed from only one parental allele. Prader-Willi syndrome (PWS) is due to the loss of expression of one or more paternally expressed genes on proximal human chromosome 15q, most often by deletion or maternal uniparental disomy. Several candidate genes and a putative imprinting centre have been identified in the deletion region. We report that the human necdin-encoding gene (NDN) is within the centromeric portion of the PWS deletion region, between the two imprinted genes ZNF127 and SNRPN. Murine necdin is a nuclear protein expressed exclusively in differentiated neurons in the brain. Necdin is postulated to govern the permanent arrest of cell growth of post-mitotic neurons during murine nervous system development. We have localized the mouse locus Ndn encoding necdin to chromosome 7 in a region of conserved synteny with human chromosome 15q11-q13, by genetic mapping in an interspecific backcross panel. Furthermore, we demonstrate that expression of Ndn is limited to the paternal allele in RNA from newborn mouse brain. Expression of NDN is detected in many human tissues, with highest levels of expression in brain and placenta. NDN is expressed exclusively from the paternally inherited allele in human fibroblasts. Loss of necdin gene expression may contribute to the disorder of brain development in individuals with PWS.      

The cytoprotective drug amifostine modifies both expression and activity of the pro-angiogenic factor VEGF-A

Background

Amifostine (WR-2721, delivered as Ethyol^®) is a phosphorylated aminothiol compound clinically used in addition to cis-platinum to reduce the toxic side effects of therapeutic treatment on normal cells without reducing their efficacy on tumour cells. Its mechanism of action is attributed to the free radical scavenging properties of its active dephosphorylated metabolite WR-1065. However, amifostine has also been described as a potent hypoxia-mimetic compound and as a strong p53 inducer; both effects are known to potently modulate vascular endothelial growth factor (VEGF-A) expression. The angiogenic properties of this drug have not been clearly defined.

Methods

Cancer cell lines and endothelial cells were used in culture and treated with Amifostine in order to study (i) the expression of angiogenesis related genes and proteins and (ii) the effects of the drug on VEGF-A induced in vitro angiogenesis.

Results

We demonstrated that the treatment of several human cancer cell lines with therapeutical doses of WR-1065 led to a strong induction of different VEGF-A mRNA isoforms independently of HIF-1α. VEGF-A induction by WR-1065 depends on the activation of the eIF2alpha/ATF4 pathway. This up-regulation of VEGF-A mRNA was accompanied by an increased secretion of VEGF-A proteins fully active in stimulating vascular endothelial cells (EC). Nevertheless, direct treatment of EC with amifostine impaired their ability to respond to exogenous VEGF-A, an effect that correlated to the down-regulation of VEGFR-2 expression, to the reduction in cell surface binding of VEGF-A and to the decreased phosphorylation of the downstream p42/44 kinases.

Conclusions

Taken together, our results indicate that amifostine treatment modulates tumour angiogenesis by two apparently opposite mechanisms - the increased VEGF-A expression by tumour cells and the inhibition of EC capacity to respond to VEGF-A stimulation.

     

Digital processing of radiographic images from PACS to publishing

Several studies have addressed the implications of filmless radiologic imaging on telemedicine, diagnostic ability, and electronic teaching files. However, many publishers still require authors to submit hard-copy images for publication of articles and textbooks. This study compares the quality digital images directly exported from picture archive and communications systems (PACS) to images digitized from radiographic film. The authors evaluated the quality of publication-grade glossy photographs produced from digital radiographic images using 3 different methods: (1) film images digitized using a desktop scanner and then printed, (2) digital images obtained directly from PACS then printed, and (3) digital images obtained from PACS and processed to improve sharpness prior to printing. Twenty images were printed using each of the 3 different methods and rated for quality by 7 radiologists. The results were analyzed for statistically significant differences among the image sets. Subjective evaluations of the filmless images found them to be of equal or better quality than the digitized images. Direct electronic transfer of PACS images reduces the number of steps involved in creating publication-quality images as well as providing the means to produce high-quality radiographic images in a digital environment.     

Comparison of radiographic image quality from four digitization devices as viewed on computer monitors

The objective of this study was to compare the quality of radiographic images digitized from commercial-grade and consumer-grade digital cameras and scanners as viewed on computer monitor. Radiographic images were digitized from hardcopy film using a commercial-grade laser scanner, a consumer-grade desktop flatbed scanner, a commercial-grade digital camera, and a consumer-grade digital camera. The quality of images without and with grayscale histogram adjustment was evaluated subjectively by 10 board-certified radiologists. Optical density response was evaluated objectively using a grayscale test pattern. There was no significant difference in subjective quality among images digitized with the commercial scanner, consumer scanner, and commercial camera. The quality of images digitized with the consumer camera was lower than the other 3. Objective tests showed the commercial scanner to have the most linear optical density response. For the purpose of viewing images on a computer monitor, a consumer-grade desktop scanner can produce images of similar quality to those produced by more expensive laser commercial-grade scanners and digital cameras and provides cost-efficient means to digitize radiographic plain films. A consumer-grade camera may not be optimal for use in this setting.     

膦肽类化合物对血管紧张素转化酶的体外抑制活性及构效关系

设计与合成了一系列膦三肽化合物。测定化合物对血管紧张素转化酶的体外抑制活性。结果表明,有些化合物的体外抑制活性已接近captopril的水平。在此基础上,探讨抑制剂的结构与抑制活性之间的关系。     

Establishment of a Modified in Vitro Cultivation of Protoscoleces to Adult Echinococcus granulosus; an Important Way for New Investigations on Hydatidosis

Background

Echinococcus granulosus, a zoonotic cestode parasite, causative agent of hydatid cyst is endemic in many parts of the world including the Middle East. Study on different aspects of this parasite is very important and valuable. However, working with adult worms which their habitat situated in the small intestine of canids, is dangerous and risky. Achieving such risky situation needs a controlled condition which is cultivation of the organisms in the laboratory. In this regard, cultivation of E. granulosus protoscoleces leading to adult worms was established in the laboratory for the first time in Iran.

Methods

Under aseptic conditions a number of protoscoleces were cultivated in diphasic S.10E.H medium using CO2 incubator to produce adult worms.

Results

Different forms of parasites including pre-segmentation stages (PS1 - PS4) and segmentation stages (S5-S8) and developing stages in segmented worms (S10-S11) were observed and evaluated in these medium. Finally adult worms contained four proglottids with a large and distinct genital pore were observed 50-55 days post cultivation. These parasites do not produce fertile eggs and conclusively do not have risk of hydatid disease transmission to the researchers.

Conclusion

The mentioned method for producing E. granulosus adult worms can open a new window for researches and facilitate working on different aspects of hydatidosis especially for diagnosis, protection and treatment studies.     

Sensitivity and specificity of typhoid fever rapid antibody tests for laboratory diagnosis at two sub-Saharan African sites

Objective

To evaluate three commercial typhoid rapid antibody tests for Salmonella Typhi antibodies in patients suspected of having typhoid fever in Mpumalanga, South Africa, and Moshi, United Republic of Tanzania.

Methods

The diagnostic accuracy of Cromotest^® (semiquantitative slide agglutination and single tube Widal test), TUBEX^® and Typhidot^® was assessed against that of blood culture. Performance was modelled for scenarios with pretest probabilities of 5% and 50%.

Findings

In total 92 patients enrolled: 53 (57.6%) from South Africa and 39 (42.4%) from the United Republic of Tanzania. Salmonella Typhi was isolated from the blood of 28 (30.4%) patients. The semiquantitative slide agglutination and single-tube Widal tests had positive predictive values (PPVs) of 25.0% (95% confidence interval, CI: 0.6–80.6) and 20.0% (95% CI: 2.5–55.6), respectively. The newer typhoid rapid antibody tests had comparable PPVs: TUBEX^®, 54.1% (95% CI: 36.9–70.5); Typhidot^® IgM, 56.7% (95% CI: 37.4–74.5); and Typhidot^® IgG, 54.3% (95% CI: 36.6–71.2). For a pretest probability of 5%, PPVs were: TUBEX^®, 11.0% (95% CI: 6.6–17.9); Typhidot^® IgM, 9.1% (95% CI: 5.8–14.0); and Typhidot^® IgG, 11.0% (6.3–18.4). For a pretest probability of 50%, PPVs were: TUBEX^®, 70.2% (95% CI: 57.3–80.5); Typhidot^® IgM, 65.6% (95% CI: 54.0–75.6); and Typhidot^® IgG, 70.0% (95% CI: 56.0–81.1).

Conclusion

Semiquantitative slide agglutination and single-tube Widal tests performed poorly. TUBEX^® and Typhidot^® may be suitable when pretest probability is high and blood cultures are unavailable, but their performance does not justify deployment in routine care settings in sub-Saharan Africa.     

Interaction of benzylidene-anabaseine analogues with agonist and allosteric sites on muscle nicotinic acetylcholine receptors

Background and purpose

Benzylidene-anabaseines (BAs) are partial agonists of the α7 nicotinic acetylcholine receptor (nAChR) but their mechanism(s) of action are unknown. Our study explores several possibilities, including direct interactions of BAs with the nAChR channel.

Experimental approach

Functional and radioligand-binding assays were used to examine the interaction of two BA analogues, 3-(2,4-dimethoxybenzylidene)-anabaseine (DMXBA) and its primary metabolite 3-(4-hydroxy-2-methoxybenzylidene)-anabaseine (4OH-DMXBA) with both agonist and non-competitive antagonist (NCA)-binding sites on muscle-type nAChRs.

Key results

Both BAs non-competitively inhibited ACh activation of human fetal muscle nAChRs and sterically inhibited the specific binding of the NCAs [piperidyl-3,4-3H(N)]-(N-(1-(2-thienyl)cyclohexyl)-3,4-piperidine ([³H]TCP) and [³H]dizocilpine to Torpedo nAChRs in the desensitized state. These compounds modulated [³H]tetracaine, [¹⁴C]amobarbital and [³H]TCP binding to resting nAChRs by allosteric mechanisms. Both BAs enhanced [³H]TCP binding when the nAChR was initially in the resting but activatable state, suggesting that both compounds desensitized the Torpedo nAChR. Although DMXBA failed to activate human fetal muscle nAChRs, 4OH-DMXBA was found to be a partial agonist. [³H]Nicotine competition-binding experiments confirmed that 4OH-DMXBA has higher affinity than DMXBA for the agonist sites, and that DMXBA is also a competitive antagonist.

Conclusions and implications

3-(4-hydroxy-2-methoxybenzylidene)-anabaseine is a partial agonist for human fetal muscle nAChRs, whereas DMXBA only has competitive and NCA activities. The NCA-binding site for BAs overlaps both the phencyclidine-and dizocilpine-binding sites in the desensitized Torpedo nAChR ion channel. The desensitizing property of BAs suggests another possible mode of non-competitive inhibition in addition to direct channel-blocking mechanisms.     

The efficacy of bismuth subsalicylate in the treatment of acute diarrhoea and the prevention of persistent diarrhoea

A controlled, randomized, double-blind study in Bangladeshi children (ages 4-36 mo) with acute diarrhoea was undertaken to determine whether bismuth subsalicylate (BSS) would prevent the development of persistent diarrhoea (PD) in young children. The children were randomized to two groups: 226 were given liquid oral BSS, (as Pepto-Bismol), 100 mg/kg/d for 5 d; 225 were given placebo of identical appearance. On admission to the study, the two groups were comparable both clinically and microbiologically. Rotavirus was found in 56% of all the children, and enterotoxigenic E. coli in 31% of a subsample studied. Children treated with BSS had less severe and less prolonged illness than those treated with placebo (p = 0.057). There was, however, no difference in the development of PD between the two groups (8% and 11%). Unexpectedly, patients treated with BSS gained significantly more weight (2.3%) than those treated with placebo (0.5%; p < 0.001) during the course of the study. No toxicity of BSS was detected. Conclusion: Treatment with BSS had a modest therapeutic effect on acute diarrhoea, as has been previously demonstrated, but with no suggestion of a therapeutic effect on the prevention of persistent diarrhoea in this group of patients.