Activation of delta-globin gene expression by erythroid Krupple-like factor: a potential approach for gene therapy of sickle cell disease

Hemoglobin A2 (HbA2; alpha 2 delta 2) is a powerful inhibitor of HbS (alpha 2 beta 2(3)) polymerization. However, HbA2 levels are normally low in sickle cell patients. We show that a major reason for low delta- globin gene expression is the defective CACCC box at -90 in the delta- globin promoter. When the CACCC box defect in delta is corrected, expression of an HS2 delta /Luciferase reporter is equivalent to HS2 beta /Luciferase. Erythroid Krupple-like factor (EKLF), which binds to the CACCC box of the beta-globin gene and activates high-level expression, does not bind to the normal delta-globin promoter. Our goal is to design a modified EKLF that binds to the defective delta-globin promoter and enhances delta-globin gene expression. To test the feasibility of this strategy, we inserted the beta-globin CACCC box at - 90 of the delta-globin gene promoter to produce an HS2 delta CAC-beta construct and quantitated human delta- and beta-globin mRNA in stably transformed murine erythroleukemia (MEL) cells. delta- Globin mRNA in these cells was 22.0% +/- 9.0% of total human globin mRNA (delta/delta + beta) as compared with 3.0% +/- 1.3% in the HS2 delta-beta control. In a second set of experiments a GAL4 DNA-binding site was inserted at - 90 of the delta-globin gene to produce an HS2 delta GAL4-beta construct. This construct and a GAL4(1-147)/EKLF expression vector were stably transfected into MEL cells. delta-Globin mRNA in these cells was 27.8% +/- 7.1% of total human globin mRNA as compared with 9.9% +/- 2.5% in the HS2 delta GAL4-beta plus GAL4(1-147) control. These results show that delta-globin gene expression can be significantly increased by a modified EKLF. Based on these results, we suggest that modified EKLFs, which contain zinc fingers designed to bind specifically to the defective delta-globin CACCC box, may be useful in gene therapy approaches to increase HbA2 levels and inhibit HbS polymerization.     

Minimally invasive techniques in common surgical procedures: Implications for training

Background Laparoscopic techniques are increasingly used in common surgical procedures. Many of these procedures are used to teach basic surgical trainees (BST) and therefore introduction of these techniques may have implications for training. Aims To establish whether the introduction of laparoscopic techniques reduced the opportunity of BSTs to perform surgical procedures. Methods Patients undergoing hernia repair or appendicectomy in 1991 (when laparoscopy was first introduced) and 1997 (when laparoscopy was readily available) were identified using the Hospital In-Patient Enquiry (HIPE) database. The principal operator and whether the procedure was open or laparoscopic were identified by chart review. Results The data showed a 50% reduction in the number of appendicectomies performed by BSTs following the introduction of laparoscopic techniques. The number of hernia repairs performed by BSTs has been preserved but the proportion by BSTs fell from 10 to 6%. The proportion of BST-performed procedures carried out laparoscopically has been reduced compared with the registrar-performed group. Conclusions The use of minimally invasive techniques has had a negative effect on surgical training. Appropriate measures must be taken to minimise this and such measures should include a structured approach to laparoscopic training and greater access to laparoscopic training facilities.     

Antifungal activities and action mechanisms of compounds from Tribulus terrestris L.

Antifungal activity of natural products is being studied widely. Saponins are known to be antifungal and antibacterial. We used bioassay-guided fractionation to have isolated eight steroid saponins from Tribulus terrestris L. , which were identified as hecogenin-3-O-beta-D-glucopyranosyl （ 1→4 ）-beta-D-galactopyranoside （ TTS-8）, tigogenin-3-O-beta-D-glucopyranosyl （ 1 →4 ）-beta-D-galactopyranoside （ TTS-9 ）, hecogenin-3-O-beta-D-glucopyranosyl （ 1 → 2 ）-beta-D-glucopyranosyl （ 1 4 ）-beta-D-galactopyranoside （ TTS-10 ）, hecogenin-3-O-beta-D-xylopyranosyl （ 1→3 ）-beta-D-glucopyranosyl （ 1→ 4 ）-beta-D-galactopyranoside （ TTS-11 ）, tigogenin-3-O-beta-D-xylopyranosyl （ 1→ 2 ）-[- beta-D-xylopyranosyl （ 1 → 3 ） ]-beta-D-glucopyranosyl （1→4）-[alpha-L-rhamnopyranosyl （1→2）]-beta-D-galactopyranoside （TTS-12）, 3-O-Ebeta-D-xylopyranosyl （1→2）-[beta-D- xylopyranosyl （1→3）]-beta-D-glucopyranosyl （ 1→4）-[alpha-L-rhamnopyranosyl （ 1→2 ）]-beta-D-galactopyranosy13-26-O-beta- D-glucopyranosyl-22-methoxy-（3beta, 5alpha, 25R）-furostan-3,26-diol （TTS-13）, hecogenin-3-O-beta-D-glucopyranosyl （ 1→ 2）-[beta-D-xylopyranosyl （ 1→ 3 ）]-beta-D-glucopyranosyl （ 1 → 4 ）-beta-D-galactopyranoside （ TTS-14 ）, tigogenin-3-O-beta-D- glucopyranosyl （ 1→ 2 ）-[ beta-D-xylopyranosyl （ 1 → 3 ）]-beta-D-glucopyranosyl （ 1 → 4 ）-beta-D-galaetopyranoside （ TTS-15 ）. The in vitro antifungal activities of the eight saponins against five yeasts, Candida albicans, Candida glabrata, Candida parapsilosis, Candida tropicalis and Cryptococcus neoformans were studied using microbroth dilution assay. In vivo activity of TTS-12 in a Candida albicans vaginal infection model was studied in particular. The results showed that TTS-12 and TTS-15 were very effective against several pathogenic candidal species and Cryptococcus neoformans in vitro. It is noteworthy that TTS-12 and TTS-15 were very active against Candida albicans （MIC50 = 10 and 2.3 microg/ml） and Cryptococcus neoformans （MIC50 = 1.7 and 6. 7 microg/ml）.     

Validation of the International Prostate Symptom Score in Chinese males and females with lower urinary tract symptoms

Objectives

To evaluate the translation of the IPSS (Hong Kong Chinese version 1) and to assess the applicability, validity, reliability and sensitivity of the instrument in both males and females with LUTS in Chinese population.

Methods

The translation of the IPSS (Hong Kong Chinese version 1) was reviewed through back translation. Modifications were made, resulting in the development of The IPSS (Hong Kong Chinese version 2). The content validity was assessed by contend validity index. 233 subjects with LUTS were recruited in Hong Kong primary care settings for pilot psychometric testing. The construct validity was assessed by corrected item-total correlation and Pearson’s correlation test against ICIQ-UI SF, IIQ-7 and SF-12 v2. The reliability was assessed by the internal consistency (Cronbach’s Alpha coefficient) and test –retest reliability (Intraclass correlation coefficient). The Sensitivity was determined by performing known group comparisons by independent T-test.

Results

The content validity index for all items could reach 1. Corrected item-total correlation scores were ≥0.4 for four symptom questions (feeling of incomplete bladder emptying, intermittency, weak stream and straining). Overall, the total symptom score moderately correlated with ICIQ-UI SF. The quality of life score moderately correlated with the IIQ-7 but weakly correlated with SF-12 v2. Overall, the reliability of the IPSS (Hong Kong Chinese version 2) was acceptable (Cronbach’s Alpha coefficient?=?0.71, ICC of the symptom questions =0.8, ICC of the quality of life question =0.7). The symptoms questions and quality of life questions of the IPSS (Hong Kong Chinese versions 2) were sensitive in detecting differences between groups.

Conclusions

The IPSS (Hong Kong Chinese version 2) is a valid, reliable and sensitive measure to assess Chinese females and males with lower urinary tract symptoms. The IPSS quality of life question is more sensitive than the generic quality of life measure to differentiate subgroups.     

Urban–rural residence and the occurrence of neural tube defects in Texas, 1999–2003

Neural tube birth defects (NTDs) affect more than 4000 pregnancies in the US annually. The etiology of NTDs is believed to be multifactorial, but much remains unknown. We examined the pattern and magnitude of urban–rural variation in anencephaly, spina bifida without anencephaly, and encephalocele in Texas in relation with urban–rural residence for the period 1999–2003. There was no evidence that urban–rural residence was associated with changes in the rate of anencephaly or spina bifida without anencephaly in unadjusted or adjusted analyses. In contrast, rates of encephalocele were statistically significantly higher in areas classified as suburban or more rural compared to urban areas using four different urban–rural residence indicators.     

Integrating multiple health behavior theories into program planning: the PER worksheet

The foundation of a logic model or any theory-based planning model is the identification of the behavioral antecedents and the relationship of these variables to the target behavior. Applying theoretical concepts to specific behaviors and populations is often challenging for practitioners and program planning students. The challenge comes from the abstract definition of theoretical constructs, the duplication of similar constructs in multiple theories, and the need to combine multiple theories. To simplify the planning process when utilizing logic models and health behavior theory, we recommend the use of the PER Worksheet. The PER Worksheet is a planning tool that provides layman-term prompts for identifying health behavior antecedents. It encompasses five common health behavior theories-health belief model, theory of planned behavior, social cognitive theory, an ecological perspective, and transtheoretical model. The PER Worksheet is organized into three columns: Predisposing, Enabling, and Reinforcing factors, terms of the PRECEDE/PROCEED model.     

姜黄素促进损伤骨骼肌修复

目的:研究姜黄素促进损伤骨骼肌修复的功能.方法:制作小鼠胫骨前肌实验性损伤模型,实验小鼠分别肌注和口服姜黄素,以组织学指标观察姜黄素体内促进损伤骨骼肌的修复功能.采用MTT和流析细胞分析技术观察姜黄素对骨骼肌成肌细胞的作用及可能的信号传导通路.结果:实验的小鼠肌肉伤口区新生肌纤维显著多于对照组.姜黄素无明显促C2C12细胞增殖,但能促进C2C12细胞融合而分化成肌纤维;姜黄素能抑制C2C12细胞NFκB反应因子的转活,而对c-Myc和CRE通路反应因子没有显著影响.大剂量使用姜黄素未见急性毒性反应.结论:姜黄素能通过促进骨骼肌干细胞的分化而促进损伤骨骼肌的修复.     

Differences in exposure assignment between conception and delivery: the impact of maternal mobility

Lupo PJ, Symanski E, Chan W, Mitchell LE, Waller DK, Canfield MA, Langlois PH. Differences in exposure assignment between conception and delivery: the impact of maternal mobility. Paediatric and Perinatal Epidemiology 2010; 24: 200–208. In studies of reproductive outcomes, maternal residence at delivery is often the only information available to characterise environmental exposures during pregnancy. The goal of this investigation was to describe residential mobility during pregnancy and to assess the extent to which change of residence may result in exposure misclassification when exposure is based on the address at delivery. Maternal residential mobility was compared between neural tube defect cases and unaffected controls from Texas participants in the National Birth Defects Prevention Study (NBDPS). Maternal residential information was obtained from the NBDPS interview. Data from the U.S. EPA National Air Toxics Assessment [Assessment System for Population Exposure Nationwide (ASPEN)], modelled at the census tract level, were used to estimate benzene exposure based on address at conception and address at delivery. Quartiles of exposure were assigned based on these estimates and the quartile assignments based on address at conception and address at delivery were compared using traditional methods (kappa statistics) and a novel application of mixed‐effects ordinal logistic regression. Overall, 30% of case mothers and 24% of control mothers moved during pregnancy. Differences in maternal residential mobility were not significant between cases and controls, other than case mothers who moved did so earlier during pregnancy than control mothers (P = 0.01). There was good agreement between quartiles of estimated benzene exposure at both addresses (kappa = 0.78, P < 0.0001). Based on the mixed‐effects regression model, address at delivery was not significantly different from using address at conception when assigning quartile of benzene exposure based on estimates from ASPEN (odds ratio 1.03, 95% confidence interval 0.85, 1.25). Our results indicate that, in this Texas population, maternal residential movement is generally within short distances, is typically not different between cases and controls, and does not significantly influence benzene exposure assessment.     

A case‐control study of airways obstruction among construction workers

Background

While smoking is the major cause of chronic obstructive pulmonary disease (COPD), occupational exposures to vapors, gases, dusts, and fumes (VGDF) increase COPD risk. This case‐control study estimated the risk of COPD attributable to occupational exposures among construction workers.

Methods

The study population included 834 cases and 1243 controls participating in a national medical screening program for older construction workers between 1997 and 2013. Qualitative exposure indices were developed based on lifetime work and exposure histories.

Results

Approximately 18% (95%CI = 2–24%) of COPD risk can be attributed to construction‐related exposures, which are additive to the risk contributed by smoking. A measure of all VGDF exposures combined was a strong predictor of COPD risk.

Conclusions

Construction workers are at increased risk of COPD as a result of broad and complex effects of many exposures acting independently or interactively. Control methods should be implemented to prevent worker exposures, and smoking cessation should be promoted. Am. J. Ind. Med. 58:1083–1097, 2015. © 2015 The Authors. American Journal of Industrial Medicine Published by Wiley Periodicals, Inc.     

Tocopherol Isomers in Intravenous Lipid Emulsions and Resultant Plasma Concentrations

Conflicting reports exist regarding the relative tocopherol isomer content of Intralipid ranging from 99% as α-tocopherol to as much as 90% as γ-tocopherol. Our direct assay of Intralipid as well as plasma levels measured in premature infants receiving Intralipid confirm the existence of a low α, high γ-tocopherol content and imply the need for α-tocopherol supplementation in patients receiving Intralipid, particularly the relatively tocopherol-deficient premature infant. Furthermore, the observation of abnormal erythrocyte hemolysis test values despite “normal” total tocopherol plasma concentrations may be explained by high plasma levels of non-α, biologically less active isomers. The quantitation of tocopherol isomers helps explain this discrepancy and suggests the need for future studies of vitamin E status to employ measurements of tocopherol isomers in reporting results. (Journal of Parenteral and Enteral Nutrition 8: 269–273, 1984)