血清UA、CysC、Lp-PLA2与急性脑梗死合并脑白质疏松症患者预后的关系研究

摘要 目的：探讨血清尿酸（UA）、胱抑素C（CysC）、脂蛋白相关磷脂酶 A2（Lp-PLA2）水平与急性脑梗死合并脑白质疏松症患者预后的关系。方法：选择2020年3月至2022年12月中国人民解放军联勤保障部队第九六0医院收治的113例急性脑梗死合并脑白质疏松症患者,检测血清UA、CysC、Lp-PLA2水平。随访1个月,根据改良Rankin量表（mRS）评分将患者分为预后良好组（0～2分,75例）和预后不良组（3分及以上,38例）。多因素Logistic回归分析急性脑梗死合并脑白质疏松症患者预后不良的危险因素,受试者工作特征曲线（ROC）分析血清UA、CysC、Lp-PLA2对急性脑梗死合并脑白质疏松症患者预后不良的预测价值。结果：预后不良组血清UA、CysC、Lp-PLA2水平高于预后良好组（P＜0.05）。多因素Logistic回归分析显示重度脑白质病变、高入院时NIHSS评分,高血清UA、CysC、Lp-PLA2水平是急性脑梗死合并脑白质疏松症患者预后不良的危险因素（P＜0.05）。联合血清UA、CysC、Lp-PLA2预测急性脑梗死合并脑白质疏松症患者预后的曲线下面积（AUC）为0.916,高于单独预测。结论：急性脑梗死合并脑白质疏松症患者血清UA、CysC、Lp-PLA2水平增高且与预后不良有关,联合血清UA、CysC、Lp-PLA2预测急性脑梗死合并脑白质疏松症患者预后不良价值较高。     

心脏彩超检查联合血清BNP、ALB、CysC在慢性心力衰竭患者预后评估中的临床价值分析

摘要 目的：分析心脏彩超检查联合血清B型钠尿肽（BNP）、白蛋白（ALB）、胱抑素C（CysC）在慢性心力衰竭（CHF）患者预后评估中的临床价值。方法：选取2017年6月-2018年5月我院收治的123例CHF患者,心脏彩超检查左心室射血分数（LVEF）、左心房内径（LAD）和左心室内径（LVD）,实验室检测血清BNP、ALB、CysC水平。按照3年随访后患者是否死亡分为死亡组35例和存活组88例,收集临床资料,采用多因素Logistic回归分析CHF患者预后的影响因素。采用受试者工作特征（ROC）曲线分析心脏彩超检查联合血清BNP、ALB、CysC对CHF患者预后的评估价值。结果：死亡组患者的LAD、LVD及血清BNP、CysC水平高于存活组患者,而LVEF及血清ALB水平低于存活组患者（P＜0.05）。心脏彩超指标LVEF、LAD、LVD及血清BNP、ALB、CysC是CHF患者预后的影响因素（P＜0.05）。心脏彩超指标LVEF、LAD、LVD联合血清BNP、ALB、CysC检测对CHF患者预后评估的ROC曲线下面积（AUC）（0.95CI）为0.857（0.771～0.938）,灵敏度及特异度分别为0.914（32/35）、0.795（70/88）,均明显高于上述各指标单独检测。结论：心脏彩超指标LVEF、LAD、LVD和血清BNP、ALB、CysC均为CHF患者预后的影响因素,且联合检测对患者预后的评估价值较高,具有一定的临床应用价值。     

逍遥散对慢性心力衰竭合并抑郁患者心功能、炎症介质和血清5-HT、NE、CORT的影响

摘要 目的：探讨逍遥散对慢性心力衰竭（CHF）合并抑郁患者心功能、炎症介质和血清5-羟色胺（5-HT）、去甲肾上腺素（NE）、皮质醇（CORT）的影响。方法：选择我院于2017年10月至2018年12月收治的94例CHF合并抑郁患者,随机分为对照组、观察组,每组47例,对照组给予常规CHF治疗及盐酸帕罗西汀治疗,研究组给予常规CHF治疗及逍遥散治疗,比较两组疗效、心功能、白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、5-HT、NE、CORT、明尼苏达心衰生活质量调查表（MLHFQ）、汉密尔顿抑郁量表（HAMD）及不良反应。结果：观察组治疗8周后的CHF总有效率、抑郁总有效率为82.98%、87.23%,高于对照组的63.83%、61.70%（P<0.05）。治疗8周后,两组MLHFQ、HAMD评分均较治疗前下降,且观察组低于对照组（P<0.05）。与治疗前相比,治疗8周后,两组CORT、IL-6、TNF-α下降,且观察组低于对照组（P<0.05）, 5-HT、NE升高,且观察组高于对照组（P<0.05）。两组与治疗前相比,治疗8周后,两组LVSD、LVED下降,且观察组低于对照组（P<0.05）,LVEF升高,且观察组高于对照组（P<0.05）。治疗期间,两组未出现严重不良反应。结论：逍遥散治疗CHF合并抑郁患者,疗效肯定,可有效改善患者心功能,减轻炎性反应,并调节血清5-HT、NE、CORT水平。     

急性心肌梗死患者新发心房颤动的危险因素及血清氨基末端脑钠尿肽前体、尿酸的预测价值分析

摘要 目的：探讨影响急性心肌梗死患者新发心房颤动的危险因素以及血清氨基末端脑钠尿肽前体（NT-proBNP）、尿酸（UA）对急性心肌梗死患者新发心房颤动的预测价值。方法：选择2019年10月至2021年5月在徐州医科大学附属医院接受诊治的急性心肌梗死患者110例,根据患者住院期间是否新发心房颤动分为房颤组（患者住院期间新发心房颤动,n=30）和无房颤组（患者住院期间未新发心房颤动,n=80）。另选择50例同期在徐州医科大学附属医院体检的健康者作为健康对照组,比较房颤组、无房颤组、健康对照组三组研究对象血清NT-proBNP、UA水平差异。收集患者各项临床资料,多因素Logistic回归分析影响急性心肌梗死患者新发心房颤动的危险因素。采用受试者工作特征（ROC）曲线分析血清NT-proBNP、UA单独以及联合检测对急性心肌梗死患者新发心房颤动的预测价值。结果：房颤组患者血清NT-proBNP、UA水平均明显高于健康对照组和无房颤组,且无房颤组高于健康对照组（P<0.05）。多因素Logistic回归分析显示,高NT-proBNP以及UA水平、高龄、左房增大、合并糖尿病、Killip心功能分级≥Ⅱ级是影响急性心肌梗死患者新发心房颤动的危险因素（P<0.05）。血清UA、NT-proBNP单独及联合预测急性心肌梗死患者新发心房颤动的曲线下面积（AUC）分别为0.730、0.737、0.840。结论：血清NT-proBNP、UA水平对急性心肌梗死患者新发心房颤动的发生具有一定预测价值,且两者联合应用的预测价值最高,除高NT-proBNP以及UA水平外,高龄、左房增大、合并糖尿病、Killip心功能分级≥Ⅱ级亦是新发心房颤动的危险因素。     

血清CysC、hs-CRP与急性脑梗死分型及并发医院感染的相关性探究

摘要 目的：探究血清胱抑素C（CysC）、超敏C反应蛋白（hs-CRP）与急性脑梗死TOAST分型及并发医院感染的相关性。方法：以2017年9月~2022年9月84例急性脑梗死患者（脑梗死组）及57例健康体检者（对照组）为研究对象，采用免疫比浊法检测受试者血清CysC水平，采用乳胶增强免疫比浊法检测其hs-CRP水平。分析血清CysC、hs-CRP水平与急性脑梗死TOAST分型及并发医院感染的关系。结果：本研究纳入病例中无不明原因性缺血性卒中（SUE）患者，主要包括大动脉粥样硬化型（LAA）患者37例、小动脉闭塞型（SAO）患者12例、心源性栓塞型（CE）患者31例、其他病因明确型缺血性卒中（SOE）患者4例。不同TOAST分型患者的血清CysC、hs-CRP水平对比，存在显著性差异（P<0.05）；CE组血清CysC、hs-CRP水平高于LAA组、SAO组、SOE组，LAA组血清CysC、hs-CRP水平高于SAO组、SOE组，SAO组血清CysC、hs-CRP水平高于SOE组（P<0.05）。根据NIHSS评分将脑梗死患者分为重度组18例、中度组29例、轻度组37例，不同严重程度脑梗死患者的血清CysC、hs-CRP水平均高于对照组，且重度组高于中度组、轻度组（P<0.05）；中度组的血清CysC水平高于轻度组（P<0.05）。血清CysC、hs-CRP水平与脑梗死患者NIHSS评分均呈正相关（P<0.05）。84例脑梗死患者伴发医院感染28例，其中上呼吸道感染13例、肺部感染8例、泌尿系统感染5例、其他2例。感染组中合并糖尿病、有侵入性操作、TOAST分型为CE型的人数比例高于未感染组，NIHSS评分及血清CysC、hs-CRP水平高于未感染组（P<0.05）。侵入性操作、NIHSS评分≥16分、TOAST分型为CE型及CysC、hs-CRP水平升高是急性脑梗死并发感染的危险因素（P<0.05）。结论：CE型急性脑梗死患者血清CysC、hs-CRP水平显著升高，与病情严重程度相关，且侵入性操作、NIHSS评分≥16分、TOAST分型为CE型及CysC、hs-CRP水平升高是急性脑梗死并发感染的危险因素。     

2型糖尿病患者血清尿酸、胱抑素C、半乳糖凝集素-3、丝氨酸蛋白酶抑制剂B1与血糖及颈动脉粥样硬化的关系研究

摘要 目的：研究2型糖尿病（T2DM）患者血清尿酸（UA）、胱抑素C（CysC）、半乳糖凝集素-3（Gal-3）、丝氨酸蛋白酶抑制剂B1（Serpin B1）与血糖及颈动脉粥样硬化（CAS）的关系。方法：选取2020年4月-2022年4月青岛市中医医院内分泌科收治的110例T2DM患者,根据有无CAS分为CAS组36例和非CAS组74例,检测并对比两组血清UA、CysC、Gal-3、Serpin B1水平。通过Pearson/Spearman相关系数分析T2DM患者血清UA、CysC、Gal3、Serpin B1水平与血糖指标的相关性。收集患者基础资料,采用多因素Logistic回归分析T2DM患者发生CAS的影响因素。结果：CAS组血清UA、CysC、Gal-3、Serpin B1水平均明显高于非CAS组（P＜0.05）。CAS组平均年龄大于非CAS组,吸烟史患者比例、空腹血糖、糖化血红蛋白（HbA1c）、胰岛素抵抗指数（HOMA-IR）、低密度脂蛋白胆固醇（LDL-C）水平高于非CAS组（P＜0.05）。相关性分析显示,T2DM患者血清UA、CysC、Gal-3、Serpin B1水平与HbA1c、HOMA-IR呈正相关（P＜0.05）。多因素Logistic回归分析显示,年龄较大、HbA1c、HOMA-IR、血清UA、CysC、Gal3、Serpin B1水平较高是T2DM患者发生CAS的危险因素（P＜0.05）。结论：血清UA、CysC、Gal-3、Serpin B1水平升高与T2DM患者血糖水平有关,同时也是T2DM患者发生CAS的影响因素。     

不同类型慢性心力衰竭患者临床特征及心功能危险因素、预后影响因素分析

摘要 目的：分析不同类型慢性心力衰竭患者临床特征及心功能危险因素、预后影响因素。方法：回顾性分析2020年1月-2022年1月我院收治的慢性心力衰竭患者80例，根据左室射血分数（LVEF）分为A组（n=25，LVEF<30%）、B组（n=25，LVEF 40%~50%）、C组（n=25，LVEF≥50%）三组，另根据随访1年后是否存活分为生存组（n=51）和死亡组（n=29）。比较不同组别临床相关指标，采用Pearson检验分析患者临床特征与慢性心力衰竭患者心功能、预后之间的相关性，采用多因素Logistic回归分析影响慢性心力衰竭患者心功能、预后的独立危险因素。结果：A组的心率及患有冠心病、心律失常1年以上、合并非心血管疾病人数占比、LAD、RAD、Scr、Hcy水平高于B组和C组（P<0.05）。死亡组的心率及患有冠心病、心律失常1年以上、LAD、RAD、Scr水平明显高于生存组（P<0.05）。Pearson相关性检验显示，心率、冠心病、心律失常、合并非心血管疾病、LAD、RAD、Scr、Hcy水平与慢性心力衰竭患者心功能之间呈正相关（P<0.05）；心率、冠心病、心律失常、LAD、RAD、Scr水平与慢性心力衰竭患者预后之间呈正相关（P<0.05）。多因素Logistic回归分析结果显示，心率、冠心病、心律失常、合并非心血管疾病、LAD、RAD、Scr、Hcy水平是影响慢性心力衰竭患者心功能的独立危险因素（P<0.05）；心率、冠心病、心律失常、LAD、RAD、Scr水平是影响慢性心力衰竭患者预后的独立危险因素（P<0.05）。结论：心率、冠心病、心律失常、LAD、RAD、Scr水平与慢性心力衰竭患者心功能、预后之间均呈正相关，是影响慢性心力衰竭患者心功能、预后的独立危险因素，可用来预测慢性心力衰竭的发生。     

甘油三酯葡萄糖乘积指数、红细胞分布宽度、血清尿酸、降钙素原与冠心病患者冠状动脉狭窄程度和短期预后的关系分析

摘要 目的：探讨甘油三酯葡萄糖乘积（TyG）指数、红细胞分布宽度（RDW）、血清尿酸（UA）、降钙素原（PCT）与冠心病患者冠状动脉狭窄程度和短期预后的关系。方法：选取2017年1月至2021年12月我院收治的150例冠心病患者,均行冠状动脉造影并以Gensini积分系统评估冠状动脉狭窄程度,分为轻度组、中度组和重度组,比较各组的TyG指数、RDW、UA、PCT水平,以Pearson相关系数分析上述指标与Gensini积分的相关性。患者出院后均随访6个月,根据有无发生主要心血管不良事件（MACE）分为预后良好组和预后不良组,以多因素Logisitc回归分析预后的影响因素,以受试者工作特征（ROC）曲线分析对预后的预测效能。结果：不同冠状动脉狭窄程度患者的TyG指数、RDW、UA、PCT水平差异均表现为,从轻度到重度依次升高（P＜0.05）。Pearson相关性分析显示,冠心病患者的TyG指数、RDW、UA、PCT水平与Gensini积分均呈正相关（P＜0.05）。预后不良组的甘油三酯（TG）、Gensini积分、TyG指数、RDW、UA、PCT水平均高于预后良好组（P＜0.05）;多因素Logistic回归分析显示,TG≥3.5 mmol/L、Gensini积分≥30分、TyG指数≥5、RDW≥12.8%、UA≥380 μmol/L、PCT≥35 μg/L是冠心病患者预后不良的危险因素（P＜0.05）。ROC曲线分析显示,TyG指数、RDW、UA、PCT单独及联合预测冠心病患者预后不良的曲线下面积分别为0.786、0.793、0.794、0.789和0.948,联合预测效能明显更高。结论：冠心病患者的TyG指数、RDW、血UA、PCT水平与冠状动脉狭窄程度呈正相关,其均为患者短期预后的影响因素,联合检测上述指标对患者短期预后有一定预测价值。     

血清Copeptin、suPAR、SIRT1与慢性心力衰竭患者心功能及短期预后的关系研究

摘要 目的：研究血清和肽素（Copeptin）、可溶性尿激酶型纤溶酶原激活物受体（suPAR）、沉默信息调节因子2相关酶1（SIRT1）与慢性心力衰竭（CHF）患者心功能及短期预后的关系。方法：选取我院2018年2月～2020年2月收治的175例CHF患者,将其按照美国纽约心功能（NYHA）分级的差异分作Ⅱ级组56例,Ⅲ级组62例,Ⅳ级组57例。另取我院同期健康体检人员50例作为对照组。比较各组血清Copeptin、suPAR、SIRT1水平及心功能指标,并进行相关性分析。此外,对所有CHF患者进行为期1年的随访,按照预后差异分作预后不良组61例及预后良好组114例。以单因素、多因素Logistic回归分析CHF患者预后的影响因素。结果：与对照组相比,CHF患者的血清Copeptin、suPAR水平及左心室舒张末期内径（LVEDD）升高,而SIRT1水平及左心室射血分数（LVEF）降低（P＜0.05）;不同心功能等级的CHF患者间比较,随着心衰程度的加重,血清Copeptin、suPAR水平及LVEDD升高,而SIRT1水平及LVEF降低（P＜0.05）。Pearson相关性分析结果显示,CHF患者血清Copeptin、suPAR水平与LVEDD均呈正相关,而与LVEF呈负相关（P＜0.05）;血清SIRT1水平与LVEDD呈负相关,而与LVEF呈正相关（P＜0.05）。单因素分析结果显示,预后不良组年龄、LVEDD、血清Copeptin、suPAR水平均高于预后良好组,而LVEF和SIRT1水平低于预后良好组（P＜0.05）。多因素Logistic回归分析结果显示,年龄偏大、LVEDD偏高、LVEF偏低、血清Copeptin、suPAR水平过高以及血清SIRT1水平过低均是CHF患者短期预后不良的危险因素（P＜0.05）。结论：血清Copeptin、suPAR、SIRT1与CHF患者的心功能及短期预后密切相关。     

基于Tilburg衰弱评估量表评价的老年慢性心力衰竭合并衰弱的危险因素探讨及其对患者生活质量和预后的影响

摘要 目的：基于Tilburg衰弱评估量表评价老年慢性心力衰竭（CHF）的衰弱状况，分析影响衰弱的危险因素，并探讨其对患者生活质量和预后的影响。方法：选取2020年3月～2022年3月我院收治的102例老年CHF患者，根据Tilburg衰弱评估量表分为衰弱组与非衰弱组。收集患者基线资料，采用明尼苏达州心力衰竭生活质量问卷（MLHFQ）评分评估患者的生活质量，采用多因素Logistic回归分析老年CHF患者合并衰弱的危险因素，Spearman相关性分析衰弱组Tilburg衰弱评估量表评分与MLHFQ评分的相关性，比较两组患者生活质量和90 d非计划再入院率、死亡率。结果：102例老年CHF患者合并衰弱的发生率为53.92％（55/102）。多因素Logistic回归分析显示：体质指数（BMI）降低、NYHA心功能分级Ⅲ～Ⅳ级、合并疾病数量增加、血红蛋白（Hb）降低、红细胞分布宽度（RDW）升高、白蛋白（Alb）降低、N末端B型脑钠肽前体（NT-ProBNP）升高为老年CHF患者合并衰弱的独立危险因素（P＜0.05）。衰弱组MLHFQ身体领域、情绪领域、其他领域评分高于非衰弱组（P＜0.05）。Spearman相关性分析显示，老年CHF患者合并衰弱患者Tilburg衰弱评估量表评分与MLHFQ身体领域、情绪领域、其他领域评分呈正相关（rs=0.505、0.424、0.526，P均＜0.001）。随访90 d，衰弱组非计划再入院率高于非衰弱组（P＜0.05）。结论：老年CHF患者衰弱状况发生率较高，BMI、NYHA心功能分级、合并疾病数量、Hb、RDW、Alb、NT-ProBNP为老年CHF患者合并衰弱的影响因素，衰弱可导致患者生活质量下降和预后不良。Tilburg衰弱评估量表能快速评价老年CHF患者合并衰弱状况，有助于指导临床及时采取干预措施改善患者生活质量和预后。     

Interactions of the DNA intercalator acridine orange, with itself, with caffeine, and with double stranded DNA

Caffeine (CAF) inhibits the intercalation of acridine orange (AO) into cellular DNA. Optical absorption and fluorescence spectroscopy were employed to determine the molecular interactions of AO with itself, with CAF, and with double stranded herring sperm DNA (dsDNA). AO dimerization was observed at concentrations >2 micromol. The sharp increase in fluorescence (lambda(em)=530 nm) at 5 micromol of AO was attributed to AO multimer formation. From 0.5 to 5.0 micromol, the AO self-association binding constant (K(assoc)) was determined to be 38620 mol(-1), however, the presence of 150 mmol NaCl increased K(assoc) to 118000 mol(-1) attributed to the charge neutralization. The K(assoc) for AO with CAF was confirmed to be 256 mol(-1). K(assoc) for the binding of AO with 20 micromol DNA ranged from, 32000 mol(-1) at 2 micromol AO, to approximately 3700 mol(-1) at 10 micromol AO, in the absence of NaCl. This AO concentration dependency of K(assoc) value with DNA was attributed to AO intercalation into dsDNA at high dsDNA/AO ratios, and electrostatic binding of AO to dsDNA at low AO ratios. The findings provide information used to explain fluorescence intensity values at lambda(em) at 530 nm from studies that combine AO, caffeine, and dsDNA.     

Interaction of tRNA with Safranal,Crocetin, and Dimethylcrocetin

Abstract

Saffron is the red dried stigmas of Crocus sativus L. flowers and used both as a spice and as a drug in traditional therapeutic. The biological activity of saffron in modern medicine is in development. Its numerous applications as an anti-oxidant and anti-cancer agent are due to its secondary metabolites and their derivatives (safranal, crocins, crocetin, dimethylcrocetin). The aim of this study was to examine the interaction of transfer RNA with safranal, crocetin, and dimethylcrocetin in aqueous solution at physiological conditions. Constant tRNA concentration (6.25 mM) and various drug/tRNA (phosphate) molar ratios of 1/48 to 1/8 were used. FT-IR and UV-Visible difference spectroscopic methods have been applied to determine the drug binding mode, the binding constants and the effects of drug complexation on the stability and conformation of tRNA duplex. External binding mode was observed for safranal crocetin and dimethylcrocetin, with overall binding constants K_safranal = 6.8 (± 0.34) × 10³ M^?1, K_CRT = 1.4 (± 0.31) × 10⁴ M^?1, and K_DMCRT = 3.4 (± 0.30) × 10⁴ M^?1. Transfer RNA remains in the A-family structure, upon safranal, crocetin and dimethylcrocetin complexation.     

Linkage relations of JK, CO, KEL and IGK with each other and with AHCY

Linkage analyses of locus pairs involving IGK, JK, CO, KEL and AHCY resulted in altogether negative lod scores, thereby dwindling the reported linkage relations.     

Carbonic anhydrase inhibitors. Inhibition of isozymes I, II, IV, V, and IX with anions isosteric and isoelectronic with sulfate, nitrate, and carbonate

The inhibition of five human carbonic anhydrase (hCA, EC 4.2.1.1) isozymes; the cytosolic hCA I and II, the membrane-bound hCA IV, the mitochondrial hCA V, and the tumor-associated, transmembrane hCA IX, with anions isosteric and isoelectronic with sulfate, nitrate, and carbonate; such as chlorate, perchlorate, bromate, iodate, periodate, silicate, bismuthate, vanadate, molybdate, and wolframate is reported. Apparently, the geometry of the inhibitor (tetrahedral or trigonal) does not influence its binding to the Zn(II) ion of the enzyme active site, but the nature of the central element is the most important factor influencing potency. Isozymes hCA I and II are best inhibited by chlorate, perchlorate, and silicate, together with the anions structurally related to sulfate, sulfamate, and sulfamidate, but sulfate itself is a weak inhibitor (inhibition constant of 74 mM against hCA I and 183 mM against hCA II). Molybdate is a very weak hCA I inhibitor (K(I) of 914 mM) but it interacts with hCA II (K(I) of 27.5mM). Isozyme IV is well inhibited by sulfate (K(I) of 9 mM), sulfamate, and sulfamidate (in the low micromolar range), but not by perchlorate (K(I) of 767 mM). The mitochondrial isozyme V has the lowest affinity for sulfate (K(I) of 680 mM) and carbonate (K(I) of 95 mM) among all the investigated isozymes, suggesting on one hand its possible participation in metabolon(s) with sulfate anion exchanger(s), and on the other hand an evolutionary adaptation to working at higher pH values (around 8.5 in mitochondria) where rather high amounts of carbonate in equilibrium with bicarbonate may be present. Metasilicate, isosteric to carbonate, is also about a 10 times weaker inhibitor of this isozyme as compared to other CAs investigated here (K(I) of 28.2 mM). Surprisingly, the tumor-associated isozyme IX is resistant to sulfate inhibition (K(I) of 154 mM) but has affinity in the low micromolar range for carbonate, sulfamate, and sulfamidate (K(I) in the range of 8.6-9.6 microM). This constitutes another proof that this isozyme best works at acidic pH values present in tumors, being inhibited substantially at higher pH values when more carbonate may be present. Bromate and chlorate are quite weak CA IX inhibitors (K(I) s of 147-274 mM).     

Kymographs,stimulators, and kymographs with stimulators

This biology investigation on Pristipomoides filamentosus larval development, survival, and aquaculture research was developed with three educational objectives: to provide high school students with (1) a scientific background on the biology and science of fisheries as well as overfishing, its consequences, and possible mitigations; (2) exposure to field and laboratory techniques in marine science; and (3) practical skills in scientific inquiry and investigation. We teach this investigation at the Hawai‘i Institute of Marine Biology, where we have access to captive broodstock of the pink snapper, P. filamentosus. During this investigation students follow several steps scientists take to study aquaculture: collecting spawn from outdoor fish pens, quantifying the number of eggs, determining the percentage of fertilisation, and estimating the time of spawning and hatching. Additionally, students perform hypothesis-driven science activities with the embryos to test the effects of water quality on their development and survival. In this paper we discuss background information of aquaculture, specifically of P. filamentosus, and thoroughly describe the several components of delivering the investigation. Lastly, we provide possible outcomes of students’ performances in the laboratory activities, and discuss how effectively the exercise met its educational objectives.     

A pheromonotropic peptide of Helicoverpa zea,with melanizing activity,interaction with PBAN,and distribution of immunoreactivity

The sequence of an 18-amino acid residue peptide was deduced from the gene encoding PBAN and other peptides with common C-termini in Helicoverpa zea. The peptide caused melanization in larvae and pheromone production in females of H. zea, and was designated pheromonotropic melanizing peptide (Hez-PMP). The peptide has a 83% sequence homology with a pheromonotropic peptide isolated from Pseudaletia separata. PMP caused melanization and mortality when injected into larvae just before molting. Whereas intense melanization was caused with a dose of 1,000 pmol, peak mortality occurred at 100 pmol, with 50% of larvae dying within 48 h after injection. Pheromonotropic activity of PMP was dose dependent. Co-injection of Hez-PMP and Hez-PBAN into a female resulted in suppression of the pheromonotropic effect of PBAN. Whole-mount immunocytochemical studies revealed PMP-like immunoreactivity in frontal ganglion, subesophageal, thoracic, and abdominal ganglia as well as the esophageal nerve.     

Interaction of tRNA with Safranal, Crocetin, and Dimethylcrocetin

Saffron is the red dried stigmas of Crocus sativus L. flowers and used both as a spice and as a drug in traditional therapeutic. The biological activity of saffron in modern medicine is in development. Its numerous applications as an anti-oxidant and anti-cancer agent are due to its secondary metabolites and their derivatives (safranal, crocins, crocetin, dimethylcrocetin). The aim of this study was to examine the interaction of transfer RNA with safranal, crocetin, and dimethylcrocetin in aqueous solution at physiological conditions. Constant tRNA concentration (6.25 mM) and various drug/tRNA (phosphate) molar ratios of 1/48 to 1/8 were used. FT-IR and UV-Visible difference spectroscopic methods have been applied to determine the drug binding mode, the binding constants and the effects of drug complexation on the stability and conformation of tRNA duplex. External binding mode was observed for safranal crocetin and dimethylcrocetin, with overall binding constants K(safranal) = 6.8 (+/- 0.34) x 10(3) M(-1), K(CRT) = 1.4 (+/- 0.31) x 10(4) M(-1), and K(DMCRT) = 3.4 (+/- 0.30) x 10(4) M(-1). Transfer RNA remains in the A-family structure, upon safranal, crocetin and dimethylcrocetin complexation.     

Titin-cap associates with,and regulates secretion of,Myostatin

Myostatin, a secreted growth factor, is a key negative regulator of skeletal muscle growth. To identify modifiers of Myostatin function, we screened for Myostatin interacting proteins. Using a yeast two-hybrid screen, we identified Titin-cap (T-cap) protein as interacting with Myostatin. T-cap is a sarcomeric protein that binds to the N-terminal domain of Titin and is a substrate of the titin kinase. Mammalian two-hybrid studies, in vitro binding assays and protein truncations in the yeast two-hybrid system verified the specific interaction between processed mature Myostatin and full-length T-cap. Analysis of protein-protein interaction using surface plasmon resonance (Biacore, Uppsala, Sweden) kinetics revealed a high affinity between Myostatin and T-cap with a KD of 40 nM. When T-cap was stably overexpressed in C(2)C(12) myoblasts, the rate of cell proliferation was significantly increased. Western analyses showed that production and processing of Myostatin were not altered in cells overexpressing T-cap, but an increase in the retention of mature Myostatin indicated that T-cap may block Myostatin secretion. Bioassay for Myostatin confirmed that conditioned media from myoblasts overexpressing T-cap contained lower levels of Myostatin. Given that Myostatin negatively regulates myoblast proliferation, the increase in proliferation observed in myoblasts overexpressing T-cap could thus be due to reduced Myostatin secretion. These results suggest that T-cap, by interacting with Myostatin, controls Myostatin secretion in myogenic precursor cells without affecting the processing step of precursor Myostatin.