Reversal of methylmercury-induced block of nerve-evoked release of acetylcholine at the neuromuscular junction

Acute bath application of micromolar concentrations of methylmercury (MeHg) blocks the nerve-evoked release of acetylcholine at the neuromuscular junction by presynaptic effects. The goal of the present study was to try to reverse this block of stimulus-evoked release. Experiments were conducted using the phrenic nerve hemidiaphragm of the rat and conventional intracellular microelectrode techniques. Myofibers were cut ("cut muscle") to prevent contractions elicited by stimulation of the phrenic nerve. End-plate potentials (EPPs) were recorded before and during MeHg application and during subsequent reversal attempts. MeHg (100 microM) blocked the EPP within 8-9 min of application. The time to block did not differ if Sr2+ (2 mM) was substituted for Ca2+ prior to exposure to MeHg. Washing the preparation with MeHg-free physiological saline at the time the EPP was blocked failed to reverse the block of synaptic transmission even during protracted washing. Increasing the extracellular [Ca2+] from 2 to 4 mM, or application of 4-aminopyridine (50 or 100 microM) failed to reverse block of the EPP. D-Penicillamine was also ineffective at reversing transmission block when applied at 0.4 mM; however, when applied at 1 mM D-penicillamine caused a return of EPPs in three of eight experiments within 5-20 min of wash. Longer periods of washing with D-penicillamine or use of higher concentrations of D-penicillamine were not effective in reversing transmission block in the refractory preparations. Increasing the intensity or duration of stimulation at the time of EPP block was uniformly successful in reversing MeHg-induced block; in 9 of the 10 preparations tested, EPPs could again be elicited from MeHg-blocked preparations merely by increasing the intensity and/or duration of stimulation, despite the continued presence of MeHg. Following reversal of transmission block by MeHg, continued exposure to MeHg resulted in a subsequent block of the EPP within approximately 3 min. During this subsequent block of the EPP by MeHg, increasing extracellular [Ca2+] or adding 4-aminopyridine did restore synaptic transmission. These results indicate that a temporary reversal of MeHg-induced block of synaptic transmission can be produced and that this effect does not require extracellular Ca2+ during the initial stages, but does seem to require extracellular Ca2+ during later stages.     

定量组织速度成像技术评价阿霉素诱导兔心肌病模型：与常规经胸超声心动图比较

目的:采用定量组织速度成像技术评价阿霉素诱导兔心肌病模型,并与常规经胸超声心动图比较其评估优势。方法:实验于2005-06/2006-08在大连医科大学完成。①实验分组及处理:取纯种新西兰白兔22只,雌雄不限,随机分成阿霉素组12只,给予阿霉素每次2mg/kg,以1g/L耳缘静脉注射,每周1次,注射8周;对照组10只每周注射2mL/kg生理盐水,共8周。②实验评估:每周应用HPSonos5500型彩色多普勒超声诊断仪(美国Agilent公司生产)对两组兔心脏进行左室收缩末期和舒张末期内径明、室间隔厚度、E峰、射血分数、左室短轴缩短率等常规超声参数测量,使用GEVivid7型彩色多普勒超声诊断仪(美国GE公司生产)进行收缩期和舒张期峰值速度、收缩期加速度定量组织速度成像参数测定。结果:22只兔进入统计。①对照组1～12周各参数与阿霉素组基础状态下比较无明显差异(P>0.05)。②第4周阿霉素组二尖瓣环运动的平均收缩期和舒张期峰值速度、收缩期加速度较基础状态明显减低(P均<0.05)。③第7周阿霉素组二尖瓣环运动的收缩期和舒张期峰值速度、收缩期加速度较基础状态明显减低(P<0.01),E峰较基础状态明显减低(P<0.05)。④第8周阿霉素组二尖瓣环运动的收缩期和舒张期峰值速度、收缩期加速度较基础状态明显减低(P<0.01),E峰较基础状态明显减低(P<0.01),左室收缩末期和舒张末期内径明显增大(P<0.05)。⑤第12周阿霉素组二尖瓣环运动的收缩期和舒张期峰值速度、收缩期加速度较基础状态明显减低(P<0.01),左室收缩末期和舒张末期内径明显增大(P<0.01),室间隔厚度、左室后壁厚度明显变薄(P<0.05),射血分数、左室短轴缩短率和E峰明显减低(P<0.01)。结论:定量组织速度成像参数可有效评价阿霉素诱导心肌病模型兔心肌的病理变化,较常规超声参数更敏感。     

Ciliary Muscle Dimension Changes With Accommodation Vary in Myopia and Emmetropia

PurposeThe purpose of this study was to determine whether accommodation-induced changes in ciliary muscle dimensions vary between emmetropes and myopes, and the effect of the image analysis method.MethodsSeventy adults aged 18 to 27 years consisted of 25 people with emmetropia (spherical equivalent refraction [SER] +0.21 ± 0.36 diopters [D]) and 45 people with myopia (−2.84 ± 1.72 D). There were 23 people with low myopia (>−3 D) and 22 people with moderate myopia (−3 to −6 D). Right eye ciliary muscles were imaged (Visante OCT; Carl Zeiss Meditec) at 0 D and 6 D demands. Measures included ciliary muscle length (CML), ciliary muscle curved length (CMLarc), maximum ciliary muscle thickness (CMTmax), CMT1, CMT2, and CMT3 (fixed distances 1–3 mm from the scleral spur), CM25, CM50, and CM75 (proportional distances 25%–75%). Linear mixed model analysis determined effects of refractive groups, race, and demand on dimensions. Significance was set at P < 0.05.ResultsMyopic eyes had greater CML and CMLarc nasally than emmetropic eyes. Myopic eyes had thicker muscles than emmetropic eyes at nasal positions, except CM25 and CMT3, and at CM75 temporally. During accommodation and only nasally, CML reduced in emmetropic and myopic eyes, and CMLarc reduced in myopic eyes only. During accommodation, both nasally and temporally, muscles thickened anteriorly (CMT1 and CM25) and thinned posteriorly (CMT3 and CM75) except for temporal CM75. Moderate myopic eyes had greater temporal CMLarc than low myopic eyes, and the moderate myopes had thicker muscles both nasally and temporally using fixed and proportional distances.ConclusionsPeople with myopia had longer and thicker ciliary muscles than people with emmetropia. During accommodation, the anterior muscle thickened and the curved nasal muscle length shortened, more in myopic than in emmetropic eyes. The fixed distance method is recommended for repeat measures in the same individual. The proportional distance method is recommended for comparisons between refractive groups.     

Interpreting DNA Evidence in Paternity Cases

The ability of jurors and juries to comprehend and utilise scientific evidence in Australian criminal trials is discussed in the context of impediments which include legislation, proce-dural law, and the inherent scientific complexity of some evidence. Existing Australian and international literature on jury performance with expert evidence is examined, and recent doctoral research utilising mock juries, real jurors and forensic scientists, is highlighted.

“Good communication with the jury is afield in which anecdote, self-assurance and self-delusion abound, within the ranks of the legal profession and the judiciary.”     

Drug enterocyte adducts: possible causal factor for diclofenac enteropathy in rats

BACKGROUND & AIMS: Enteropathy is a frequent complication of diclofenac and other nonsteroidal anti-inflammatory drugs, yet little is known about the underlying mechanism. One possibility is that reactive metabolites of diclofenac form adducts with enterocyte macromolecules, as previously shown for liver. We addressed this possibility by using immunohistochemistry to detect diclofenac adducts. METHODS: Rats were treated orally with diclofenac (10-100 mg/kg) and killed after 1-24 hours, and their gastrointestinal (GI) tracts were evaluated for ulcer number and area. Adduct distribution and intensity were assessed by immunohistochemistry by using a technique to simultaneously process and stain multiple intestinal rings. RESULTS: Drug treatment led to dose-dependent formation of both adducts and ulcers only in small intestine and only in animals with intact enterohepatic circulation. Adducts formed within enterocytes by 1 hour, translocated to the brush border, preceded ulceration and vascular protein leakage, and were intense at sites of ulceration. Adducts and ulcers exhibited a parallel distribution within intestinal quintiles: 3rd > 5th > 1st. CONCLUSIONS: Diclofenac treatment resulted in the formation of drug adducts in enterocytes. Because this molecular change occurred before ulceration, was dose dependent, and exhibited concordant distribution with extent of ulceration, the results suggest a causal role for drug adduct formation in diclofenac enteropathy.     

Small basic proteins of myelin from central and peripheral nervous systems are encoded by the same gene.

Peripheral nervous system (PNS) and central nervous system (CNS) rodent myelins, which are produced by different cell types, share common morphological and functional characteristics although their major integral membrane proteins are completely different. Both types of myelin however, contain sets of four myelin basic proteins (MBPs), which share similar immunochemical and electrophoretic properties. We have isolated and characterized cDNA clones corresponding to the rat mRNAs encoding the small MBPs (SMBPs) found in both CNS and PNS myelin. Sequence analysis of these clones indicate that SMBPs in both divisions of the nervous system are encoded by the same nucleotide sequences, which suggests that they are the products of the same gene expressed in both oligodendrocyte and Schwann cells. In dot-blot hybridization experiments with the CNS SMBP cDNA as a probe, it was shown that there is a 20-fold higher level of MBP mRNA in a CNS myelin fraction than in total brainstem mRNA. It also was found that in optic and sciatic nerves, which contain oligodendrocytes and Schwann cells respectively, there are higher levels (4-fold and 2-fold, respectively) of MBP mRNA than in brainstem. Blot-hybridization experiments showed that a probe derived from the coding region of the rat SMBP cDNA hybridizes to an homologous mRNA (approximately equal to 2.6 kilobases) present in human optic nerve, which is not detectable with a probe derived from the 3' untranslated region. This conservation of coding-region sequences is in accord with the highly homologous amino acid sequences reported for the MBPs in the two species.