鄂西苗族氨基糖甙类抗生素致聋患者的mtDNAl555位点突变研究

目的 为探讨遗传因素在鄂西苗族氨基糖甙类抗生素致聋(AAID)发生中的作用，从分子水平研究该病的发病机制。方法 对鄂西苗族部分家系及医学散发病例通过问卷调查和进行听力测试，确诊为AAID的82名患者。采外周血作PCR—RFLP分析，对照组为正常苗族50名。结果 82名患者中27例具有mtDNAl555^A→C。异质性突变，10例为均质性突变．该位点突变率为45．1％。其中异质性突变占72．9％，而对照组均无此突变。结论 mtDNAl555^A→C变是鄂西苗族个体对氨基糖甙类抗生素易感致聋的分子基础，异质性突变在苗族AAID中发生率较高。     

七个非综合征型耳聋家系患者的mtDNA A1555G突变性质与特点

目的 探讨非综合征型耳聋家系患者mtDNA A1555G突变性质及其特点,探索临床表型多样性的分子遗传学基础.方法 应用聚合酶链反应-限制性片段长度多态和实时荧光-扩增阻碍突变系统-定量PCR(real time-amplification refractory mutation system-quantitative PCR,RT-ARMS-qPCR)检测7个非综合征型耳聋家系71个成员的mtDNA A1555G突变,并收集、分析其临床资料.结果 7个家系中所有受检的母系成员mtDNA A1555G突变均为阳性,突变性质含同质性和异质性两种;非母系成员及配偶该突变为阴性.7个家系mtDNA A1555G同质性突变的拷贝数与耳聋轻重程度相关(R=0.341,P=0.022);mtDNA A1555G异质性突变的拷贝数与耳聋轻重程度相关(R=0.85,P=0.015).结论 mtDNA A1555G突变可导致非综合征型耳聋和氨基糖甙类抗生素致聋,其突变性质含同质性和异质性两种,且含mtDNA A1555G位点的突变型与野生型的比例与耳聋的严重程度密切相关.     

线粒体基因组A1555G突变在中国非综合征性聋患者中的流行病学分析

目的检测线粒体基因组12SrRNA、基因A1555G突变在中国非综合征性聋患者中的携带频率,探讨中国非综合征性聋的分子病因的流行病学意义。方法提取中国人群中22例氨基糖甙类药物致聋患者、158例散发的非综合征性聋患者以及60例非综合征性聋家系先证者的DNA,以聚合酶链反应结合限制性内切酶酶解分析法检测线粒体基因组A1555G突变的发生情况。结果线粒体基因组A1555G阳性患者占所有耳聋患者的4.2%,散发病例组中A1555G阳性率为1.3%,非综合征性聋家系组中A1555G阳性率为13.3%,22例氨基糖甙类药物致聋患者中未发现A1555G突变。结论线粒体基因组A1555G的突变发生率略高于以往报道,是非综合征性聋家系中致聋的主要病因之一,这对于中国人群耳聋的病因学研究有积极意义。     

线粒体DNA突变与氨基糖甙类抗生素致聋的关系

目的探讨线粒体ＤＮＡ突变与氨基糖甙类抗生素致聋的关系。方法应用ＰＣＲ－ＢｓｍＡＩ酶切及ＰＣＲ产物直接银染测序,对４８例氨基糖甙类抗生素致聋散发患者外周血标本的线粒体ＤＮＡ进行分析。结果发现６例患者线粒体ＤＮＡ１２ＳｒＲＮＡ基因核苷酸第１５５５位点发生了Ａ→Ｇ突变。结论氨基糖甙类抗生素致聋有明显的个体差异,带有１５５５Ｇ的个体对氨基糖甙类抗生素的耳毒作用有高度易感性。     

中国人非综合征性耳聋患者线粒体基因组C1494T突变筛查

目的进行非综合征性聋患者的线粒体DNA C1494T突变的分子流行病学调查。方法对中国人群中20例氨基糖甙类药物致聋患者、136例散发的非综合征性聋患者以及50例非综合征性聋家系先证者，以聚合酶链反应结合限制性片段长度多态性分析法检测线粒体DNA C1494T突变的发生情况。结果全部受检者无线粒体DNA C1494T突变的发生。结论中国人群中非综合征性聋患者的线粒体DNA C1494T突变的发生率较低，且明显低于线粒体DNA A1555G的突变发生率。通过聋病分子流行病学调查，提示线粒体DNA C1494T突变不是氨基糖甙类药物致聋的主要病因。     

修饰因子对线粒体DNA突变致聋的影响

线粒体DNA突变是引起感音神经性耳聋的重要原因之一,这些突变主要位于线粒体12SrRNA和tRNA基因上.其中12S rRNA基因上的同质性A1555G和C1494T突变与氨基糖甙类抗生素造成的耳聋相关.携带这两个突变的个体对耳毒性药物高度敏感,导致临床上常见的"一针致聋"现象.但携带A1555G或C1494T突变的个体在没用药的情况下也能产生非综合征型耳聋,而且同一家系内和不同家系间的母系成员在听力损失程度、发病年龄及听力曲线上存在很大差异.这些数据表明A1555G或C1494T突变是导致非综合征型耳聋发生的首要因子,其他修饰因子包括氨基糖甙类抗生素、线粒体DNA单倍型和核修饰基因等,在线粒体12S rRNA A1555G或C1494T突变相关的耳聋表型表达上起协同作用.作者简要介绍了这些因素对线粒体DNA突变致聋的影响以及母系遗传性耳聋发生的可能致病机制.     

修饰因子对线粒体DNA突变致聋的影响

线粒体DNA突变是引起感音神经性耳聋的重要原因之一,这些突变主要位于线粒体12SrRNA和tRNA基因上.其中12S rRNA基因上的同质性A1555G和C1494T突变与氨基糖甙类抗生素造成的耳聋相关.携带这两个突变的个体对耳毒性药物高度敏感,导致临床上常见的"一针致聋"现象.但携带A1555G或C1494T突变的个体在没用药的情况下也能产生非综合征型耳聋,而且同一家系内和不同家系间的母系成员在听力损失程度、发病年龄及听力曲线上存在很大差异.这些数据表明A1555G或C1494T突变是导致非综合征型耳聋发生的首要因子,其他修饰因子包括氨基糖甙类抗生素、线粒体DNA单倍型和核修饰基因等,在线粒体12S rRNA A1555G或C1494T突变相关的耳聋表型表达上起协同作用.作者简要介绍了这些因素对线粒体DNA突变致聋的影响以及母系遗传性耳聋发生的可能致病机制.     

线粒体DNA A1555G突变在新生儿大规模筛查的临床意义

目的 探讨在新生儿中进行线粒体DNA(mitochondria DNA,mtDNA)A1555G突变基因大规模筛查在预防药物性耳聋的必要性.方法 随机取2008年在深圳市出生的1000名新生儿的血滤纸标本,用Chelex-100树脂提取DNA,PCR扩增,变性高效液相色谱法(denaturing hig-performance liquid chromatography,DHPLC)进行mtDNA A1555G突变基因筛查,计算出阳性突变频率.结果 1000名新生儿血滤纸样本中,共检测出2例样本存在mtDNA A1555G突变,突变率为0.2%.结论 mtDNA A1555G突变在新生儿中出现的频率较高,对其进行mtDNA A1555G突变大规模筛查发现氨基甙类抗生素敏感个体,能有效地对新生儿及其家族高危人群进行合理性指导用药,从而更好地预防药物性耳聋.     

7个非综合征型耳聋家系患者的mtDNA A1555G突变分析

目的探讨非综合征型耳聋家系患者mtDNA A1555G突变及其临床特征。方法应用聚合酶链反应、限制性核酸内切酶酶切和DNA测序技术对7个非综合征型耳聋家系112个成员的mtDNA A1555G突变进行检测，并分析听力临床资料。结果7个家系中所有受检的母系成员mtDNA A1555G突变均为阳性，突变性质含同质性和异质性二种；非母系成员及配偶该突变为阴性。突变的性质与临床表型的有关。结论mtDNA A1555G突变可导致非综合征型耳聋和氨基糖苷类抗生素致聋，其突变性质含均质性和异质性两种，且与临床表型相关。     

氨基糖甙类抗生素耳毒性相关的线粒体DNA突变

线粒体12S rRNA基因是引起氨基糖甙类抗生素耳毒性和非综合征型耳聋的突变热点区域。其中,位于高度保守的12S rRNA基因解码区的同质性1555A＞G和1494C＞T突变,可导致部分患者对氨基糖甙类抗生素超敏感。当发生1555A＞G或1494C＞T突变时,12S rRNA高度保守的A位就会形成新的1494C-G1555或1494U-A1555碱基配对,使得人类线粒体核糖体的结构与细菌核糖体更加相似,以致氨基糖甙类抗生素与12S rRNA的结合更加容易,从而解释了为何携带这些突变的个体在使用了氨基糖甙类抗生素后会出现或加重耳聋表型。相关功能研究表明,无论是否存在氨基糖甙类抗生素的作用,携带1555A＞G或1494C＞T突变的细胞均会出现线粒体蛋白合成缺陷,并随之引发细胞呼吸功能障碍。此外,携带这些突变的家系,其母系成员听力损失程度、发病年龄和外显率均存在较大差异,提示核修饰基因、线粒体单体型以及氨基糖甙类抗生素等对12S rRNA 1555A＞G和1494C＞T突变的表型表达起着修饰作用。这些研究成果为以下三个方面提供了科学依据：①预测个体耳毒性风险;②提高氨基糖甙类抗生素治疗的安全性;③降低耳聋发生率。     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

The dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for $\text{primates}$. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.