Apoptosis in squamous cell carcinoma of the lung: correlation with survival and clinicopathological features

AIMS: Apoptosis is recognised as a physiological mechanism for controlling cell numbers and its subversion is thought to contribute to carcinogenesis. The aims of this study were to measure the apoptotic index (AI) in a series of squamous cell carcinomas (SCCs) of the lung using standard histological staining and confirm this by immunohistology using an antibody to an apoptosis specific protein (ASP), and to seek to correlate the AI with clinicopathological parameters. METHODS: Sections of 134 SCCs were stained by haematoxylin and eosin (H&E) for counting apoptotic bodies of determine the AI (number of apoptotic bodies/10,000 tumour cells); 26 of these were also stained with anti-ASP antibody and the proportion of ASP positive cells counted. Clinical data were obtained from hospital notes. RESULTS: The mean AI obtained by H&E staining of all 134 SCCs was 30.3 (SD, 24.75). Anti-ASP staining allowed easy identification of apoptotic bodies, and generated a somewhat higher index (mean, 51.4; SD 39); this was not a result of the selection of tumours because the AI by H&E in the subset stained with anti-ASP was 31.1. Regression analysis showed that the correlation between the two values of AI was highly significant (Rs = 0.9760; p < 0.001), indicating that the two methods were both reliable measures of apoptosis but that the anti-ASP staining is the more sensitive method. The tumours were grouped into high AI (> 50) and low AI (< 50) and survival analysis was carried out. The mean survival of the high AI group was 109 weeks and of the low AI group 72 weeks (p = 0.036). CONCLUSIONS: Anti-ASP staining is a reliable, easy, and sensitive method for assessing apoptosis in tumour sections and confirms the validity of the AI obtained by H&E staining. AI is a guide to the behaviour of SCCs of the lung.     

Comparative clinico-morphological characteristics of peace-time anaerobic infections

Altogether 26 cases of anaerobic infection (AI) of various etiology were analysed. Local tissue ischemia and operations on gastrointestinal organs in patients with secondary immunodeficit conditions are the factors facilitating the development of AI. The distinction is made between clostridial AI with a rapidly progressing gaseous gangrene and the non-clostridial AI with a slower course of a serous-purulent phlegmoma. Anaerobic myositis is observed in all forms of AI. Depending on the localisation of AI, anaerobic cellulitis and fasciitis are mentioned which occur mainly in non-clostridial AI. The treatment of AI and the mechanisms of death are discussed.     

Expression of the liver form of arginase in erythrocytes

Arginase I (AI) has a critical function in mammalian liver as the final enzyme in the urea cycle responsible for the disposal of ammonia from protein catabolism. AI is also expressed in various extrahepatic tissues and may play a role in regulating arginine levels and in providing ornithine for biosynthetic reactions that generate various critical intermediary metabolites such as glutamate, glutamine, GABA, agmatine, polyamines, creatine, proline, and nitric oxide. AI is expressed in red blood cells (RBCs) only in humans and certain higher primates. Macaca fascicularis has been identified as an evolutionary transition species in which RBC-AI expression is co-dominantly regulated. The M. fascicularis AI gene was analyzed to understand AI expression in erythrocytes. Erythroid progenitor cells [nucleated red blood cells (nRBCs)] isolated from cord blood were utilized to demonstrate AI expression by immunocytochemical staining using anti-AI antibody. Introduction of EGFP reporter vectors into nRBC showed that the proximal 1.2 kbp upstream of the AI gene is sufficient for AI expression. Expression of a second arginase isoform, AII, in nRBCs was discovered by cDNA profiling. This contrasts with mature fetal or adult RBCs which contain only the AI protein. In addition, an alternatively spliced AI (AI(')) variant was observed from erythroid mRNA analysis with an alternative splice acceptor site located within intron 2, causing the insertion of eight additional amino acids yet retaining significant enzymatic activity.     

Representation of cochlea within primary auditory cortex in the cat.

The representation of sound frequency (and of the cochlear partition) within primary auditory cortex has been investigated with use of microelectrode-mapping techniques in a series of 25 anesthetized cats. Among the results were the following: 1) Within vertical penetrations into AI, best frequency and remarkably constant for successively studied neurons across the active middle and deep cortical layers. 2) There is an orderly representation of frequency (and of represented cochlear place) within AI. Frequency is rerepresented across the mediolateral dimension of the field. On an axis perpendicular to this plane of rerepresentation, best-frequency (represented cochlear place) changes as a simple function of cortical location. 3) Any given frequency band (or sector of the cochlear partition) is represented across a belt of cortex of nearly constant width that runs on a nearly straight axis across AI. 4) There is a disproportionately large cortical surface representation of the highest-frequency octaves (basal cochlea) within AI. 5) The primary and secondary field locations were somewhat variable, when referenced to cortical surface landmarks. 6) Data from long penetrations passing down the rostral bank of the posterior ectosylvian sulcus were consistent with the existence of a vertical unit of organization in AI, akin to cortical columns described in primary visual and somatosensory cortex. 7) Responses to tonal stimuli were encountered in fields dorsocaudal, caudal, ventral, and rostral to AI. There is an orderly representation of the cochlea within the field rostal to AI, with a reversal in best frequencies across its border with AI. 8) Physiological definitions of AI boundaries are consistent with their cytoarchitectonic definition. Some of the implications of these findings are discussed.     

The effect of oxytocin antagonist on uterus in response to exogenous oxytocin

This study was performed to determine the action mode of oxytocin antagonist. In Study 1, the duration of in vivo action of oxytocin antagonist I (AI) was examined. After infusing AI, oxytocin was given and repeated every hour for 5 hr. Uterine activities were monitored with a polygraph. Study 2 determined the effect of AI on uterine oxytocin receptor number (Rn) and binding affinity (Kd). AI treated rats were sacrificed at 0.5 and 4 hr later for receptor assay. In Study 1, the uterine contractile response to oxytocin was significantly inhibited (p<0.05) compared to controls at five min, 1 and 2 hr after injection of AI. No differences in response were detected compared to controls (p>0.05) at later hours. In Study 2, no differences (p>0.05) between the AI and control animals in either oxytocin receptor number or binding affinity was found. These data suggest that the major mode of AI action is via competitive inhibition at the uterine oxytocin receptor and not by altering receptor number or binding affinity. AI is suggested to have the potential of being a potent and specific tocolytic agent for prevention of preterm labor in human.     

Comparative studies on tree pollen allergens. VIII. Immunological properties of the alder (Alnus incana) pollen extract

The immunological properties of the aqueous crude alder pollen extract (AI crude) and gel filtration fractions AI 3, AI 4 and AI 34 (pool of fractions AI 3 and AI 4) were examined by immuno- and radioimmuno-electrophoretic techniques, RAST titration, RAST inhibition and skin prick tests (SPT). In CIE, the AI crude extract and AI 34 displayed reference precipitate patterns consisting of 27 and 24 visible Coomassie brilliant blue stained lines, respectively. The CRIE allergogram performed by incubation with 18 individual reaginic sera detected three IgE-binding antigens characterized by different IgE-binding properties. Antigen No. 7 (Ag 7) was demonstrated to be the major IgE antibody-binding antigen of alder pollen, while Ag 1 and Ag 11 were classified as intermediate allergens. The allergens of alder pollen were located in fractions AI 3 and AI 4 of the gel filtration chromatogram. Ag 7 was present in both fractions as demonstrated by FRIE with autoradiography (FRIEWA) on the gel filtration fractions and tandem-CRIE of AI 3 and AI 4. The CRIE allergogram, RAST, RAST inhibition and SPT demonstrated fraction AI 34 to be allergenically representative of the AI crude extract both qualitatively and quantitatively. Thus, fraction AI 34 was considered an optimal purified allergen extract of alder pollen, a suitable material for further biochemical characterization and trials on purification of the allergenic reactive antigens.     

鼻咽癌细胞凋亡指数与p73表达及患者临床预后的关系

探讨鼻咽癌组织的细胞凋亡情况与p73基因表达、患者临床预后的关系。采用脱氧核糖核酸末端转移酶介导的缺口末端标记技术 (TUNEL)和免疫组化方法检测鼻咽低分化鳞状细胞癌和正常鼻咽黏膜组织。 4 8例鼻咽癌和正常鼻咽黏膜组织的凋亡指数 (AI)无差异。癌组织p73表达与癌细胞AI间无相关关系。癌组织AI与患者性别、年龄、有无复发、临床分期间无明显关系 ,但与患者生存时间呈负相关 (P <0 0 1)。经COX回归分析发现 ,癌组织AI和有无复发均属风险因素。鼻咽低分化鳞状细胞癌不属于高凋亡的肿瘤。p73在鼻咽低分化鳞状细胞癌组织中的表达与癌组织的细胞凋亡无相关关系。该癌的凋亡指数有判断患者预后的价值     

Introduction of autopsy imaging redefines the concept of autopsy: 37 cases of clinical experience

A new autopsy imaging (AI) system was introduced at the Research Center Hospital for Charged Particle Therapy (RCCPT) in January 2000. Autopsy imaging is a postmortem and preautopsy diagnostic procedure using magnetic resonance imaging (MRI). Scanning is performed with a 1.5 Tesla MRI system before autopsy. The AI results are reported to the pathologist and, in light of this information, autopsy is performed with minute precision. Autopsy imaging was performed on 37 cancer cases. In seven cases, AI was less informative than the autopsy, but in 30 cases, more precise reports on the final diagnosis were available with the combined application of autopsy and AI than autopsy alone, particularly in eight limited autopsy cases. Thus, AI provides critical and supplementary information for autopsy; furthermore, AI itself is a unique imaging system of great importance.     

Smoking may cause genetic alterations at 5q22.2 approximately q23.1 in clear-cell renal cell carcinoma

The aim of this study was to examine the relationship between allelic imbalance (AI) on chromosome 5q and clinico-pathologic parameters, including cigarette smoking. We examined the AI on chromosome 5q in 119 clear-cell renal cell carcinomas (CRCC) by a fluorescent polymerase chain reaction technique using nine microsatellite markers. AI of one or more loci was found in 43 cases (36.1%). The most frequent AI was observed at chromosome 5q23.1 (D5S467), where the LOX gene is located. The minimally involved region ranged from 5q22.1 to 5q23.2, with a possible breakpoint between 5q22 and 5q22.1. Regarding the relationships between the frequency of AI and the clinico-pathologic and environmental backgrounds, smokers had a significantly higher frequency of AI of 5q22.2 approximately q22.3 (D5S471) than nonsmokers (P = 0.013). No other significant associations were found between AI of specific loci and other parameters. Our results suggest that AI at 5q plays an important role in the genesis of CRCC. In addition, smoking may cause genetic alterations at 5q22.2 approximately q23.1, where the LOX gene is located.     

Interfacial interactions of apolipoprotein AI and high density lipoprotein: overlooked phenomena in blood-material contact

Apolipoprotein AI (apo AI) is the major protein component of high density lipoprotein (HDL), and represents ～1% of the total protein content of plasma. In previous work, apo AI was identified as a major component of the protein layers adsorbed from plasma to biomaterials having a wide range of surface properties. Notwithstanding such indications of the major contribution of lipoprotein interactions, these phenomena have been largely overlooked in the blood-contacting biomaterials area. In this communication, detailed quantitative data on the adsorption of apo AI to typical "biomedical" segmented polyurethane (PU) are reported. Using radiolabeled apo AI, adsorption levels from buffer and plasma were found to be ～0.5 and ～0.2 μg/cm(2), respectively. Albumin adsorption from plasma was comparable at about ～0.17 μg/cm(2). Since, it is unknown how much of the adsorbed apo AI is associated with HDL versus how much is in the free state, the corresponding molar quantities cannot be determined with certainty. However, if it is assumed that all of the adsorbed apo AI is free, the molar quantities adsorbed from plasma were in the ratio apo AI:albumin = 2.77, compared with 0.00063 in plasma. Immunoblot data showed similar trends with respect both to the variation in adsorbed quantity with plasma concentration and to the relative adsorbed quantities of apo AI and albumin. These data show unequivocally the very strong surface activity of apo AI and suggest that a key future focus for blood compatibility research should be lipoprotein interactions.     

鼻咽癌细胞凋亡与细胞毒淋巴细胞和LMP—1表达之间的关系

目的 :探讨鼻咽癌细胞凋亡与肿瘤内细胞毒淋巴细胞和LMP 1表达之间的关系。方法 :收集未经治疗的鼻咽癌活检组织 38例 ,采用TUNEL法定量检测鼻咽癌细胞凋亡程度 ,用免疫组化法检测肿瘤细胞LMP 1表达情况和肿瘤内TIA 1+细胞毒淋巴细胞数。结果 :(1) 38例鼻咽癌组织学分为角化性鳞癌 7例 ,非角化性癌 8例 ,未分化癌 2 3例。平均凋亡指数分别为72 3、74 6和 30 7。角化性鳞癌和非角化性癌与未分化癌相比 ,差异有显著性 (P <0 0 5 ) ;(2 ) 38例中LMP 1阳性病例 19例 ,占 5 0 %。除外角化性鳞癌后 ,LMP 1阳性组平均凋亡指数为 5 1 7,高于LMP 1阴性组的 37 3,差异有显著性 (P <0 0 5 ) ;(3)每 10 0 0个癌细胞范围内浸润的TIA 1+细胞数为 3～ 75个 ;在角化性鳞癌中平均 12 ,低于非角化性癌和未分化癌 (平均2 1 4) ,差异有显著性 (P <0 0 5 ) ;除外角化性鳞癌后 ,TIA 1+细胞数多者凋亡指数较高 (r =0 40 47,P <0 0 5 )。结论∶(1)鼻咽癌细胞凋亡程度在不同组织学类型中差异有显著性 ;(2 )鼻咽癌组织内TIA 1+细胞数在不同的组织学类型中有差别 ;(3)非角化性鼻咽癌组织中 ,LMP 1阳性者凋亡指数高 ,TIA 1+细胞数多者凋亡指数高。说明细胞毒淋巴细胞和LMP 1的表达具有促进鼻咽癌细胞凋亡的作用。     

Methods of separate heart section of fetuses and newborns

Modified Avtandilov's method of separate heart section of fetuses aged from 13 to 27 weeks of gestation obtained at artificial and spontaneous pregnancy interruption (newborn of 28 to 42 weeks of gestation) provided data of the atrial index (AI) and ventricular index (VI) in each age groups and distributed according to percentil method. Thus in the artificial fetuses normal AI is 1.118-1.353; VI-0.767-1.066; in spontaneous fetuses AI is 1.066-1.500; VI-0.772-1.083; in premature newborns AI is 1.116-1.538. Separation of heart compartments hypertrophy according to the degree is suggested.     

Left heart pressures can be the key to know the limitation of left ventricular assist device support against progression of aortic insufficiency

Aortic insufficiency (AI) is a worrisome complication under left ventricular assist device (LVAD) support. AI progression causes LVAD-left ventricular (LV) recirculation and can require surgical intervention to the aortic valve. However, the limitations of LVAD support are not well known. Using an animal model of LVAD with AI, the effect of AI progression on hemodynamics and myocardial oxygen metabolism were investigated. Five goats (Saanen 48?±?2 kg) underwent centrifugal type LVAD, EVAHEART, implantation. The AI model was established by placing a vena cava filter in the aortic valve. Cardiac dysfunction was induced by continuous beta-blockade (esmolol) infusion. Hemodynamic values and myocardial oxygen extraction ratio (O₂ER) were evaluated while changing the degree of AI which was expressed as the flow rate of LVAD-LV recirculation (recirculation rate). Diastolic aortic pressure was decreased with AI progression and correlated negatively with the recirculation rate (p?=?0.00055). Systolic left ventricular pressure (LVP) and mean left atrial pressure (LAP) were increased with AI progression and correlated positively with the recirculation rate (p?=?0.010, 0.023, respectively). LVP and LAP showed marked exponential increases when the recirculation rate surpassed 40%. O₂ER was also increased with AI progression and had a significant positive correlation with the recirculation rate (p?=?0.000043). O₂ER was increased linearly, with no exponential increase. AI progression made it difficult to reduce the cardiac pressure load, worsening myocardial oxygen metabolism. The exponential increase of left heart pressures could be the key to know the limitation of LVAD support against AI progression.     

Reduction in apoptosis relative to mitosis in histologically normal epithelium accompanies fibrocystic change and carcinoma of the premenopausal human breast.

The aims of this study in 227 premenopausal women were (a) to determine the mitotic index (MI), the thymidine labelling index (LI), and the apoptotic index (AI) within the epithelial cells of histologically 'normal' human breast biopsy material removed away from the site of either a fibroadenoma or a carcinoma; and (b) to relate differences in the kinetic indices of the 'normal' epithelium to the pathology in the same breast diagnosed as fibroadenoma alone (125 patients), fibroadenoma with accompanying mild fibrocystic change (79 patients), or carcinoma (23 patients). Ratios of the average indices (AI/MI, AI/LI, MI/LI) were also calculated to minimize uncertainties related to the total cell population counted, the denominator in the LI, MI, and AI. All indices and ratios of indices were corrected for age, averaged over the cycle, and expressed as log-transformed values for analysis. Significant (P less than 0.001) reductions in AI and in apoptosis relative to mitosis (reduced AI/MI) were found in 'normal' epithelium from breasts having fibrocystic change (AI = 0.17 +/- 0.02; AI/MI = 1.01 +/- 0.18) and carcinoma (AI = 0.19 +/- 0.04; AI/MI = 0.88 +/- 0.29), compared with breast with fibroadenoma alone (AI = 0.27 +/- 0.03; AI/MI = 1.29 +/- 0.39). In the absence of significant differences in MI and LI between the 'normal' tissue groups, this finding raises the possibility that reduced epithelial cell apoptosis might be causally associated with the development of fibrocystic change and with an increased risk of development of carcinoma.(ABSTRACT TRUNCATED AT 250 WORDS)     

Molecular analysis of neonatal IgA expression: Implications for class switching,allelic polymorphism and somatic mutation

This work was supported by National Institutes of Health Grants AI18016 (PWT), AI20313, AI19896, HD21693 (JMT) and AI15797 (NRK) and grant RA75 from the Morrison Trust (JMT). SCR was supported by an American Cancer Society Postdoctoral Fellowship.     

胃癌细胞凋亡与增殖的观察

目的 :探讨胃癌细胞凋亡与增殖的临床病理意义。方法 :采用切口末端标记法 (TUNEL)、电镜观察显示凋亡细胞 ,免疫组化S P法显示PCNA及 p5 3蛋白表达 ,利用图像系统分析癌细胞DNA倍体。 结果 :观察 60例胃癌细胞凋亡的TUNEL染色及超微结构形态 ;发现高分化型癌凋亡指数 (ApoptosisIndex ,AI)、AI/PI(PCNAIndel,PI)均高于低分化型癌 (P <0 0 5 ) ,而PI值、DNA >5C值则相反 ,高分化型低于低分化型癌 (P <0 0 5 ) ;癌侵及黏膜层、肌层、浆膜层时AI、AI/PI值逐层递减 ,而PI值、DNA >5C值则逐层递增 (P <0 0 5 ) ,肿瘤直径 <3cm、3～ 5cm、>5cm 3组AI、AI/PI值随肿瘤增大而递减 (P <0 0 1) ,其PI值、DNA >5C值则随肿瘤增大而递增 (P <0 0 5 )、淋巴结阳性组PI值较阴性组高 (P <0 0 5 ) ,p5 3阴性组较p5 3阳性组AI值高 (P <0 0 5 )。结论 :(1)分化高、体积小的早期及早期胃癌其AI值较高 ,分化低、体积大的晚期胃癌AI值较低 ,PI值增高 ;AI与PI呈负相关 ;PI值与淋巴结转移有关。显示癌瘤进展以细胞增殖为主而凋亡受抑。 (2 )p5 3基因突变可显著降低癌细胞凋亡     

Celiac disease and endocrine autoimmune disorders in children: an update

Celiac disease (CD) is a life-long inflammatory condition of the gut that occurs in genetically susceptible individuals. Several autoimmune diseases (AI) are associated with CD. To date, no conclusive evidence is available that proves if the relationship between CD and AI is mediated by gluten exposure, or if CD and AI could co-occur due to other causes, in particular the loss of the intestinal barrier function and the common genetic background. Furthermore, it is not clear yet if CD needs a regular screening program for AI. This review will cover the key studies on both the pathogenetic and clinical evidence explaining this association. We will review the reports including patients aged <18 years with CD and endocrine AI.     

Drinking in goats as effect of simultaneous intravenous infusions of angiotensin (I or II) and hypertonic NaCl or mannitol

Drinking during the simultaneous intravenous infusion of angiotensin I (Al) or II (AII) and hypertonic NaCl or mannitol was studied in the goat, and was compared to the dipsogenic responses to the separate infusion of each of these four factors. Approximately the same amount of water was drunk during the infusion of AI/NaCl, AI/mannitol and AII/NaCl. The amount was roughly equal to the sum of the amounts taken when each of two paired stimuli was infused separately. Significantly less water was drunk in response to AII/mannitol. Somewhat more water was drunk during the separate AI than during the separate AII infusion. Administration of an AI converting enzyme inhibitor completely abolished the AI contribution to drinking during the AI/NaCl infusion but did not reduce AII/NaCl drinking, indicating that the response to Al was entirely due to its conversion into AII. The possibility is discussed that the considerable difference between AI/mannitol and AII/mannitol drinking might have been the result of choroidal and/or ependymal AI converting enzyme activity.     

Apoptosis as an independent prognostic indicator in squamous cell carcinoma of the esophagus

Apoptosis plays a crucial role in determining net cell proliferation and cell turnover in various tumors. The rate of apoptosis in tumor cells has been reported to be a useful prognostic indicator in colorectal carcinoma. We examined apoptosis in 72 specimens of esophageal squamous cell carcinoma, by the terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) digoxigenin-nick end labeling (TUNEL) method. We examined correlation of apoptosis with outcome, clinicopathological features, and expression of the apoptosis-related proteins p53 and Bcl-2. The percentage of apoptotic cells, or apoptotic index (AI), ranged from 0.8 to 9.4 (mean: 3.47; SD: 2.02). Overall, 5-year survival of patients with high AI (AI > or = 5.0; n = 18) tumors was significantly higher than that of patients with low AI tumors (AI < 5.0; n = 58; 76.9% versus 44.9%; P = 0.042). AI did not correlate significantly with the clinicopathological features of patient age and sex, depth of tumor and histological differentiation, lymph node metastasis, lymphatic invasion, or venous invasion. In p53-negative tumors, the AI was significantly higher than in p53-positive tumors. We concluded that AI may be a useful prognostic indicator in esophageal squamous cell carcinoma following curative surgery, and that apoptosis in this tumor is related to relative underexpression of p53 protein.     

The genome sequence analysis of H5N1 avian influenza A virus isolated from the outbreak among poultry populations in Thailand

In this report, the genome of the Thai avian influenza virus A (H5N1); A/Chicken/Nakorn-Pathom/Thailand/CU-K2/04, isolated from the Thai avian influenza A (AI) epidemic during the early of 2004 was sequenced. Phylogenetic analyses were performed in comparison to AI viruses from Hong Kong 1997 outbreaks and other AI (H5N1) isolates reported during 2001-2004. Molecular characterization of the Thai AI (H5N1) HA gene revealed a common characteristic of a highly pathogenic AI (HPAI), a 20-codon deletion in the neuraminidase gene, a 5-codon deletion in the NS gene and polymorphisms of the M2 and PB2 genes. Moreover, the HA and NA genes of the Thai AI displayed high similarity to those of the AI viruses isolated from human cases during the same epidemic. Finally, our results demonstrated that the Thai AI emerged as a member of 2000's AI lineage with most of the genetic sequences closely related to the Influenza A/Duck/China/E319.2/03 (H5N1).