Radiation-induced changes in oral carcinoma cells – a multiparametric evaluation

The aim of this study was to see whether serial cytological evaluation of various cellular abnormalities in tumours from patients receiving fractionated radiotherapy can predict radio-response in oral carcinoma. Cytological assessment was carried out in scrape smears collected prior to and during the course of radiotherapy in 68 patients with squamous cell carcinoma of the oral cavity planned for radical radiotherapy with accelerated fraction schedule. Smears were evaluated for a set of 15 radiation-induced cellular abnormalities. The relationship between the cellular alterations and the cumulative radiation dose was analysed by Kruskal-Wallis one-way anova. The results showed that among the various quantifiable changes that occur in irradiated cancer cells, karyolysis, karyorrhexis, pyknosis, cytolysis, multinucleation, micronucleation and nuclear budding show significant increase depending on the dose of radiation. The radio-resistant group of patients exhibited a lesser degree of change compared with the radio-sensitive group. This suggests that radio-resistance may be due to the defective induction of cell damage and that these cytological features may have potential use as predictive markers of radio-sensitivity in oral carcinoma.     

Genetic metabolic polymorphisms and the risk of cancer: a review of the literature

The purpose of this paper is to systematically analyse the design and results of epidemiological studies on the association between various types of cancer (lung, bladder, breast, colon, stomach) and four genetically-based metabolic polymorphisms, involved in the metabolism of several carcinogens (glutathione-S-transferase M1, debrisoquine hydroxylase, N acetyltransferase, aryl hydrocarbon hydroxylase). These inherited polymorphisms usually cause modifications in the quality or quantity of the relevant enzymes. Such enzymes are involved in the activation/inactivation of known carcinogens and seem to modify the extent to which carcinogens interact with DNA in target tissues. Two enzymes, debrisoquine hydroxylase and aryl hydrocarbon hydroxylase, activate procarcinogens to carcinogens (phase I enzymes). The other two, glutathione-S-transferase M1 and N-acetyltransferase, mainly detoxity carcinogenic substances (phase II enzymes). Because of their role as host factors (modulating the action of carcinogens), it has been hypothesized that subjects presenting a specific phenotype for such polymorphisms could be at a greater risk of developing various types of cancer. A number of epidemiological studies have investigated such associations, often with discordant results. We examine and discuss the design of the studies, and present a meta-analysis of the available data.     

Upregulation of Fas-induced apoptosis by interferon-γ accompanied by increased ICE expression in renal cell cancer cells

The integration of Fas/Apo-1 (CD95) by Fas ligand or anti-Fas antibody induces apoptosis, and this system plays a pivotal role for the lysis of target cells by cytotoxic T lymphocytes. Fas-mediated apoptosis is also increased by a prior incubation of Fas-bearing cells with interferon(IFN)-. Interleukin-1- converting enzyme (ICE) and/or CPP32, or other members of ICE family act as direct cell death executors downstream of this mechanism, and a tetrapeptide inhibitor of these cysteine proteases blocks Fas-mediated apoptosis. In this study, we examined the effect of IFN- on Fas-mediated apoptosis in ACHN cells. IFN- augmented apoptosis in a dose dependent manner and reached a plateau at 400 U/ml when exposed for 48 h before the end of culture. The kinetics revealed a significant increase in apoptosis after 24 h. Exposing ACHN cells to IFN- increased pro-ICE expression accompanied with a decrease of pro-CPP32. These results suggest that direct enhancement of ICE expression and/or upregulation of conversion of pro-CPP32 to active form increases Fas-mediated apoptosis by IFN- in ACHN cells.     

生长特性差别的胃癌MGC803细胞亚系间细胞骨架、细胞通讯与癌基因表达的比较研究

肿瘤细胞表型差异是临床药物敏感性、抗原性、转移性、潜伏性和进展速度等差异的主要原因。为了弄清恶性表型差别的表现、相关性及调节机理、我们从胃癌MGC 803系分离了单细胞亚系,各亚系按克隆过程的时间差别归纳分组,选快组M 17、中组M 6和慢组M 3进行比较。软琼脂生长实验表明三者的恶性程度有显著差别;M17恶性度最高,M3最低,M6居中。与其它表型联系比较,证明细胞生长速度、细胞微丝、微管骨架破坏及细胞通讯功能抑制都与锚着不依赖性生长的恶性特征有平行相关性。同时,M 17和M 3原癌基因表达有明显差别。     

Interlaboratory Reproducibility In Reporting Inadequate Cervical Smears—A Multicentre Multinational Study

A set of 300 vaginal smears was interpreted by 13 cytologists from six European laboratories, who were requested to report inadequate and suboptimal smears. the set had been appropriately seeded to reach approximately 10% inadequate and 20% suboptimal smear frequency. According to the majority report, 230 smears were classified as adequate (76.7%), 43 as inadequate (14.3%), and 27 as suboptimal (9.0%). Agreement with the majority report ranged from 52% to 91% (average 78%). Kappa statistics for reporting inadequate smears showed a high level of agreement for five cytologists, and fair to good agreement for eight. In contrast, K statistics for reporting suboptimal smears showed fair to good agreement with the majority report only in five instances, whereas agreement was poor for eight cytologists. ‘Inadequate smear’rates may be used to compare the quality of smears received in different laboratories, as there is a high level of agreement among cytologists as to what constitutes an inadequate smear. However, this is not true for ‘suboptimal smear’ rates, and more precise reporting criteria must be defined and tested if an intermediate category is to be retained to report poor quality smears: more precise reporting criteria must be defined and tested if an intermediate category is to be retained to report poor quality smears. Une série de 300 frottis cervico-vaginaux a été examinee par 13 cytologistes appartenant à 6 laboratoires européens auxquels il avait été demandé de signaler les frottis inadéquats et sub-optimaux. Cette série de frottis a été constituée de façon à atteindre approximativement une fréquence de 10% de frottis inadéquats et de 20% de frottis sub-optimaux. Si I'on considère les réponses faites par la majorité des cytologistes, 230 frottis ont été classés adéquats (76,7%), 43 classés inadéquats (14,3%), et 27 comme sub-optimaux (9,0%). La concordance avec les réponses de la majorité des cytologistes varie de 52%à 91% (moyenne 78%). Concernant le signalement des frottis inadéquats, le test Kappa est excellent pour 5 cytologistes et moyen à bon pour les 8 autres. Au contraire, pour le signalement des frottis sub-optimaux, le test Kappa montre un accord moyen à bon avec les réponses de la majorité seulement dans 5 cas alors que I'agrément est mauvais pour 8 cytologistes. La fréquence des frottis inadéquats peut être utilisée pour comparer la qualité de prélèvement du frottis dans différents laboratoires, alors que les comparaisons de la fréquence des frottis sub-optimaux sont discutables, du fait d'un biais de diagnostic évident. Des critéres plus précis pour le signalement des frottis sub-optimaux doivent étre définis et testés si cette catégorie doit être retenue pour rapporter les frottis de mauvaise qualité. 300 Abstriche wurden durch 13 Zytologen aus 6 europäischen Laboratorien hinsichtlich der Kriterien ‘unbrauchbar’und ‘suboptimal’beurteilt. Die Sammlung enthielt etwa 10% unbrauchbare und 20% suboptimale Präparate. Mehrheitlich wurden 230 Abstriche als auswertbar (76,7%), 43 als unbrauchbar (14,3%) und 27 als suboptimal (9,0%) bezeichnet. Die Statistik ergab hinsichtlich der unbrauchbaren Abstriche eine ausgezeichnete Ûbereinstimmung für 5 und eine gute Ûbereinstim-mung für 8 Zytologen. Demgegenüber zeigte sich hinsichtlich der Beurteilung suboptimal nur 5mal eine gute und 8mal eine schlechte Ûbereinstimmung. Damit ist die Kategorie suboptimal ungeeignet zum Vergleich und bedarf einer genaueren Definition wenn sie für die Qualitätskontrolle eingesetzt werden soll.     

Correlation of endoscopic cytology and histology in oesophageal cancer: results in Porto Alegre,RS—Brazil

A retrospective study of oesophageal cytopathology at the Hospital de Clínicas de Porto Alegre (HCPA), RS, Brazil, from 1989 to 1992 was made to assess the sensitivity, specificity, predictive values and accuracy of endoscopic cytology and biopsy; and study the correlation between cytopathological and histopathological diagnosis. Specimens from 94 patients were available for review. The final diagnosis was based on surgical pathology and follow up. The 81 patients with cancer of the oesophagus had the following sex distribution: 64 males and 17 females (a 3.7–1 ratio). No tumour was found in 13 patients. The following conclusions were made: (i) there is excellent correlation between cytology and histology in oesophageal lesions sampled by endoscopy; (ii) a correct positive cytologic report was obtained in 77 (95%) of the 81 proven oesophageal cancers; a false-negative or unsatisfactory result was given in four patients. A false-positive diagnosis of cancer was not made. There were 13 true-negative reports. These findings result in a sensitivity of 95% with 95% confidence intervals (CI) of 90.26–99.74%; a specificity of 100% (CI of 98.5–100%); a positive predictive value of 100% (CI of 99.3–100%); a negative predictive value of 76% (CI of 55.7–96.3%); (iii) a correct positive histological report was obtained in 67 (83%) of the 81 proven oesophageal cancers; a false-negative or unsatisfactory result was given in 14 patients. A false-positive diagnosis of cancer was not made. There were 13 true-negative reports. These findings result in a sensitivity of 83% with 95% CI of 74.82–91.18%; a specificity of 100% (CI of 98.5–100%); a positive predictive value of 100% (CI of 99.25–100%); a negative predictive value of 48% (CI of 29.16–64.84%); (iv) of 81 patients with proven cancer, in 79 (98%) at least one of the methods was positive. In only two patients with cancer were both methods negative. These findings result in a combined sensitivity of 98% (CI of 94.92–100%); a specificity of 100% (CI of 98.5–100%); a positive predictive value of 100% (CI of 99.31–100%); and a negative predictive value of 87% (CI of 70–100%). Our series confirms the value of the combined use of cytology and biopsy for the investigation of oesophageal lesions. However, it should be remembered that even with the combined use of cytology and biopsy there are some tumours that will be negative by both procedures: we had only two such cases, confirming the rarity of such an event.     

人乳腺癌cAMP结合蛋白

建立了测定人乳腺癌胞浆ｃＡＭＰ结合蛋白（ｃＡＭＰｂ．ｐ．）方法。综合研究了其温度、保温时间、配体浓度、稳定性等条件。ｃＡＭＰｂ．ｐ．的Ｋ_Ｄ值为２．９０×１０~（－８）ｍｏｌ／Ｌ．并测定了６０例雌激素受体（ＥＲ）Ｆｕ性乳腺癌标本的ｃＡＭＰｂ．ｐ．含量。此组病人术后均接受系统的内分泌治疗,ＥＲ／ｃＡＭＰｂｐ,比值范围为７．７～３６２×１０~（－３）,ＥＲ／ｃＡＭＰｂ．ｐ．比值≥４０×１０~（－３）的五年生存率明显高于比值＜４０×１０~（－３）组,（ｐ＜０．００５）．表明测定ＥＲ／ｃＡＭＰｂ．ｐ．比值对预测患者内分泌治疗疗效,优于单独测定ＥＲ．     

Reconstitution of a pentameric complex of dimeric transforming growth factor beta ligand and a type I,II, III receptor in baculoviral-infected insect cells

Summary Two transmembrane serine-threonine kinases (type I and II receptors), a membrane-anchored proteoglycan (type III), and a homodimeric ligand participate in the transforming growth factor beta type on (TGFβ1) signal transduction complex. The expression of recombinant receptors in insect cells co-infected with up to three recombinant baculoviruses was employed to study interactions among the ectodomains of the three types of receptors and the TGFβ1 ligand in absence of uncontrollable extrinsic factors in mammalian cells. Multi-subunit complexes were assembled in intact cells and purified on glutathione-conjugated beads for analysis by tagging one of the subunits with glutathione S-transferase (GST). Intrinsic ligand-independent interactions were observed among receptor subunits as follows: type III–III type I–I, type III-I, and type II-I. The homeotypic complex of type II–II receptors and the heterotypic type III-II interaction was ligand dependent. The type I, but not the type III, subunit displaced about 50% of the type II component in either ligand-dependent homomeric type II-type II complexes or heteromeric type III-type II complexes to form type II-I or type III-II-I oligomers, respectively. The type II subunit displaced type I subunits in oligomers of the type I subunit. Specificity of type I receptors may result from differential affinity for the type II receptor rather than specificity for ligand. A monomeric subunit of the TGFβ1 ligand bound concurrently to type III and type II or type III and type I receptors, but failed to concurrently bind to the type II and type I subunits. The binding of TGFβ1 to the type I kinase subunit appears to require an intact disulfide-linked ligand dimer in the absence of a type III subunit. The combined results suggest a pentameric TGFβ signal transduction complex in which one unit each of the type III, type II, and type I components is assembled around the two subunits of the dimeric TGFβ1 ligand. An immobilized GST-tagged subunit of the receptor complex was utilized to assemble multi-subunit complexesin vitro and to study the phosphorylation events among subunits in the absence of extrinsic cell-derived kinases. The results revealed that (a) a low level of ligand-independent autophosphorylation occurs in the type I kinase; (b) a high level of autophosphorylation occurs in the type II kinase; (c) both the type III and type I subunits aretrans-phosphorylated by the type II subunit; and (d) the presence of both type I and II kinases complexed with the type III subunit and dimeric TGFβ1 ligand in a pentameric complex causes maximum phosphorylation of all three receptor subunits.     

双功能抗体介导的胃癌杀伤效应

将诱变的αＣＤ３杂交瘤（ＴＫ~－）与ＰＤ４杂交瘤（ＨＧＰＲＴ~－）融合,获得分泌双功能抗体（ＢｓＡｂ）的四体杂交瘤Ｃ３．ＢｓＡｂＣ３可分别与ＣＤ３分子及胃癌相关抗原Ｐ４０反应．体外杀伤试验证实,当效靶比为４０：１,ＢｓＡｂＣ３浓度为１ｍｇ／Ｌ时,其杀伤效应可达７７．６％．该杀伤效应具有明显的特异性,仅Ｐ４０阳性表达的靶细胞可被溶解,体内杀伤试验证实,裸鼠接种胃癌细胞后５ｄ,以ＢｓＡｂＣ３活化的外周血淋巴细胞（ＰＢＬｓ）经局部皮下注射处理,可使移植胃癌完全消退（５／５）．这一明显的治疗作用可能与局部注射途径有关,可供临床应用参考．     

The Nucleotide Sequence of Human Acylamino Acid-Releasing Enzyme

The nucleotide sequence of a cDNA coding for the human acylaminoacid-releasing enzyme (AARE, also known as acylpeptide hydrolase)[EC 3.4.19.1] subunit has been determined. The amino acid sequenceof human AARE subunit deduced from its cDNA nucleotide sequenceshowed a high degree of identity (91.5%) with both the correspondingproteins from the pig and the rat. The AARE cDNA shows 99.2%identity with a 3.3 kb cDNA transcribed from a locus (DNF15S2)on the short arm of human chromosome 3, whose deletion is associatedwith small cell lung cancer, taking into consideration thatthe sequence of the 3.3-kb cDNA previously reported was causedby misreading.