Directly observed therapy short course (DOTS) is the internationally recommended strategy to ensure cure of tuberculosis; it has become the standard for the diagnosis, treatment and monitoring of tuberculosis worldwide and has been implemented in 182 of 211 countries, covering more than 77% of world's population(The present study included patients with pulmonary tuberculosis (previously treated within RNTCP then included under CAT II) who failed DOTS CAT II treatment at various centers mainly from eastern and north-eastern parts of Uttar Pradesh and some even from surrounding states like Madhya Pradesh, Bihar and Uttarakhand. The study period extended from June 2004 to December 2005. Informed consent was taken from all subjects. All cases were critically evaluated by detailed clinical history including history of antitubercular treatment (ATT) taken before inclusion in DOTS. History regarding their socioeconomic background, smoking, alcohol intake and drug abuse was recorded and cause for their inclusion in CAT II was evaluated. Pretreatment sputum bacillary count was noted from their records. Leading questions were asked regarding non-adherence to CAT II treatment and the causes were sought into. They underwent thorough clinical examination, radiological and bacteriological evaluation. The radiographs were classified based on the report of American Thoracic Society., in CAT III , default in previous DOTS therapy  and relapse  [The reason for inclusion of these 95 cases in CAT II was treatment failure in CAT I  . Thirty-8, 8.4%) . Since tThe pretreatment sputum bacillary load was 1+ in 22 cases (23%), 2+ in 31 (33%) and 3+ in 42 (44%) cases . MaximumThere were 35 cases (37%) who had taken CAT II treatment irregularly and finally became CAT II treatment failure cases (18 cases had interrupted treatment in the intensive phase while 17 in continuation phase and 80% of the non-adherence occurred for more than 1 month). The most important causes of interruption were the lack of relief of symptoms (37%), intolerance/toxicity (17%), inability to come for drug administration during intensive phase due to loss of earning and poor general condition (15%) and migration (14%).The study included 95 cases of failed CAT II (previously treated within RNTCP), who reported to our hospital for further treatment. The majority of patients (80%) were from economically productive age group (15-44 years) and from lower socioeconomic status. There were 32 patients who were initially wrongly categorized in CAT I since they had already received ATT for >1 month from local practitioners before inclusion in CAT I; they should have been categorized in CAT II directly, which could have been one of the important reason for the ultimate CAT II failure.n = 51) belonged to already failed CAT I cases and this could be one of the important reason for CAT II failure., drug intolerance (17%) and migration (14%). Another important factor was non-willingness to come thrice weekly for the drug administration (9%) during the intensive phase and most of the cases were from low socioeconomic strata and to them earning was more important than spending time at DOTS center for treatment. This factor becomes still more important when the symptoms of the disease are alleviated by initial treatment.The average contact of tuberculosis patients was as high up to seven to eight in our study and as nucleus of Indian household is very compact and transmission of disease is easier, presence of a resistant case in such a situation increases chances of primary drug resistance.The above study was prompted by large number of DOTS failure coming to our department and it throws some light on some of the problems of DOTS CAT II. The important causes of CAT II failure were previous CAT I failure, initial wrong categorization in CAT I, initial heavy bacillary load and non-adherence to treatment.


Mice were injected with tritiated misonidazole (750 mg/kg-1), killed after 24h and the excised tissues prepared for autoradiography (ARG) to identify sites of accumulation. The previously reported high grain count associated with bound misonidazole metabolite(s) was observed in the liver. The ratio of grain count in the emulsion above the centrilobular hepatocytes to the count over connective tissue (stroma) was 12. A higher count ratio for 'target' cells to stroma was observed in the following cells/tissues: meibomian gland , oesophagus (keratinised layer 60), incisor (enamel organ 17), nasal septum . For some of these tissues the explanation might appear to lie with localised hypoxia, but for others which were probably normoxic there is as yet no obvious reason for these findings.


Serum alpha-foetoprotein (AFP) and serum carcinoembryonic antigen (CEA) levels were measured, serially whenever possible, in 70 patients attending the Institute of Radiotherapy, Rotterdam, on account of testicular (65) or ovarian (4) germ cell tumours or, in one case, an endodermal sinus (yolk sac) tumour in the mediastinum. In 15 patients the disease was active; in the others it was in remission. Patients with active disease had raised serum AFP levels which correlated well with disease activity; no patient without evidence of active disease had raised serum AFP levels. None of the patients with active disease was found to have raised serum CEA levels. There was no correlation between serum AFP and CEA levels in patients with germ cell neoplasms, but good correlation between serum AFP levels and disease activity. Serum CEA levels did not correlate with disease activity, and serial determinations would therefore not be useful in monitoring progress in this group of diseases.


Serial determinations of serum alpha-foetoprotein (AFP) concentrations are well established in monitoring the response to therapy of malignant germ cell tumours. Using a radioimmunoassay (RIA) with a sensitivity down to 2kul-1 the majority (57%) of 28 patients with non-AFP producing germ cell tumours had measurable immunologically-reactive AFP in their serum while on treatment. Follow-up for 11-43 months (mean 27) without evidence of tumour activity indicated that this immunologically-reactive AFP was unlikely to be produced by tumour. In patients where the initial serum AFP was raised prior to chemotherapy the AFP concentration did not fall to the normal range at the end of the treatment in 16 (32%) of 41 patients. Follow-up of these patients for 9-48 months (mean 27) has resulted in 5 (12%) relapses in this group. Serum AFP greater than 20kul-1 three months after stopping chemotherapy was a good indicator of residual active tumour and 4 (57%) of 7 patients in this group relapsed. The production of detectable serum AFP is probably related to the type of chemotherapy used and only 7 (14%) of 51 patients treated for gestational choriocarcinoma had detectable AFP concentrations while on cytotoxic chemotherapy. The problem of interpretation of serum AFP concentration in patients with malignant germ cell tumour stresses the need to determine whether there are differences between AFP produced by germ cell tumours and that produced at other sites as a basis for a sensitive assay system able to discriminate between them.


Human dialysable Transfer Factor (TFd) extracted from lymphocytes of patients with transitional cell carcinoma of bladder (TCCB) was replicated in culture by lymphoblastoid cell lines. The effectiveness of two such TFdLs produced in vitro in transferring sensitivity to TCCB was assessed in the lymphocyte migration test (LMT) using formalin-treated TCCB cells as antigen. The results, showed that one TFdL transferred sensitivity in 5/14 cases and the other in 12/15, not only to leucocytes of healthy individuals but also to leucocytes of TCCB patients. Preliminary results showing an in vivo transfer of sensitivity are discussed.


RNA extracted from the spleens of tumour-bearing (TLRNA) and tumour-immune (ILRNA) mice was shown to transfer to normal lymphocytes (NL) the ability to produce factors that blocked specific tumour-cell cytotoxicity and mediated specific antibody-dependent cell cytotoxicity (ADCC). Aliquots of normal C3H mouse lymphocytes were treated with TLRNA or ILRNA and cultured in vitro in the absence of tumour antigen. Supernatants were collected at 24h intervals and tested in a microcytotoxicity assay for blocking and ADCC activities. Factors that inhibited tumour destruction by specifically sensitized lymphocytes at the level of both the tumour cells and effector cells were demonstrable in culture supernatants of NL pretreated with TLRNA (50 or 100 microgram/4 X 10(6) cells) but not ILRNA. However, treatment of NL with either RNA resulted in the production factors that mediated tumour-specific ADCC. Cytotoxicity testing and absorption studies of the tumour cell and a control cell (LM) indicated that factors mediating ADCC and blocking at the target-cell level were specific for the tumour. Suppressor activity at the effector-cell level was not absorbed by tumour cells and represents a separate and distinct mechanism of immunosuppression. These data indicate that RNA faithfully transfers "suppressive" as well as "positive" types of immune responses that have been reported previously for lymphocytes obtained directly from tumour-bearing and tumour-immune animals.


Itis suggested that the lower biological effects of IRIM compared to ICR are a consequence of the larger kineticinertness of the iridium(III) center with respect to ruthenium(III).Some biological aspects of the new complex imidazolium bisimidazole tetrachloro iridate(III)-IRIM- the iridium(III) analogue of ICR, were considered. More in detail the conformational effects produced by IRIM on DNA and the cytotoxic properties of IRIM on some selected human cell lines were measured.Dialysis experiments and DNA thermal denaturation studies are suggestive of poor binding of IRIM to DNA;formation of interstrand crosslinks is not observed. In any case CD measurements suggest that addition ofincreasing amounts of IRIM to calf thymus DNA results into significant spectral changes, that are diagnosticof a direct interaction with DNA. A number of experiments carried out on the A2780 human ovarian carcinoma,B16 murine melanoma, MCF7 and TS mammary adenocarcinoma tumor cell lines strongly point outthat IRIM does not exhibit significant growth inhibition effects within the concentration range 10


Local free cell (eosinophils and macrophages) and extracellular (acid mucopolysaccharides) reactions were studied by histochemical techniques in 72 primary lung cancers (Group A), 17 pulmonary metastases and 5 lung tumours of unknown origin (Group B). Strong cellular reactions were more frequent in Group A than in Group B, whereas extracellular reactions were more frequent in Group B than in Group A. In both groups the degrees of eosinophilic infiltration and accumulation of acid mucopolysaccharides tended to be negatively correlated. In contrast, macrophages and eosinophils showed a positive correlation in the primary cancers, but none in the other tumours. The extracellular material had physical and chemical properties  of the sulphated acid mucopolysaccharides. From its distribution it appeared to be derived from fibroblasts and/or tumour cells.


The occurrence of aberrations involving chromosomes 11 and 17 in malignant tissues of breast cancer patients has not yet been studied systematically. Using fluorescence in situ hybridisation (FISH) with centromere-specific probes, we determined chromosome 11 and 17 status in interphase nuclei from primary and/or metastatic breast cancer cells. In all cancerous specimens obtained from 30 patients, FISH identified cells with clonal chromosomal abnormalities, with aneuploidy rates ranging from 6% to 92% (median 59%). There was a gain of centromeric signals for chromosome 11, most likely corresponding to hyperploidy; aberrations of chromosome 17 in specimens from 26 patients (87%) were hyperploid as well; however, four cases (13%) showed loss of chromosome 17 centromeres. All specimens contained heterogeneous aneuploid cell populations with excessive gain of signals in some cases. The pattern of aneuploidy did not appear to correlate with tumour grade/stage and was comparable in primary tumours and corresponding metastatic axillary lymph nodes, indicative of genetic instability early in tumour development. Screening with a panel of FISH probes may lead to enhanced sensitivity and specificity in detecting malignant cells, as demonstrated in this study with effusions which could not be conclusively interpreted as being malignant by cytological criteria.


To elucidate the role of ras gene mutations during the early stage of colorectal tumour progression, K-ras gene mutations were analysed in 32 benign adenomas and 36 adenomas with focal carcinoma in the colorectum by microscraping of histologically pure regions from tissue sections, polymerase chain reaction-restriction fragment length polymorphism and in part by direct sequencing. Several regions were scraped out and analysed when an adenoma contained areas with different grades of dysplasia. The frequencies of K-ras gene mutation in mild dysplasia, moderate dysplasia and focal carcinoma were 19% (7/36), 51% (25/49) and 39% (14/36) respectively. The K-ras gene status was heterogeneous in 4 of the 11 benign adenomas from which multiple samples were obtained, and mutations were always found in the regions with more advanced dysplasia in these adenomas. Thirteen of the 36 adenomas with focal carcinoma showed heterogeneity of mutations between the adenoma region and the focal carcinoma. Seven of which had mutations only in the adenoma region. These findings indicated that the K-ras gene mutations occur during the late stage of adenoma progression and may confer a more advanced morphological phenotype of adenoma, but these mutations are not mainly involved in malignant transformation from adenoma to carcinoma.


Methotrexate (MTX) shows consistent cytotoxicity for melanoma cells in vitro but it is ineffective in clinical use at equivalent concentrations in vivo. This apparent paradox has been investigated by cell culture techniques and results quantified by cell number. In an in vitro model of high dose MTX therapy followed by leucovorin rescue (HD-MTX-LCR) there was survival of both melanoma and choriocarcinoma cell lines but not of an acute lymphocytic leukaemia cell line. The 70H metabolite of MTX was identified by HPLC in plasma samples of melanoma patients treated by HD-MTX-LCR, in which MTX concentrations approximately 10(-5) M were maintained for 24 h. However, metabolism per se is unlikely to account for the lack of response to MTX clinically. In vitro 70H MTX (10(-7) - 10(-6) M) was two orders of magnitude less cytotoxic for melanoma than MTX (10(-9) - 10(-8) M). The cellular accumulation of [3H]-MTX, using a rapid gradient centrifuge technique for separation of melanoma cells from medium, was reduced in the presence of 70H-MTX. The results suggest that reduced cellular uptake of MTX combined with biochemical rescue of tumour cells may partially explain the paradoxical lack of clinical response of melanoma to the drug.


The extracellular pH (pHe) of solid tumours is often lower than in normal tissues, and this may influence the uptake and/or activity of anti-cancer drugs. The cytotoxicity of mitoxantrone, paclitaxel and topotecan was therefore assessed at low pHe and after manipulation of intracellular pH (pHi) in murine EMT6 and in human MGH-U1 cells. The cytotoxic efficacy of all three agents was reduced at pHe 6.5 as compared with pHe 7.4. The ionophore nigericin and inhibitors of membrane-based ion exchange mechanisms that regulate pHi  were used to cause intracellular acidification. Combined use of the cytostatic drugs with pHi modifiers reduced their cytotoxicity under both physiological and low-pHe conditions. The uptake into cells of mitoxantrone (a weak base) was inhibited at pHe 6.5 as compared with pHe 7.4, and smaller effects of low pHe to inhibit uptake of topotecan were also observed. DNA analysis of cell cycle distribution revealed that intracellular acidification, as observed during incubation at low pHe and/or using pHi modifiers, resulted in accumulation of cells in G1 phase, where they may be more resistant to these drugs. Reduced uptake of weak bases (mitoxantrone) at low pHe and altered cell cycle kinetics upon acidification are the postulated causes of reduced cytotoxicity of the agents investigated.


Nuclear and cytoplasmic oestrogen receptors (REN and REC) were sought in 5 normal cervices and in 43 specimens of squamous carcinoma of the cervix. All 3 tissues components of the 5 normal cervices contained both REN and REC. Thirty-five (81%) of the tumours contained receptors, but in only 9 (21%) were they found in both subcellular compartments. Twenty-four tumours (56%) had only REC and 2 had only REN. The potential therapeutic significance of these findings is not yet known, but it seems possible that tumours with an intact receptor mechanism might benefit from oestrogen therapy and have a more favourable prognosis.


We evaluated West Nile (WN) virus seroprevalence in healthy horses, dogs, and cats in New York City after an outbreak of human WN virus encephalitis in 1999. Two (3%) of 73 horses, 10 (5%) of 189 dogs, and none of 12 cats tested positive for WN virus-neutralizing antibodies. Domestic mammals should be evaluated as sentinels for local WN virus activity and predictors of the infection in humans.


In order to identify the possible role of the DCC gene in neoplasms of the human female reproductive tract, messenger RNA expression of the DCC gene was examined by reverse transcriptase-polymerase chain reaction, and expression of the DCC gene product was detected immunohistochemically. While histologically normal endometrium, cervical epithelium and ovary expressed detectable mRNA of the DCC gene, three of eight (37%) endometrial carcinomas, one of two (50%) cervical carcinomas and 9 of 22 (41%) ovarian malignant tumours had significantly reduced or negligible DCC expression, and another endometrial carcinoma and two other ovarian tumors underexpressed DCC when compared with histologically normal endometrial or ovarian tissues. Impaired DCC mRNA expression was detected more frequently in grade 3 ovarian epithelial tumours than in grade 1 tumours (P = 0.002). Loss of expression of the DCC gene product detected by immunohistochemistry significantly correlated with the loss of mRNA expression in ovarian carcinomas (P = 0.01 by chi-square test) or in both endometrial and ovarian carcinomas combined (P = 0.001). Loss of heterozygosity of the DCC gene was also evaluated by restriction fragment polymorphism analysis of the polymerase chain reaction-amplified DNA fragment. Loss of heterozygosity of the DCC gene was detected in one of seven (14%) informative cases of endometrial carcinomas, 1 of 11 (9%) informative cases of cervical carcinomas and two of six (33%) informative cases of ovarian tumours. These results demonstrate that inactivation of the DCC gene, especially by the loss of expression, plays a significant role in the aetiology of neoplasms of the human reproductive tract.


Signal transduction pathways activated by injury play a central role in coordinating the cellular responses that determine whether a cell survives or dies. GADD153 expression increases markedly in response to some types of cellular injury and the product of this gene causes cell cycle arrest. Using induction of GADD153 as a model, we have investigated the activation of the cellular injury response after treatment with taxol and cisplatin (cDDP). Activation of the GADD153 promoter coupled to the luciferase gene and transfected into human ovarian carcinoma 2008 cells correlated well with the increase in endogenous GADD153 mRNA after treatment with taxol but not after treatment with cDDP. Following treatment with cDDP, the increase in endogenous GADD153 mRNA was 10-fold greater than the increase in GADD153 promoter activity. Likewise, at equitoxic levels of exposure (IC80), cDDP produced a 5-fold greater increase in endogenous GADD153 mRNA than taxol. The tyrosine kinase inhibitor tyrophostin B46 had no significant effect on the ability of taxol to activate the GADD153 promoter, but inhibited activation of the GADD153 promoter by cDDP in a concentration-dependent manner. Tyrphostin B46 synergistically enhanced the cytotoxicity of cisplatin; however, the same exposure had no significant effect on the cytotoxicity of taxol. We conclude that (1) taxol and cDDP activate GADD153 promoter activity through different mechanisms; (2) the signal transduction pathway mediating induction by cDDP involves a tyrosine kinase inhibitable by tyrphostin B46; and (3) that inhibition of this signal transduction pathway by tyrphostin synergistically enhances cDDP toxicity.


In 104 patients with gastric cancer the serum proteins carcinoembryonic antigen (CEA), C-reactive protein (CRP), alpha 1-acid glycoprotein (AGP) (orosomucoid) and alpha 1-antichymotrypsin (ACT) were measured pre-operatively. The estimated median survival of patients with both raised CEA and ACT was only 5 weeks in contrast to 64 weeks for those with both proteins normal. An intermediary group with one of these proteins raised and the other normal had an estimated median survival of 15 weeks. Similar results were obtained by considering a combination of CEA with either AGP or CRP. For these data the results were not explicable in terms of associations between survival time and patient's age, stage, operative procedure, histological classification or site of primary tumour.


The expression of complex carbohydrates recognised by Helix pomatia lectin  has been shown to predict unfavourable prognosis in breast and other cancers. It has been suggested that the prognostic significance of HPA binding may be through recognition of either Tn epitope  or blood group A antigen  GlcNAc-->R). In this study, the expression of glycoproteins terminating in alpha-GalNAc residues was investigated immunohistochemically using HPA and two monoclonal antibodies--BRIC 66  and BRIC 111 (anti-Tn). In paraffin sections, 74/87 (85%) of breast cancers expressed HPA-binding ligands, while 28/87 (32%) were positive for BRIC 66 binding and 25/87 (29%) expressed Tn. Distribution of staining patterns were distinctive and different with the three markers. BRIC 66, BRIC 111 and HPA binding to glycoproteins derived from breast cancer homogenates and to blood group A and Tn positive glycoproteins in Western blots confirmed the immunohistochemistry data. The results suggest that the prognostic significance of HPA binding in breast cancer is unlikely to be simply through recognition of blood group A antigen or Tn epitope on cancer cells. Breast cancers may express a complex profile of related but distinct glycans sharing similar terminal immunodominant sugar GalNAc, which may be implicated in aggressive biological behaviour.


The transformation of Wistar rat embryo cells in vitro has been studied in passaged cultures using two criteria for transformation: (1) the ability of cells to form colonies in soft agar and (2) the ability of cells to form tumours in young syngeneic animals. In general there was good correlation between the two criteria. Spontaneous transformation was observed in all untreated cultures by 44 weeks although, by not allowing the cells to become confluent, the tendency was for cultures to transform earlier (i.e. 15-21 weeks). It was noticeable that despite untreated cultures having been in vitro for different lengths of time, most cultures transformed after a similar number of passages (42-50). Treatment of the embryo cells with the alkylating agent nitrosomethylurea (NMU) or benzopyrene (BP) sometimes resulted in transformation after a shorter period in vitro than the controls (minimum 12 weeks) although some treated cultures took longer. Transformed cells produced transplantable fibrosarcomata in syngeneic hosts and those arising from NMU transformed cells were histologically different from those arising from spontaneously transformed cells. The significant of spontaneous transformation in in vitro rat cell transformation systems is discussed.


P < 0.0001). However, TIMP-1 expression was not increased in NPA and SW579 cells, both of which are derived from poorly differentiated thyroid tumours. Immunohistochemical study showed strong TIMP-1 staining in the stroma cells of advanced stages of carcinomas. Overexpression of TIMP-1 by gene transfer resulted in a significant suppression of the malignant phenotype of NPA cells as judged by an in vitro tumour invasion assay. These results suggest that high levels of TIMP-1 transcripts in advanced stages of thyroid carcinoma likely come from stroma rather than thyroid cancer cells, and TIMP-1 may function as a thyroid tumour invasion/metastasis suppressor. © 1999 Cancer Research CampaignTumour cell invasion and metastasis is a multistep process that involves the degradation of extracellular matrix proteins by matrix metalloproteinases (MMPs). Tissue inhibitors of metalloproteinases (TIMPs) act as negative regulators of MMPs and thus prevent tumour cell invasion and metastasis by preserving extracellular matrix (ECM) integrity. In the present study we examined the expression of one member of TIMPs, TIMP-1, in 39 thyroid tumour specimens and two thyroid carcinoma cell lines (NPA and SW579). We also investigated the effect of high TIMP-1 expression on the invasive potential of NPA cells. Northern blot analysis showed that TIMP-1 mRNA levels correlated directly with tumour aggressiveness: the highest number of TIMP-1 transcripts was found in stages III and IV vs benign goitre (


Rh2(tfacam)4 was tested both in vitro (U937 and K562 human leukemia cells and Ehrlich ascitic tumor cells) and in vivo for cytostatic activity and lethal dose determination, respectively. This is the first rhodium tetra-amidate to have its biological activity evaluated. The LD50 value for Rh2(tfacam)4 is of the same order as that of cisplatin, and it was verified that the rhodium complex usually needs lower doses than cisplatin to promote the same inhibitory effects.The rhodium (II) complexes Rh


Equimolar concentrations of cysteamine and reduced glutathione protected against the cytotoxicity of 5 mM misonidazole (MISO), whereas 5mM ascorbate enhanced its toxicity to hypoxic CHO and HeLa cells in vitro. Protection (reappearance of a shoulder region) could also be seen when cysteamine was added at later incubation times. These changes in toxicity were accompanied by changes in drug metabolism, as evidenced by radiochromatograms of cell extracts obtained after treatment with 14C-labelled MISO. In contrast, radiochromatograms obtained from cells treated with toxic levels of MISO (75 mM) under aerobic conditions indicated no drug metabolism. Both toxicity and drug metabolism could be immediately halted by introducing O2 during hypoxic exposure to MISO. These observations are discussed in terms of a possible model for the metabolism-mediated toxicity of MISO and the roles which sulphydryls and O2 may play.


Optimized functional results are difficult to achieve following hand injuries. This prospective study was undertaken to evaluate the functional outcome after surgical stabilization of metacarpal and phalangeal fractures.Forty-five fractures of digits of hand in 31 patients were managed by surgical stabilization. Five fractures were fixed with closed reduction and percutaneous Kirschner wire fixation; 10 with external fixator; 26 with open reduction and Kirschner wire fixation; and four with open reduction and plate and screw or screw fixation.Final evaluation of the patients was done at the end of three months. It was based on total active range of motion for digital functional assessment as suggested by the American Society for surgery of hand. Overall results were excellent to good in 87%. Better total active range of motion (excellent grade) was observed in metacarpal fractures (47%) versus phalanx fractures (31%); closed fractures (57%) versus open fractures (27%); and single digit involvement (55%) versus multiple digits (29%). Excellent total active range of motion was observed with all four plate and screw/ screw fixation technique (100%) and closed reduction and percutaneous kirschner wire fixation (60%). Twenty-two complications were observed in 10 patients with finger stiffness being the most common.Surgical stabilization of metacarpal and phalangeal fractures of hand seems to give good functional outcome. Closed fractures and fractures with single digit involvement have shown a better grade of total active range of motion. Fractures of metacarpals and phalanges constitute between 14-28% of all visits to the emergency department.Forty-five metacarpal and phalangeal fractures of the hand in 31 patients aged 14 yrs or more were included in the prospective study conducted during the period 2004-2005. Unstable metacarpal and phalangeal fractures, intraarticular fractures, avulsion fractures, fracture dislocations and open fractures with sharp and clean wounds were included in the study. An unstable fracture was defined as one in which the patient was able to move the adjacent joints by less than 30% of the expected normal range of motion.The fractures were reduced and fixed using various implants including Kirschner wires, external fixators, plates or screws depending on fracture site, configuration and associated soft tissue damage. The fractures were divided into four groups depending on the type of internal fixation.Group I (n=5): The closed reduction and percutaneous Kirschner wire fixation was done in patients with fractures of the middle phalanx, proximal phalanx and metacarpal when closed reduction was possible.Group II (n=10): Open/closed reduction and external fixation was performed for open fractures with sharp and clean wounds.Group III (n=26): Open reduction and Kirschner wire fixation was done in fractures where closed reduction was not possible Figures –4. RetroGroup IV (n=4): Here open reduction with plate and/or screw fixation was performed for long oblique or spiral fractures of both metacarpals and phalanges Figures –8. The aThe hand was kept elevated and patients were encouraged to perform movements of the fingers and hand to prevent edema and stiffness. Antibiotics were given for 48h and prolonged in cases of open fractures. The Kirschner wires and external fixation devices were removed between 3 to 6 weeks. Active assisted mobilization was started after removal of Kirschner wire / external fixator usually at three to six weeks. Wax bath and contrast bath were given. Continuous passive motion (CPM) was not used. Patients were followed weekly for the first month and fortnightly for the next two months before final evaluation at three months.The mean age of the patients was 35.6 years (range 14-74 years). The most common mode of trauma was assault and roadside accidents (60%). In all, 43 digits were involved and there were a total of 45 fractures. Twenty-two patients had only a single digit involvement while two digits were involved in six patients and three digits in three patients. Fracture site and configuration in metacarpal and phalanges is shown in The functional outcome after fracture treatment was assessed by calculating total active range of motion (TAM).Metacarpal fractures (n=32) had an excellent n=15, 47%); and good  TAM in 94% of fractures [Figures 5, 47%; aOverall end results of hand fractures in 31 patients managed by surgical stabilization were excellent in 14 (45.16%); good in 13 (41.93%); fair in three (9.68%); and poor in one(3.23%).A total of 22 complications were observed  in 10 paResults in various groups according to TAM are tabulated in Many factors, such as delicate handling of tissues, preservation of gliding planes for tendons, prevention of infection and early and appropriate physiotherapy other than accurate reduction and fixation affect recovery of good mobility.5et al.et al.A higher incidence of excellent TAM n=13, 57%) was observed in closed fractures as compared to open fractures n=6, 27%). Pun , 27%. Pu% was obset al.Closed reduction and percutaneous Kirschner wire fixation was done where fracture could be reduced and maintained by closed means. Of the five fractures treated, three achieved excellent TAM (60%). Belskey et al.et al.External fixator was used for surgical stabilization of 10 open fractures. Advantage of external fixation is that no joint transfixation is required and hence patient does not develop any stiffness of the adjacent joints. Sixty per cent (n=6) of the fractures treated by this technique achieved excellent to good TAM. The results are comparable to those reported by FreelandOpen reduction with Kirschner wire fixation was done for stabilization of 26 fractures using three different techniques. (a) Retrograde insertion of Kirschner wire (n=16) with transfixation of joint was first advocated by Vomsaal.Open reduction and plate fixation was done in two fractures and two patients were treated with screw fixation. Excellent TAM was obtained in all four fractures treated by these techniques. Bosscha and SnellenFinger stiffness was the most commonly observed complication in the present series. Only one case of open fracture developed superficial infection. None of the cases developed osteomyelitis. Two patients with multiple fractures developed angulation at fracture site as the Kirschner wire was removed four weeks postoperatively before any radiological union. The movements, however, were almost full at all joints. Two cases of multiple fractures developed hypertrophic nonunion after Kirschner wire fixation. The cause was thought to be inadequacy of Kirschner wire fixation, which cannot check movements at fracture site completely. Ikuta and Tsugeet al.The results of the present study following surgical stabilization of fractures of metacarpals (94% excellent to good) and phalanges (54% excellent to good) were observed to be superior to those reported by Duncan Limitation of the study: The sample size is very small in some groups for statistical comparison.Surgical stabilization of metacarpal and phalangeal fractures of the hand seems to give good functional outcome. Closed fractures and fractures with single digit involvement are important determinants to achieve a better grade of total active range of motion.


The concentration of bound sialic acid in the sera of 56 normal subjects and 65 subjects with breast cancer was measured, in order to determine (1) whether serum sialic acid concentrations are raised in breast cancer and (2) whether the concentration of sialic acid in serum reflects tumour stage. The amount of sialic acid in serum was compared to serum carcinoembryonic antigen (CEA) values. Urinary hydroxyproline and serum alkaline phosphatase concentrations were used as indicators of bone and liver involvement. Erythrocyte sedimentation rate (ESR) was also measured. Significantly elevated serum sialic acid concentrations were found in breast cancer, and showed correlation with tumour stage. Serum sialic acid values did not correlate with CEA values. The results suggest that measurement of serum sialic acid concentrations may be of adjunctive value in assessing tumour stage.


We have studied the incidence of microsatellite instability at three trinucleotide repeats and seven dinucleotide repeats from five chromosomal regions, in a group of 30 mammographically detected 'early' invasive breast cancers and correlated its occurrence with clinicopathological parameters. The myotonic dystrophy (DM-1) trinucleotide repeat was analysed in 48 additional cases. In 4 out of 78 (5%) paired tumour-normal DNA samples we found evidence of somatic microsatellite instability at DM-1: a novel allele of a different size was seen in the tumour DNA which was not present in the normal DNA sample. All four tumours that showed evidence of instability were from the core group of 30 cases (13%) and were well or moderately differentiated, oestrogen receptor-positive, infiltrating ductal carcinomas. Two of these tumours were unstable at nine of ten loci studied, both trinucleotide and dinucleotide repeats. DNA prepared from different normal tissues showed no evidence of instability, for all four instability cases. These data indicate that microsatellite instability is specific to the tumour DNA and is an early event in the genesis of some sporadic breast cancers.


To date, cytogenetic studies on pancreatic carcinoma are rare, and little is known about the frequency of cytogenetic aberrations in primary carcinomas compared with metastatic tumour cells. We therefore evaluated the frequency of chromosomal aberrations in 12 primary pancreatic carcinomas and in effusion specimens from 25 patients with pancreatic cancer by using interphase fluorescence in situ hybridization (FISH) and a panel of four centromeric probes. Hyperdiploidy and chromosomal imbalances, predominantly affecting chromosome 8, were a constant finding in metastatic effusion cells, whereas concordant gain of chromosomes or relative loss of chromosome 18 characterized primary pancreatic carcinomas. The potential role of oncogenes located on chromosome 8 for pancreatic cancer progression was further investigated by double-hybridization studies of aneuploid effusion cells with a probe to 8q24 (MYC) and a centromeric probe to chromosome 8, which demonstrated amplification of the MYC oncogene in two of ten cases (20%). Finally, a potential application of basic findings in the clinical setting was tested by searching for micrometastatic cells in effusions from pancreatic cancer patients primarily negative by FISH. Two-colour FISH in combination with extensive screening  seems to be a useful tool to unequivocally identify micrometastatic cells by demonstrating hyperdiploidy and intranuclear chromosomal heterogeneity.


Diabetic maculopathy, the leading cause of vision loss in patients with type 2 diabetes, is characterized by hyper-permeability of retinal blood vessels with subsequent formation of macular edema and hard exudates. The degree of hyperglycemia and duration of diabetes have been suggested to be good predictors of retinal complications. Intervention studies have determined that while intensive treatment of diabetes reduced the development of proliferative diabetic retinopathy it was associated with a two to three-fold increased risk of severe hypoglycemia. Thus we hypothesized the need to identify downstream glycemic targets, which induce retinal vascular permeability that could be targeted therapeutically without the additional risks associated with intensive treatment of the hyperglycemia. Betacellulin is a 32 kD member of the epidermal growth factor family with mitogenic properties for the retinal pigment epithelial cells. This led us to hypothesize a role for betacellulin in the retinal vascular complications associated with diabetes.A disintegrin and metalloproteinase (ADAM)-10 which plays a role in the cleavage of betacellulin was seen in the retinas of diabetic mice and humans.In this study, using a mouse model of diabetes, we demonstrate that diabetic mice have accentuated retinal vascular permeability with a concomitant increased expression of a cleaved soluble form of betacellulin (s-Btc) in the retina. Intravitreal injection of soluble betacellulin induced retinal vascular permeability in normoglycemic and hyperglycemic mice. Western blot analysis of retinas from patients with diabetic retinopathy showed an increase in the active soluble form of betacellulin. In addition, an increase in the levels of These results suggest that excessive amounts of betacellulin in the retina may contribute to the pathogenesis of diabetic macular edema. The incidence of diabetes world-wide, is expected to reach epidemic proportions by 2025. Progression of diabetic retinopathy often results in diabetic macular edema, which is a consequence of the breakdown of the blood-retinal barrier, increased retinal vascular permeability and leakage of plasma from small blood vessels in the macula leading to loss of central vision. While epidemiological studies have suggested that glycemic control plays a major role in the development of vascular complications of diabetesBetacellulin (BTC) is a 32 kD member of the epidermal growth factor (EGF) family that was originally isolated from the conditioned medium of a mouse pancreatic β-cell tumor lineRecombinant human VEGF was a kind gift from Genentech, CA and recombinant betacellulin was purchased from R&D. Antibodies: polyclonal anti-mouse and anti-human betacellulin antibody (R&D), monoclonal anti-ADAM-10  and HRP-conjugated anti-rabbit, anti-mouse IgG antibodies (Amersham Pharmacia Biotech) and anti-goat IgG (Santa Cruz). Post mortem human eyes were obtained from he National Disease Research Interchange (NDRI), Philadelphia. Postmortem time ranged from 3–10 hours.Lysates from mouse or human retina prepared using sonication on ice with RIPA buffer (Boston Bioproducts) containing EDTA free protease inhibitors (Roche) were subjected to SDS-PAGE. Proteins were probed with antibody and detected with either a HRP-conjugated anti-rabbit or anti mouse IgG antibody  followed by ECL. The blots were restripped with Restore™ solution (Thermo Scientific) for 30 minutes and reprobed as indicated.6–8 week old C57BL6 mice  were used in this study. The animals were cared for in accordance with the ARVO statement for the Use of Animals in Ophthalmic and Vision research. All experimental procedures used aseptic sterile techniques and were approved by the Animal Care and Use Committee of the Cleveland Clinic (ARC08792).6–8 week old mice were rendered diabetic with 3 consecutive daily intraperitoneal injections of STZ (55 mg/Kg) freshly dissolved in citrate buffer (pH 4.5). Control mice were given injections of buffer alone. Development of diabetes (defined by blood glucose greater than 250 mg/dl) was confirmed 1 week after the initial injection (One Touch®ltra®Test Strips and One Touch®UltraMini™glucometer).Vascular permeability in the retina was quantitated after 3–4 weeks of diabetes using an Evans blue quantitation technique as described previously Steptozotocin-induced diabetic mice with hyperglycemia (fasting blood sugar ≥200 mg/ml) were examined for increases in retinal vascular permeability using an Evans Blue leakage assay. As described previously To determine if the increase in soluble active betacellulin seen in diabetic mice could contribute to increased vascular permeability we examined whether increased concentration of betacellulin in the retina could induce retinal vascular leakage in normal mice. Intravitreal injections of increasing doses of recombinant soluble betacellulin (0–300 ng) resulted in increased retinal hemorrhage  and diabetic macular edema (DME), we wanted to determine if the effect of betacellulin was VEGF-dependent. No increase in VEGF was observed in mouse retinas injected with betacellulin  indicatiBetacellulin is expressed as a membrane anchored precursor protein (pro-betacellulin) containing an extracellular N-terminal ectodomain, a transmembrane domain and a cytoplasmic domainadisintegrin and metalloproteinase-10) can mediate the constitutive and activated shedding of pro-betacellulinRecent studies have determined that ADAM-10 , characterized by an absolute insulin deficiency as a consequence of destruction of the insulin secretory pancreatic β cells of the islets of Langerhans; and type II diabetes  and type II (30% incidence)ADAMs and ADAM-10 in particular have been recently reported to be novel regulators of vascular permeabilityWe have examined betacellulin levels in the plasma and found no significant increase in betacellulin in the circulating plasma of patients with PDR (n = 23) . While tNeurod-Btc gene therapy to be a promising approach to induce islet regeneration for the treatment of insulin-dependent diabetesRecent studies have suggested Figure S1Betacellulin induces retinal vascular leakage independent of VEGF. Retinas from mice injected intravitreally with PBS or betacellulin (200 ng) or no injection (control) were evaluated for VEGF protein 24 hours post injection.(0.74 MB TIF)Click here for additional data file.Figure S2No significant increase in circulating plasma betacellulin in patients with proliferative diabetic retinopathy. Plasma was prepared from whole blood from patients with no diabetes, diabetes with no evidence of retinopathy and proliferative diabetic retinopathy and analyzed by ELISA for betacellulin.(0.54 MB TIF)Click here for additional data file.


It is universally acknowledged that if cancer is to be controlled, prevention and down staging are essential. In the year 2000 about 10 million new cases were registered, while 6.3 million people died from cancer worldwide. Stamps are regarded as a very useful and educative tool in fighting cancer by creating awareness and raising money for treatment and research. This year (2003) is the seventy-fifth anniversary of the issue of the first anticancer stamps in 1928, so an up-to-date review of the field of oncophilately is timely. Cancer is posing a serious social and economic problem globally since it was estimated by various organisations including It is necessary to take effective preventive measures and cancer awareness among the general population is essential. It is universally recognised that the anticancer stamps can play an excellent role in various ways as to create awareness and to collect money for treatment and research. This is because philately or stamp collection is still one of the most popular hobbies. Due to their universal circulation and appeal, stamps are powerful and effective messengers. It is also possible to use stamps issued for some other purposes in describing various aspects of cancer as will be exemplified later on. Only one report  and two Five Swedish stamps on King Gustaf V issued in 1928 are regarded as the first anticancer stamps because the entire amount collected from sales was donated for cancer control (stamp 2). This was followed by a set of three Danish stamps issued in 1929 (stamp 3). Norway is the third country with a semipostal issue in 1931 (stamp 4) in the aid of Norwegian Radium Hospital. Since then, numerous countries have published several regular or semipostal stamps (stamps 5–8). Record shows that by the year 1964, 46 countries had issued 110 stamps  and by tStamps are available on lung cancer (stamp 13), which is the most common cancer in terms of both incidence  and mortality  in 2000 . It is tBreast cancer is the most prevalent cancer in the world among females  in 2000 . Many stFigure 1is stamp .Stamps have also been published on other cancers like prostate cancer . Passive smoking is also very dangerous. Evidence suggests that nonsmoking women married to smokers experience an excess risk of developing lung cancer in the order of 20%. A Japanese stamp may be used to illustrate this aspect (stamp 24).Excessive drinking causes liver, oral, pharynx, larynx and oesophageal cancers, and may increase the risks of colorectal and breast cancers. Simultaneous drinking and smoking habits are much more dangerous. Alcoholic beverages consumption potentiates the effects of tobacco smoking on cancers of the mouth, pharynx, oesophagus and larynx and has been estimated to account for about 3% of all cancer deaths. Stamps issued by many countries warn people about the use of alcohol (stamps 25 and 26).The British Surgeon Dr Percivall Pott demonstrated the earliest example of occupational cancer in 1775, showing a greater incidence of scrotal cancer among young chimneysweepers who were exposed to tar. Lord Shaftesbury, the great social reformer, worked for the well-being of these poor lads. A British stamp is available in his memory, depicting the hand of a chimneysweeper holding the brush (stamp 27). The stamp showing a picture of aniline (stamp 28) illustrates the need for care to be taken to minimise exposure to this chemical, because as early as in 1895, Rehn was the first to observe increased rate of bladder cancers among aniline dye workers. The stamp 29 describing the structure of benzene may be similarly used to show the need to take care in handling benzene, since it is proved to be a potent human carcinogen causing leukaemia with a latency period of about 15–20 years. It is widely used as a solvent in leather, petroleum and other industries. Polyaromatic hydrocarbons (PAH) are found in exhaust gases from automobiles run by petrol and diesel, from railway locomotives and in smoke, dust and exhaust gases from various factories. It is advisable to use protection to prevent inhaling of exhaust gases from automobiles as described in a Swedish stamp (stamp 30).92235U (stamp 31) and development of cancer dates from early observations on survivors of the atomic bombs in Japan.The relation between ionising radiation from various sources including radioactive substances such as Ultraviolet radiation from sunlight is the major risk factor for skin cancers and that of lip. Stamps showing sunrays and flares (stamp 32) may be used to make people aware of the dangers of overexposure to sunrays.Figure 3Links have already been established between some viral infections and the development of cancers such as Epstein–Barr virus and Burkitt lymphoma, hepatitis B and C and hepatocellular (liver) carcinoma, human papilloma virus (HPV) and cervical cancer in females, human immunodeficiency virus (HIV). AIDS patients infected with HIV have higher incidence and death rates from malignancies like Kaposi's sarcoma, non-Hodgkin's lymphoma, etc. Hence awareness regarding HIV infection is very important from a cancer preventive perspective also. So far about 65 countries have issued over 120 anti-AIDS stamps (e.g. stamps 33–36).Catharanthus rosea) may be used to point out that some important alkaloidal drugs like vincristine and vinblastine have been isolated from the plant (stamp 40). Taxol has been isolated from Taxus brevifolia (Pacific Yew tree) and Taxus baccata  (stamp 41).Figure 4Nowadays, three chief treatment modalities – surgery, radiotherapy and chemotherapy – are used in all sorts of combinations. Bangladesh has issued a beautiful stamp depicting the picture of a crab attacked with these three modes of treatment (stamp 37). One can find individual stamps on surgery (stamp 38), radiotherapy (stamp 39) and chemotherapy. Anticancer drugs are obtained by chemical synthesis or from natural sources such as plants. A French stamp on the Madagascar periwinkle plant  and using ‘smokeless’ tobacco products like crude leaves, snuff, etc.Limit alcohol consumption to a minimum, if at all (stamps 26 and 43).β-Carotene present in various yellow, green or orange fruits and vegetables is converted to vitamin A in the liver. Hence it is recommended to eat fresh vegetables and fruits rich in these substances and vitamins like carrot, beans, gourd, broccoli, cauliflower, tomato, etc. (stamp 47) and a variety of fruits (stamp 48).Both vitamin A (stamp 45) and vitamin C (stamp 46) are considered as antioxidants and possess potential cancer chemopreventive properties by inhibiting precancerous changes and damages of DNA leading to cancer. The protective effects of dietary fibre present in vegetables against a number of cancers like colorectal, oesophageal and stomach have been demonstrated. Hence it is advisable to eat vegetables rich in roughage like jackfruit (stamp 49), beans (stamp 47), ladies finger, etc.Curcuma longa (stamp 50) is the source of turmeric, which contains the yellow natural colouring agent namely curcumin which may have chemopreventive properties  have been shown in lab studies to have antitumour as well as tumour promotion inhibitory activities. Onions and ginger have also been shown to possess similar properties.Constituents of Camellia sinensis (stamp 52). Tea is one of the most ancient and next to water the most widely consumed liquid in the world. Many phytochemicals including polyphenols catechin compounds like epigallocatechin gallate (EGCG), etc. present in tea leaves have shown potential chemopreventive and antioxidant properties.Tea is a beverage made from the leaves of the plant species Figure 5Diet has a vital role in the prevention of about 30–40% of all cancers (Being very underweight or overweight increases the risk of cancer. Obesity (stamp 53) is associated with the endometrium, breast, kidney, colorectal and gallbladder cancers. Thus it is advisable to maintain a healthy body weight.An active lifestyle is recommended. Physical activity decreases the risk of colon and breast cancers. Walking, cycling, gardening, household chores, climbing the stairs, practising yoga (stamp 54), etc. are recommended.Early detection of cancer is very important to save lives (stamp 55). Annual check-up (stamp 56) of oral cavity, skin, lymph nodes, routine examinations of blood for detection of any abnormalities in its constituents (stamp 57) and chest X-ray (stamp 58) are internationally accepted tests  for earlFor women: Breast awareness is important (Indian stamp 59) and clinical breast examination by a physician may be helpful where there is no national screening programme. It is also suggested to carry out mammography (stamp 60). Cervical PAP test (stamps 61 and 62) should be carried out in adult females at regular intervals.For men: Population-based prostate-specific antigen (PSA) testing can reduce mortality in countries where urological follow-up and treatment are available. US Postal Service issued a semifiscal stamp in 1998 on this aspect , electron microscope (stamp 64), X-ray instruments (stamp 65), radiotherapy instruments (stamps 66 and 67), ultrasonography (USG) (stamp 68), computed topographic (CT) scanning (stamp 69), magnetic resonance imaging (MRI) (stamp 70), nuclear medicine imaging (scintigraphy) (stamp 71), mammography, etc. In 1992, Macedonia had brought out a set of four stamps with the message of early cancer detection in one of them and with the picture of diagnostic equipment in the other three (stamps 72–75).Many scientists and physicians appear on stamps for their outstanding contribution in the fight against cancer. WC Roentgen (1845–1923) – discoverer of X-rays in 1895 and the first recipient of the Nobel prize in 1901 (stamp 76). Henri Becquerel (1852–1908) (stamp 77) – discoverer of the phenomenon of spontaneous emanations later termed radioactivity by Madame Marie Curie. Radiotherapy has originated from this discovery. Madame Marie Curie (1867–1934) and Pierre Curie (1859–1906) – world-renowned French scientists who discovered radium in 1898, which was the first radioactive substance used for the treatment. Madame Curie appeared in about 100 stamps either alone  or with her husband Pierre Curie (stamps 81 and 82). In 1938, France and its 21 colonies issued the latter stamps (81 and 82) as a tribute to the 40th anniversary of this discovery. The surtax of these stamps was given to the International Union Against Cancer. Pierre Curie was also remembered alone in some stamps. Irene Joliot Curie (1897–1956), elder daughter of Madame Curie, and Frederic Joliot (1900–1958) son-in-law of Madame. Curie – joint discoverers of artificial radioactivity (stamp 83). Based on their discovery, a number of artificial radioisotopes have been prepared some of which find use in cancer. George N Papanikolaou (1883–1962) (stamps 62 and 84) – discoverer of PAP test for early diagnosis of cervical cancer that has saved the lives of millions of women worldwide. Moritz Kaposi (1837–1902) (stamp 85) – famous physician who identified in 1872 a special type of sarcoma that was named after him as Kaposi's sarcoma. Rudlof Virchow (1821–1902) (stamp 86) – founder of cellular pathology. He discovered and named gliomas and giant cells. He had also named many common tumours.Postage stamps and cancellations are available on world-famous organisations like International Union Against Cancer (UICC) (stamps 81 and 82), World Health Organization (WHO) (stamp 87), etc.Starting from 1933, UICC organises International Cancer Congress every fourth year where scientists, clinicians, nurses, etc. from all over the world meet. Special commemorative stamps have been issued on these occasions by various countries: 8th held in Russia in 1962 (stamp 88), 9th held in Japan in 1966 (stamps 39 and 89), 12th held in Argentina in 1978 (stamp 90), 14th held in Hungary in 1986 (stamp 85) and 16th held in India in 1994 (stamp 91).Thus our fight against cancer will continue till we achieve success (stamp 92). Research is ongoing globally, and newer concepts, improvements in therapeutic modalities and new drugs are being added continuously. Recent global efforts in DNA decoding (stamp 93) and human genome mapping leading to genome genetic engineering (stamps 94 and 95) will help to identify and repair the defective genes (gene therapy of cancer). The message that should reach everybody is that cancer will be defeated – sooner or later – as is beautifully demonstrated in a French stamp issued in 1970 picturing a smiling lady cured of cancer overcoming the threat of death displayed underneath (stamp 96).It is therefore convincingly demonstrated that, in addition to specific anticancer stamps, various other stamps can be used to describe different aspects of cancer. The significant role of stamps in raising awareness among people regarding this dreadful disease has been highlighted.


The ultrastructure of 16 tumours derived from spontaneously transformed cell lines established from young and old C57 and C3H mouse organs is described. Three types of tumour were found: myxoid (fibrosarcomatous), consisting of cells with long processes and much interstitial material; leiomyomatous, consisting of a bundle of smooth muscle-like cells with less interstitial material; or epithelial-like consisting of closely packed round cells with little interstitial material. The cell types in the tumours were similar to those found in the tissue culture cell lines from which they were derived.


The Src family kinases possess two sites of tyrosine phosphorylation that are critical to the regulation of kinase activity. Autophosphorylation on an activation loop tyrosine residue (Tyr 416 in commonly used chicken c-Src numbering) increases catalytic activity, while phosphorylation of a C-terminal tyrosine (Tyr 527 in c-Src) inhibits activity. The latter modification is achieved by the tyrosine kinase Csk , but the complete inactivation of the Src family kinases also requires the dephosphorylation of the activation loop tyrosine. The SH3 domain of Csk recruits the tyrosine phosphatase PEP, allowing for the coordinated inhibition of Src family kinase activity. We have discovered that Csk forms homodimers through interactions mediated by the SH3 domain in a manner that buries the recognition surface for SH3 ligands. The formation of this dimer would therefore block the recruitment of tyrosine phosphatases and may have important implications for the regulation of Src kinase activity. Members of the Src family of protein tyrosine kinases, such as c-Src, Lck, Fyn and Hck, are key players in many signalling pathways that regulate cell growth, proliferation and motility The kinase domain of Csk is responsible for the recognition of the Src family kinases as its specific substrates, resulting in phosphorylation of the C-terminal tail The SH3 domain of Csk interacts with several tyrosine phosphatases Pursuing the observation that Csk is dimeric at high protein concentration  in vitro FL) migrates as a dimer by size exclusion, the removal of the SH3 domain (CskSH2KD) or the SH3 and SH2 domains (CskKD) resulted in proteins that no longer appear to dimerize and instead elute as apparent monomers by gel filtration , CskSH2KD (residues 68–450) and CskKD (residues 187–450) were expressed in bacteria and purified as described The constructs of human Csk, CskPurified Csk proteins were subjected to analytical gel filtration using a Superdex S200 column  or SMART S200 column . Column calibration was performed using molecular weight standards .


We analysed oestrogen receptor (ER) and progesterone receptor (PR) expression in a retrospective series of 21 low-grade endometrial stromal sarcomas (LGSSs). Archival formalin-fixed and paraffin-embedded material was analysed by immunohistochemistry. ER and PR were measured with monoclonal antibodies and the peroxidase-antiperoxidase method and a score was calculated as for breast carcinoma based on both the percentage of positive tumour cell nuclei and the staining intensity. ER were seen in 15 (71%) and PR in 20 (95%) of tumours respectively. ER expression was scored as high in three (14%), moderate in four (19%), and low in eight (38%) tumours. Six (29%) tumours did not stain for ER and all of these were positive for PR. PR expression was scored as high in eight (38%), moderate in ten (47%) and weak in two (10%) LGSSs. Only one (5%) LGSS did not stain for PR (this tumour was positive for ER). ER and PR expression in LGSS is heterogeneous. This may have implications for hormone therapy in the management of these tumours. These results suggest that ER and PR should be routinely quantified in LGSSs by immunohistochemical methods. © 2000 Cancer Research Campaign


The objective of the study was to review the natural history of patients with a malignant pleural effusion but without obvious evidence of a primary, to assess the value of investigations used to look for a primary and to assess the response to palliative chemotherapy. This was done by a retrospective study of patients' notes at the Lung Unit, Royal Marsden Hospital, Sutton, Surrey. Improvement in tumour-related symptoms (and duration) on chemotherapy was assessed by the patient before the first course of chemotherapy and following each course using simple descriptive criteria as follows: (1) complete disappearance of symptoms (CR); (2) good improvement in symptoms (PR); (3) minor or no change in symptoms (NC); (4) worse symptoms (PD). Pleural effusion objective response (and duration) according to Hamed definition: success defined as a continued absence of reaccumulation of pleural fluid on all follow-up radiographs; any reaccumulation was regarded as a treatment failure. Overall survival was measured from the date of histological/cytological diagnosis to death. The study included 42 patients, 27 males and 15 females with a median age of 55 years. A primary was found in 15 patients (36%), and considered to be lung cancer. A total of 11/32 (34%) had a thoracic computed tomography (CT) scan with abnormalities compatible with a diagnosis of lung primary. When thoracic CT scan was negative, fibre optic bronchoscopy was always negative (0/13). Abdominal and pelvic CT scan, abdominal ultrasound, pelvic ultrasound and mammograms failed to reveal the primary. Twenty-three patients underwent local treatment and 37 received systemic chemotherapy. A total of 29/37 (78%) patients achieved symptomatic improvement  and 32/37 (86%) an objective response of their pleural effusion on chemotherapy . The median survival of the whole group was 12 months (3-60+ months). In this series the thoracic CT led to a diagnosis of lung primary in 34% of the cases. Other radiological examinations and bronchoscopy were unhelpful. Chemotherapy achieved symptom relief in 78% of patients.


Analysis of human tumour-derived cell lines has previously resulted in the identification of novel transformation-related elements and provided a useful tool for functional studies of different genes. To establish the utility of such cell lines as indicators of change relevant to urothelial cancer, we have characterised the expression of five genes  within a panel of human bladder carcinoma cell lines. Using single-strand conformation polymorphism (SSCP) and direct sequencing, p53 mutations were identified in 7/15 (47%) cell lines reflecting events reported in bladder tumours. Immunohistochemical analysis of p53 in cultured cells and in paraffin-embedded sections of xenografts from the cell line panel revealed discordant results. An absence of p53 nuclear staining was associated with an exon 5 mutation in EJ and with multiple p53 mutations found in J82. Two cell lines positive for p53 staining in the absence of detectable mutation displayed overexpression of MDM2  in Western blot analysis. Loss or aberrant Rb expression was recorded in 5/15  cell lines. Absence of E-cadherin was recorded in 5/15 cell lines  with loss of alpha-catenin in immunoprecipitated E-cadherin complexes of CUBIII. Western blot analysis of APC revealed a truncated protein in 1/15 (CUBIII) cell lines. The characterisation of oncogenic events within this panel of human bladder carcinoma cell lines establishes a representation of change observed in bladder tumours and better defines the genotypic background in these experimental human cell models of neoplastic progression.


Two human deaths caused by Australian bat lyssavirus (ABL) infection have been reported since 1996. Information was obtained from 205 persons (mostly adults from south Brisbane and the South Coast of Queensland), who reported potential ABL exposure to the Brisbane Southside Public Health Unit from November 1,1996, to January 31, 1999. Volunteer animal handlers accounted for 39% of potential exposures, their family members for 12%, professional animal handlers for 14%, community members who intentionally handled bats for 31%, and community members with contacts initiated by bats for 4%. The prevalence of Lyssavirus detected by fluorescent antibody test in 366 sick, injured, or orphaned bats from the area was 6%. Sequelae of exposure, including the requirement for expensive postexposure prophylaxis, may be reduced by educating bat handlers and the public of the risks involved in handling Australian bats.


Furthermore, strong correlations were found between the levels of both uPA  and PAI-1  in the cytosols and their levels previously measured with ELISAs based on commercial reagents. In both Cox univariate and multivariate analysis, high cytosolic levels of uPA or PAI-1 were significantly associated with increased rates of relapse and death. The levels of uPA and PAI-1 in the pellet extracts also provided prognostic information, although to a lesser extent compared with the cytosolic extracts. The prediction of prognosis on the basis of uPA and PAI-1 assessed by an alternative ELISA once again emphasizes the established prognostic role and usefulness of these parameters in selection of breast cancer patients at high or low risk of recurrence. © 1999 Cancer Research CampaignTo evaluate the clinical relevance of urokinase-type plasminogen activator (uPA) and its type-1 inhibitor (PAI-1) measured by a recently developed enzyme-linked immunosorbent assay (ELISA), we analysed both components in samples derived from 892 patients with primary breast cancer (median follow-up 99 months). The assays were performed in cytosolic extracts as well as in corresponding detergent extracts of pellets obtained after ultracentrifugation, which was carried out when preparing the cytosolic fractions for routine steroid hormone receptor determination. Statistically significant correlations were found between the cytosolic levels and those determined in the pellet extracts (Spearman correlation coefficient


Recent studies have shown that various gastrointestinal tumours express substantial amounts of vasoactive intestinal peptide (VIP) receptors. Based on these observations, we have developed a receptor scintigraphy using [123I]VIP as a radioligand. An initial series performed at our institution showed promising potential for visualization of various gastrointestinal adenocarcinomas by means of [123I]VIP. In this article, we now report the results obtained in 80 consecutive patients with colorectal adenocarcinoma. Eighty consecutive patients with histologically verified colorectal cancer underwent scanning by means of [123I]VIP . Thirteen patients were free of tumour after complete resection of Dukes' C cancer, eight patients presented with primary and 14 with locally recurrent tumours but were free of metastases. Ten patients had locally recurrent disease and liver, lung or lymph node metastases. Disease confined to organ metastases  was present in 35 patients. The size of the primary or recurrent tumours ranged between 3 and 6 cm, and the size of metastases was between 1 and 13 cm in diameter. Scan results were evaluated independently by two nuclear medicine physicians in a blinded way, and results were then compared with computerized tomography (CT)scans not older than 4 weeks. Seven out of eight primary (87%) and 21 out of 24 (82%) locally relapsing cancers were imaged with [123I]VIP. Negative VIP scans were obtained in all 13 patients in whom the cancers had been curatively resected. All patients with lymph node metastases showed positive VIP scans (four out of four), and positive scans were obtained in 25 out of 28 (89%) patients with liver metastases and in two out of three cases with lung metastases. In four patients with relapsing cancer, the VIP scan indicated the presence of disease before CT, and in two patients the diagnosis of scar tissue instead of a local recurrence of rectal cancer as suggested by CT could be established. We conclude that [123I]VIP receptor scanning is a sensitive method for radioimaging of colorectal cancer with the potential to provide valuable additional information to conventional radiological methods.


To study pathophysiological roles of mesothelial barrier functions in protection against cancer cell invasion, we isolated mesothelial cells from the rat abdominal cavity and then cultured them with 10(-6)M all-trans-retinoic acid (RA) for 10 days. Mesothelial barrier function assessed by measuring transcellular electrical resistance (TER) and the expression of 7H6 tight junction-associated antigen at the cell border were induced by the treatment . Then we quantified the attachment and penetration of rat mammary cancer cells (SST-2 cells) into the mesothelial cell monolayer by prelabelling of the cancer cells with fluorescent dye and by observing optical sections at different heights using a laser confocal scanning microscope. When SST-2 cells were overlaid onto the mesothelial cell monolayer treated with RA, the number of cancer cells found at the basal level of the monolayer was significantly reduced. These results showed that enhanced mesothelial barrier function at least partially prevents the penetration of cancer cells into mesothelial cells and suggested that 7H6 antigen serves as a reliable immunocytochemical marker for monitoring mesothelial barrier function.


Anthracenyl-amino acid conjugates represent a novel chemical class of topoisomerase (topo) inhibitor. NU/ICRF 505 is a lead compound that stabilises topo I cleavable complexes and is actively cytotoxic at low microM concentrations. In this study, endonucleolytic DNA cleavage was used as a marker of apoptosis to investigate mechanisms of cell death produced by this compound. NU/ICRF 505 (5 microM) induced a substantial increase in the level of DNA fragmentation in HL60 cells  but only after a 48 and 72 h drug exposure (compared with 6 h after treatment with camptothecin), as determined qualitatively by conventional gel electrophoresis and quantitatively by spectrofluorimetry. This effect was substantially reversed by co-treatment with zinc (1 mM). Subsequent studies with the human lung (NX002), ovarian (A2780) and colon (HT29) cancer cell lines yielded evidence of formation of higher molecular weight DNA fragments in NX002 and A2780 cells in response to NU/ICRF 505 (5 microM). Co-treatment with zinc (1 mM) caused a small decrease in DNA fragmentation. These data suggest that the induction of apoptosis may play an important role in the mechanism of cytotoxicity of NU/ICRF 505 in HL60 cells and that other pathways of cell death may also be operative in NX002 and A2780 in conjunction with apoptosis.


The relationship between the bioenergetic status of human glioma xenografts in nude mice and morphometric parameters of the perfused vascular architecture was studied using (31)P magnetic resonance spectroscopy (MRS), fluorescence microscopy and two-dimensional digital image analysis. Two tumour lines with a different vascular architecture were used for this study. Intervascular distances and non-perfused area fractions varied greatly between tumours of the same line and tumours of different lines. The inorganic phosphate-nucleoside triphosphate (P(i)/NTP) ratio increased rapidly as mean intervascular distances increased from 100 microm to 300 microm. Two morphometric parameters - the percentage of intervascular distances larger than 200 microm (ivd200) and the non-perfused area fraction at a distance larger than 100 microm from a nearest perfused vessel (area100), - were deduced from these experiments and related to the P(i)/NTP ratio of the whole tumour. It is assumed that an aerobic to anaerobic transition influences the bioenergetic status, i.e. the P(i)/NTP ratio increased linearly with the percentage of ivd200 and the area100.


The characteristics of lung cancers induced by inhaled chromate were studied in 13 consecutive autopsies on male ex-chromate workers. In addition to histopathology, we examined: (1) the relationship between the occurrence of lung cancer and the amount of chromium (Cr) deposited in the lung as determined by atomic absorptiometry and (2) the chronological changes in five precancerous lung lesions followed by bronchoscopy till death. Twenty-one cancers were identified, including 16 lung tumours observed either during follow-up or at autopsy. Of these 16 tumours, 13 were found in six subjects, implying a high frequency of multiple cancers. Eleven (69%) out of the 16 tumours were of squamous cell type (including carcinoma in situ), this being twice as frequent as in age-matched controls. A further characteristic was predominance in the central part of lung (69%). The lung Cr burden was very much higher  in patients with lung tumours than in those without (8-28 micrograms g-1). Five of the precancerous lesions followed by bronchoscopy originated at bronchial bifurcations. Four of these cases showed a return to normal histology at autopsy even without therapy, and the other did not progress.


Background: Morbidity and mortality after surgical treatment of bileduct stones increase with age and associated diseases. A proposed alternative therapy is endoscopic sphincterotomy (ES) with the gallbladder left in situ, and we elected to compare this option with standard open surgery in high-risk patients.Methods: 98 patients (mean age 80 years) with symptoms likely to be due to bileduct stones or a recent episode of biliary parcreatitis were randomised to be treated either by open cholecystectomy with operative cholangiography and (if necessary) bileduct exploration (n=48) or by endoscopic sphincterotomy alone (n=50).Findings: The procedure was accomplished successfully in 94% of the surgery group and 88% of the ES group, and there were no significant differences in immediate morbidity (23% vs 16%) or mortality (4% vs 6%). During mean follow-up of 17 months biliary symptoms recurred in three surgical patients, none of whom underwent repeat surgery, and in 10 ES patients, seven of whom had biliary surgery. By multivariate regression analysis endoscopic sphincterotomy was an independent predictor of recurrent biliary symptoms .Interpretation: In elderly or high-risk patients, surgery is preferably to endoscopic sphincterotomy with the gallbladder left in situ as a definitive treatment for bileduct stones or non-severe biliary pancreatitis.


Microsatellite instability (MSI) was examined at 36 loci, and found in 9 (43%) of the 21 prostatic cancers. A loss of heterozygosity had occurred in five cases (24%). MSI did not correlate with clinical stage, but might play a role in the development of a subset of prostate cancers.


Development of incisional hernia after open abdominal surgery remains a major cause of post-operative morbidity. The aim of this study was to determine the current practice of surgeons in terms of access to and closure of the abdominal cavity in elective open surgery.Twelve surgical departments of the INSECT-Trial group documented the following variables for 50 consecutive patients undergoing abdominal surgery: fascial closure techniques, applied suture materials, application of subcutaneous sutures, subcutaneous drains, methods for skin closure. Descriptive analysis was performed and consensus of treatment variables was categorized into four levels: Strong consensus >95%, consensus 75–95%, overall agreement 50–75%, no consensus <50%.157 out of 599 patients were eligible for analysis (85 (54%) midline, 54 (35%) transverse incisions). After midline incisions the fascia was closed continuously in 55 patients (65%), using slowly absorbable , braided  sutures with a strength of 1 . In the transverse setting the fascia was closed continuously in 39 patients (72%) with slowly absorbable  braided sutures  with a strength of 1 .In the present evaluation midline incision was the most frequently applied access in elective open abdominal surgery. None of the treatments for abdominal wall closure (except skin closure in the midline group) is performed on a consensus level. Approximately 700.000 open abdominal procedures are performed annually in Germany and 4.000.000 in the United States . DevelopThere is still controversy about the best strategy for abdominal wall closure due to inconsistent and incomplete evidence provided by several randomized controlled trials (RCT) -8 as wel1. Is midline incision still the most frequently applied access to the abdominal cavity in elective situations?2. Has consensus for abdominal wall closure already been established in surgical routine?The primary objective of this cross sectional study was therefore to identify current practice of abdominal cavity access and closure in elective surgery.Twelve of the 25 participating centres of the INSECT-Trial group agreed in contribution to data acquisition (see acknowledgment). The local ethics committees of participating centres granted ethical approval for the study These include hospitals of all levels of health care , thus ensuring representative study results. Surgeons at these hospitals were asked to assess surgical access and closure methods of 50 consecutive patients who underwent surgery at their service. Patients who underwent more than one operation during the study period were excluded.The study was designed as a cross-sectional cohort study and performed between July and August 2005. A standardized data entry form was designed in order to collect the following information: patient's gender and age, setting of surgery (elective vs. emergency surgery), surgical approach (open vs. laparoscopic surgery), history of previous abdominal surgery, type of applied abdominal incision (transverse vs. midline), suture material used for fascia closure , the technique used for fascia closure , the use of subcutaneous suture and subcutaneous drainage, and the method of skin closure. Classification of suture strengths was reported according to the United States Pharmacopeia XXII (1990).All data entry forms were sent in an anonymous fashion to the Study Centre of the German Surgical Society and analyzed accordingly using Statistical Analysis System (SAS) version 9.1. Descriptive statistics were used in univariate analyses.The treatment of patients were categorized into four levels of consensus according to a classification of Hoffmann et al. :1. Strong consensus: treatment applied in >95% of cases2. Consensus: treatment applied in 75–95% of cases3. Overall agreement: treatment applied in 50–75% of cases4. No consensus: treatment applied in <50% of casesA total of 599 patient data entry forms out of 600 with an equal distribution of genders  and a mean age of 57 years (SD 19 years) were available for analysis. An open approach was chosen in 375 patients (63%), a laparoscopic in 224 patients (37%). After exclusion of patients with previous open abdominal surgery, an emergency procedure, and laparoscopic procedures, a total of 157 patients with primary elective laparotomy were available for further analysis , midline incisions were performed in 85 patients (54%), transverse incisions (crossing the midline) in 54 patients (35%) and miscellaneous incisions  in 13 patients (8%).Fascia closure for midline incisions was performed with a continuous suture in 55 patients (65%) and interrupted sutures in 16 patients (19%), respectively. In 13 (15%) of these patients a combination of both techniques was applied. Monofilament sutures were applied for fascia closure in 51 patients (60%), braided sutures in 32 patients (38%). Non-absorbable sutures were chosen in four cases (5%), slowly absorbable sutures in 47 patients (55%), and moderately rapid absorbable sutures in 33 patients (39%). Sutures with the strength of 0 were used in two patients (2%), with strength of 1 in 48 patients (57%), and with strength of 2 in 31 patients (36%).Subcutaneous suture was performed in 48 patients (57%) and a subcutaneous drain was placed in 19 patients (22%). Skin closure of patients who received midline incisions was performed with staples in 67 cases (79%).In terms of consensus levels all technical features for closure of midline incisions met the level of "overall agreement", except the item of skin closure for which the level of "consensus" was reached Table .Abdominal fascia of patients who had a transverse incision (n = 54) was sutured continuously in 39 patients (72%), interruptedly in eleven patients (20%), and with a combination of both techniques in two patients (4%). Suture material with the strength was 0 were chosen in 6 patients (11%), with a strength of 1 in 30 patients (56%) and with a strength of 2 in 17 patients (31%). Monofilament sutures were applied for fascia closure of 25 patients (46%) and braided sutures for 27 patients (50%). In two patients (4%) non-absorbable sutures, in 22 patients (41%) slowly absorbable sutures, in 29 patients (54%) moderately rapid absorbable sutures, and in one patient (2%) rapidly absorbable sutures were applied.Thirty-nine patients (72%) received a subcutaneous suture; a subcutaneous drain was placed in 12 patients (22%). Skin closure of patients who received midline incisions was performed with staples in 39 cases (72%).All technical features for closure of transverse incisions reached the level of "overall agreement" Table .In the present analysis midline incision is still the most frequent access to the abdominal cavity in elective surgery. No consensus for fascia closure either for midline or transverse incisions in daily practice of abdominal surgery was detected (>75% of surgeons acting similarly). Use of staples for skin closure after midline incisions was the only item that reached the level of consensus.Our finding that midline incisions are the most frequently applied access in open abdominal surgery is in accordance with previous reports . While tThe lack of consensus for abdominal wall closure strategies after midline incisions demonstrates persistent uncertainty within the surgical community. Several RCTs -8 and meAccording to our data, there is no consensus in closure of transverse incisions either. Closure technique of the abdominal fascia with a continuous suture and skin closure with staples were the only items that almost reached the level of consensus (both performed in 72%). In contrast to midline incisions, however, so far no randomized trial has been solely conducted in order to investigate closure techniques in this setting. Hence, the ideal closure strategy is still to be determined. From some trials comparing closure techniques in midline incisions ,18,19 paThe results of our study might be affected due to bias. The participating hospitals of the INSECT-Trial who agreed to contribute to this study may be a subgroup of hospitals dedicated to clinical research and therefore act differently to other hospitals (e.g. surgeons agree to randomize their patients). However, it is unlikely that such an effect might have systematically shifted the results in favour of consensus.A cross-sectional consecutive data collection of real patient data seems to be the optimal design to answer our initial study questions. Selection bias within the hospitals is prevented due to a consecutive recruitment and measurement bias due to standardized data collection. Furthermore, surgeons of the INSECT-Trial group were trained in data collection during specific preparatory INSECT-Trial meetings . Other sCurrently, no clinical guidelines are available for abdominal wall closure after any type of incision. Further well designed pragmatic randomized controlled trials are necessary to support a development of such evidence-based guidelines and to change the current practice. Otherwise a reduction of incisional hernias seems unfeasible and treatment remains on the level of "overall agreement". The results of the recently published INSECT-Trial certainly contribute to the body of evidence on abdominal fascia closure techniques. FurtherIn conclusion, midline incisions were the predominant type of abdominal access in the present study of participating centers of the INSECT-Trial. There is no consensus regarding the strategy for closure of the abdominal fascia after midline and transverse incisions.The authors declare that they have no competing interests.NNR: Study design, data analysis and interpretation, drafting of the manuscript PK: Study design, data acquisition, data analysis, revsion of the manuscript. MKD: Data acquisition, analysis and interpretation, revsion of the manuscript. CS: Study design, data acquisition, data interpretation, revision of the manuscript. KR: Study design, data acquisition, data interpretation, revision of the manuscript. HS: Study design, data acquisition, data interpretation, revision of the manuscript. CMS: Study design, data acquisition, data analysis and interpretation, drafting of the manuscript.The pre-publication history for this paper can be accessed here:


In the recent past, compliance with 21CFR11 has become a major buzzword within the pharmaceutical and biotechnology industries. While commercial solutions exist, implementation and validation are expensive and cumbersome. Frequent implementation of new features via point releases further complicates purchasing decisions by making it difficult to weigh the risk of non-compliance against the costs of too frequent upgrades. This presentation discusses a low-cost interim solution to the problem. While this solution does not address 100% of the issues raised by 21CFR11, it does implement and validate: (1) computer system security; (2) backup and restore ability on the electronic records store; and (3) an automated audit trail mechanism that captures the date, time and user identification whenever electronic records are created, modified or deleted. When coupled with enhanced procedural controls, this solution provides an acceptable level of compliance at extremely low cost.


The transcription factor, milk protein binding factor (MPBF/Stat5), is a member of the STAT family of signalling molecules which mediates prolactin signal transduction in lactating mammary gland by binding to GAS (gamma-interferon activation site) DNA elements. We have determined the levels of STAT factors in nuclear extracts from a variety of human breast tissues including carcinoma and normal 'resting' breast by electrophoretic mobility-shift assay. The results show that the level of STAT binding activity is low in normal 'resting' breast and benign lesions while carcinoma samples have significantly higher (P < 0.01) amounts of STAT binding activity. Supershift analysis suggests that Stat1 and possibly other members of the STAT family of signalling factors, including Stat3, are activated in breast cancer tissues.


The main results of these studies can besummarized as follows: (1) exogenous or endogenous catecholamines suppress PBL functions through alpha-2-receptor-mediatedmechanisms, lymphocytes of the spleen are resistant to adrenergic in vivo stimulation,(2) direct or indirect cholinergic treatment leads to enhanced ex vivo functions of splenic and thymic lymphocytesleaving PBL unaffected, (3) cholinergic pathways play a critical role in the “talking back” of the immune system to the brain,(4) acetylcholine inhibits apoptosis of thymocytes possibly via direct effects on thymic epithelial cells, and maythereby influence T-cell maturation, (5) lymphocytes of the various immunological compartments were found to beequipped with the key enzymes for the synthesis of both acetylcholine and norepinephrine, and to secrete theseneurotransmitters in culture supernatantsFor several years, our group has been studying the


Autoimmunity and inflammation are associated with marked changes in lipid and lipoprotein metabolism in SLE. Autoantibodies and cytokines are able to modulate lipoprotein lipase (LPL) activity, a key enzyme in lipid metabolism, with a consequent “lupus pattern” of dyslipoproteinemia characterized by elevated levels of very low-density lipoprotein cholesterol (VLDL) and triglycerides (TG) and lower high-density lipoprotein cholesterol (HDL) levels. This pattern favors an enhanced LDL oxidation with a subsequent deleterious foam cell formation. Autoantibodies and immunocomplexes may aggravate this oxidative injury by inducing accumulation and deposition of oxLDL in endothelial cells. Drugs and associated diseases usually magnify the close interaction of these factors and further promote the proatherogenic environment of this disease.


The isoenzyme Mu of glutathione S-transferase (GSTM1) is dominantly inherited, and the prevalence of this isoenzyme in the population is about 60%. The lack of GSTM1 has been linked with cancer risk. The frequency of the phenotypes of this isoenzyme in melanoma (MM) patients (n = 197) is reported here. A significantly higher proportion of individuals in the control group (n = 147) had measurable GSTM1 than MM patients ; there was a higher proportion of positive phenotypes in general among women than among men. Odds ratio analysis indicated that individuals with this polymorphic variant have an approximately 2-fold risk of developing these cancers. GSTM1 phenotype distribution depends on age, smoking habit and tumour pathology. A group of MM patients with dysplastic naevi was also studied.


A study was made of the relationship between the intrinsic radiosensitivity of human cervical tumours and the expression of the DNA repair enzyme human apurinic/apyrimidinic endonuclease (HAP1). The radiosensitivity of clonogenic cells in tumour biopsies was measured as surviving fraction at 2 Gy (SF2) using a soft agar assay. HAP1 expression levels were determined after staining of formalin-fixed paraffin-embedded tumour sections with a rabbit antiserum raised against recombinant HAP1. Both measurements were obtained on pretreatment biopsy material. All 25 tumours examined showed positive staining for HAP1, but there was heterogeneity in the level of expression both within and between tumours. The average coefficients of variation for intra- and intertumour heterogeneity were 62% and 82% respectively. There was a moderate but significant positive correlation between the levels of HAP1 expression and SF2 . Hence, this study shows that there is some relationship between intrinsic radiosensitivity and expression of a DNA repair enzyme in cervical carcinomas. The results suggest that this type of approach may be useful in the development of rapid predictive tests of tumour radiosensitivity.


Between January 1977 and January 1988, 19 patients with non-Hodgkin's lymphoma (NHL) involving the ileocaecal region were cared for by the CRC Wessex Medical Oncology Unit. Fifteen of these patients had primary ileocaecal NHL (stages IE or IIE) and four had secondary involvement of this region (stage IV). The commonest clinical presentation was with abdominal pain and a palpable mass in the right iliac fossa. Bulky (greater than 10 cm) disease was a particularly common feature, and complete surgical removal was possible in only seven patients. All patients had intermediate (18) or high grade (one) NHL using the Working Formulation. The commonest histological subtype was diffuse large cell. Seventeen patients received postoperative therapy, comprising local radiotherapy in one and combination chemotherapy in the remaining 16. Eleven of the 19 patients remain disease-free 6-60 months from diagnosis. Because of the high incidence of bulky disease at this site, postoperative therapy may be indicated, even for patients with apparently completely excised stage I disease.


Sir,We read with great interest the article by The diagnosis of chronic pancreatitis was based on the radiologic data obtained by means of either endoscopic retrograde cholangiopancreatography or computed tomography.DNA was extracted from 20 ml of the serum by using the QIAmp Blood Kit (Qiagen) and the mutations in codon 12 of the K-ras gene were searched as described previously (BstNl restriction enzyme (PCR-RFLP).As positive controls, we used DNA from neoplastic tissues of 10 patients with pancreatic carcinoma by using the DNeasy Tissue Kit (Qiagen). For molecular analysis, DNA was amplified in the codon 12 region introducing a restriction site (GACCT) for digestion with DNA from peripheral blood resulted not mutated in the 40 patients with chronic pancreatitis and in the 30 with pancreatic carcinoma, while DNA from pancreatic neoplastic tissue resulted mutated in 70% of the samples.To verify our results, all the samples were analysed by direct sequencing using Big Dye terminator v 1.1 cycle sequencing Kit and performing runs on ABI Prism 310 genetic analyzer (Applied Biosystem)Despite what was mentioned in Maire's article, we failed to find any mutations in all patients analysed, as well as we failed to correlate K-ras mutations with the levels of tumour markers such as Ca 19.9, CA242, CA50, CEA.The results of the present investigation lead us to these conclusions: (1) the eventual presence of cancer cells in peripheral blood may be a rare event, even if numerous reports support the detection of K-ras abnormalities in the serum, (2) neoplastic cells are supposed to circulate in clusters, and consequently their cognition could be hampered by a single blood sample extraction. (3) Large amounts of nonmutated DNA, coming from leucocytes held in the buffy coat layer, might also mask some vestiges of the mutant type of K-ras gene.


Saccharomyces cerevisiae, has been established as part of our new drug discovery programme. Use of this assay has provided the first direct evidence that TOP1 and RAD52 proteins are involved in the mode of action of bisdioxopiperazine ICRF compounds, which thus express a mode of action quite distinctive from the other known TOP2 inhibitors evaluated. The functional assay is based on a comparison of pairs of yeast differing in their phenotypes by specific traits: the expression or lack of expression of ectopic human DNA topoisomerase I, with or without that of the RAD52 gene. Amongst a series of anticancer agents, inhibitors of topoisomerase I (camptothecin) were identified as such in yeast expressing human topoisomerase I, whilst the presence or absence of RAD52 protein permitted the discrimination of compounds generating double-stranded DNA breaks, either directly (bleomycin) or involving DNA adduct formation (cisplatin), or indirectly with DNA damage mediated via inhibition of the topoisomerase II enzyme (etoposide). Notably, however, both the RAD52 protein and the lack of TOP1 enzyme appeared implicated in the cytotoxic activities of the series of bisdioxopiperazine ICRF compounds tested. This functional assay in a living organism therefore appears to provide a valuable tool for probing distinctive and specific mode(s) of action of diverse anticancer agents. © 1999 Cancer Research CampaignA screening procedure which permits identification of compounds based on their activities against specific biological targets directly in a living organism,


The uptake, retention and effects of aluminium chloro sulphonated phthalocyanine (AlSPc) were measured in two cell lines, UV-2237 a murine fibrosarcoma and the non-tumorigenic NIH/3T3 fibroblast line. The behaviour of cells treated with AlSPc was compared with that of those treated with haematoporphyrin derivative (HpD), a photosensitizer often used in photodynamic therapy (PDT) of cancer. AlSPc absorbs light strongly in the red region, is taken up by cells in a dose dependent fashion and is retained in vitro over a period of days (5 days after exposure greater than 40% remains cell-associated versus less than 25% of HpD). Additionally AlSPc was less cytotoxic to cells, maintained in darkness or exposed to room light, compared to HpD (100% viability versus 0% viability 3 days after 60 min exposure to room light). However red light (approximately 600-700 nm) caused greater toxicity in AlSPc-treated cells (100%) than in similarly exposed HpD-treated cells (less than 60%). No significant differences were detected between the responses of the fibrosarcoma and the fibroblast cell lines. These characteristics of AlSPc suggest that it may prove to be a useful photosensitizer for PDT of cancer and this possibility is discussed.