In 2016, CDC saw an increase in cases of acute flaccid myelitis (AFM); 144 persons in 37 states and the District of Columbia were confirmed to have AFM. After investigations in California (3 when corrected for red blood cells). A confirmed case must have an MRI demonstrating lesions restricted primarily to the gray matter of the spinal cord, in addition to acute onset of flaccid limb weakness , but MRI results indicated alternative etiologies . The case that met the probable case definition had pleocytosis (CSF WBC = 7/mm3), but MRI findings were inconsistent with AFM, and no other plausible diagnosis was identified.The 2015 Council of State and Territorial Epidemiologists and CDC case definition for probable AFM requires acute onset of flaccid limb weakness and cerebrospinal fluid (CSF) pleocytosis  or gastrointestinal illness (one patient), with onset dates for those illnesses occurring during August 14–September 13. Their illness began a median of 2 days (range = 2–5 days) before onset of focal limb weakness; three patients experienced tactile or measured fever preceding onset of neurologic symptoms . Among p3 (range = 50–207), and initial viral testing at the hospital included CSF reverse transcription–polymerase chain reaction (RT-PCR) assays for enterovirus (three patients) and West Nile virus (WNV) (two patients), polymerase chain reaction (PCR) assay for herpes simplex virus (two patients), and enzyme immunoassay to detect immunoglobulin M (IgM) or immunoglobulin G (IgG) for WNV (three patients). All results were negative. All four CSF specimens were negative on TGen amplicon sequencing assay . Serum collected from one patient at initial evaluation was negative for WNV IgM and IgG on a hospital immunoassay; serum collected from the same patient 47 days after onset of focal limb weakness and from two additional patients  were negative for WNV IgM and St. Louis encephalitis IgM at the Arizona State Public Health Laboratory.CSF was collected from all four patients with confirmed AFM. Median CSF WBC count was 133/mmThree of the four patients had nasopharyngeal (NP) swabs available from initial evaluation that were forwarded to CDC; one specimen was positive for enterovirus/rhinovirus on a panviral respiratory PCR panel at the admitting hospital laboratory and for EV-D68 at CDC. RNA extracted from NP swabs from all three patients was positive by the TGen amplicon sequencing test for EV-D68 (GenBank Bioproject); an NP specimen from a patient who did not meet the AFM confirmed or probable case definitions also was positive for EV-D68 by the same assay.Stool specimens were collected from two patients at the time of initial evaluation; vital cultures of these specimens were negative on viral. One available specimen and three additional specimens, collected 28, 47, and 63 days after onset of focal limb weakness, were sent to CDC for four enterovirus/parechovirus RT-PCR assays. A stool specimen, collected at day 28 from the patient who did not have an NP swab available, was positive for coxsackievirus A10.This cluster of AFM at one children’s acute care hospital is the largest cluster identified to date in Arizona and is part of a nationally identified increase in AFM cases. Although no statewide surveillance system specific to AFM is available, this cluster was detected by physician reporting, highlighting the need for physicians to remain vigilant for this emerging disease and to report cases that fit the AFM case definition to their local health department. Metagenomic analyses identified EV-D68 in NP swabs from the three patients for whom specimens were available, along with a specimen from a patient who did not meet the AFM case definition; therefore, no single etiology or risk factor was associated with only confirmed cases. Patient and family history of asthma was the most common comorbidity reported among confirmed AFM cases and should be considered in future case investigations. Expanded analysis of infectious, postinfectious, and noninfectious etiologies might provide further insight into the mechanism of AFM.


Epidemiology of drug resistant HIV has focused on trends and less attention has been given to identification of factors, especially behaviors including substance use, in acquisition of drug-resistant HIV. From 2009 to 2012 The Metromates Study enrolled and followed for one year men who have sex with men (MSM) seeking testing for HIV in a community clinic in Los Angeles assessing those testing positive for acute and recent HIV infection. Behavioral data were collected via Computer-Assisted Self-Interview from 125 classified as newly HIV infected and 91 as chronically infected (newly HIV-diagnosed); specimens were available and viable for resistance testing for 154 of the 216 HIV positives with new diagnoses. In this community clinic we found prevalence of resistance among MSM with new HIV-diagnosis was 19.5% (n = 30/154) with no difference by recency of HIV infection. Sexual partnership characteristics were associated with resistance; those who reported transgendered sex partners had a higher prevalence of resistance as compared to those who did not report transgendered sex partners , while those who reported having a main partner had a lower prevalence of drug resistance . In multivariable analyses adjusting for HIV recency and antiviral use, reporting a main partner decreased odds , reporting a transgendered partnered increased odds ; and being African American increased odds of drug resistance . This suggests African American MSM and TG individuals in Los Angeles represent pockets of exceptional risk that will require special approaches to prevention and care to enhance their own health and reduce their likelihood to support transmission of drug resistance in the US. Among those with a new diagnosis of HIV infection, some are infected by a strain of HIV-1 resistant to one or more drugs, known as primary or transmitted drug resistance (TDR)  but evidBetween 2009–2012 321 MSM enrolled in the NIDA funded Metromates Cohort Study conducted at The Los Angeles LGBT Center, a community-based organization providing sexual health services, social services and HIV care in Los Angeles. Men at least 18 years of age reporting sex with a man in the past 12 months who sought HIV testing at the LGBT Center were offered an opportunity to undergo a consent process including a written consent with a full description of the study with choices about if and how to participate in specimen donation, and then were provided a copy of their written consent form . Those who tested HIV-positive provided a specimen for testing of recency of infection using the Vironostika detuned assay (BioSystems Laboratory in San Francisco) a validated assay for use of detection of early HIV infection, 14; negA Computer-Assisted Self-Interview (CASI) assessed sexual partner type ; types of sexual activity ; substance use . Also assessed was use of ARV drugs for treatment, pre-exposure prophylaxis (PrEP) or post-exposure prophylaxis (PEP), substance use, and sexual behavior. “Treatment status” was determined from reports of date starting ARVs as some men started ARVs before enrollment due to a lag between communication of an HIV positive test result and study enrollment. Initiation of care was not an exclusion criteria for this analysis. The questions and scales used in this study were developed from two previous studies of men with acute HIV infection or previous diagnosis; with careful attention to included comparable questions/measures to the previous cohort that identified behavioral factors associated with resistant HIV –17. The http://hivdb.stanford.edu/). Samples were classified as “resistant” if results indicated reduced sensitivity to at least one class of ARVs.HIV genotyping was performed using stored plasma samples collected at study enrollment. The testing of those assessed as acutely infected (n = 104) was conducted at the HIV Genotyping Laboratory at Johns Hopkins University (JHU) and the other new diagnoses at University of California Los Angeles (UCLA) School of Medicine (n = 69). Testing at JHU was conducted using the ViroSeq HIV Genotyping System v2.8 . This system provides HIV sequences for the region encoding HIV protease (amino acids 1–99) and HIV reverse transcriptase (amino acids 1–335). Testing at UCLA was done using the UltraSens Viral Isolation kit (Qiagen) according to the manufacturer’s protocol. The Reverse Transcriptase (RT) and Protease (PRO) genes were amplified and sequenced according to the previously published protocol of Snoeck et al. using SuperScript III RT (Invitrogen) for RT and KOD high fidelity polymerase (Novagen) for PCR. PCR products were directly sequenced using the BigDye v3.1 Kit (Applied Biosystems) according to the manufacturer’s protocol using the primers described in Snoeck et al. SequencDifferences in ARV drug resistance to at least one class of HIV drugs by demographics and behaviors were evaluated using t-tests, Wilcoxon rank sum, chi-square methods, Fisher’s exact tests and multivariable logistic regression analysis. Behaviors included in the analyses were selected based on findings from the research team’s previous work with this population and this topic, 17 as wAmong the 154 recently HIV-diagnosed MSM with testable plasma the mean age was 30 years, nearly half identified as Hispanic (48%), followed by white (29%) and African American (17%); 58% were identified as having an acute/recent HIV-infection. The median number of sexual partners in the past 12-months was eight [interquartile range (IQR) 4–20], with a median of two anonymous partners and one one-time partner. More than half the participants  reported substance use in the past 3 months, including marijuana (41%), methamphetamine (20%), cocaine (15%), and opioids (3%). Eleven (7%) reported sex with both men and women; and 30 (19.5%) reported a transgender partner in the last 12 months; 30 (19.5%) reported transactional sex in the past 3 months. Between screening and enrollment 30 men (19%) had initiated ARV therapy. Among these HIV positive men more methamphetamine use was reported among newly infected than chronically infected .Drug resistance to at least one class of ARVs was identified in 30 (19.5%) of the 154 participants. Most of the resistance was to NNRTIs: 25/30 cases of resistance. The prevalence of ARV resistance did not vary significantly by HIV diagnosis group  althoughReporting a main partner decreased odds of drug resistance , reporting a transgendered partnered increased odds ; and being African American increased odds of drug resistance .The prevalence of drug resistance of 19.5% among MSM in Los Angeles with new HIV diagnosis is relatively high compared to other reports  even wheThe increased risk of resistance among men who have sex with transgendered (TG) partners is a new finding. TGs represent a marginalized group of individuals among whom adherence to HIV treatment is particularly poor and who Our findings point to the role of the main partner in providing protection against acquiring resistant virus and the increased risk with having casual partners. Clearly the more exposures, the more likely one of these is to someone with a resistant virus. Yet this also suggests that sexual networks are playing an increasingly important role in the transmission of resistance. It points to the need to counsel men without main partners that when they have unprotected sex with casual partners whose HIV status or use of therapy is unknown they may be at higher risk of acquiring resistant virus and that this has the potential to affect their therapeutic options.Our finding that African American MSM were more likely than other MSM in Los Angeles to acquire resistance raises continuing concerns about the epidemic among African American MSM. These men are most likely to acquire HIV in general in the US–with African Americans representing half of all new infections in 2009 and AfriGiven the challenges of enrolling individuals into studies at the time of their HIV diagnosis and identifying those with acute or recent infection in any community, the sample size for this study was relatively small. This limited the power to more sensitively detect factors associated with resistance that were low prevalence. Although the study used computer-assisted interviewing and emphasized the conduct of the study in the most sensitive way for individuals with new HIV diagnosis, behavioral reports are always prone to mis-reporting and that could result in misclassification of participants in the analysis. Finally, the study and was initiated and tools developed in 2008 and earlier, when awareness of and well-developed tools for studying transgender individuals were not yet evolved fully limiting our ability to better study how them.Our findings support the rationale for examining behavioral characteristics of resistance because we could not confirm findings from earlier studies that identified strong use, notably methamphetamine use, as a driver of multi-resistant HIV suggestiThese findings indicate trends in HIV transmission patterns among MSM in Los Angeles. As a case of multi-drug resistant HIV has been documented among an individual on pre-exposure prophylaxis with emtricitabine (FTC)–tenofovir disoproxil fumarate (TDF) that was identified as transmitted and not acquired there cl


The mechanistic data presented in this interesting review suggests that long-term exposure to safe levels of ultra-violet radiation (UVR) has protective effects against the development of obesity and cardiovascular dysfunction beyond those induced by the cutaneous synthesis of vitamin D3 through factors such as the induction of cutaneous NO secretion . Evidenc


This article contains one NVivo 10® file with the complete 432 projects design documents (PDD) of seven waste management sector industries registered as Clean Development Mechanism (CDM) under United Nations Framework Convention on Climate Change (UNFCCC) Kyoto Protocol Initiative from 2004 to 2014. All data analyses and sample statistics made during the research remain in the file. We coded PDDs in 890 fragments of text, classified in five categories of barriers (nodes): technological, financial, human resources, regulatory, socio-political. The data supports the findings of author thesis [4] Specifications TableValue of the data•The UNFCCC raw data source is mixed up, disconnected, fragmented, and laborious.•This data is all inclusive . Therefore, any other CAQCA can be immediately made for the WM sector.•The data is standardized by UNFCCC, which means that it can be easily appended, replaced, compared or transposed to the others 14 sectors and 4.000 projects .•The dataset is an instructive example of a Computer Assisted Qualitative Content Analysis (CAQCA) using the features of a CAQCA Software1Despite a clear indication that waste management research needs a more system-oriented approach This data article shares, in a single NVivo 10® software file, 432 large waste management projects design documents (PDD) from 2004 to 2014 1)the entire PDD documents;2)a qualitative data analysis (QDA) aiming the study of the project's designers declared barriers;3)results of a primary search for the stated barriers (890 text fragments);4)we code (hermeneutic) fragments into five barriers categories: technological, financial, human resources, regulatory, and socio-political.The sample selected from CDM Project Search site of UNFCCC The data is standardized following the main UNFCCC guidelines for any project 2The computer assisted qualitative content analysis (CAQCA) drives the data file design. CAQCA is a well-known methodology largely used in social applied and human sciences The use of software  to treat a large volume of data produced almost in real time had a significant impact on research institutions and doctoral programs because part of those research without the QDAS would be impractical and impossible. This fact explains why the more Universities offers the QDAS at an institutional level The search criteria used to the content selection (raw data) from the UNFCCC Project Search site The projects have many possible statuses . We choose the registered status because no other guarantees UNFCCC consent for the registered operation. At last, the sort by reference number is strategic considering that the computer database key-index system is more reliable than registration date.The results by the host country and year are in the original article There is also an abnormal concentration of 124 projects in 2012 After the simple search for the term ‘barrier’ that returned more than a thousand occurrences, took place the coding second part by semantic-hermeneutic analysis of fragments. Some of these fragments proof meaningless and discarded. The rest classified in categories for future interpretation and pattern identification.


Functional neurological changes after surgery combined with diffusion tensor imaging (DTI) tractography can directly provide evidence of anatomical localization of brain function. Using these techniques, a patient with dysgraphia before surgery was analyzed at our hospital in 2011. The patient showed omission of kana within sentences before surgery, which improved after surgery. The brain tumor was relatively small and was located within the primary sensory area (S1) of the inferior parietal lobe (IPL). DTI tractography before surgery revealed compression of the branch of the superior longitudinal fasciculus (SLF) by the brain tumor. These results suggest that the left SLF within the S1 of IPL plays a role in the development of dysgraphia of kana omission within sentences.


Prostaglandins E2 and F2 alpha  were measured by gas liquid chromatography--mass spectrometry (glc-ms) in extracts of primary tumours from 78 patients with early breast cancer. These levels have been related to factors of established prognostic value and the patients disease-free interval. Although there was a wide variation in amounts of both prostaglandins extracted from different tumours, no significant relationship was observed between levels of prostaglandins and oestrogen receptors (ER), tumour size, presence of lymph node involvement and disease-free interval following primary treatment. It therefore seems unlikely that the level of these particular prostaglandins within breast carcinomas plays a fundamental role in the prognosis of the disease.


The present study tests whether a combined treatment of acupuncture and transcutaneous electrical nerve stimulation (TENS) is more effective than acupuncture or TENS alone for treating knee osteoarthritis (OA).Thirty-two patients with knee OA were randomly allocated to four groups. The acupuncture group (ACP) received only acupuncture treatment at selected acupoints for knee pain; the TENS group (TENS) received only TENS treatment at pain areas; the acupuncture and TENS group (A&T) received both acupuncture and TENS treatments; the control group (CT) received topical poultice (only when necessary). Each group received specific weekly treatment five times during the study. Outcome measures were pain intensity in a visual analogue scale (VAS) and knee function in terms of the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC).The ACP, TENS and A&T groups reported lower VAS and WOMAC scores than the control group. Significant reduction in pain intensity (P = 0.039) and significant improvement in knee function (P = 0.008) were shown in the A&T group.Combined acupuncture and TENS treatment was effective in pain relief and knee function improvement for the sampled patients suffering from knee OA. Osteoarthritis (OA) is the most common form of joint disease and most often affects the knee ,2. OA ofThe present study aims to test whether TENS or a combined treatment of acupuncture and TENS is more effective than acupuncture or TENS alone for treating knee OA in older patients.Outpatients aged 60 years or older with knee OA were recruited from the Meiji University of Oriental Medicine Hospital. The patients had been clinically and radiologically diagnosed of knee OA according to the criteria of the American College of Rheumatology. Further inclusion criteria for the present study were: (1) knee pain lasting for six months or longer; (2) no radiation of knee pain; (3) radiographic evidence of at least one osteophyte at the tibiofemoral joint (Kellgren-Lawrence grade ≥ 2); (4) normal neurological functions of lumbosacral nerve, including deep tendon reflexes, voluntary muscle action and sensory function; and (5) not receiving acupuncture treatment for more than six months. Exclusion criteria were: (1) major trauma or systemic disease; and (2) receiving conflicting or ongoing co-interventions. Patients under drug treatment were included if there had been no change in medicine and its dosage for one month or longer. This study was approved by the Ethics Committee of the Meiji University of Oriental Medicine.All enrolled patients gave their written informed consent. According to a block randomised allocation table , the enrolled patients were allocated to (1) the control (CT) group, (2) the acupuncture (ACP) group, (3) the transcutaneous electrical nerve stimulation (TENS) group or (4) the acupuncture and TENS (A&T) group.The design of this study was a randomly controlled clinical trial using a block randomised procedure. Each patient received a total of five treatments, once per week, and follow-up was measured for ten weeks after the first treatment.The CT group patients did not receive any specific treatment, but when necessary, were allowed to use topical poultice containing methylsalicylic acid.Liangqiu (ST34), Dubi (ST35), Zusanli (ST36), Yinlingquan (SP9), Xuehai (SP10) and Yanglingquan (GB34). Disposable stainless steel needles  were inserted into the muscle to a depth of 10 mm using the 'sparrow pecking' acupuncture technique  by acupuncturists who had four years of acupuncture training and three to eight years of clinical experience. When the subject felt dull pain or the acupuncture sensation (de qi) was achieved, the needle manipulation was stopped and the needle was left in place for ten more minutes.The ACP group patients received acupuncture treatment at selected acupoints for 15 minutes on the OA affected knee. The selected acupoints are widely accepted for treating knee pain -13, name2 and 5688 mm2 were respectively placed on the site with the most tenderness and the opposite side of the site. Two electrodes were different in size (ratio: 1:7). The smaller one was placed on the site of tenderness. The intensity of TENS stimulation was adjusted so that a tingling sensation 2–3 times of the subject's sensory threshold was produced.The TENS group patients received treatment at the OA affected knee for 15 minutes from a single-channel portable TENS unit , which sends between two electrodes a premixed amplitude-modulated frequency of 122 Hz (beat frequency) generated by two medium frequency sinusoidal waves of 4.0 and 4.122 kHz (feed frequency). Surface disposable electrodes of 809 mmThe treatment for the A&T group combined the treatments for the ACP and TENS groups. The patients received 15 minutes of TENS, and then 15 minutes of acupuncture treatment at the OA affected knee.We confirmed that the patients in all groups did not receive any other co-interventions including analgesics, anti-inflammatory agents or topical hyaluronic acid injection during the study period.Primary outcome measures were: (1) pain intensity, quantified with a 10 cm visual analogue scale  and (2) pain disability measured with the Western Ontario and McMasters Universities Osteoarthritis Index  . The WOMThe VAS scores were measured immediately before the first treatment and subsequently at one, two, three, four, five and ten weeks after the first treatment. The WOMAC scores were measured immediately before the first treatment and subsequently five and ten weeks after the first treatment. Each VAS and WOMAC score was measured immediately before treatment of the specified week.Repeated measures analysis of variance (ANOVA) was used to study the changes in the VAS and WOMAC scores in the three groups. Changes in the time course among groups were considered significant when the interaction was significant at a level of 0.013 (0.05/4). After detection of significant changes in the overall time course with repeated measures ANOVA, pair comparisons were detected with Bonferroni correction. StatView for Windows (version 5.0) or SYSTAT 10 (SYSTAT Inc) was used for the statistical analysis. The results with P values of less than 0.05 were considered statistically significant.A total of 32 patients  were randomly allocated to four groups for specific treatment. Before the treatments, no significant difference was found in baseline variables including age, disease, pain duration, and pain intensity VAS scores among the four groups.i.e. deterioration of symptoms). The total dropout rate was the same in every group. The analyses were performed on the 24 patients who completed the study and provided required information  compared to the CT group during treatment Table . The treThe present study demonstrated that acupuncture and TENS treatments were effective in pain relief. TENS is a common modality for treating musculoskeletal pain . TENS exOur other studies consistently showed that combined treatment of acupuncture and TENS was more effective than separate treatments ,25. TENSThe present study clearly demonstrated that combined acupuncture and TENS treatment is effective for pain relief in terms of VAS and knee function improvement in terms of WOMAC in patients suffering from knee OA. Large scale clinical trials are warranted.ACP: acupuncture group; ANOVA: analysis of variance; OA: osteoarthritis; QOL: quality of life; ROM: range of motion; TENS: transcutaneous electrical nerve stimulation; VAS: visual analogue scale; WOMAC: Western Ontario and McMaster Universities Osteoarthritis IndexThe author(s) declare that they have no competing interests.KI was responsible for study design, acupuncture treatment, manuscript preparation and submission. SH was responsible for patient recruitment. HO was responsible for all statistical design and data analysis. YK was responsible for manuscript review. HK was responsible for study design, critical manuscript review and patient recruitment. All authors read and approved the final manuscript.


Unique germline mutations in BRCA1 and BRCA2 account for inherited predisposition to breast and ovarian cancer in high-risk families. In Jewish high-risk individuals of Ashkenazi (east European) descent, three predominant mutations, 185delAG and 5382insC (BRCA1) and 6174delT (BRCA2), seem to account for a substantial portion of germline mutations, and two of these mutations (185delAG and 6174delT) are also found at about 1% each in the general Jewish-Ashkenazi population. We identified a novel BRCA1 mutation in two Jewish-non-Ashkenazi families with ovarian cancer: a thymidine to guanidine alteration at position 3053, resulting in substitution of tyrosine at codon 1017 for a stop codon (Tyr1017Ter). The mutation was first detected by protein truncation test (PTT) and confirmed by sequencing and a modified restriction digest assay. Allelotyping of mutation carriers using intragenic BRCA1 markers revealed that the haplotype was identical in these seemingly unrelated families. No mutation carrier was found among 118 unselected Jewish individuals of Iranian origin. Our findings suggest that this novel mutation should be incorporated into the panel of mutations analysed in high-risk families of the appropriate ethnic background, and that the repertoire of BRCA1 mutations in Jewish high-risk families may be limited, regardless of ethnic origin.


An ascitic form of plasmacytoma (MP-5563) in C3H mice has proved stable on transplantation. A simple linear relationship was demonstrated between the serum level of the paraprotein produced and the total number of plasmacytoma cells within the ascitic fluid.


The kinetics, specificity and morphology of cytolethal responses have been studied in human glucocorticoid-sensitive and -insensitive lymphoid cell lines (HLCL) and fibroblasts following treatment with high (10(-3)M) and low (10(-6)M) doses of steroid. The high dose cytolethal response appears non-specific occurring in all cell lines with every steroid tested. By contrast, the low dose  cytolethal response requires an active glucocorticoid and a sensitive HLCL. However, both high and low concentrations of steroid induce virtually identical morphological changes in dying cells and similar changes can be induced in cells killed by deliberate feed exhaustion. Although the morphological features in each case resemble apoptosis, the "programmed" physiological form of cell death, the intracellular events leading to cytolysis seem likely to differ. The earliest morphological changes presaging cell death comprise rounding up of cells and condensation of nuclear chromatin. Nuclear changes progress rapidly thereafter and appear to result from detachment of chromatin from the nuclear matrix. The low dose cytolethal response requires the continuous presence of glucocorticoid for periods in excess of 24h, prior to which cell growth appears unaffected. The constancy of this latent interval suggests glucocorticoids may influence some replication control mechanism unrelated initially to macromolecular biosynthesis.


We examined the localization of basic fibroblast growth factor (bFGF) in a series of human breast carcinomas using immunohistochemistry. Staining was observed in tumour cells in 15 out of 54 (28%) tumours and in the adjacent stroma in 34 out of 54 (63%) tumours examined. No correlation was observed between positive staining of these two compartments. The relationship between bFGF staining and expression of the metalloprotease stromelysin-3, and between bFGF and microvessel density, was examined. A statistically significant correlation (P < 0.003) was observed between bFGF staining of the stromal compartment and high expression of stromelysin-3  metalloprotease mRNA by stromal cells. In contrast, no correlation was observed between bFGF and intratumour microvessel density (IMD). These results raise the possibility that bFGF may be involved in the induction of stromelysin-3 mRNA expression in breast cancer stroma.


Patients treated with tamoxifen (TAM) for primary breast cancer often manifest de novo or acquired resistance, possibly through changes in drug metabolism. Using solid-phase extraction methods and reversed-phase high-performance liquid chromatography separations, levels of TAM and metabolites 4-hydroxytamoxifen (4OH) and desmethyltamoxifen (DMT) have been measured in plasma and tumour tissue from breast cancer patients treated with TAM for at least 3 months. Patients were categorized into those with tumours responding to TAM and those showing de novo or acquired resistance. Levels of TAM, 4OH and DMT in both plasma and tissue samples were correlated with clinical response, length of treatment and patient weight. Interesting results included accumulation of 4OH in tumour tissues over time in all patients, with significance reached in the acquired resistance group. In addition, significantly lower levels of 4OH and DMT were found in plasma taken from responding patients after 3 months of treatment when compared to non-responding patients, and a small group of ER-poor patients showed significantly lower levels of all three species in plasma when compared to other patients. Whilst not explaining TAM resistance in all cases, these differences could account for the development of resistance to TAM treatment in certain subgroups of patients. © 2000 Cancer Research Campaign


A possible variable is the influence of growth conditions on discerning candidates. The present study examined the influence of growth conditions on the expression of chromosome 3 genes in the tumorigenic EOC cell lines, OV-90, TOV-21G and TOV-112D using Affymetrix GeneChipChromosome 3 gene expression profiles  were extracted from U133A expression microarray analyses of the EOC cell lines OV-90, TOV-21G and TOV-112D that were grown as monolayers, spheroids or nude mouse xenografts and monolayers derived from these tumors. Hierarchical cluster analysis was performed to compare chromosome 3 transcriptome patterns of each growth condition. Differentially expressed genes were identified and characterized by two-way comparative analyses of fold-differences in gene expression between monolayer cultures and each of the other growth conditions, and between the maximum and minimum values of expression of all growth conditions for each EOC cell line.An overall high degree of similarity (> 90%) in gene expression was observed when expression values of alternative growth conditions were compared within each EOC cell line group. Two-way comparative analysis of each EOC cell line grown in an alternative condition relative to the monolayer culture showed that overall less than 15% of probe sets exhibited at least a 3-fold difference in expression profile. Less than 23% of probe sets exhibited greater than 3-fold differences in gene expression in comparisons of the maximum and minimum value of expression of all growth conditions within each EOC cell line group. The majority of these differences were less than 5-fold. There were 17 genes in common which were differentially expressed in all EOC cell lines. However, the patterns of expression of these genes were not necessarily the same for each growth condition when one cell line was compared with another.in vivo and in vitro growth conditions of tumorigenic EOC cell lines appeared to modestly influence the global chromosome 3 transcriptome supporting the notion that the in vitro cell line models are a viable option for testing gene candidates.The various alternative  The molecular genetic analysis of ovarian cancer has been facilitated by the establishment and characterization of spontaneously immortalized epithelial ovarian cancer (EOC) cell lines that have been derived from malignant cells by long-term growth in cell culture . In our in vitro EOC cell line models. Recently, our group has reported on global differences in gene expression between EOC cell lines that were cultured as monolayers, spheroids, or nude mouse xenografts suggesting that microenviroment could impact the transcriptome [in vitro model with the potential for identifying and testing chromosome 3 tumor suppressor genes because of extensive loss of heterozygosity of the 3p arm [A possible variable in the application of EOC cell line models is the influence of growth conditions on discerning and then characterizing gene candidates which initially exhibit differential gene expression in criptome . To furtcriptome . We havecriptome ,8,14,15.criptome . Both OVcriptome ,16. Karycriptome ,5. In pae 3p arm  but has The present study was focused on addressing the magnitude and extent of transcriptome modifications for different EOC cell line models that may be influenced by tumor microenvironment. As each cell line exhibits unique molecular genetic characteristics comparisons of chromosome 3 transcriptome modifications were made with respect to gene expression profiles generated with each growth condition within each experimental cell line model.The EOC cell lines were derived from a stage III/grade 3 clear cell carcinoma TOV-21G), a stage IIIc/grade 3 endometrioid carcinoma (TOV-112D), and from the ascites fluid of a stage IIIc/grade 3 adenocarcinoma (OV-90), all from chemotherapy naïve patients, as described [1G, a sta® HG-U133A microarray analyses of the OV-90, TOV-21G and TOV-112D EOC cell lines that were each grown under different conditions as described previously [. These conditions include monolayer cultures (L), spheroid cultures (S), nude mouse xenografts at subcutaneous (TSC) or intraperitoneal (TIP) sites, and monolayer cultures of subcutaneous (LSC) and intraperitoneal (LIP) xenografts, as described previously [® experiments, was achieved from the raw expression data using the MAS5 software (Affymetrix Microarray Suite®) by multiplying the value for an individual probe set by 100 and dividing by the mean of the raw expression values for the given sample data set as described previously [Chromosome 3 gene expression profiles were extracted from normalized Affymetrix GeneChipeviously . The sof and the UniGene Homo sapiens database, based on UniGene Build 198 . Additional mapping information was obtained from the University of California Santa Cruz (UCSC) Human Genome Browser database, March 2006 (NCBI Build 36.1) hg 18 assembly . Based on these databases, 1147 probe sets were identified that mapped to chromosome 3, representing 735 genes and ESTs. Chromosome 3 alignment of probe sets (represented genes) was determined based on the UCSC Human Genome Browser database, where 535 probe sets mapped to genes on the chromosome 3p arm and 612 probe sets mapped to genes on the chromosome 3q arm.The normalized data set was then used to extract the expression profiles associated with probe sets representing chromosome 3 genes. Probe sets corresponding to chromosome 3 genes were identified using the Affymetrix NetAffx Batch Query tool The normalized expression data sets were also rescaled to eliminate systematic biases due to low expression values. Low values with A-calls are considered to be technical noise, which may influence fold-difference comparisons and overestimate expression differences that result from high variability of low expression values. Probe sets containing A-calls may also reflect either absent expression or poorly designed probe sets . To redu. In order to determine the significance of the differential expression, modified t-tests were performed with Bioconductor's limma package, where p-values from the resulting comparison were adjusted for multiple testing as described [genefilter package was used to filter out probe sets with insufficient variation in gene expression across all tested samples for the analysis of each EOC cell line data set. In the remaining expression values, a log base 2 scale of at least 0.5 for the interquartile range was required across all tested samples for each EOC cell line group as described. Hierarchical clustering analysis was performed with R's cluster package, using the Pearson correlation distance.Hierarchical cluster analysis was performed on normalized and rescaled gene expression data analyzed using Bioconductor, an open-source software library for the analyses of genomic data  based onescribed . This meTwo-way comparative analyses based on fold differences of expression values were performed on normalized and rescaled gene expression data derived from each EOC cell line. The expression values with at least one high-reliability score or P call for each EOC cell line sample set  were evaluated in two-way comparative analyses. Differentially expressed genes were defined as those which exhibited at least a 3-fold difference in two-way comparative analyses with expression value for monolayer culture and each growth condition, or between the maximum and minimum value of expression observed within a set of data for each EOC cell line.Hierarchical cluster analysis of chromosome 3 gene expression data from each EOC cell line grown in monolayer cultures (L) and alternative growth conditions such as spheroid cultures (S), nude mouse xenografts at subcutaneous (TSC) or intraperitoneal (TIP) sites, and monolayer cultures of subcutaneous (LSC) and intraperitoneal (LIP) xenografts is shown in Figures To further characterize the gene expression profiles and identify the genes that may be influenced by the growth conditions, we applied a two-way fold-difference comparative analysis approach. As monolayer cultures are often used in molecular genetic assays of cancer derived cell lines, we compared gene expression values of the EOC cell lines grown as monolayer cultures with each of the alternative growth conditions comprised of spheroid cultures (S), nude mouse xenografts at subcutaneous (TSC) or intraperitoneal (TIP) sites, and monolayer cultures of subcutaneous (LSC) and intraperitoneal (LIP) xenografts. We performed two-way comparison analysis based on fold-differences using the expression values which contained at least one high reliability score  per probe set for each EOC cell line. Using this criterion, the expression values of 692 (60.3%), 739 64.4%), and 693 (60.4%) probe sets from the total of 1147 chromosome 3 probe sets for OV-90, TOV-21G and TOV-112D, respectively, were evaluated for fold-differences  of the expression values exhibited by all alternative growth conditions. Two-way comparative analysis was performed between the maximum and minimum values of expression observed for probe sets for each EOC cell line group. A minimum 3-fold cut-off was used to characterize differences in gene expression. Overall less than 23% of probe sets exhibited greater than 3-fold differences in gene expression in this comparative analysis in any EOC cell line group  and both subcutaneous (TSC) and intraperitoneal (TIP) xenografts exhibited the lowest values of expression of this gene  xenograft sample and the lowest value was observed with intraperitoneal (TIP) xenograft  in gene expression in each EOC cell line group tested irrespective of growth condition. It was also reflected in the two-way comparative analyses where a 3-fold cut-off was applied. Although we have previously shown that replicates of Affymetrix GeneChip® expression data derived from the EOC cell lines grown as monolayer cultures were highly reproducible [® expression microarrays of the EOC cell lines, we have used a lower threshold level of 15 rather than 50 or 100 depending on the GeneChip® used [In this study, we described chromosome 3 transcriptome changes for three well characterized EOC cell lines  that each responded differently in relation to various growth conditions such as in three dimensional spheroid culture and nude mouse xenograft models, relative to the conventional monolayer culture. However, the alternative oducible ,6, a lowoducible ,17. Unliip® used -10. A loip® used .TP53, whereas TOV-21G harbors a somatic mutation in KRAS and exhibits microsatellite instability. OV-90 also is monoallelic for the 3p arm, however this gross genomic anomaly did not significantly impact on global patterns of gene expression of the chromosome 3p arm as assayed by Affymetrix expression analyses of this cell line and the other EOC cell line used in the present study [The EOC cell line lines exhibited unique patterns of gene expression as shown by the hierarchical cluster analysis. These unique differences are also reflected in a previous global analyses of gene expression from the entire Affymetrix U133A microarray . These Ent study ,15. The ARPC4 and NCK1), cell growth and division (CDC25A and SKIL), and extracellular signaling/cell-cell junctions  (Additional file TMEM158, HEG1, PLOD2 and ATP13A3, were recently found differentially expressed greater than 3-fold in a comparative analysis of primary cultures of normal ovarian surface epithelial cells and malignant serous ovarian tumors [Given the molecular genetic differences in the EOC cell lines, it is not surprising that there were few similarly differentially expressed genes found in common with all of EOC cell lines. Indeed there were only 17 genes in common in all three EOC cell lines which exhibited differential expression greater than 3-fold in all comparative analyses (Additional file in vitro and in vivo growth conditions may all still be significantly different when each is compared with normal cells [S100A6, was found differentially expressed relative to culture conditions and further validated by RT-PCR and immunohistochemistry [in vitro model systems for studying gene candidates, our results in the present study show that a high correlation of gene expression in the transcriptomes generated from ovarian cancer cell lines propagated in different contexts. Overall these results attests to the validity of the EOC cell lines as an in vitro model for studying gene candidates but point out that some genes may be influenced by microenvironment, a factor that should be taken into consideration when investigating the molecular biology of specific genes. As our EOC cell lines are amenable to propagation in alternative growth conditions one could assay and further investigate the magnitude of transcriptional effects for specific candidate genes of interest and their consequences at the protein level to further understand the biological relevance gene expression differences associated with microenvironment.Future experiments are required to determine if the differences observed in the EOC cell lines grown in alternative conditions are biologically relevant or a reflection of experimental design. The magnitude of the differences in gene expression observed in the EOC cell lines grown under the various al cells . The EOChemistry . While tin vivo and in vitro growth conditions affords the opportunity to study gene expression of candidates in contexts that more closely mimic tumor growth in vitro. However, the analyses of chromosome 3 transcriptomes are highly comparable within each EOC cell line context. These observations would argue that gene expression studies using monolayer cultures of ovarian cancer lines is still a viable option for initial studies involving the characterization of gene expression pattern of candidate genes.The ability to culture tumorigenic EOC cells under different The authors declare that they have no competing interests.NALC participated in the study design of expression analysis, performed the data analysis, and drafted the manuscript. MZ performed experiments involving the growth of the EOC cell lines in different conditions. DMP participated in the study design. A-MM-M supervised the growth conditions assays and participated in the study design. PNT conceived the chromosome 3 study and study design, and drafted the manuscript. All authors have read the manuscript and approved the final manuscript.The pre-publication history for this paper can be accessed here:Table S1: Genes exhibiting greater than 3-fold difference in gene expression in common in all EOC cell lines.Click here for file


Cytotoxic drug resistance developing after chemotherapy is thought to be the main cause of treatment failure in several human tumours, including small cell lung cancer (SCLC). Cell lines showing drug resistance following prolonged exposure to a single agent frequently acquire resistance to several functionally unrelated drugs, the phenomenon of multi-drug resistance (MDR). Classical MDR is thought to arise from changes effecting a reduction in intracellular availability of cytotoxic drugs. We describe a flow cytometry (FCM) technique to monitor the MDR phenotype in drug resistant variants of SCLC and non-SCLC cell lines. The technique is based on a multiparametric analysis of the nuclear binding of a model chemotherapeutic agent, the fluorescent dye Hoechst 33342 (Ho342), which is capable of supra-vital staining of DNA in intact, viable cells. A laboratory derived drug resistant SCLC cell line, H69/LX4, showed a significant (30%) reduction in nuclear binding compared to the parental line H69/P. Exposure to verapamil (VPL) rapidly increased (within 2 min) nuclear binding of Ho342, and the new equilibrium of nuclear staining, attained within 20 min, remained lower than the level achieved in the parental cell line, suggesting some ability of H69/LX4 to limit the effect of the drug efflux blocker. A drug resistant large cell carcinoma line showed only a small reduction (10%) in nuclear binding when compared to the parent line, and this difference was not altered by VPL. A drug resistant adenocarcinoma line showed less than 10% difference in nuclear binding compared with the parental line and neither line was significantly affected by VPL treatment. Our findings suggest that different mechanisms of resistance may occur in lung tumours of different tissue types. This technique may be extended to the rapid and direct examination of biopsy specimens of human solid tumours for evidence of multi-drug resistance.


Several cobalt(II), copper(II) and nickel(II) complexes of nicotinoylhydrazine-derived compounds were prepared and characterised by physical, spectral and analytical data. These compounds and their complexes have proven to be antibacterial. The screening data show the metal complexes to be more potential/bactericidal than the uncomplexed compounds against one or more bacterial species.


From 1980 through 1988, biliary surgery was performed in 197 patients with acute gallstone disease andconcomitant elevation of serum glutamic oxalacetic transaminase (SGOT) or serum glutamic pyruvictransaminase (SGPT) of over 300 Karmen units. In 137 patients, anatomic inspection and liver biopsywere performed during the acute stage of the disease. Impacted and floating bile duct stones were foundin 69 (50%) and in 43 (32%) of the 137 patients, respectively. The main liver histology was necrosis ofliver cells. After surgery, high serum transaminase fell rapidly with immediate recovery in 99% of thepatients. In the remaining 60 patients, their signs and symptoms settled soon after initial conservativetreatment and surgery was performed after an average time of 21 days. At laparotomy, impacted bileduct stones were found in 2 (3%) and liver histology revealed regeneration of liver cells.		These findings suggest that marked elevation of serum transaminase in patients with acute gallstonedisease might be due to an acute inflammatory liver cell injury caused by impacted bile duct stones ormigrating stones, which would be transient and reversible after early resolution of the bile ductobstruction.


As these complexes are candidates for a novel class anti-HSV drugs, further studies on their mutagenicity are of particular interest. In the present paper we present data showing that Zn(pic)2 and Zn(asp)2 do not express mutagenic effect in both prokaryotic  and eukaryotic (Saccharomices cerevisiae) test systems.Former studies showed that complexes of Zn(II) with picolinic and with aspartic acids, Zn(pic)


Radiotherapy of tumours proximal to normal CNS structures is limited by the sensitivity of the normal tissue. Prior to the development of prophylactic strategies or treatment protocols a detailed understanding of the mechanisms of radiation induced CNS toxicity is mandatory. Histological analysis of irradiated CNS specimens defines possible target structures prior to a delineation of cellular and molecular mechanisms. Several lesions can be distinguished: Demyelination, proliferative and degenerative glial reactions, endothelial cell loss and capillary occlusion. All changes are likely to result from complex alterations within several functional CNS compartments. Thus, a single mechanism responsible cannot be separated. At least four factors contribute to the development of CNS toxicity: (1) damage to vessel structures; (2) deletion of oligodendrocyte-2 astrocyte progenitors (O-2A) and mature oligodendrocytes; (3) deletion of neural stem cell populations in the hippocampus, cerebellum and cortex; (4) generalized alterations of cytokine expression. Several underlying cellular and molecular mechanisms involved in radiation induced CNS toxicity have been identified. The article reviews the currently available data on the cellular and molecular basis of radiation induced CNS side effects.   http://www.bjcancer.com © 2001 Cancer Research Campaign


To evaluate visual outcome following pars plana vitrectomy (PPV) and intraocular foreign body (IOFB) removal through the sclerocorneal tunnel combined with simultaneous cataract extraction and sulcus-fixated intraocular lens (IOL) implantation as a single procedure in penetrating ocular trauma with IOFB and traumatic cataract.Eighteen cases of penetrating ocular trauma with retained IOFB and traumatic cataract who underwent PPV, IOFB body removal and cataract extraction with posterior chamber IOL (PCIOL) implantation in the same sitting, between June '04 and December '05 were retrospectively analyzed. All the foreign bodies were removed through the sclerocorneal tunnel.All the 18 patients were young males, with an average follow-up period of 12 months. In 12 cases the foreign body was intravitreal and in six cases it was intraretinal but extramacular. Thirteen cases had a best corrected visual acuity ranging from 20/20 to 20/60 at their last follow-up. Five cases developed retinal detachment due to proliferative vitreoretinopathy (PVR) changes postoperatively and were subsequently managed by surgery.Primary IOL implantation with combined cataract and vitreo-retinal surgery is a safe option reducing the need for two separate surgeries in selected patients with retained IOFB and traumatic cataract. This combined procedure provides good visual outcome with early rehabilitation in young working patients. Open globe injury with or without intraocular foreign body (IOFB) often has a poor visual outcome owing to possible endophthalmitis and/or vitreo retinopathy with the varied and unsterile wounds associated with it.–3 ManageMethods for removal of the traumatic cataract include lensectomy, conventional extra-capsular cataract extraction (ECCE), manual small-incision cataract surgery (SICS) and phacoemulsification. Unlike lensectomy other methods of cataract extraction offer a better visual rehabilitation because of implanted intraocular lens (IOL).–8 Most sTo avoid such complications, in this series we performed cataract extraction by manual SICS , removed the foreign body through the same tunnel and implanted a posterior chamber IOL (PCIOL) over the anterior capsular rim. The aim of the study was to evaluate visual outcome following pars plana vitrectomy (PPV) and IOFB removal through the sclerocorneal tunnel combined with simultaneous cataract extraction and sulcus-fixated IOL implantation as a single procedure in penetrating trauma with IOFB and traumatic cataract.Medical records of patients with penetrating eye injury and IOFB attending the retina clinic between June 2004 and December 2005 were reviewed. Out of 28 consecutive patients, 10 patients were excluded from this study because of absence of cataract, presence of associated endophthalmitis, severely injured lens with vitreous in the anterior chamber and evidence of zonular dehiscence.Eighteen patients with clinically significant lens opacification and IOFB with or without concomitant vitreoretinal pathology were included in the study. Primary repair was done in all cases except three, where the wound was self-sealed and they were taken up for vitreoretinal procedure directly. All patients underwent a complete general ophthalmologic examination prior to the surgical procedure. Preoperative visual acuity, site of entry of foreign body (FB), capsular and zonular integrity was assessed. Ultrasound B-Scan and/or X-ray of orbit were performed in all patients to locate and evaluate the IOFB. Keratometry and biometry of the injured eyes were done. If keratometry and biometry was not possible on the injured eye, fellow eye measurements were used. In all cases manual SICS was performed before the vitreoretinal procedure. A 6 mm frown-shaped sclerocorneal tunnel was made, followed by continuous curvilinear capsulorrexis and hydrodelineation. The lens nucleus was prolapsed into the anterior chamber and removed through the tunnel by sandwich method.–11 Rest 12Sclerotomies and conjunctiva were closed with 6-0 vicryl and 2 mg dexamethasone and 2 mg gentamicin sulphate were injected subconjunctivally. Postoperatively all patients received topical antibiotics and steroid for four weeks with gradual tapering. Oral ciprofloxacin 500 mg twice daily along with nonsteroidal anti-inflammatory agents was given in all cases with addition of systemic steroids when necessary. In each case recording of best corrected visual acuity (BCVA), intraocular pressure measurement, slit-lamp biomicroscopy and indirect ophthalmoscopy was done postoperatively. Cause of decreased vision was assessed considering site of entry of FB, site of lodgment of FB and postoperative complications like RD in each case.All our patients were young males with mean age of 24.5 years. The minimum follow-up period was four months, the maximum being 18 months with median follow-up time of 12 months. In 12 out of 18 cases the FB was intravitreal and in six cases it was intraretinal but extramacular. Thirteen out of 18 cases had BCVA between 20/20 and 20/60. Anatomical success was achieved in 13 out of 18 cases with 11 patients having visual acuity 20/30 or better with first vitreoretinal procedure. With the second vitreoretinal procedure four out of five patients had anatomical success .et al.[We reviewed the visual outcome and complications in 18 patients with penetrating intraocular injury, significant cataract and retained IOFB, who underwent manual SICS, PPV, removal of IOFB and implantation of PCIOL. This simultaneous procedure was chosen to get rid of cataract, which diminished the visualization of posterior segment and for prevention of lens-induced uveitis due to a ruptured lens capsule. We found no significant correlation in ‘duration between injury and vitreoretinal procedure’ which is well supported by other studies.14–19 ButSclerocorneal tunnel and sclerotomy have their advantages and disadvantages as a route for removal of IOFB. Enlargement of sclerotomy carries risk of hypotony, vitreous hemorrhage, peripheral vitreous incarceration into the wound intraoperatively, and RD postoperatively. Removal of the FB through sclerocorneal tunnel may damage or sacrifice the integrity of the posterior lens capsule and anterior capsular rim, but large FBs are easily removed through the sclerocorneal tunnel.–19et al.[et al.[et al.[et al.[Except few isolated case reports with large IOFBs, so far there is no data on removal of cataract and FB through the same route. In our experience, IOFB can be safely removed through the sclerocorneal tunnel and IOL can easily be implanted into the cilliary sulcus. In our study, the final BCVA was 20/30 or better in seven out of 18 cases, between 20/40 and 20/60 in six cases and counting finger (CF) at 1 meter (1 m) or less in five cases. This was very well comparable with similar studies.–81314 Az813et al. reportedl.[et al. reportedLate onset of RD in our series was seen in five out of 18 cases. The incidence of RD in the intraretinal FB group in this series was seen in five out of six cases. The possible causes of this overwhelming incidence of RD in the intraretinal FB group might be increased manipulation of FB and retinal penetration increasing the chance of proliferative vitreoretinopathy (PVR) changes postoperatively. These ch3Combined cataract and vitreous surgery with FB removal and IOL implantation offers not only faster visual rehabilitation but also24According to our clinical experience, the surgery that combines cataract extraction, IOFB removal and IOL implantation is a safe and desirable operation in patients with IOFB and significant lens opacities. The main advantage of this procedure is rapid visual rehabilitation with a single surgery, reducing the cost and patient discomfort. Randomized controlled studies involving larger number of patients with longer follow-up period may be needed to establish these facts.


A population-based case-control study of oral contraceptive use and breast cancer was carried out among young women (less than 43 years of age) at Group Health Cooperative of Puget Sound, Seattle, Washington. Use of oral contraceptives before first pregnancy did not materially differ between cases or controls. The rate ratio estimate of breast cancer incidence in women who had used oral contraceptives before first pregnancy compared to those who had not was 0.9 . There were no meaningful patterns of association between breast cancer and duration of use or formulation of oral contraceptive used before first pregnancy.


Connective tissue growth factor (CTGF) is a major chemotactic and mitogenic factor for connective tissue cells. The amino acid sequence shares an overall 28–38% identity to IGFBPs and contains critical conserved sequences in the amino terminus. It has been demonstrated that human CTGF specifically binds IGFs with low affinity and is considered to be a member of the IGFBP superfamily (IGFBP-rP2). In the present study, the expression of CTGF (IGFBP-rP2) in human leukaemic lymphoblasts from children with acute lymphoblastic leukaemia (ALL) was investigated. RNA samples from tumour clones enriched by ficoll separation of bone marrow or peripheral blood mononuclear cells (MNC) from 107 patients with childhood ALL at diagnosis and 57 adult patients with chronic myeloid leukaemia (CML) were studied by RT-PCR. In addition MNC samples from children with IDDM and cord blood samples from healthy newborns were investigated as control groups. Sixty-one percent of the patients with ALL (65 of 107) were positive for CTGF (IGFBP-rP2) expression. In the control groups, no expression of CTGF (IGFBP-rP2) in peripheral MNC was detected, and in the group of adult CML patients only 3.5% (2 of 57) were positive for this gene. The role of CTGF (IGFBP-rP2) in lymphoblastic leukaemogenesis requires further evaluation, as does its potential utility as a tumour marker. © 2000 Cancer Research Campaign


The aim of this study was to examine the relationship between the pharmacokinetics of daunorubicin (DNR), overexpression of P-glycoprotein (Pgp) and treatment response in acute leukaemia. Twenty-seven patients with acute leukaemia received DNR as part of induction therapy. The plasma and cellular levels of DNR and its metabolite daunorubicinol (DOL) were determined using high-performance liquid chromatography. There were no significant differences between patients who went into complete remission (12/23) compared with those who did not respond for the following pharmacokinetic parameters: DNR and DOL plasma AUC (area under the curve) and DNR plasma half-life and clearance. There was a significant difference in the cellular DNR and DOL AUC between responders and non-responders (P < 0.02). Seven patients were Pgp positive and 18 Pgp negative. There was no correlation between patient response and the presence of Pgp (P > 0.1), nor was there any correlation between the cellular concentration of DNR or DOL and Pgp (P > 0.3). To our knowledge this is the first report examining the relationship between DNR pharmacokinetics, patient response and Pgp expression. Our data indicated that acute leukaemia patients responding to chemotherapy had higher cellular DNR and DOL than non-responders; also, overexpression of Pgp appeared not to be the sole explanation for the lower cellular DNR levels as expected from in vitro studies.


Immunohistochemical, cytochemical and ultrastructural data showing vivid angiogenesis and numerous mast cells (MCs) in the bone marrow of 24 patients with active multiple myeloma (MM) compared with 34 patients with non-active MM and 22 patients with monoclonal gammopathy of undetermined significance (MGUS) led us to hypothesize that angiogenesis parallels progression of MM, and that MCs participate in its induction via angiogenic factors in their secretory granules. © 1999 Cancer Research Campaign


The performance of the Lung Imaging Fluorescence Endoscope (LIFE) system was compared with conventional bronchoscopy in 158 patients: 68 patients with invasive cancer, 42 patientswith abnormal sputum cytology findings (12 early cancer and 26 dysplasia), 17 cases withresected lung cancer and 31 smokers with symptoms. The respective results of conventionalbronchoscopy and LIFE for detection of dysplasia were; sensitivity 52% and 90% (biopsybasis), 62% and 92% (patient basis). Fluorescence bronchoscopy may be an importantadjunct to conventional bronchoscopy to improve the localization of subtle lesions ofbronchus.


Preoperative bronchoscopic findings of thyroid carcinoma with tracheal invasion wereexamined and compared with the histopathological findings. Tracheal sleeve resectionwas performed in 20 cases. The bronchoscopic findings were classified into 5 types:confirmed tumor in the tracheal lumen (5 cases), extramural compression of the tracheaplus mucosal change (9 cases), extramural compression of the trachea (2 cases), mucosalchanges only (3 cases), normal findings (1 case). Pathological findings revealed that theextent of invasion in the tracheal wall varied according to each of the above bronchoscopictypes. The number of tracheal rings with adventitial invasion averaged 0.8 (maximum2) more than preoperative bronchoscopic findings of the number of tracheal ringswith mucosal invasion. This study demonstrated the necessity of resective 2 more trachealrings than is indicated by the bronchoscopic findings.


Recently trans-dicarboxylato platinum(IV) complexes with mainly acetate,trifluoracetate or short-chain carboxylate groups (<11 carbons) in the axial position have beendescribed in the literature. In this paper we report on the synthesis and characterization of a new class of ethylenediamineplatinum(IV) compounds that have high lipophilic long-chain carboxylate ligands either in the axialor equatorial position. The platinum(IV) compounds with the lipophilic trans-carboxylate ligands inthe axial position were prepared by acylation of the trans-dihydroxo platinum(IV) species using anacyl halide in the presence of pyridine. In contrast to previous publications[1] the yields wereexcellent (up to 94%!).A series of new ethylenediamine (en) platinum(IV) complexes of the type Pt


A slowly progressive Heymann nephritis (SPHN) was induced in three groups of rats by weekly injections of a chemically modified renal tubular antigen in an aqueous medium. A control group of rats received the chemically unmodified version of the antigen in an aqueous solution. One group of SPHN rats were pre- and post-treated with weekly injections of IC made up of rKF3 and rarKF3 IgM antibody at antigen excess (MIC) . One group of SPHN rats were post-treated with MIC 3 weeks after the induction of the disease and one group of SPHN animals received no treatment. The control group of rats received pre- and post-treatment with sonicated u/c rKF3.            The incidence of immune-complex glomerulonephritis (ICGN) in the untreated SPHN rats was 87%, in the pre- and post-treated animals 13%, and in the post-treated-only rats 20%. Rats receiving sonicated ultracentrifuged rKF3 antigen did not develop ICGN.            The present experiment demonstrates that the development of SPHN can be not only prevented but also effectively terminated by our newly developed modified vaccination             technique.


In the present study we describe the effect of Zn(pic)2 and Zn(asp)2 on thereplication of VZV and on the infectivity of free virions. The experiments are done using BHK-21cells, a clinical isolate of VZV and Zn-complexes in concentration of 10 μM. When Zn-complexesare present during the whole period of infection, the yield of infectious virus progeny decreases upto 98%. The infectivity of VZV is completely restored after the removal of zinc. The virucidal effect ismanifested at the 2nd h of contact, when 90% of the virions are inactivated. The results show thatboth Zn(pic)2 and Zn(asp)2 specifically inactivate free VZV virions with no effect on viral replication.Zn(II) picolinate and aspartate, Zn(pic)


In vivo and in vitro antitumour activity of Htbh,Thiiobenzyhdrazide (Htbh) and its Cu(II) complexes,


Evidence in the literature suggests that hyperthermia (HT) or inhibitors of calmodulin can increase the sensitivity of rodent cells to bleomycin (BLM) by interfering with DNA repair functions. In an attempt to explore methods of improving the efficacy of thermochemotherapy we have investigated the individual and combined effects of HT (44 degrees C) and the calmodulin inhibitor trifluoperazine  on early plateau phase monolayer cultures of mouse EMT6 tumour cells for simultaneous exposures to BLM. Early plateau phase cultures are relatively resistant both to HT and to BLM. The selected HT and TFP regimens  were non-toxic. Comparing the sensitizing effect (given by the ratio: Do BLM/Do BLM + modifier) of the various regimens on BLM-treated cells, we found that: TFP alone had a marginal effect (ratio 1.3), HT alone showed significant potentiation (ratio 19) and the combination of HT and TFP strongly sensitized (ratio greater than 110) cells to BLM cytotoxicity. We propose that the use of calmodulin inhibitors in thermochemotherapy is worthy of further evaluation.


To the Editor: Verotoxin-producing Escherichia coli (VTEC) of serogroup O157 causes severe gastrointestinal and renal illness; clinical signs may be mild diarrhea, hemorrhagic colitis, or hemolytic uremic syndrome (HUS). Typically, 10%–15% of reported VTEC infections quickly progress to HUS  identified 18 cases of verotoxin-producing SF-O157 infection in Scotland, 13 of which were associated with a nursery. HUS developed in 8 of the 18 patients; those with thrombotic microangiopathy were admitted to the renal unit of a specialist pediatric hospital, which immediately reports cases of HUS to HPS as part of national surveillance (t test showed that nadirs for serum albumin were significantly higher for children with SF-O157 HUS (p = 0.03;  Statistical analysis by  Our study highlights a number of lessons. Medical practitioners rarely have the opportunity to recognize patients at such an appreciable and predictable risk of progressing rapidly to anuric renal failure as they do when they see children with early O157 infection. Failure to appreciate the potential gravity of O157 infection and the possible development of HUS may result in avoidable illness and even death. Our investigation of the prehospital management of SF-O157 and non–SF-O157 in this cohort found no difference in pharmacologic intervention or duration of delay in admission to hospital.Our study has limitations. A number of patients in the cohort were prescribed antimicrobial drugs and/or antimotility drugs or were sent home from the local hospital without hospital admission or further monitoring; such actions potentially exacerbate clinical outcomes (Our data suggest that infection with SF-O157 results in less severe colitis than does the more common non–SF-O157 infection. Less severe colitis could result in a lower risk for renal disease because less verotoxin would be translocated into the bloodstream and bound to the kidneys. However, patients infected with SF-O157 had anuria for longer periods and consequently had longer sessions of peritoneal and hemodialysis. Although unknown bacterial or host inflammatory cytokines may contribute to enhanced disease progression, this observation is surprising and requires further investigation. Additional research is needed to learn more about the virulence of SF-O157 strains and establish other host factors that contribute to disease progression.


A total of 85 spontaneous rat fibrohistiocytic tumours were evaluated histologically and assessed for the presence or absence of metastases. The overall incidence in controls from 2-year carcinogenicity studies was 2.7%. The tumours occurred principally in the subcutaneous and deep soft tissues, and generally appeared after 18 months of age. Four histological types were recognized: histiocytic (17%), pleomorphic (33%), cellular (17%) and very fibrous (33%). Histiocytic tumours were highly malignant, and most produced metastases. Pleomorphic and cellular neoplasms occasionally produced metastases and must be regarded as potentially malignant. Very fibrous lesions were essentially benign. The close resemblance, both histologically and biologically, between rat and human fibrohistiocytic neoplasms supports the use of the fibrohistiocytic concept in laboratory-animal pathology. Study of these rat tumours may provide insight into the development of human fibrohistiocytic neoplasms.


Serum carcinoembryonic antigen (CEA) concentrations were found to be raised in 28 of 72 black patients (39%) with hepatocellular cancer (HCC). The degree of elevation was slight or moderate, except in 3 patients in whom values greater than 20 ng/ml were recorded. No significant correlation could be demonstrated in individual patients between the serum CEA concentration and various tests of liver function. The mean CEA value in the patients with cirrhosis in the non-tumorous liver was slightly higher than that in those without cirrhosis, but the difference did not reach statistical significance. There was no correlation between serum CEA and alpha-foetoprotein (AFP) levels.


The effects of histone deacetylase inhibitor (HDACi) treatment on SV40 transcription and replication were determined by monitoring the levels of early and late expression, the extent of replication, and the percentage of SV40 minichromosomes capable of transcription and replication following treatment with sodium butyrate (NaBu) and trichostatin A (TSA).The HDACi treatment was found to maximally stimulate early transcription at early times and late transcription at late times through increased numbers of minichromosomes which carry RNA polymerase II (RNAPII) transcription complexes and increased occupancy of the transcribing minichromosomes by RNAPII. HDACi treatment also partially relieved the normal down-regulation of early transcription by T-antigen seen later in infection. The increased recruitment of transcribing minichromosomes at late times was correlated to a corresponding reduction in SV40 replication and the percentage of minichromosomes capable of replication.These results suggest that histone deacetylation plays a critical role in the regulation of many aspects of an SV40 lytic infection. Structural and biological changes in chromatin structure are brought about by changes in the activity of histone acetyltransferases (HATs) and histone deacetylases (HDACs) which add or remove acetyl groups from lysine residues on histone tails respectively. It has been well established that inhibition of HDAC activity is characterized by two important changes within the cell (i) an increase in the amount of hyperacetylated histones  and (ii)Streptomyces and a potent inhibitor of HDAC activity. NaBu has been extensively used as a HDAC inhibitor (HDACi), though it is far less efficient (required in millimolar quantities) in its inhibition capabilities as compared to TSA (required only in nanomolar quantities). DNA micro array studies have shown that almost 7 % to 10% of genes undergo altered gene expression on HDACi using TSA   for one hour. The cells were then washed twice in chilled PBS and extracted using the Hirt method of viral DNA extraction  or RPA 70 [replicating Minichromosomes]). ΔCT was expressed either as a percentage of minichromosomes containing the epitope for a particular antibody. Statistical analysis was performed using two tailed Student's t test.® polyacrylamide gel  under denaturing conditions and electroblotted onto PVDF membrane . Antibodies used for western blotting were goat polyclonal anti GAPDH (sc-20357); Santa Cruz Biotechnology; rabbit GST-fusion anti VP1 [Protein extracted from untreated, NaBu treated and TSA treated 48 hour wild type 776 SV40 virus infected cells were harvested and lysed using 1× RIPA buffer. Protein levels were determined with the Micro BCA™ Protein Assay kit . The proteins were separated on a 4–20% PAGEranti VP1  and antiRNAP II: RNA Polymerase II, NaBu: sodium butyrate; TSA: trichostatin A; HDACi: HDAC inhibitors; ISF: immune selection and fragmentation; ISFIP: ISF followed by a second immunoprecipitation; ReChIP: re-chromatin immunoprecipitation; GAPDH: glyceraldehydes-3-phosphate dehydrogenase; VP1: viral protein 1; P.I.: post-infectionThe author(s) declare that they have no competing interests.LB performed the experiments and wrote the manuscript. BM edited the manuscript and provided input into the experiments. Both authors read and approved the final manuscript.


To test the role of serotonin in chemotherapy-induced nausea and emesis, ten cancer patients were pretreated with the serotonin synthesis inhibitor para-chlorophenylalanine (PCPA). PCPA (2 g 8 hourly for 2 or 3 days prior to cisplatin) reduced the spontaneous urinary excretion of 5-hydroxyindoleacetic acid (5-HIAA), inhibited the increase in urinary 5-HIAA induced by cisplatin and markedly attenuated the acute period of nausea and vomiting associated with the cytotoxic drug. These results indicate that gastrointestinal serotonin mediates cisplatin-induced emesis and that the amount of serotonin released by cisplatin is a major factor in determining the severity of the acute period of emesis experienced by the patient.


Between January 1981 and December 1985, 122 patients with non-seminomatous germ cell tumours (NSGT) were seen at a regional referral centre. Of these, a total of 98 patients received chemotherapy for metastatic disease. Treatment was given within collaborative EORTC Urology group studies, all of which involved cis-platin-containing schedules. Ninety patients had tumours of testicular origin, and their 2 year actuarial survival rate is 91%; 8 had tumours of extragonadal origin and their 2 year actuarial survival is 25%. Patients with testicular tumours were subdivided by volume of metastatic disease using the recommendations of the Testicular Cancer Subgroup of the MRC Urological Cancer Working Party and survival was significantly worse in the group with very large volume metastatic disease  compared with the groups with large volume metastases  and small volume metastases . There were 31 patients with Stage I disease at presentation; of these 6 were treated by prophylactic abdominal radiotherapy and 25 were managed by a policy of surveillance only. Seven of these Stage I patients (23%) relapsed with metastatic disease (median 8 months); all have been successfully treated with chemotherapy. These data confirm that the majority of patients now presenting with metastatic NSGCT are curable with chemotherapy, but that a small proportion with very large volume metastases or extragonadal tumours require alternative chemotherapy schedules.


Cytological studies in the light microscope showed that thioacetamide (TAA) depressed the mitotic index in cultures of skin fibroblasts at the lowest concentrations used (100 μg/ml). At high concentration (1 mg/ml), TAA tended to cause aberration in nuclear morphology. Ethylenethiourea (ETU) had no effect on either mitotic index or nuclear morphology at 1 mg/ml. Fibroblast cultures treated with 1 mg/ml TAA and cultures grown in the presence of 2 mg/ml ETU were studied by electron microscopy. In some TAA-treated cells there was unfolding of the nuclear membrane and enlargement and granulation of the nucleolus, but these effects were not correlated. In all cells, TAA caused severe and characteristic damage to the majority of mitochondria, whether or not there were nuclear aberrations. The organelle showed extensive swelling of the cristae of the inner membrane and an increase in matrix density. Ultrastructure of other cell components appeared to be unaffected by this treatment. In ETU-treated cells some less severe swelling of inner mitochondrial membranes was seen and only in a minority of cells, whilst all other cell structures appeared normal. Similar membrane swelling and increase in matrix density was seen in isolated rat liver mitochondria after incubation with TAA, indicating a direct antimitochondrial effect of the carcinogen.When yeast cells were treated with TAA and ETU, primary antimitochondrial activity of these compounds was apparent from (1) inhibition of growth in non-fermentable medium, (2) selective blockage of mitochondrial protein synthesis and (3) induction of mitochondrial mutations. TAA was much more effective than ETU in all these respects.


Sir,In a recent letter criticising one of several IARC Interphone studies investigating possible associations between mobile phone use and brain tumour risk, Editors NoteThe Editor-in-Chief of British Journal of Cancer now considers the correspondence published in Volume 94 Issue 9 and above closed.


Purpose: Soft tissue sarcomas (STS) account for less than 1% of all malignancies and constitute a heterogeneous tumor entity in which malignant fibrous histiocytomas (MFH) represent one-third and are characterized by a lack of type-specific differentiation. A defective mismatch repair (MMR) system cause the familial cancer syndrome hereditary non-polyposis colorectal cancer (HNPCC), and since occasional MFH have been described in HNPCC patients we assessed the contribution of defective MMR to the development of MFH.Methods: MMR status was characterized in a series of 209 histopathologically reviewed MFH. Tissue microarray sections from the tumors were immunohistochemically stained for the MMR proteins MLH1, MSH2 and MSH6, and cases with aberrant staining were further characterized for microsatellite instability.Results and Discussion: Two of the 209 STS–a storiform-pleomorphic MFH and a myxofibrosarcoma–showed concomitant loss of MSH2 and MSH6, but retained staining for MLH1 on both cases. The myxoid tumor also had a microsatellite unstable phenotype. These findings, together with previous observations of defective MMR in pleomorphic STS, indicate that these tumors may be part of the HNPCC-associated tumor spectrum and demonstrate that MMR defects occur in a small subset of STS.


Girls who engage in strenuous physical activity are often amenorrheic and have recently been reported to be at a reduced risk of breast cancer. To determine whether moderate amounts of exercise affect menstrual cycle patterns and ovulatory frequency in young postmenarcheal girls, the menstrual cycles and physical activity patterns of 168 high school girls were monitored for a 6 month period. Anovulatory cycles were associated with later age at menarche, fewer elapsed years since menarche and greater levels of energy expended per week in physical activity. After adjusting for age at menarche and years since menarche, there was a significant dose-related trend in the risk of anovular menstrual cycles associated with increasing levels of physical activity (1-sided P = 0.03). Major determinants of average cycle length were weekly average energy expenditure , age at menarche (an increase of 0.7 days per year of age) and race (Asians having cycles about 1.9 days longer than Caucasians). Because a major determinant of breast cancer risk may be the cumulative number of ovulatory cycles, these data suggest that regular participation in moderate physical activity, by reducing the frequency of ovulatory cycles in adolescence, may provide an opportunity for the primary prevention of breast cancer.


The structure of circulating chromogranin A (CgA) of phaeochromocytoma patients was characterised and compared with that of CgA extracted from tumours. Size exclusion chromatography experiments provided evidence that CgA is present in the blood of different patients, as well as in tumour extracts, as multiple forms having different hydrodynamic sizes of 600 kDa (CgA-I), 100 kDa (CgA-II) and 55 kDA (CgA-III). The amount of each CgA form as a proportion of the total antigenic material was different in different patients. Western blot analysis of chromatographic fractions indicated that these forms are made up by polypeptides of similar molecular weight (about 60-70 kDa). All CgA forms express the epitopes recognised by two monoclonal antibodies (A11 and B4E11), directed against residues 68-70 and 81-90 of human CgA. However, their relative immunoreactivity was markedly different. No evidence for the presence of multimeric complexes in the CgA-I fraction was obtained by various immunological and biochemical methods. These results suggest that circulating CgA in phaeochromocytoma patients consists of at least three forms that appear to be made up by polypeptides with similar molecular weight and different hydrodynamic properties and immunoreactivity. We hypothesise that different conformations and shapes contribute to the heterogeneity of circulating CgA.


After the 1999 West Nile (WN) encephalitis outbreak in New York, 2,300 overwintering adult mosquitoes were tested for WN virus by cell culture and reverse transcriptase-polymerase chain reaction. WN viral RNA and live virus were found in pools of Culex mosquitoes. Persistence in overwintering Cx. pipiens may be important in the maintenance of WN virus in the northeastern United States.


The prevalence of primary liver cancer (PLC) varies throughout the world. It has been attributed to variations in incidence of the predominant histological type, hepatocellular carcinoma (HCC). The incidence of PLC types other than HCC such as cholangiocellular carcinoma (CCC) is far less known, especially in low-incidence areas. The aetiology of HCC and other PLC types is obscure, with the exception of the association between HCC and cirrhosis as well as chronic viral hepatitis. The present retrospective incidence and aetiology study concerns a well-defined population from a period with a high autopsy frequency. Preserved biopsy specimens were re-evaluated histopathologically and patient records were studied. Among 590 histologically verified cases of PLC, HCC constituted 90%, CCC 8% and a mixed form of these types 1%. At the end of the study period the annual age-standardised incidence rate of HCC was 3.6 cases per 100,000 inhabitants. Other PLC types were hepatoblastoma (n = 3), fibrolamellar carcinoma (n = 2), angiosarcoma (n = 1) and infantile haemangioendothelioma (n = 1), each constituting less than 1% of the PLC cases. Comparing HCC with CCC we found that cirrhosis (70%) and alcoholism (21%) was significantly more frequent in HCC, and cholelithiasis was significantly more common (60%) in patients with CCC. In the majority of the PLC cases with liver cirrhosis this disorder was unknown before diagnosis of the tumour.


Malignant pericarditis, when associated with massive pericardial effusion, presents a critical condition in lung cancer patients. Because this condition often arises in terminally ill patients, intensive therapy such as multi-drug combination chemotherapy is rarely appropriate. This study evaluated the clinical relevance of intrapericardial administration of carboplatin for controlling malignant pericardial effusions associated with non-small-cell lung carcinoma (NSCLC). The method used for 10 eligible patients consisted of draining the pericardial effusion and infusing 300 mg/body of carboplatin in 50 ml of saline through an in-place catheter into the pericardial space and clamping the catheter for 40 min. Nine of the 10 patients showed satisfactory results, and 8 experienced complete regression of the effusion. No major or minor adverse effects were observed. Pharmacokinetics analysis revealed that the concentration of free platinum in the pericardial fluid was very high while that of total platinum in the circulating plasma was very low, assuring the usefulness of the intrapericardial instillation of carboplatin in terminally ill patients for controlling malignant pericardial effusion when the systemic delivery of cytotoxic agents is inappropriate. © 2000 Cancer Research Campaign


The present study evaluated serial serum measurements of tissue polypeptide-specific antigen (TPS) in comparison with prostate-specific antigen (PSA) for assessment of tumour progression in patients with advanced prostate cancer receiving intermittent androgen suppression therapy (IAS). Twenty-three men were recruited into an IAS trial consisting of an initial 8 months of androgen suppression, followed by cycles of treatment cessation and resumption of therapy upon increases of PSA > 20 ng ml(-1) to prolong the hormone responsiveness of the tumour cells. Periods of androgen suppression resulted in reversible reduction in serum testosterone (< 1.8 nmol I(-1)) and PSA (< 4 ng ml(-1)) and decreases in tumour volume (mean reduction for first cycle 24 +/- 10%), indicating partial growth arrest and apoptotic regression of the tumours. In contrast to PSA values, non-specifically elevated TPS values were found in 8 of 23 patients. In 15 of 23 patients, TPS fell during periods of apoptotic tumour regression and increased simultaneously with testosterone and preceded the increases in PSA by 2 months during the period of treatment cessation. Although TPS represents a highly sensitive marker of tumour proliferation in this IAS clinical model of controlled tumour regression and regrowth, its low specificity compared with PSA limits its usefulness to monitoring of prostate cancer patients with proven absence of non-specific elevations of this marker.


The presence of tumour cells in the blood circulation may predict disease recurrence and metastasis. We have evaluated the specificity and sensitivity of detecting hepatoma cells in blood using nested polymerase chain reaction with primers specific for the alphafetoprotein (AFP) gene. The nested polymerase chain reaction amplified a 270-base pair AFP DNA fragment from cDNA of Hep 3B hepatoma cells. In a reconstitution experiment, AFP mRNA was detected from peripheral mononuclear cells isolated from 10 ml of blood containing as few as ten Hep 3B cells. Peripheral mononuclear cells from the blood of 20 hepatoma patients were analysed, and 19 patients showed positive AFP mRNA expression. Seven of 13 samples from hepatitis patients also showed positive AFP mRNA expression. All five paired samples of peripheral blood or umbilical cord blood from pregnant mothers and their babies, respectively, showed positive AFP expression. None of 22 control samples was positive. The presence of AFP mRNA in the blood of hepatitis or hepatoma patients suggests the presence of circulating hepatoma cells or hepatocytes in the circulation. The high incidence of AFP mRNA in the blood of hepatoma patients supports the notion of early haematogenous spreading of the disease.


The cytoreductive effects of anti-transferrin receptor (anti-TfnR) immunotoxins (ITs) and of ricin toxin against tumour micromasses have been evaluated in a multicellular tumour spheroid (MTS) model. More than 600 (656) MTSs obtained with human breast carcinoma (MCF7) or rat glioblastoma (9L) cell lines were treated individually with ITs or toxin and the effects induced by the treatment were measured for each MTS as volume variation vs time by applying the Gompertz growth model. Two dose-dependent patterns of MTS growth were observed in MTSs of both cell lines in response to IT or toxin treatment: (1) complete inhibition of MTS growth ('sterilisation'); and (2) partial/complete inhibition ('heterogeneous response'). Within the range of IT or toxin concentrations resulting in partial inhibition of MTS growth, the sensitivity of treated MTSs was extremely heterogeneous (the cytoreductive effects varying between 0.1 and 4 logs of cells killed for a given IT or toxin concentration). Analysis of the post-treatment regrowth kinetics indicated that treated non-sterilised and control MTSs reached the same final limiting volumes. However, the doubling time estimated for the surviving cells of treated MCF7 and 9L MTSs ranged between 15 and 50 h, indicating that each MTS had individual growing potential. In conclusion, our results indicate that at substerilising IT concentrations individual heterogenicity of MTSs may greatly influence the cytoreductive potential of ITs. An implication of our study is that the efficacy of an IT treatment in eradicating disseminated micrometastases may not be predictable a priori. The MTS model that we describe in this paper may help in dissecting out factors limiting the effect of ITs in three-dimensional tumours.


Twenty-seven patients with Hodgkin's (n = 19) and non-Hodgkin's (n = 8) lymphomas underwent cytological and colposcopic screening of the uterine cervix. Colposcopically directed cervical punch biopsies were taken from all patients in whom a colposcopic abnormality was detected. Lymphoma patients were compared with 79 controls with normal cervical cytology and no known haematological abnormality. Colposcopically directed punch biopsies were taken from the cervical transformation zone of all controls. Significantly more lymphoma patients (19%) than controls (3%) had CIN II or III (P less than 0.01) and cervical human papillomavirus infection, as judged by the presence of koilocytes . All six lymphoma patients with CIN had Hodgkin's disease (HD), and five had received combination chemotherapy. Half of the cases of CIN in lymphoma patients and all the cases of CIN in control patients were not detected by cervical cytology. This study suggests that female patients with HD are at increased risk of CIN, and that cervical cytology alone may be an inadequate form of screening for these patients.


Ontogenetic study on the expression of cytokeratin (CK) polypeptides within particularsubsets of rat thymic epithelial cells (TEC) has been performed by a large panel of anti-CKmonoclonal antibodies (mAbs) using the streptavidin-biotin immunoperoxidase method.Simultaneous presence of two or more CK subunits in the same TEC has been demonstratedby double immunoflouorescence labeling. The obtained results showed that the expressionof CK polypeptides in fetal and neonatal thymus differed from the adult patterns. The maindifference was observed in expression of CK10, 18, and 19 polypeptides. During fetalontogeny, CK10 and 18 are markers for most medullary TEC or a subset of medullary TEC,respectively, whereas CK19 is mainly a pan-TEC marker. In the adult animals, they arelocalized in the cortical and a subset of medullary TEC (CK18), subcapsular/perivascular andsome medullary TEC (CK19), or in a subset of medullary TEC and Hasall’s corpuscles (HC)(CK10). The switch in their expression in the cortex was observed during the first two weeksof postnatal life.


In the present study, we investigated telomerase activation and its implications in plasma cell dyscrasias including monoclonal gammopathy of undetermined significance (MGUS), multiple myeloma (MM) and plasma cell leukaemia (PCL). All 5 patients with MGUS exhibited normal levels of telomerase activity in their plasma cells. Elevated telomerase activity was found in the samples from 21/27 patients with MM and 4/4 with PCL. In addition, 4 myeloma cell lines all expressed high levels of telomerase activity. The expression of telomerase reverse transcriptase (hTERT) and telomerase RNA template (hTER) was positively associated with the levels of telomerase activity in MM/PCL. Tankyrase expression was upregulated, concomitant with the induction of hTERT and activation of telomerase in MM/PCL. The present findings indicate that MGUS cells may not be immortalized and that activation of telomerase plays a role in the malignant transformation from MGUS to MM. © 2001 Cancer Research Campaign


Two P388 cell lines with acquired resistance to daunorubicin have been shown to exhibit cross-resistance to anthrapyrazolone (NSC 357885). The degree of cross-resistance observed in these cell lines (58 and 150 fold) is similar to those observed towards daunorubicin (34 and 142 fold). However the decreased drug accumulation observed for daunorubicin in the resistant cell lines (4-6 fold) is not observed for anthrapyrazolone. Similarly, verapamil can increase daunorubicin accumulation in resistant cells but has no effect on anthrapyrazolone accumulation. It is concluded that in contrast to daunorubicin, decreased anthrapyrazolone accumulation is not the resistance mechanism operative in daunorubicin resistant cell lines towards anthrapyrazolone.