LM985 has been shown previously to hydrolyse to flavone acetic acid (LM975) in mouse plasma and to produce significant anti-tumour effects in transplantable mouse colon tumours (MAC). It has undergone Phase I clinical trials and dose limiting toxicity was acute reversible hypotension. Substantially higher doses of LM975 can be given clinically without dose limiting toxicity. We have investigated the activity of LM975 against a panel of MAC tumours and also the in vitro cytotoxicity of both LM985 and LM975 in two cell lines derived from MAC tumours. LM985 is considerably more cytotoxic than LM975 in vitro but increased length of exposure to LM975 results in improved activity. Single in vivo injection of LM975 showed no activity against the ascitic tumour MAC 15A, moderate activity against the s.c. poorly differentiated tumour MAC 13 and produced a significant growth delay in the well differentiated MAC 26. These latter responses were considerably enhanced by repeated injection 7 days later. Pharmacokinetic studies in mice following i.p. injection of LM985 demonstrated rapid degradation of LM985 to LM975 in the peritoneum. Length of exposure as well as drug concentration appear important factors in determining anti-tumour responses.


The effects of butyrate and retinoic acid in combination with catecholamines or histamine on the HN-1 human head and neck squamous carcinoma cell line were investigated analysing cell proliferation, placental alkaline phosphatase (PLAP) activity, and relative cytokeratin content. Butyrate inhibited cell proliferation in agar, whereas retinoic acid induced a small inhibitory effect. Butyrate enhanced PLAP activity in a time related manner in contrast to retinoic acid, which had no significant effect. However, retinoic acid inhibited the efficacy of butyrate to induce PLAP activity. A synergistic enhancement of PLAP activity was demonstrated after treatment of butyrate pretreated cells with catecholamines or histamine. The beta-adrenergic antagonist propranolol partly inhibited the aforementioned enhancement of PLAP activity, whereas the alpha-adrenergic antagonist phentolamine further enhanced PLAP activity. Indirect labeling of keratins with a polyclonal antibody showed that cytokeratin content was enhanced by butyrate but not by retinoic acid. Further analysis of cytokeratin content using four monoclonal antibodies showed that labeling of cytokeratins (5 + 8) was increased by butyrate. PLAP activity could be modulated by a concerted action of either butyrate plus retinoic acid or butyrate plus catecholamines or histamine, indicating a possible role for PLAP in tumour cell proliferation.


Potential anti-cancer agents have classically undergone clinical assessment in Phase I, II and III trials. This paper examines the role of these trials and pre-clinical studies in the light of improving cancer chemotherapy. Many patients must now be treated with standard therapy before investigational drugs can be ethically used. The introduction of combined modality trials will require a very prolonged follow-up to demonstrate improved survival and recognize late onset of chronic toxicity.


Arthropods are infected by a wide diversity of maternally transmitted microbes. Some of these manipulate host reproduction to facilitate population invasion and persistence. Such parasites transmit vertically on an ecological timescale, but rare horizontal transmission events have permitted colonisation of new species. Here we report the first systematic investigation into the influence of the phylogenetic distance between arthropod species on the potential for reproductive parasite interspecific transfer.We employed a well characterised reproductive parasite, a coccinellid beetle male-killer, and artificially injected the bacterium into a series of novel species. Genetic distances between native and novel hosts were ascertained by sequencing sections of the 16S and 12S mitochondrial rDNA genes. The bacterium colonised host tissues and transmitted vertically in all cases tested. However, whilst transmission efficiency was perfect within the native genus, this was reduced following some transfers of greater phylogenetic distance. The bacterium's ability to distort offspring sex ratios in novel hosts was negatively correlated with the genetic distance of transfers. Male-killing occurred with full penetrance following within-genus transfers; but whilst sex ratio distortion generally occurred, it was incomplete in more distantly related species.This study indicates that the natural interspecific transmission of reproductive parasites might be constrained by their ability to tolerate the physiology or genetics of novel hosts. Our data suggest that horizontal transfers are more likely between closely related species. Successful bacterial transfer across large phylogenetic distances may require rapid adaptive evolution in the new species. This finding has applied relevance regarding selection of suitable bacteria to manipulate insect pest and vector populations by symbiont gene-drive systems. Reproductive parasites represent a diverse assemblage of maternally inherited microorganisms that induce aberrations in their hosts' reproductive biology in order to proliferate within arthropod populations. Their vertical transmission route greatly limits opportunities for interspecific transfer. However, phylogenies of hosts and parasites are generally not congruent, indicating that interspecific horizontal transfer events characterise the evolutionary history of these selfish genetic elements .Wolbachia is the best known, a variety of other microbial taxa has also evolved to manipulate arthropod reproduction, including microsporidia, gamma proteobacteria and members of the genera Cardinium, Rickettsia and Spiroplasma  – [AM779613]). We discarded the data from the 3' region of the 16S fragment which spanned the 5' end of the ND1 gene and the leucine tRNA and combined the remaining sequence from the two rDNA genes. A sequence alignment was constructed using CLUSTALW and manually edited, then a tree was constructed using the neighbour joining method in MEGA version 4.0 [A. bipunctata were calculated in MEGA using the parameter options above. Correlations of the species means for bacterial transmission rate and offspring sex ratio with genetic distance were determined non-parametrically using Spearman rank tests due to non-normality of dependent variables.For phylogenetic analysis we investigated two conserved sections from the 3' ends of the 16S and 12S mitochondrial ribosomal subunits of all host species. These were amplified using primer pairs N1-J-12585 – LR-N-13398 (16S) and SR-J-14233 – SR-N-14588 (12S) AM7798 – [AM77sion 4.0 ). RobustMCT collected insect samples from the field, performed injections and conducted molecular work. MCT and MENM collaboratively designed the study, reared ladybirds in the laboratory and interpreted the results. MCT drafted the paper. Both authors read and approved the final manuscript.


A panel of doxorubicin-resistant sublines of the human small-cell lung carcinoma cell line GLC4 displays decreasing DNA topoisomerase II alpha  mRNA levels with increasing resistance. In the present study we describe how this decrease may be regulated. No significant differences in TopoII alpha mRNA stability or gene arrangement were found, using mRNA slot-blotting and Southern blotting, in the most resistant cell line compared with the parental cell line. To investigate if TopoII alpha gene copy loss contributed to the mRNA decrease, fluorescence in situ hybridisation using a TopoII alpha-specific probe was performed. During doxorubicin resistance development, the composition of the population in each cell line shifted with increasing resistance, from a population in which most cells contain three TopoII alpha gene copies (GLC4) to a population in which most cells contain only two copies. A partial revertant of the most resistant cell line displayed a shift back to the original situation. We conclude that the TopoII alpha gene copy number decrease per cell line is in good agreement with the decreased TopoII alpha mRNA and protein levels, and TopoII activity levels in these cell lines which were described previously.


Of the 2 patients with left-sided portal hypertension, one died after splenectomy and one rebled 6 months after left colectomy. This study suggests that extrahepatic portal hypertension is a series complication after liver transplantation that could be prevented by meticulous portal anastomosis and closure of portal tributaries or portasystemic shunts to improve the  portal venous flow. However, any ligation has to be performed under ultrasound guidance to avoid inadventent venous ligations.This study reports our experience of 8 cases of extrahepatic portal hypertension after 273 orthotopic liver transplantations in 244 adult patients over a 10- year period. The main clinical feature was ascites, and the life-threatening complication was variceal bleeding. Extrahepatic portal hypertension was caused by portal vein stenosis in 6 patients, and left-sided portal hypertension in 2 patients after inadventent ligation of portal venous tributaries or portasystemic shunts. All patients with portal vein stenosis had complete relief of portal hypertension after percutaneous transhepatic venoplasty (


The predicted folded structures of these twenty H/ACA snoRNA-like miRNA precursors reveal molecules which resemble the structures of known box H/ACA snoRNAs. The genomic regions surrounding these predicted snoRNA-like miRNAs are often similar to regions around snoRNA retroposons, including the presence of transposable elements, target site duplications and poly (A) tails. We further show that the precursors of five H/ACA snoRNA-like miRNAs  bind to dyskerin, a specific protein component of functional box H/ACA small nucleolar ribonucleoprotein complexes suggesting that these molecules have retained some H/ACA snoRNA functionality. The detection of small RNA molecules that share features of miRNAs and snoRNAs suggest that these classes of RNA may have an evolutionary relationship.MicroRNAs (miRNAs) and small nucleolar RNAs (snoRNAs) are two classes of small non-coding regulatory RNAs, which have been much investigated in recent years. While their respective functions in the cell are distinct, they share interesting genomic similarities, and recent sequencing projects have identified processed forms of snoRNAs that resemble miRNAs. Here, we investigate a possible evolutionary relationship between miRNAs and box H/ACA snoRNAs. A comparison of the genomic locations of reported miRNAs and snoRNAs reveals an overlap of specific members of these classes. To test the hypothesis that some miRNAs might have evolved from snoRNA encoding genomic regions, reported miRNA-encoding regions were scanned for the presence of box H/ACA snoRNA features. Twenty miRNA precursors show significant similarity to H/ACA snoRNAs as predicted by snoGPS. These include molecules predicted to target known ribosomal RNA pseudouridylation sites  The major functions known for RNA were long believed to be either messenger RNAs, which function as intermediates between genes and proteins, or ribosomal RNAs and transfer RNAs which carry out the translation process. In recent years, however, newly discovered classes of small RNAs have been shown to play important cellular roles. These include microRNAs (miRNAs), which can regulate the production of specific proteins, and small nucleolar RNAs (snoRNAs), which recognise and chemically modify specific sequences in ribosomal RNA. Although miRNAs and snoRNAs are currently believed to be generated by different cellular pathways and to function in different cellular compartments, members of these two types of small RNAs display numerous genomic similarities, and a small number of snoRNAs have been shown to encode miRNAs in several organisms. Here we systematically investigate a possible evolutionary relationship between snoRNAs and miRNAs. Using computational analysis, we identify twenty genomic regions encoding miRNAs with highly significant similarity to snoRNAs, both on the level of their surrounding genomic context as well as their predicted folded structure. A subset of these miRNAs display functional snoRNA characteristics, strengthening the possibility that these miRNA molecules might have evolved from snoRNAs. Small nucleolar RNAs (snoRNAs) and microRNAs (miRNAs) are two classes of abundant non-coding regulatory RNAs that carry out fundamental cellular activities but that have only been comprehensively investigated in recent years. SnoRNAs are small RNA molecules of approximately 60–300 nucleotides in length which generally serve as guides for the catalytic modification of selected ribosomal RNA nucleotides MiRNAs are ∼18–24 nucleotide-long RNAs that are processed out of ∼70 nucleotide-long hairpin structures  Many members of the snoRNA and miRNA classes are well conserved throughout evolution Recent reports have also described some miRNAs as being derived from TEs, suggesting a possible mechanism for the rapid generation of miRNAs and their corresponding target sites. In the first such report, Smalheiser and Torvik identified six miRNAs that are derived from TEs A recent report investigating the small RNAs present in human cells has demonstrated the existence of specific small RNA fragments derived from larger known non-coding RNA molecules A comparison between the genomic positions of reported miRNA genes from miRbase In addition to these known miRNAs encoded in box H/ACA snoRNAs, ten small fragments matching exactly to portions of eleven box H/ACA snoRNAs have been detected Apart from HBI-100, all box H/ACA snoRNAs from The UCSC Genome Browser mammalian conservation track shows that for all snoRNAs listed in The box H/ACA snoRNA/miRNA relationship described above was further investigated by studying all known miRNAs to determine whether they might be encoded within genomic regions predicted to harbour H/ACA snoRNAs. Indeed, if some miRNAs have evolved from H/ACA snoRNA encoding regions, they might still display snoRNA features. The mammalian version of the snoGPS program predicts pseudouridylation guides in human, mouse and rat genomes by scoring weakly conserved primary and secondary structure motifs using a deterministic search algorithm In order to investigate whether the number of snoGPS predicted hits above a certain threshold was significant, we used snoGPS to scan 100 sets of 676 randomly generated sequences of same length distribution as the miRNAs under study, as described in the 148 distinct extended miRNA molecules were predicted to encode at least one hit above a score of 35.0, which is the threshold that is ‘typically’ used when predicting new candidate snoRNAs by snoGPS-C The position of the predicted H/ACA snoRNA was compared to the position of the miRNA hairpin, taking coordinates downloaded from the UCSC Table Browser as described in the SnoGPS predicts guide sequences and corresponding rRNA pseudouridylation sites within snoRNAs. For all the best snoGPS hits with scores above 40 listed in The miRNAs reported in miRbase have not all been validated to the same extent. While the mature forms of some of the miRNAs have only been identified with a very small number of sequence reads, others have been identified by larger numbers of reads, display characteristic miRNA signatures . While the H box is located between the two hairpins, the ACA box is located at the 3′ end of the molecule. One or both of the hairpins contain bulges, allowing base-pairing with the target RNA, in complex pseudo-knot structures. In order to better characterise the predicted snoRNAs encoding miRNAs and visualise the position of the mature miRNA within these molecules, all predicted snoRNA sequences were folded using RNAstructure http://www.repeatmasker.org). Sixteen of the twenty miRNAs originally considered have repeat elements either overlapping the predicted snoRNA encoding the miRNA or within 400 nucleotides. The position of the repeat elements with respect to the position of the miRNA and predicted snoRNA is shown in Because numerous snoRNAs and miRNAs have been described as being derived from TEs, all extended miRNA molecules predicted to have box H/ACA snoRNA features surrounding them from  were furin silico predictions that some miRNA precursors sufficiently resemble box H/ACA snoRNAs to be bound by dyskerin.To explore further potential snoRNA-like features of these miRNA precursors and to investigate whether they have retained some H/ACA snoRNA functionality, we tested whether they can bind dyskerin, a protein component of the functional box H/ACA snoRNPs. Dyskerin serves as the pseudouridine synthase Two of the dyskerin-bound miRNA precursors were further characterised by fractionated northern analysis to investigate where the predicted snoRNAs and smaller fragments localise in the cell. As shown in Numerous miRNAs have been previously shown to be repeat-derived Due to the inherent similarity between miRNAs and snoRNAs, such a relationship is easy to overlook as once a region is categorized as belonging to one molecular class, it is often no longer considered when searching for other types of molecules. The human genome has been scanned previously for the presence of box H/ACA snoRNAs using mammalian snoGPS Although no significant sequence similarity is detected between predicted snoRNA molecules encoding miRNAs and the known snoRNAs that target the same pseudouridylation sites, it is interesting to note that three snoRNAs  share their target pseudouridylation sites with eight of the predicted snoRNAs encoding miRNAs . This siin silico data presented here support these ideas. We tested the predictions by experimentally showing that the precursors of five of the predicted snoRNA-like miRNAs, mir-664, mir-151, mir-605, mir-215 and mir-140, interact with dyskerin, a protein component of functional H/ACA snoRNPs. While a lack of interaction to dyskerin could not rule out an evolutionary relationship between these miRNAs and snoRNAs as the miRNAs might have evolved sufficiently to no longer interact with functional protein components of snoRNPs, the detection of such an interaction considerably reinforces such claims. These results show that the snoRNA-like miRNA precursors sufficiently resemble box H/ACA snoRNAs to bind dyskerin, strengthening the possibility of an evolutionary relationship between these molecules. Further experiments will be necessary to investigate whether these molecules also retain the capability of targeting rRNA in vivo. It will also be necessary to experimentally test whether the remaining fifteen predicted snoRNA-like miRNA precursors also display aspects of H/ACA snoRNA functionality. The fact that three of these molecules  display the sequence AGA instead of the canonical ACA box could indicate that they have evolved sufficiently to no longer retain H/ACA snoRNA functionality. We do note that while identifying molecules that display both miRNA and snoRNA functionality supports our evolutionary hypothesis, we also expect to find a larger number of molecules that display features of both molecules but do not represent completely prototypical examples.This redundancy in rRNA complementarity may suggest some box H/ACA snoRNAs and snoRTs are not under as much selective pressure to avoid mutations. We hypothesize that some of these snoRNA encoding regions might be in the process of evolving from functional snoRNAs to miRNA-like precursors. This process might be facilitated by the fact that box H/ACA snoRNAs have a structure (2 hairpins) that is probably favourable to the formation of miRNAs. The Saccharomyces cerevisiae has snoRNAs but no reported miRNAs, consistent with a relationship where primordial snoRNAs may have given rise to certain classes of miRNAs. This idea is supported by a recent article by Saraiya and Wang reporting that the primitive parasitic protozoan Giardia lamblia, which does not have RNA interference capabilities but has miRNA processing machinery, uses box C/D snoRNAs as miRNA precursors Arabidopsis and Schizosaccharomyces pombe generate small RNAs , which are associated with argonaute proteins GiardiaIt is interesting to note that In addition to investigating whether some of the H/ACA snoRNA-like miRNA precursors display functional H/ACA snoRNA capability by binding to dyskerin , we haveWhile all the miRNAs characterised in A recent large-scale study defining an expression atlas for mammalian miRNAs, by Landgraf and colleagues, classifies known miRNAs into four different groups: prototypical, repeat-derived, repeat-clustered and unclassified http://www.repeatmasker.org). MiRNA and box H/ACA snoRNA genes as well as repeat-elements and sequence features were visualised with the UCSC Genome Browser The genomic positions, sequences and flanking regions of human miRNA genes and box H/ACA snoRNA genes were downloaded from the UCSC Table Browser 676 distinct human miRNA encoding regions (including the miRNA hairpins and 175 flanking nucleotides on either side) were scanned for the presence of box H/ACA snoRNAs using a locally-installed copy of the snoGPS program RNA secondary structures were predicted using RNAstructure 4.5 YFP-Dyskerin and HeLaGFP stable cell lines. Purified nuclei were resuspended in RIPA buffer to solubilise proteins. Fluorescence proteins were immunoprecipitated using GFP binder (ChromoTek) covalently coupled to NHS-activated Sepharose 4 Fast Flow beads  using the SuperScript one-step RT-PCR kit (Invitrogen). To decide linearity of cycles, we performed real time PCR using the Superscript III Platinum SYBR Green one-step qRT-PCR Kit (Invitrogen) and Rotor-Gene RG-3000 system (Corbett Research). The same amount of RNA for input and immunoprecipitated RNA (IP) was used as templates for RT-PCR reactions. Each experiment was repeated three times independently.Immunoprecipitations were prepared as previously described 32P 5′ end-labelled oligoribonucleotide probes specific for the following RNA species; . High sensitivity RNA blots were prepared as previously described HeLa cell extracts were fractionated using sucrose gradients, as previously described Figure S1Mammalian conservation of box H/ACA snoRNAs that encode experimentally detected smaller fragments(1.63 MB PDF)Click here for additional data file.Figure S2Predicted secondary structures of H/ACA snoRNA-like miRNA precursors(0.07 MB PDF)Click here for additional data file.Figure S3Repeat elements in proximity of H/ACA snoRNA-like miRNA genomic regions(3.58 MB PDF)Click here for additional data file.Figure S4Sequence elements surrounding selected miRNAs(0.03 MB PDF)Click here for additional data file.Figure S5Northern blot showing the subcellular localization of H/ACA snoRNA-like miRNA precursors (same as (0.31 MB PDF)Click here for additional data file.


The secreted MIA protein is strongly expressed by advanced primary and metastatic melanomas but not in normal melanocytes. Previous studies have shown that MIA serum levels correlate with clinical tumour progression in melanoma patients. To provide direct evidence that MIA plays a role in metastasis of malignant melanomas, A-mel 3 hamster melanoma cells were transfected with sense- and antisense rhMIA cDNA and analysed subsequently for changes in their tumorigenic and metastatic potential. Enforced expression of MIA in A-mel 3 cells significantly increased their metastatic potential without affecting primary tumour growth, cell proliferation or apoptosis rate in hamsters, compared with control or antisense transfected cells. Additionally, MIA overexpressing transfectants showed a higher rate of both tumour cell invasion and extravasation. Cells transfected with MIA antisense generally exerted an opposite response. The above changes in function attributed to the expression of MIA may underlie the contribution of MIA to the malignant phenotype. © 2000 Cancer Research Campaign


Herpes viruses by constructing a graph alignment, which is based jointly on the similarity of their protein interaction networks and on sequence similarity. This hybrid method provides functional associations between proteins of the two organisms that cannot be obtained from sequence or interaction data alone.Sequence alignment is a prolific basis of functional annotation, but remains a challenging problem in the 'twilight zone' of high sequence divergence or short gene length. Here we demonstrate how information on gene interactions can help to resolve ambiguous sequence alignments. We compare two distant We find proteins where interaction similarity and sequence similarity are individually weak, but together provide significant evidence of orthology. There are also proteins with high interaction similarity but without any detectable sequence similarity, providing evidence of functional association beyond sequence homology. The functional predictions derived from our alignment are consistent with genomic position and gene expression data.Our approach shows that evolutionary conservation is a powerful filter to make protein interaction data informative about functional similarities between the interacting proteins, and it establishes graph alignment as a powerful tool for the comparative analysis of data from highly diverged species. With the advent of genome-wide functional data, cross-species comparisons are no longer limited to sequence information. A classic extension of sequence alignment is structural alignment, which has been used to compare evolutionary distant RNAs  and protvaricella-zoster virus (VZV), causing chicken pox and shingles, and the Kaposi's sarcoma-associated herpesvirus (KSHV), responsible for cancer of the connective tissue. The two viruses have diverged approximately 200 million years ago. Their sequence dynamics is characterised by a high rate of point mutations  and a hulations ). As a rgraph alignment establishes a mapping between genes of two species ; ORF 23: [GenBank: NP 572075.1]; ORF 41: [GenBank: NP 572094.1]; ORF 29b: [GenBank: NP 572081.1]. VZV ORFs: ORF25: VOCs ID 59436; ORF65: 59475; ORF39: 59450; ORF60: 59470; ORF42: 59453.Supplementary Text. The Supplementary Text [Additional file Click here for fileSupplementary Animation. The Supplementary Animation [Additional file Click here for file


To administer medication in high concentration:for severe inflammationfor treating infectionat the end of an operationfor dilating the pupil.local anaesthetic dropssterile 2 ml syringeprescribed medicationsterile 21G needlesterile 25G needleeye padtapebandageclean cotton wool or gauze swabs.Remember!This procedure can be very distressing for the patient and needs to be explained sensitively to ensure maximum cooperation.Position the patient lying comfortably with his/her head supported on a pillow.Reassure the patient that adequate anaesthetic drops will be instilled before the injection is given.Instill anaesthetic drops at five-minute intervals over half an hour - a minimum of six drops.Away from the patient's view, draw up the prescribed medication using the 21G needle  may be given for pain relief.


Significant correlations betweenBOM and operation time and blood loss were found(p<0.01). Conclusions Bilirubin appears to act as anantioxidant for invasive surgery in humans. UrinaryBOM could be a reliable marker for the degree ofsurgical stress.Background Bilirubin has been recognized as an antioxidant.The purpose of this study was to examinewhether bilirubin would act as an antioxidant forsurgical stress in humans. Materials and MethodsSerum bilirubin and urinary bilirubin oxidativemetabolites (BOM) were measured in 96 patientswho underwent surgery. The antioxidant activityof bilirubin was assessed using BOM measuredby enzyme-linked immunosorbent assay with ananti-bilirubin monoclonal antibody. Results Serumbilirubin levels increased after surgery in all 96patients (


Multislice computed tomography (MSCT) scanners offer newperspectives for cardiac kinetics evaluation with 4D dynamicsequences of high contrast and spatiotemporal resolutions. A newmethod is proposed for cardiac motion extraction in multislice CT.Based on a 4D hierarchical surface-volume matching process, itprovides the detection of the heart left cavities along theacquired sequence and the estimation of their 3D surface velocityfields. A Markov random field model is defined to find, accordingto topological descriptors, the best correspondences between a 3Dmesh describing the left endocardium at one time and the 3Dacquired volume at the following time. The global optimization ofthe correspondences is realized with a multiresolution process.Results obtained on simulated and real data show the capabilitiesto extract clinically relevant global and local motion parametersand highlight new perspectives in cardiac computed tomographyimaging.


Survival from colorectal cancer has been analysed in relation to marital status in a nationwide Danish study of 9596 patients with complete follow-up of 22-26 years. After exclusion of 2294 patients with missing information adjusted five-year survival among married patients was significantly longer .


We have measured the serum concentration of the acute phase reactant, C-reactive protein (CRP), in 20 patients with histologically proven small cell lung cancer undergoing their first pulse of induction cytotoxic chemotherapy. Baseline CRP concentrations were raised in 16 of 20 patients . CRP levels more than doubled in 11 of 20 patients during induction chemotherapy. This acute phase reaction was seen in seven of the 10 chemosensitive patients, but was not observed in any of the five non-responding patients. Five patients were non-evaluable for chemoresponse. These data indicate that there is a previously undescribed quantifiable acute phase response during chemotherapy for small cell lung cancer which has potential for predicting chemoresponse.


A 45-year-old alcoholic male patient presented with hypovolemic shock and intense anemia (Hemoglobin 04.7 g/dl), and was operated on. A bleeding retroperitoneal varix located near the right colon was responsible for the clinical picture and was sutured. After operation the patient developed haemodynamic instability and pneumonia a situation which was reverted with intensive medical therapy. The patient is now doing well.


Lymph node pathologic status is the most important prognostic factor in vulvar cancer; however, complete inguinofemoral node dissection is associated with significant morbidity. Lymphoscintigraphy associated with gamma-probe guided surgery reliably detects sentinel nodes in melanoma and breast cancer patients. This study evaluates the feasibility of the surgical identification of sentinel groin nodes using lymphoscintigraphy and a gamma-detecting probe in patients with early vulvar cancer. Technetium-99m-labelled colloid human albumin was administered perilesionally in 37 patients with invasive epidermoid vulvar cancer (T1–T2) and lymphoscintigraphy performed the day before surgery. An intraoperative gamma-detecting probe was used to identify sentinel nodes during surgery. A complete inguinofemoral node dissection was then performed. Sentinel nodes were submitted separately to pathologic evaluation. A total of 55 groins were dissected in 37 patients. Localization of the SN was successful in all cases. Eight cases had positive nodes: in all the sentinel node as positive; the sentinel node was the only positive node in five cases. Twenty-nine patients showed negative sentinel nodes: all of them were negative for lymph node metastases. Lymphoscintigraphy and sentinel-node biopsy under gamma-detecting probe guidance proved to be an easy and reliable method for the detection of sentinel node in early vulvar cancer. This technique may represent a true advance in the direction of less aggressive treatments in patients with vulvar cancer. © 2000 Cancer Research Campaign


Natural killing (NK) by lymphocytes from normal individuals against primary and established ovarian carcinoma cell lines was tested in short-term chromium release assays. Two established cell lines and 5/6 primary cell lines tested showed significant susceptibility to NK. Primary cell lines are, in general, less sensitive to NK than long-term cultured target cells. A common NK recognition determinant on ovarian carcinoma cells and on the erythroleukaemic K562 cells was demonstrated by cold target inhibition assays. The recognition structure was also present on an ovarian cell line resistant to NK but not on the insensitive leukaemic cell line, SB. The activity against ovarian carcinoma cells was associated with the presence of large granular lymphocytes (LGL) previously shown to be the major effector cells against K562 targets. In fractions obtained by Percoll gradient centrifugation of lymphocytes, only fractions with a high content of LGL demonstrated good NK activity. LGL mediated NK against both non-adherent K562 and the adherent ovarian carcinoma target cells independent of monocytes.


Total upper body drainage via left superior vena cava into coronary sinus (i.e. absent right superior vena cava) is a rare anomaly and distorts the anatomy of coronary sinus and triangle of Koch. Herewith we are reporting the first report of ablation in a patient with superior vena cava draining into coronary sinus totally. This patient with atrioventricular nodal re-entrant tachycardia, baseline left bundle branch block, transient complete heart block during electrophysiological study and total upper body venous drainage into coronary sinus had successful slow pathway ablation using anatomical approach. Anatomical approach to atrioventricular nodal re-entrant tachycardia (AVNRT) ablation based on the triangle of Koch is the standard technique now, with a success rate close to 99%. Anatomy of the coronary sinus is important to guide anatomy guided ablation. Left superior vena cava (LSVC) draining into coronary sinus is not an uncommon anomaly that enlarges the coronary sinus . The enlA 49 years old female presented with history of recurrent palpitations for last 10 years. During last year, she had 5 episodes of palpitations which required hospitalizations. The tachyardia was repeatedly terminated by intravenous adenosine and verapamil. She was nondiabetic and nonhypertensive. General physical and cardiovascular examination were normal. Her baseline electrocardiogram (ECG) showed left bundle branch block with normal PR interval. The electrocardiogram  of tachycardia revealed similar morphology, with the heart rate of 210 beats/min. . Her traThe approach for the ablation of slow pathway in patients with anomalous drainage of superior vena cava into coronary sinus is not clear because of distorted triangle of Koch due to dilated coronary sinus -5. This


Lymphocytes obtained from hilar and bronchial lymph nodes from 23 patients undergoing radical surgery for carcinoma of the bronchus were fused with established rat or mouse myeloma lines. 62% of the resultant hybrids were found to be secreting human Ig detected by a sensitive staphylococcal Protein A-coupled SRBC assay. Immunoglobulins synthesized by such hybrids were internally labelled with 3H-lysine and their antibody activity against a variety of membrane preparations determined. Nine monoclonal antibodies were found which bound to molecules on lung-cancer membranes and not on normal lung membranes from the same patient.


Mice were exposed to plutonium dioxide (PuO2) aerosols 2 weeks before or after urethan injection. Both exposures reduced the number and size of adenomas. The incidence of arrested metaphases showed no consistently significant differences between plutonium-exposed and mock-exposed animals. The results are discussed in relation to recent electron microscopic evidence of degenerative changes in the type II epithelial cells of the mouse lung following PuO2 inhalation. It is concluded that damage at the cellular level may account for the observed reduction in growth of pulmonary adenomas in mice whose lungs contained plutonium particles.


Methotrexate (MTX) is a folate antagonist widely used both as an anticancer drug and as an immunosupressant. Administration of an 8-day methotrexate and folinic acid regime may be associated with pleuritic chest pain and pneumonitis. We have reviewed the toxicity seen in 168 consecutive patients treated with low-dose MTX for persistent trophoblastic disease. Twenty-five per cent of patients developed serosal symptoms, pleurisy was the commonest complaint. The majority of patients had mild to moderate symptoms which were controlled with simple analgesia and did not necessitate a change in treatment; 11.9% had severe symptoms which necessitated a change in treatment. One patient developed a pericardial effusion and a second patient developed severe reversible peritoneal irritation. The possible aetiology and pathophysiology of methotrexate-induced serosal toxicity is discussed. © 1999 Cancer Research Campaign


Its ability to suppress DNA synthesis contributes little to the inhibition of cell cycling.Bleomycin inhibits cell division in HeLa S-3 and other mammalian epithelial cell lines. Its effect on suspension cultured cells is far greater than on monolayer cells; this was found to be due partly to the absence of divalent ions. Other mammalian epithelial lines in suspension culture are very sensitive but fibroblastic cells (BHK 21/C13/DWS-3) grown under identical conditions are relatively insensitive. This study has established that bleomycin is a powerful anti-mitotic agent during the G


The plus/minus symbol appears incorrectly as an upside-down Latin capital letter A in the Results subsection "Depth-sampling reveals multiple copy RXLR effectors are prevalent," as well as in a number of other instances highlighted throughout the article.


These studies were undertaken to assess the relative expression and autocrine activation of the epidermal growth factor receptor (EGFR) in normal and transformed prostatic epithelial cells and to determine whether EGFR activation plays a functional role in androgen-stimulated growth of prostate cancer cells in vitro. EGFR expression was determined by Western blot analysis and ELISA immunoassays. Immunoprecipitation of radiophosphorylated EGFR and evaluation of tyrosine phosphorylation was used to assess EGFR activation. The human androgen-independent prostate cancer cell lines PC3 and DU145 exhibited higher levels of EGFR expression and autocrine phosphorylation than normal human prostatic epithelial cells or the human androgen-responsive prostate cancer cell line LNCaP. PC3 and DU145 cells also showed higher levels of autonomous growth under serum-free defined conditions. Normal prostatic epithelial cells expressed EGFR but did not exhibit detectable levels of EGFR phosphorylation when cultured in the absence of exogenous EGF. Addition of EGF stimulated EGFR phosphorylation and induced proliferation of normal cells. LNCaP cells exhibited autocrine phosphorylation of EGFR but did not undergo significant proliferation when cultured in the absence of exogenous growth factors. A biphasic growth curve was observed when LNCaP cells were cultured with dihydrotestosterone (DHT). Maximum proliferation occurred at 1 nM DHT with regression of the growth response at DHT concentrations greater than 1 nM. However, neither EGFR expression nor phosphorylation was altered in LNCaP cells after androgen stimulation. In addition, DHT-stimulated growth of LNCaP cells was not inhibited by anti-EGFR. These studies show that autocrine activation of EGFR is a common feature of prostatic carcinoma cells in contrast to normal epithelial cells. However, EGFR activation does not appear to play a functional role in androgen-stimulated growth of LNCaP cells in vitro.


An 18-year-old male patient presented with chronic nonspecific pain of three months located at his left proximal tibia. The patient was admitted to our department for plain X-ray, computed tomography and magnetic resonance imaging examination. Plain X-ray and computed tomography revealed a geographic lytic lesion at the medial aspect of the proximal tibia. Biopsy of the lesion showed telangiectatic osteosarcoma. Image findings of all modalities are presented. Telangiectatic osteosarcoma is an uncommon histopathologic subtype that represents 2.5 -12% of all osteosarcomas . These histologic features are the cause of telangiectatic osteosarcoma often having been described as showing non-specific bone lysis on radiographs and simulating aneurysmal bone cyst. The soft tissue counterpart of telangiectatic osteosarcoma is uncommon in young patients and usually affects adults older than 50 years . It presThe knowledge of radiologic findings of telangiectatic osteosarcoma is important as its often much less aggressive appearance may lead to underdiagnosis of this lesion and delay in therapeutic management of the patient.


We report the first case of endocarditis caused by a Gordonia species genetically related to G. sputi but exhibiting some atypical biochemical features in a 31-year-old woman with a central venous catheter. This unusual pathogen may be a new cause of opportunistic infections in patients with severe underlying diseases.


A 75 year old lady presented with acute central retinal artery occlusion and contralateral cotton wool spots.General physical examination and investigations led to a diagnosis of CREST syndrome; however, association of central retinal artery occlusion with CREST syndrome is not well known. While diabetes, systemic hypertension, carotid atherosclerosis and cardiac pathology are common causes of CRAO it is always important to rule out giant cell arteritis.This case highlights that inflammatory causes of central retinal artery occlusion other than giant cell arteritis should also be considered as a possibility to spare unnecessary use of excessive systemic corticosteroids. We present an unusual case of acute central retinal artery occlusion and cotton wool spots as the presenting feature in a patient subsequently diagnosed with systemic sclerosis, in particular with features of CREST syndrome.A 75-year old Caucasian lady presented with acute onset of painless visual loss in her left eye, which she noted on waking up that morning. She denied any associated headaches, jaw claudication or any clinical symptoms suggestive of temporal arteritis. Past ophthalmic history was not significant apart from one episode of transient visual loss in the left eye of about 5 minutes duration with zigzag lines three months earlier. This was accompanied by spontaneously resolving short-duration headache. The general practitioner diagnosed an episode of migraine with no further investigations or recurrence. She had treated essential hypertension and suffered especially in cold weather with Raynaud's phenomenon for 20 years; this had worsened in severity in the last few months.On examination visual acuity of 20/20 in the right and CF (counting fingers at 0.5 m) was noted in the left eye with a left relative afferent pupillary defect. Anterior segment examination was unremarkable with normal intraocular pressures. Dilated fundal examination showed a cherry-red spot in the left foveal area. No visible arteriolar embolus was noted Fig . Apart fFurther investigations revealed a positive ANA's at 8.1 U (0–1.0 reference range) with a positive anti-centromere antibody and a negative anti-Scl-70 and anti-ds DNA antibody. X-rays of the hands did not reveal any significant calcinosis or erosions. Barium swallow was unremarkable but echocardiography showed mild concentric hypertrophy of ventricular walls consistent with long-standing systemic hypertension.Mild non-flow limiting plaque was also detected on coronary angiography. Her pulmonary hypertension was attributed to her cardiac status with atrial fibrillation and systemic hypertension rather than connective tissue disorder related. Carotid duplex scans revealed minor calcific plaques with no significant stenosis in either carotid arterial system. She remains under joint rheumatology and ophthalmology review on immunosuppression with methotrexate 10 mg per week, prednisolone 7.5 mg daily and anti-platelet therapy with no further visual symptoms after 12 months of follow up.Various authors -5 have nCREST syndrome can be quite variable and may or may not show all the features in a single patient at one time point. Our case highlights the importance of thorough clinical examination including review of systems and appropriate referral as the appearance of her hands, telangiectasias & presence of Raynaud's phenomenon lead to well-directed investigations and subsequent clinical diagnosis. While temporal arteritis was a reasonable diagnosis to entertain initially, we stopped the oral steroids immediately with unremarkable blood and angiogram results. The patient was thus saved a significant systemic corticosteroid load with avoidance of possible adverse effects including scleroderma renal crisis which may be seen in scleroderma patients treated with high dose steroids.The presence of cotton wool spots in her right eye strongly suggested an inflammatory cause for CRAO and although microvasculopathy is a more consistent feature of the systemic sclerosis spectrum, macrovascular disease has been reported. It should be noted that in general CRAO is rarely caused by inflammation with atherosclerotic disease being the usual aetiology -7. In ouIn a review  listing Ophthalmologists should be aware of this unusual association between central retinal artery occlusion and CREST syndrome. While the prognosis for central retinal artery occlusion is extremely guarded, systemic workup to rule out life threatening co-morbid conditions remains the cornerstone of management.CREST: ; CRAO: ; ESR: Erythrocyte sedimentation rate; CRP: C Reactive Protein; ANA: (Anti-Nuclear antibodies); Anti-ds: (Anti-double strand).The authors declare that they have no competing interests.MR analyzed and reviewed the literature and drafted the case report. TM and BJB provided valuable input to the manuscript. All authors read and approved the final manuscript.Written informed consent was obtained from the patient for the publication of this case report and accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.


P=0.119).The authors have performed this study to explore the effectiveness of neoadjuvant androgen suppression in patients of localized prostate cancer treated with high-intensity focused ultrasound (HIFU). From January 1999 to January 2005, they had treated 250 patients with HIFU. One hundred and fifty-four patients received neoadjuvant hormonal therapy and 96 were not treated with androgen suppression. Treatment failure was defined by presence of prostate cancer on biopsy taken six months after HIFU. In this unrandomized trial, the authors found a slightly lower treatment failure rate in patients who received neoadjuvant androgen suppression (31% vs. 34%), but this was not statistically significant (Use of neoadjuvant/concurrent hormonal therapy for localized prostate cancer treated by radical radiotherapy is currently getting studied in many trials. Initial results have been encouraging with significant benefit in the relapse-free /PSA-free interval in these patients. There ha


Pulmonary tumor thromoembolism is well described in literature especially in patients known to have cancer. We are presenting this report of a patient presenting with Acute Cor-pulmonale secondary to Occult Gastric carcinoma causing pulmonary tumor embolism. This is an unusual presentation of occult neoplasm. Pulmonary tumor embolism is an important but ‘uncommon’ cause of dyspnea in patients with cancer. It is ‘uncommon’ as it is under-recognized! Autopsy reports show that up to 26% of patients who die with cancer have tumor cells in their pulmonary vasculature. By this article we aim to increase the awareness of this condition, thereby leading to appropriate management.A 77-year-old Caucasian male who was admitted in the surgical ward with complaints of abdominal pain and altered bowel habits of 3-week duration was referred to the medical team due to increasing dyspnea. He had sustained an injury to knee 4 weeks back and had been taking regular codeine and paracetamol tablets for pain. His abdomen X-ray had shown fecal loading, rigid sigmoidoscopy was normal and the provisional diagnosis was constipation secondary to codeine. He gave a history of gradually progressive dyspnea over 2 weeks. There was no other significant past medical history. On examination he had features of right-sided heart failure with pulmonary hypertension. Arterial blood gases showed hypoxia. ECG showed S1Q3T3 pattern. The diagnosis of pulmonary embolism was made and he was started on full-dose low- ‘molecular weight’ Heparin. Echocardiogram showed dilated right ventricle with impaired function and right ventricular systolic pressure >80 mmHg. CT pulmonary angiogram showed no evidence of pulmonary embolism but did show multiple lymph node enlargement. Patient had cardiac arrest and resuscitation attempts were unsuccessful.Postmortem showed multiple lymph node enlargements inside mediastinum and abdomen, malignant-looking ulcer in stomach and tumor infiltration throughout small bowel mesentery. Microscopy of gastric ulcer revealed adenocarcinoma.Right ventricle did not show any features of hypertrophy.Macroscopically, lungs were edematous but pulmonary arteries showed no evidence of pulmonary embolism. Microscopy of pulmonary parenchyma showed pulmonary hypertensive changes and extensive tumor thromboemboli involving pulmonary microvasculature.Final diagnosis: Widespread carcinamatosis due to carcinoma stomach.Acute cor pulmonale due to pulmonary tumor thromboembolism.The entity of pulmonary tumor thromboembolism has been described as early as in 1897 (Schmidt).The most common presenting complaint is dyspnea. Other feThe incidence of pulmonary tumor thromboembolism is between 3 and 26% in patients with cancer, but therThe pulmonary arteries are grossly normal but microscopy reveals fibrocellular intimal proliferation with smooth muscle colonization of luminal neoplastic lesions and associated microthrombi. This cauHypoxemia with normal chest radiograph is the common finding. Ventilation perfusion scanning characteristically shows multiple subsegmental mismatched defects. CT pulmonary angiography, which is emerging as the investigation of choice in pulmonary embolism, is not a helpful test in this case. Pulmonary angiography, which is the gold standard to diagnose pulmonary embolism, has poor sensitivity and specificity. Right heThe median survival of these patients reported in literature is 3 days. Thrombol


Ventricular tachycardia (VT) occurring late after myocardial infarction is often due to reentry circuit in the peri-infarct zone. The circuit is usually located in the sub-endocardium, though subepicardial substrates are known. Activation mapping during VT to identify target regions for ablation can be difficult if VT is non inducible or poorly tolerated. In the latter, a substrate based approach of mapping during sinus rhythm in conjunction with pace mapping helps to define the reentry circuit and select target sites for ablation in majority of patients with hemodynamically unstable VT. Percutaneous epicardial catheter ablation has been attempted as an approach where ablation by a conventional endocardial access has been unsuccessful. We report a case of post myocardial infarction scar VT which could be successfully ablated with a substrate based approach from the epicardial aspect. A 62 year old male presented with recurrent ventricular tachycardia (VT). He had suffered an inferior wall myocardial infarction with resultant mild left ventricular (LV) dysfunction (LV ejection fraction 40%) and had an ICD implanted 3 years back. The patient was now hospitalised with recurrent appropriate ICD shocks. He was loaded upfront with intravenous amiodarone followed by oral amiodarone and beta blockers. However, VT continued to recur resulting in frequent ICD shocks despite maximal tolerated drug therapy.Patient was taken up for radiofrequency ablation. An initial endocardial approach was used using electro-anatomic mapping system (EAM) . The equipment used for recording was an EP-TRACER . An endocardial voltage map of the LV was created using a Navistar catheter under EAM guidance by a retrograde transfemoral approach. While mapping, a sustained monomorphic VT was induced with a LBBB, superior axis morphology, QS complexes in V1-V6; tachycardia cycle length (TCL) of 318 msec . The endThe pericardial space was exposed using a percutaneous subxiphoid approach. Pericardial puncture was performed with a 17-gauge epidural needle. The puncture was performed at the left lower edge of the xiphoid, at a 45 degree angle toward the left scapula. Two milliliters of contrast was injected via the needle to delineate the heart silhouette to confirm intra-pericardial location. A standard 8F sheath mounted over a guidewire was then introduced into the pericardium through a small incision. An externally irrigated RF catheter  was then introduced through the sheath and carefully pushed into the pericardial space. The catheter was constantly irrigated with saline (1 mL/min flow rate). Programmed electrical stimulation (PES) from the epicardium failed to induce the VT. Following isoprenaline infusion VT could be induced on PES. However the VT was hemodynamically unstable and had to be promptly terminated.A substrate based voltage map of the LV epicardium in sinus rhythm was constructed which revealed presence of scar (defined by voltages < 0.5 mV) in posterobasal LV. Multiple areas of low voltage fractionated activity in the peri-scar area associated with late diastolic potentials (LDP) during sinus rhythm could be identified . These aA review EP study was performed 24 hours later which did not induce any tachycardia on PES with isoprenaline. Oral amiodarone was stopped and the patient was discharged on beta blockers. Patient remained symptom free and at a follow up of three months, the ICD did not reveal  any new tachyarrhythmia by telemetry.Catheter ablation has an increasingly important role in controlling ventricular tachycardia (VT) episodes in patients with structural heart disease who have implanted cardioverter defibrillators. Substrate for reentrant VT is often subendocardial and is accessible for catheter ablation by a transvenous approach. However, epicardial ablation is required for 10-30% of postinfarct VTs, and for more than 30% of VTs due to nonischemic cardiomyopathy [The substrate for postinfarction VT is well characterized. Surviving muscle bundles commonly located in the subendocardium but also in the mid myocardium and epicardium, traverse the borders and penetrate the deeper scar . The locCiccasio et al  used sinPercutaneous, pericardial instrumentation via subxiphiod approach permits characterization and imaging of the epicardial substrate in a fashion similar to that for the endocardium. A substrate based ablation reduces or eliminates the need for mapping during prolonged periods of tachycardia and extends the potential benefits of ablation to the large majority of VTs that are hemodynamically unstable.


Background: Although maternal infections with coxsackievirus during pregnancy are relatively common, fetal infections are quite rare. Coxsackievirus infection in utero has been associated with myocarditis, but has not been proven a teratogen.Case: A patient whose fetus had structural cardiac anomalies and hydrops was found to have an intrauterine infection with Coxsackie B-1 virus, proven by virus isolation from the amniotic fluid. This infection led to increasing intrauterine hydrops and subsequent neonatal death. Conclusion: This interesting association of intrauterine infection with Coxsackie B virus and structural cardiac anomalies in the fetus warrants further investigation.


A promising new treatment for glioma involves Auger electron emitters such as 125I or 123I conjugated to deoxyuridine (IUdR). However, the presence in tumour deposits of non-proliferating cells with clonogenic potential poses a major limitation to this cycle-specific therapy. We have used multicellular tumour spheroids derived from the human glioma cell line UVW to study [125I]IUdR-targeted radiotherapy in aggregates containing cells in different proliferative states. Autoradiographic identification of labelled cells indicated that nuclear incorporation of [125I]IUdR decreased markedly with increasing size of spheroid. IUdR incorporation was maximal in the surface layer of cells and decreased with depth within spheroids. Radiopharmaceutical uptake corresponded closely to the regions of cell cycling as indicated by staining for the nuclear antigen Ki67. The uptake of drug was enhanced by increasing the duration of incubation from 52 h to 104 h. These observations suggest that significant sparing of non-cycling malignant cells would result from treatment delivered as a single injection of radiolabelled IUdR. To achieve maximal therapeutic effect. IUdR should be administered by multiple injections, by slow release from biodegradable implants or by slow-pump delivery.


Biventricular (BiV) is extensively used in the treatment of congestive heart failure but so far no recommendations for optimized programming of atrioventricular-delay (AVD) settings have been proposed. Can AVD optimization be performed using a simple formula based on non-invasive doppler-echocardiography?25 patients (ejection fraction 30±8%) received BiV ICDs. Doppler-echocardiographic evaluation of diastolic and systolic flow was performed for different AVDs (30ms to 150ms) and different stimulation sites (left ventricular (LV), right ventricular and BiV). The optimal atrioventricular delay was calculated applying a simple formula based on systolic and diastolic mechanical delays determined during doppler-echocardiography.The mean optimal AVD was calculated to be 112±29ms (50 to 180ms) for BiV, 95±30ms (65 to 150ms) for LV and 75±28ms (40 to 125ms) for right ventricular pacing with wide interindividual variations. Compared to suboptimal AVDs diastolic optimization improved preejection and ejection intervals independent to pacing site. Optimization of the AVD significantly increased ejection time during BiV pacing . Compared to LV or right ventricular pacing BiV pacing produced the shortest mean pre-ejection and longest ejection intervals as parameters of improved systolic ventricular contractile synchrony. Diastolic filling times were longest during BiV pacing compared to LV or RV pacing.Individual programming of BiV pacing devices increases hemodynamic benefit when implementing the inter-individually widely varying electromechanical delays. Optimization applying a simple formula not only improves diastolic ventricular filling but also increases systolic functional parameters. Systolic resynchronization using biventricular (BiV) pacing has evolved as an established add-on therapeutic option in patients with symptomatic congestive heart failure. When pacing the left ventricle via the coronary sinus the contraction pattern of the interventricular septum and the contraction sequence of both ventricles is improved. This therapy has proven to increase systolic function (dP/dt) and cardiac function -5.In patients with left ventricular pump failure conventional dual chamber pacemakers have been shown to alter hemodynamics in regard to the programmed atrioventricular delay (AVD). A long AVD leads to atrial contraction too early for optimal ventricular filling . On the other hand a short AVD may lead to atrial contraction after closure of the atrioventricular valve (due to systolic increase of ventricular pressure) -10.Different methods have been proposed to determine the most favorable  AVD aiming at either optimizing systolic or diastolic cardiac function. Diastolic optimization of the AVD aims at restoring the atrial kick by coincidental timing of the end the left atrial systole and the mitral valve closure ,6,11-17.We studied the feasibility and efficacy of AVD optimization adapting a modification of a formula for optimal AVD programming in patients under biventricular stimulation with intrinsic atrioventricular conduction and left ventricular insufficiency. The effects on non-invasively determined parameters of cardiac systolic and diastolic function     were assessed. 25 consecutive patients were included after implantation of biventricular implantable defibrillators (ICD) (Medtronic InSync™ ICD) for chronic heart failure class II to IV in between 1999 and 2001. All patients gave informed consent; the protocol was evaluated by the institutional ethics review board.In all patients a standard ECG (12-lead) and Doppler-echocardiographic studies were performed 30 days after implantation. Different AVDs were programmed  for the 3 programmable stimulation sites . Fusion beats were excluded on the basis of QRS morphology. For each programming mitral- and tricuspid-valve and aortic valve Doppler-echocardiograms were performed using our Hewlett Packard Sonos 5500 echocardiography system with continuous display of ECG at a paper speed of 100mm/sec. Measurements were performed after a resting period of 1 minute in intrinsic rhythm. Prior to each measurement an adapting period of 1 minute was established. 3 consecutive atrioventricular and aortic valve flow profiles were analyzed and different time intervals were measured see  and the Mitral valve diastolic filling time: Measure of diastolic left ventricular filling.Tricuspid valve diastolic filling time:Measure of diastolic right ventricular filling.Pre-ejection time:Indicating the intra- and interventricular conduction and contraction synchrony.Ejection time:Measure of left ventricular ejection.As a next step the AVD producing the most favorable diastolic left ventricular inflow  was calculated applying a simple formula see : Two steLong AVD pacing (150 to 200ms) to determine the atrial electromechanical delay:The atrial electromechanical delay constitutes the time interval in between the right atrial sensed electrical impulse to the end of the active mitral valve flow. This includes the interatrial conduction time and the electromechanical coupling of the left atrium. This interval is intraindividually fixed and has a wide interindividual variation.Short AVD pacing (30ms as the shortest programmable AVD) to determine the isovolumic contraction time:The isovolumic contraction time is the time interval in between the ventricular electrical stimulation and the closure of the mitral valve due to the left ventricular systolic pressure increase. This interval can be measured when the atrial contraction is attenuated by the ventricular systole as documented in Doppler-echocardiography. This interval is intraindividually fixed and includes the conduction time from the pacing electrode to the left ventricle, the electromechanical coupling of the left ventricle and the duration from the beginning of the left ventricular systole to increasing intraventricular pressure above atrial pressure.Calculation of the optimal AVD when subtracting the atrial electromechanical delay and the isovolumic contraction time .When the calculated optimal AVD was different from the before programmed AVDs measurements were performed after optimized programming.In each patient, measurements during optimized pacing  were compared to the mean of the three AVDs  (= control) excluding the optimal AVD .The means of the 3 consecutive measurements were calculated and the optimal AVD was calculated for each patients. Non-categorical variables of the different pacing site groups  and different AVDs  were compared using Student t-test. Treatment effects within the groups (optimized pacing versus control settings) were assessed by ANOVA analysis. A significant difference was proposed when p < 0.05.25 patients with a mean ejection fraction pre-implant of 30%  and mean end diastolic diameter (echocardiography) of 66mm (± 7) were analyzed. Mean age was 65 (±10)years, 22 had left bundle branch block morphology whereas 3 had right bundle branch block morphology and mean intrinsic QRS width was 191 . Position of the LV pacing lead was posterolateral in 16 (64%), lateral in 6 (24%) and anterior in 3 (12%).All patients were in sinus rhythm . Mean QRS width was significantly shortened during BiV pacing to 162ms (± 23) (p = 0.02), significantly longer during LV-pacing (238ms ±35) (p = 0.001) and significantly longer during right ventricular pacing (231ms ±39) (p = 0.006).Heart rate did not significantly differ intraindividually (± 5 bpm) during any of the Doppler-echo studies. A consistent finding was the consecutive shortening of the diastolic filling times  when prolonging the programmed AVD see . LV paciLongest mean ejection period resulted during BiV pacing compared to the monoventricular pacing modes independent to the programmed AVD. LV pacing seems to produce a longer ejection time compared to right ventricular pacing except at the shortest programmed AVD (30ms) see .1 of the 25 patients (4%) had no atrial contraction documented during transthoracic Doppler-echo. This patient was conclusively not eligible for optimization of the AVD. 24 of the 25 underwent Doppler-echocardiographic AVD optimization. The optimal AVD during BiV pacing was found to range in between 50ms to 180ms with a mean of 112ms (± 30). When correlating the optimal AVD to the intrinsic PQ-interval  it was found to translate into 23% to 80%, indicating a shortening of the PQ-interval in between 20% to 77% to gain optimized BiV pacing. The optimal AVD during LV pacing was found to be 95ms (± 30) (p = 0.95 vs. BiV) (65ms to 150ms) and during right ventricular pacing was significantly shorter compared to BiV pacing at 75ms (± 28) (p = 0.003) (40ms to 125ms) see .BiV optimized pacing produced the most favorable systolic and diastolic functional Doppler parameter compared to LV and right ventricular pacing see . StatistAVD optimization compared to control settings during BiV pacing significantly increased ejection time from 266ms (± 30) to 279ms (± 25) (p = 0.03) correlating with an improved systolic ejection see . The preDuring LV-pacing AVD optimization increased ejection time from 266ms (± 30) to 268ms (± 33) (p = 0.96) and pre-ejection time was shortened from 177ms (± 31) to 166ms (± 35) (p = 0.32) see . DiastolDuring optimized right ventricular pacing ejection period stayed constant at 263ms (± 35)  but pre-ejection time was shortened from 189ms (± 27) to 182ms (± 25) (p = 0.32) see . DiastolThere is still controversy about the effects of different programmings of the AVD in patients with pacing devices for chronic heart failure. Studies have emphasized the influence of the optimal pacing site in patients treated with biventricular pacing and there seems to be a beneficial effect of different AVD programmings modulating systolic ventricular function ,16,23.This study demonstrates improved systolic and diastolic function after non-invasive diastolic optimization on patients with implanted biventricular pacing devices. The optimal AVD was determined using a simple formula derived from findings in patients with complete heart block and is based on Doppler-echocardiographically measured electromechanical time intervals. Applying calculated optimal AVD programming diastolic filling is optimized and systolic function is improved. Improvements in systolic function may be due to a leftward shift in the Frank-Starling curve initiated by improved diastolic function. Although, optimizing the AVD may not lead to chronic increase in systolic function. Only a single report exists indicating functional improvement in biventricular pacing patients due to AVD optimization more than one month after optimization .Different studies have evaluated the effects of different AVD optimization strategies. In most cases, the optimal AVD was determined by echocardiographic parameters of systolic function (e.g. left ventricular outflow). A recent study on 215 patients undergoing cardiac resynchronization therapy indicated the usefulness and safety of AVD optimization using Doppler mitral inflow data . ControvWe documented beneficial effects on systolic functional Doppler parameters indicating improved ventricular ejection and beneficial effects on intra- and interventricular electrical synchronization when coincidentally timing the end of the left atrial contraction and the beginning of the left ventricular systole. The optimal AVD can easily and non-invasively be calculated using a formula integrating two time intervals measured during long AVD and short AVD pacing. This formula was proposed by Ritter et al. in 1995 for patients with complete AV-block to resynchronize left atrial and ventricular systole enabling optimal ventricular filling and pre-load. However this formula has not yet been validated invasively -22. In oApplying the calculated optimal AV-delay not only led to optimized timing of the atrial contraction just prior to the left ventricular systole but also produced changes in systolic functional Doppler parameters. During BiV pacing non-invasive diastolic AVD optimization significantly increased ejection time (5% increment compared to control settings). This implements significantly improved left ventricular systolic function only by modulating the AVD settings. Even though the correlation between ejection time and ejection volume is weak it is a measure of systolic ejection function indicating increased contractile function. Also AVD optimization led to shorter pre-ejection times documenting improved intra- and interventricular synchronicity. These findings are consistent with the documented changes of systolic function by Kindermann et al. when tailoring diastolic flow ,21. The 22It seems possible to improve the beneficial effect of electric resynchronization therapy by careful and Doppler-echo guided individual programming of biventricular pacing devices. Even though, no guidelines for programming parameters for AVD optimization are available ,16,24-28It is well known that the effect of pacing therapies for chronic heart failure depend on selection of the pacing site. Whereas right ventricular pacing was shown to have controversial effects on left ventricular performance left ventricular based pacing seems to have beneficial effects on systolic function due to electrical resynchronization of the left ventricular contraction pattern ,16,18-23As indicated in our study not only the filling volume but the preload which can be increased by optimal timing of the left atrial contraction and its contribution to ventricular filling is crucial for left ventricular performance. During optimized pacing the diastolic filling time is shorter than during short AV-delay pacing but correct timing of the left atrial contraction leads to optimized ventricular filling and readjusted mitral valve Doppler flow. These considerations explain the shorter diastolic filling times during optimized pacing compared to the suboptimal AVD programmings but still improved cardiac function during AVD optimization ,18-22,28This study is limited by the small number of patients included making it impossible to determine any variables leading to changes in optimal AV-delay settings like LV-electrode position, cardiac disease or ejection.As control the mean of AVD settings of 80ms, 100ms and 150ms were calculated and compared to optimized AVD pacing. This artificially constructed control setting is usually close to the range of the optimal AVD and therefore differences in between AVD settings may be marginal. On the other hand this control may resemble manufacturer's AVD settings implemented in the BiV pacing devices and therefore the analysis indicates the incremental benefit of AVD optimization compared to baseline settings.We did not perform any invasive hemodynamic studies to validate the applied formula for AV-delay optimization. Even though the documented improvements in Doppler echocardiographic functional parameters indicate hemodynamic benefit this has not been clinically tested. In addition, no comparative study to other methods of AVD optimization was performed. It remains unclear, whether the proposed formula leads to the highest benefits or if other strategies (e.g. tailoring systolic Doppler-echo parameters) may further increase hemodynamics.Especially in the latest pacing devices for biventricular resynchronization not only atrioventricular delay but also interventricular delays are programmable. Therefore there is a definitive need for guidelines or recommendations how to program different parameters of diastolic and systolic synchronization in these devices.Individual programming in patients under biventricular stimulation can increase the benefit of the resynchronization therapy when implementing the differences in electromechanical delays. Optimization of the AVD integrating these individual intervals can easily and non-invasively be performed using Doppler echocardiography applying a simple formula. Optimized pacing cannot only improve diastolic but also optimize systolic functional Doppler parameters.


Immunoreactive collagenase has been demonstrated in 5/14 specimens of cutaneous secondary melanomas. In contrast, very little enzyme was seen in 10 specimens of normal human skin. All specimens were fixed within minutes of excision. These findings support the hypothesis that collagenase facilitates connective-tissue breakdown which is associated with tumour invasiveness and metastatic spread.


Dear Editor,A 70-year-old lady was seen in the retina clinic following a referral for suspected bilateral occult choroidal neovascular membranes (CNV). Her symptoms included intermittent blurred vision. Best corrected visual acuity was 20/60 in the right and 20/30 in her left eye. Fundus examination revealed a central area of cystoid macular edema surrounded by exudates and hemorrhages in her right eye and an area of serous pigment epithelial detatchment in the left eye. She had a healthy optic nerve with distinct disc margins . FluoresThere were no identifiable risk factors for NAION in her case apart from the age. Intraocular pressures were found to be normal during pre and post-treatment phase. One possible explanation can be a compromise in the vascular integrity of the optic nerve vessels as vascular endothelial growth factor (VEGF) participates in the maintenance of vascular systems in adults. To date,2


Human papillomavirus DNA was detected in cervical specimens from 366 sexually active young women with cytomorphologically normal cervices using the polymerase chain reaction. In 93% (25/27) of initially infected women, the same viral type was not detected upon re-examination four menstrual cycles later. These results suggest that the majority of HPV infections in young women are transient.


Neural network breakdown is a key issue in neurodegenerative disease, but the mechanisms are poorly understood. Here we investigated patterns of brain atrophy produced by defined molecular lesions in the two common forms of genetically mediated frontotemporal lobar degeneration (FTLD). Nine patients with progranulin (GRN) mutations and eleven patients with microtubule-associated protein tau (MAPT) mutations had T1 MR brain imaging. Brain volumetry and grey and white matter voxel-based morphometry (VBM) were used to assess patterns of cross-sectional atrophy in the two groups. In a subset of patients with longitudinal MRI rates of whole-brain atrophy were derived using the brain-boundary-shift integral and a VBM-like analysis of voxel-wise longitudinal volume change was performed. The GRN mutation group showed asymmetrical atrophy whereas the MAPT group showed symmetrical atrophy. Brain volumes were smaller in the GRN group with a faster rate of whole-brain atrophy. VBM delineated a common anterior cingulate–prefrontal–insular pattern of atrophy in both disease groups. Additional disease-specific profiles of grey and white matter loss were identified on both cross-sectional and longitudinal imaging: GRN mutations were associated with asymmetrical inferior frontal, temporal and inferior parietal lobe grey matter atrophy and involvement of long intrahemispheric association white matter tracts, whereas MAPT mutations were associated with symmetrical anteromedial temporal lobe and orbitofrontal grey matter atrophy and fornix involvement. The findings suggest that the effects of GRN and MAPT mutations are expressed in partly overlapping but distinct anatomical networks that link specific molecular dysfunction with clinical phenotype. The role of neural networks in neurodegenerative disease has attracted much recent interest  age at baseline scan 53.5 (5.2)) and nine patients with a GRN mutation (mean (standard deviation) age at baseline scan 62.9 (6.1)) were included in the study: the MAPT mutation group comprised 8 patients with a 10 + 16 mutation and single patients with 10 + 14, S320F and G389R mutations; the GRN mutation group . T1-weighted volumetric brain images were obtained using an IR-prepared fast SPGR sequence  with a 24-cm field of view and 256 × 256 matrix to provide 124 contiguous 1.5-mm-thick slices in the coronal plane. We initially calculated whole-brain and hemisphere volumes in all patients. The brain region was created by segmenting the whole brain using a semi-automated technique in the MIDAS software package . For rathttp://www.fil.ion.ucl.ac.uk/spm) with default settings for all parameters. The images underwent an initial segmentation process in SPM5 which simultaneously estimated transformation parameters for warping grey matter (GM), white matter (WM) and cerebrospinal fluid (CSF) tissue probability maps (TPMs) onto the images. The native space GM and WM segments were then rigidly spatially normalised, using just the rotations and translations from the inverse of the TPM transformation, and resampled to 1.5 mm isotropic resolution. These “imported” images were then iteratively warped to an evolving estimate of their group-wise GM and WM average template using the DARTEL toolbox  at a 0.001 significance level. For disease group comparisons maps were generated uncorrected at a 0.001 significance level. Statistical parametric maps were displayed as overlays on a study-specific template, created by warping all native space whole-brain images to the final DARTEL template and calculating the average of the warped brain images. In order to visualise hemispheric asymmetries, we performed two VBM analyses: firstly with all images in their native space; and secondly, with native images flipped in the midsagittal plane within SPM5, such that the most severely affected cerebral hemisphere was on the same side in each patient. An image was selected for flipping if it had a hemispheric asymmetry index outside the control range and more severe right hemisphere atrophy : four images from the GRN group and three from the MAPT group met criteria for flipping.Voxel-based morphometry (VBM) was performed on the baseline images using SPM5 software  we performed non-linear registration of each follow-up image to its corresponding baseline using a multi-scale viscous fluid algorithm  with a NBaseline brain volumes were smaller in both disease groups compared to healthy controls and mean brain volume was significantly smaller in the GRN group compared to the MAPT group . Rates op = 0.07)  , Fig. 2APatterns of grey matter atrophy differed in the MAPT and GRN groups compared to healthy controls. The GRN group analysis on unflipped images showed an overall pattern of symmetrical brain atrophy including frontal, temporal and parietal lobes, cingulate cortex and thalamus. However, this result obscures any asymmetries in favour of left or right hemisphere at individual subject level: after flipping of images so that all patients had the most affected hemisphere in the same orientation the true asymmetry of GRN disease was apparent . The mosIn the GRN group compared to controls, the white matter VBM analysis showed most significant involvement of intrahemispheric long association tracts including inferior longitudinal fasciculus, superior longitudinal fasciculus, inferior fronto-occipital fasciculus and cingulum. There was additional involvement of the corpus callosum and brainstem tracts, particularly in the pons . In the Comparing the two mutation groups directly in the flipped VBM analysis  revealedDifferential patterns of longitudinal volume loss in the two mutation groups were similar to those seen in the cross-sectional VBM analysis . The GRNUsing convergent imaging techniques, we have shown that GRN and MAPT mutations are associated with distinct profiles of neuronal loss affecting distributed cortical areas and their white matter connections. GRN mutations are associated with asymmetric atrophy of frontal, insular, cingulate, parietal and temporal areas linked by intrahemispheric long association tracts, while MAPT mutations are associated with more restricted but bi-hemispheric atrophy of anteromedial temporal and orbitofrontal areas linked via the fornices and uncinate fasciculi. Disease evolution is more rapid in GRN- than MAPT-associated FTLD, as predicted from the more distributed cerebral damage associated with GRN mutations. Moreover, the degree of asymmetry increases over time in the GRN- (but not the MAPT) associated cases: in conjunction with the evidence presented here for asymmetric longitudinal intrahemispheric volume loss, this increasing asymmetry implies that the pathological process in GRN-associated FTLD is focused within the maximally affected hemisphere. The profiles of grey matter involvement demonstrated here help to integrate previous evidence concerning anatomical signatures and disease evolution in GRN and MAPT mutation cases. In particular, our findings suggest an anatomical substrate for the shorter disease duration  and greaWhite matter involvement in FTLD has been little studied but long association tracts including the anterior cingulum and superior longitudinal fasciculus have been implicated . A diffuNeural network dysfunction has been proposed to underpin phenotypic features of neurodegenerative disease including FTLD , and in A crucial unsolved question concerns the mechanisms whereby different molecular lesions may produce strikingly dissimilar patterns of neural network breakdown. There are three interrelated pathophysiological issues here: firstly, how one mutation produces asymmetrical cerebral damage and another more symmetrical damage; secondly, how these distinctive patterns of atrophy are maintained or amplified as the disease evolves; and finally, how phenotypic variation arises such that a particular mutation may selectively damage different cerebral hemispheres even between members of the same family . The varThe concept of large-scale neural network breakdown linked to specific molecular lesions may be relevant to the pathogenenesis of a number of neurodegenerative pathologies. Specific mutations within a general category of genetically mediated neurodegeneration (such as GRN or MAPT-associated FTLD) may mediate more fine-grained profiles of brain damage, as suggested by recent evidence within the MAPT spectrum . The sma


We are reporting an extremely prolonged sinus arrest documented by Holter monitoring. A 77-year-old woman was admitted to the geriatric unit at our institution. She complained of recurrent episodes of a sudden loss of consciousness at rest followed by 'seizures' lasting several minutes with complete spontaneous recovery after a few seconds. The patient had suffered multiple episodes in the previous weeks. She had a moderate degree of senile dementia and was not taken any medication. A blood analysis was unremarkable and the ECG showed normal sinus rhythm at 60 bpm with normal PR interval and QRS length. An echocardiogram revelead no significant cardiac structural abnormalities. Twenty-four hours ECG Holter recording was performed and the patient presented a syncopal episode while she was sitting on the armchair. Patient recovered completely after the episode. Holter examination revealed predominant sinus bradycardia (mean of 50 bpm) with no significant atrial or ventricular arrhythmia. At 19:34 pm, when she presented the syncopal attack, a gradual slowing of the sinus beats was observed followed by a sudden drop of ventricular activity due to extremely prolonged sinus arrest (2 minutes and 18 seconds) with two escape beats in between. The episode was terminated by 4 idioventricular beats followed by an accelerated junctional rhythm. Although a malignant vasovagal syncope can not be ruled out, lack of significant heart rate acceleration before ventricular asystole was in favour of sinus node dysfunction. A permanent pacemaker was implanted and no recurrences of syncopal episodes occurred during a year follow-up.


Method. We performed a two-stage vascularized ipsilateral fibular            graft transfer for segmental tibial defect following excision of malignant bone tumours.            Results. We report 10 patients who had this procedure with an average            follow-up of 116 months.The graft was transposed medialy on its vascular pedicle by            two-stage surgery. Full weight bearing was achieved in six patients at 8–43 months            postoperatively, but every patient had a significant complication.            Discussion. The use of this method in isolation is not recommended             for reconstruction of the tibia following tumour excision.


The population pharmacokinetics of51Cr-EDTA were analysed using NONMEM to evaluate the influence of each covariate. The formulae derived were then validated using a further 38 patients and compared with those obtained using existing formulae51Cr-EDTA clearance (GFR) was positively related to Dubois surface area, negatively related to age, and inversely related to serum creatinine (SCr). Females had lower51Cr-EDTA clearance than males. The enzymatic method of SCr assay gave more reliable results than the Jaffe colorimetric method. A measure of creatine kinase significantly improved the estimation of GFR. The new formula produced estimates of GFR which were less biased (Mean Prediction Error = –3%) and more precise (Mean Absolute Prediction Error = 12%) than Cockcroft and Gault (–8% and 16%) or Jelliffe (–15% and 19%) estimates. The formulae developed here can be used to provide reliable estimates of GFR, particularly in regard to targeted dosing of carboplatin. © 2001 Cancer Research Campaign http://www.bjcancer.comThe frequent need to obtain an estimate of renal function in cancer patients, not least for targeting carboplatin dose, has led to a number of approaches to estimate glomerular filtration rate (GFR). This study aimed to develop a simple and reliable method to estimate GFR using readily-available patient characteristics. Data from 62 patients with estimates of


Background: A concomitant infection with human immunodeficiency virus (HIV) may alter the natural history of other infections. Several reports indicate that syphilis may behave more aggressively when HIV infection is present.Case: A woman presented with a rash involving her hands and feet and progressive loss of the vision in her right eye. Her serologic tests for syphilis and HIV infection were positive. A diagnosis of neurosyphilis was confirmed by an analysis of cerebrospinal fluid (CSF). She was treated with high-dose intravenous (IV) penicillin. Her skin lesions resolved, but her vision did not improve.Conclusion: The incidence of HIV infection among women is rising. A patient with HIV and syphilis may develop neurosyphilis in a much shorter time than a patient without HIV infection.


Vascular endothelial growth factor (VEGF) plays a critical role in normal development as well as retinal vasculature disease. During retinal vascularization, VEGF is most strongly expressed by not yet vascularized retinal astrocytes, but also by retinal astrocytes within the developing vascular plexus, suggesting a role for retinal astrocyte-derived VEGF in angiogenesis and vessel network maturation. To test the role of astrocyte-derived VEGF, we used Cre-lox technology in mice to delete VEGF in retinal astrocytes during development. Surprisingly, this only had a minor impact on retinal vasculature development, with only small decreases in plexus spreading, endothelial cell proliferation and survival observed. In contrast, astrocyte VEGF deletion had more pronounced effects on hyperoxia-induced vaso-obliteration and led to the regression of smooth muscle cell-coated radial arteries and veins, which are usually resistant to the vessel-collapsing effects of hyperoxia. These results suggest that VEGF production from retinal astrocytes is relatively dispensable during development, but performs vessel stabilizing functions in the retinal vasculature and might be relevant for retinopathy of prematurity in humans. Retinal astrocytes play an important role in the development of the mammalian retinal vasculature. They invade the retina from the optic nerve head as a proliferating population of cells and spread across the inner surface of the retina, creating a template for the developing retinal vasculature which follows in their wake Vegf mRNA in the not yet vascularized peripheral portion of the retina Vegf mRNA stabilization Numerous studies have shown that during retinal vascularization retinal astrocytes produce high levels of Vegf mRNA near arteries and consequently for the formation of so-called capillary free spaces along arteries Apart from driving angiogenesis, VEGF may also influence vessel remodelling, stabilization and differentiation during vascular plexus maturation. The stability of vessels plays an important role in the pathogenesis of retinopathy of prematurity (ROP), where exposure of premature infants to therapeutic hyperoxia can cause vaso-obliteration in the developing retinal vasculature It has been shown that VEGF secreted from pericytes can also contribute to vessel stabilization However, it is less clear to which degree retinal astrocytes and pericytes contribute to vessel stabilization and how important VEGF secretion from these cells is during the critical period of network maturation in the first 2–3 weeks after birth. We therefore investigated the role of astrocyte-derived VEGF in retinal vascular development by genetically abolishing VEGF expression using the Cre-lox system. Surprisingly, this only had a minor impact on the physiological development of the retinal vasculature. However, under hyperoxic conditions deletion of astrocyte-derived VEGF led to a pronounced destabilization of the retinal vasculature.Vegf mRNA is expressed by retinal astrocytes, retinal ganglion cells and cells in the inner nuclear layer  promoter (Gfap-Cre mice) In the developing retina ar layer  [7], [28 hypoxic . To testGfap-Cre allele into a homozygous background containing loxP sites flanking exon 3 of the Vegfa gene (Vegfc/c mice) Gfap-Cre allele was compared against littermates without the transgene. Cre recombinase activity in Gfap-Cre positive animals was confirmed by PCR of genomic DNA isolated from the retina with primers spanning a 2.1kb region across exon 3, which demonstrated deletion of a 1.3kb region in the Vegfa gene with the appearance of an additional shorter band of 800bp in Cre-positive animals  has been linked to sprouting angiogenesis at the leading edge of the expanding vascular plexus. We therefore assessed whether the absence of VEGF in astrocytes affected the morphology of angiogenic sprouts at the leading edged of the growing vascular plexus, but found no noticeable differences . To furtdeletion , confirmdeletion .Vegf mRNA, the in situ hybridization signal was still detectable within the vascular plexus, particularly along veins  Hif1ac/c) All animals were handled in accordance with the UK Animals (Scientific Procedures) Act 1986. The project was approved by the UK Home Office and the UCL Institute of Ophthalmology Ethics Sub-Committee . Transgenic mice expressing Cre recombinase under the control of promoter segment from the human CAG GCT TCG GTG GGG TGT GA and reverse GAG CAG GGA TAG GTG GTG GAG ATA. To assess VEGF isoforms reverse-transcribed cDNA obtained from P5 retinas was amplified using Taq polymerase (Promega). The VEGF isoform primer sequences were: forward GAA GTC CCA TGA AGT GAT CCA G and reverse TCA CCG CCT TGG CTT GTC A, as described previously To confirm Cre recombinase activity genomic DNA was isolated from P5 retinas and amplified by PCR using Megamix (Cambio Ltd.) containing 2% DMSO with the following primers: forward Retinal whole mount preparations were dissected from whole eyes after brief fixation in 2% (w/v) paraformaldehyde (PFA) in phosphate-buffered salt solution (PBS) and then processed for wholemount immunohistochemistry or in situ hybridization as previously described Network morphology of the retinal vasculature was analyzed in retinal whole mount preparations from 6 P5 and 5 P10 conditional VEGF litters and in 5 P5 conditional HIF1α litters. We also analyzed 5 further litters after hyperoxia exposure for 5 days at P12 and 5 litters after 1 day of hyperoxia at P8. The surface area of the developing vascular network, width of capillary free spaces and vascular depleted areas in hyperoxia-exposed animals were measured using ImageJ. BrdU positive cells and artery side branches were counted by hand over fixed distances from the retinal centre. Intact and degenerating radial arteries and veins were counted by hand in each retina. Measurements were averaged for control and mutant animals in each litter and normalized against the control values. Statistical analysis was then performed across values obtained from each litter.


Eight children treated between 1977 and 1983 with the UK Children's Cancer Study Group's non-Hodgkin lymphoma (NHL) and T-cell protocols have developed second malignancies within 7 years of commencing treatment. Five developed acute non-lymphoblastic leukaemia and a sixth died from infection while pancytopenic with a pre-leukaemic marrow. The other malignancies were cerebral astrocytoma and an undifferentiated low grade sarcoma. These eight children were included among 261 children studied in the first UKCCSG NHL and T-cell trials giving an actuarial incidence of 7.8% second malignancy at 7 years. Six had received adjuvant radiotherapy which may have contributed to the high incidence of second malignancy.


A significant correlation  was found between the extent of residual DNA dsbs and clonogenicity for all 11 fibroblast strains. The parameter showing the highest correlation with fibroblast cell killing (D-) for the nine normal fibroblasts alone was the ratio of initial/residual DNA dsb dose–response slope . A significant correlation  with clonogenic radiosensitivity was also found for all 11 cell strains when using the ratio of initial/residual DNA dsb damage at a single dose of 180 Gy. This study shows that fibroblast radiosensitivity measured using the neutral comet assay correlates with clonogenic radiation survival parameters, and therefore may have potential value in predictive testing of normal tissue radiosensitivity. © 1999 Cancer Research CampaignA study was made of the neutral comet assay as a potential method for measuring normal cell radiosensitivity. Eleven fibroblast strains were studied comprising nine derived from vaginal biopsies from pretreatment cervical cancer patients and two strains from radiosensitive individuals. DNA double strand break (dsbs) dose–response curves for both initial and residual (20-h repair time) damage were obtained over the dose range 0–240 Gy, with slopes varying 3.2 and 8-fold respectively. Clonogenic cell survival parameters were available for all the cell strains following both high- and low-dose rate irradiation. There were no correlations between the dose–response slope of the initial level of DNA dsbs and parameters that mainly describe the initial portion of clonogenic radiation survival curves (SF


L -lactate and L -deprenyl prevented bystander factor associated cell kill. The lactate analogue, oxamate, was less effective. Data from experiments where media harvested from the different cell lines was exchanged suggest that signal production and cellular response may involve different mechanisms. The effects on exposed cells were transmitted to progeny which also showed excessive levels of cell death for several generations. The results suggest that energy/REDOX metabolism may be involved in the expression of a radiation induced bystander response. Given the aberrant energy metabolism in tumour cells, this may have implications for dose escalation in radiotherapy. © 2000 Cancer Research CampaignThese experiments were done to determine if interference with energy metabolism and REDOX biochemistry during low LET radiation exposure would alter the ability of medium harvested from the irradiated cells to induce a bystander effect in unirradiated cells. Human keratinocyte cells and CHO-K1 mutant cell lines were irradiated using cobalt 60. Clonogenic assays were used to determine the reproductive death of the cells exposed to direct irradiation or medium from irradiated cells. The persistence in progeny was also examined. Use of apoptosis inhibitors or medium from the LDH or G6PD null cell lines, reduced or prevented the bystander effect. Transfection with G6PD recovered the effect. Treatment with anti-oxidant substances,


Attempts were made to induce tumour-specific immunity in syngeneic mice by treatment with x-irradiated  tumour or surgical excision of developing subcutaneous tumour grafts. Resistance was evoked against 6 tumours and this was relatively weak. With the remaining tumours, no resistance against the immunizing tumour could be demonstrated, even though the transplantation tests were made highly sensitive by the use of inocula of as few as 2 × 103 cells in pre-irradiated (400 rad) hosts. Sera from mice immunized against each of the tumours were tested against viable cells of the immunizing tumour by indirect immunofluorescence. In no instance did tumour antisera give a convincing reaction with tumour cells although alloantisera raised by hyperimmunization of H-2 identical and H-2 different donors with osteosarcomata consistently gave strongly positive reactions. The results are interpreted as illustrating the weak tumour specific antigenicity of radiation-induced murine osteosarcomata. The possibility that antigenic deficiency is a consequence of immunosurveillance in this tumour system is discussed.The immunogenicities of 15 murine osteosarcomata induced with a bone seeking radioisotope (