Recent data has shown a consistent but modest association between hearing impairment and poor mobility; both are strongly associated with cognition. Cognitive function may moderate the relationship between hearing and mobility. We analyzed 601 cognitively normal older participants from the Baltimore Longitudinal Study of Aging who had concurrent data on cognition , hearing , and mobility . We performed multivariable-adjusted linear regression to test two-way interactions between each cognitive measure and PTA. There were significant PTA interactions with all cognitive measures on 400-meter time. There was a significant interaction between PTA and sensorimotor function on 6-meter gait speed. Among cognitively normal older adults, poorer hearing is more strongly associated with poor mobility in those with low cognition, especially sensorimotor function. Future studies are needed to understand how cognition may moderate the relationship of hearing impairment with mobility decline over time.


Pituitary adenomas (PA) are one of the most common primary brain tumors and comprise approximately 15% of brain neoplasms. Most PA are histologically benign, but can cause significant morbidity. Previous studies utilized whole genome and exome sequencing to identify a few somatic variants, but no recurrent mutations were observed. Further studies are warranted to identify driver mutations occurring at low frequencies. We used single-cell RNA sequencing (10X Genomics) to investigate cellular heterogeneity in 12 non-functioning pituitary adenomas. Our analysis identified discrete clusters of cells associated with specific functional pathways. One of these clusters corresponded to cells expressing genes related to metabolic pathways, primarily lipid metabolism. Another cluster consistent amongst the three patients comprised cells involved in antigen presentation and processing. In addition, the copy number variation analysis highlighted distinct chromosomal alterations within our samples. Interestingly, we were able to identify clonal variations within each tumor based on chromosomal aberrations. For example, in our first patient we observed a gain of chromosome 19 and loss of chromosome 2. Our analysis showed three different clonal populations within this tumor. All three populations harbored the loss of chromosome 2, one population exhibited gain of chromosome 19, while a third population exhibited loss of chromosome 19. These early results indicate the loss of chromosome 2 as an early event in tumorigenesis and gain/loss of chromosome 19 as late events. We are currently in a process of identifying somatic variations within these tumors by variant calling. Currently we are expanding our analysis to 20 non-functional PA. Mapping the single-cell gene expression profiles with mutational phylogeny will reveal the differences in clonal evolution within the tumor subtypes. This study will help us define the molecular fingerprint of pituitary adenomas and provide insights which could be utilized in the clinic for better management of these tumors.


Miño et al.). After receiving an extensive counseling, more women opted for an invasive prenatal testing while less women opted for the first trimester combined test and cell free DNA testing, as compared to those without extensive counseling .There has been a geometric explosion in genetic capabilities such that currently a wide variety of prenatal screening and diagnostic testing for fetal chromosomal abnormalities are available. Ideally, all woman should be counseled in each pregnancy about the benefits and limitations of available testing . In counCheng et al.). Although more non-pregnant women accepted ECS compared to pregnant women (70.7 vs. 61.2%), fewer non-pregnant women heard about ECS than pregnant women (32.3 vs. 42.4%). The majority of them showed a lack of understanding about ECS despite being given pamphlets . It is important to improve women’s understanding of reproductive risk before making an informed decision, especially the issue related to false-negatives, of which the readers can refer to a recent excellent review  recommends a consistent and equitable approach for offering carrier screening to all women during pregnancy or before conception . Next-get review .While prenatal chromosomal microarray analysis (CMA) is a current standard genetic testing, prenatal whole exome sequencing and eventually whole genome sequencing are emerging technologies that will become front line tests in the future. CMA can detect major chromosomal imbalances as well as copy number variations that are too small to be detected by traditional karyotyping. Diagnostic sequencing is useful for evaluation of fetuses for whom CMA is uninformative. It may be offered concurrently according to accepted practice guidelines. CMA is less optimal than sequencing for the presenting fetal phenotype according to expert genetic opinion . The authors used an extended segregation analysis including the parents and their relatives to provide further useful information for genetic counseling.The prenatal identification of a baby with a MECP2 missense mutation and 15q11.2 microduplication in a family with a child having developmental epileptic encephalopathy associated with a Lin et al.). A compound heterozygous mutation with c.856 + 1G > T and c.857-619_1269 + 243delinsTTGCCTTGC changes were found in the first family while homozygous c.857-619_1269 + 243delinsTTGCCTTGC mutations were found in the second family. These mutations are associated with autosomal recessive renal tubular dysgenesis which is a severe disorder with an unfavorable outcome. Notably, a few earlier reports from Taiwan had pointed out that the mutant allele c.857-619_1269 + 243delinsTTGCCTTGC of the AGT gene is very likely to be a founder variant in the Chinese population, which may aid in the prompter identification of the molecular pathology in that rapid trio-WEs case .Another limitation of prenatal sequencing is long turn-around time. In the present issue, prenatal diagnosis of two families with recurrent oligohydramnios by rapid trio-whole-exome sequencing (WES) which revealed mutations in the AGT gene within 1 week was reported . GS enabled accurate identification of all breakpoints in these ten cases, revised the conventional karyotype results in nine cases, and provided additional information and changed the interpretation of the BCAs in four cases . These results show us the importance of precisely delineation of breakpoints as BCAs are associated with neurodevelopmental disorders including intellectual disability and autism spectrum disorder.Recently, whole-genome sequencing (WGS) has been used to delineate the breakpoints of balanced chromosomal abnormalities (BCA) which have no visible gain or loss of genetic material at cytogenetic resolution . The usePauta et al.). This new molecular work-up included two quantitative fluorescence PCR (QF-PCR) rounds, and a high-resolution SNP-array in those cases with normal QF-PCR results. QF-PCR and CMA can overcome the pitfalls of conventional karyotyping including culture failure and submicroscopic abnormalities. Besides, the authors also proposed transcervical chorionic villus sampling (CVS) during surgical or before medical uterine evacuation to avoid maternal contamination. They found that transcervical CVS was more effective in the retrieval of embryonic tissue for chromosome analysis than examining POC after evacuation .Chromosome analysis of both couples with recurrent pregnancy loss (RPL) and products of conception (POC) is most informative in the investigation of possible genetic causes of RPL . ParentaChuang et al.). The authors explored whether the incidence of mosaicism for individual chromosome in blastocysts is correlated with chromosome length, highlighting the complex mechanisms of causing mosaicism in blastocysts.NGS after trophectoderm (TE) biopsy have been used for preimplantation genetic testing for aneuploidies (PGT-A) because of its high sensitivity . Compare


Cells of osteogenic sarcoma line 791T were treated in vitro with a selectively cytotoxic methotrexate-human serum albumin-monoclonal antibody conjugate at concentrations which were toxic but allowed the "escape" of a small number of tumour cell colonies (less than 0.3% compared with controls). These colonies were propagated as clones in order to test their expression of the monoclonal antibody ( 791T /36)-defined antigen and their resistance to methotrexate (MTX) by comparison with parental cells. Most of the conjugate-treated clones were incapable of prolonged growth and died out, in contrast to untreated 791T clones which virtually always grow progressively. Only four treated clones grew at rates comparable with the parental line. Flow cytofluorometric analysis indicated that the surviving clones expressed normal or enhanced amounts of 791T /36-defined antigen and clonogenic assays demonstrated that they were sensitive to cytotoxicity by MTX. As could be predicted from these results, further exposure to the conjugate inhibited growth of the clones at doses comparable with those active against parental 791T cells. It is concluded that tumour cell clones emerging after exposure to a toxic concentration of a drug-antibody conjugate are not necessarily modified resistant clones, but may have severely impaired long-term growth potential or be susceptible to further contact with the same conjugate.


Iodo(14C-)antipyrine autoradiography and imaging bioluminescence have been combined to obtain pixel-to-pixel correlations between absolute values for local blood flow and ATP concentrations at a microscopical level within designated areas in hamster melanomas. Positive pixel-to-pixel correlations were obtained in 4 of 6 tumours. Both flow and ATP values were less in mostly necrotic than in mostly viable tumour regions. The data provide evidence for the energetic state of cancer cells being strongly influenced by the efficiency of tumour microcirculation in several but not in all malignancies investigated.


Production of colony-stimulating factor (CSF) was examined in three patients with lung cancer associated with neutrophilia. All three patients presented a marked increase in neutrophil count  that continued at least for 3 weeks and rapidly disappeared after surgical removal of the tumours. Culture media (CM) incubated with the excised tumour tissues stimulated the colony formation of bone marrow myeloid progenitor cells in vitro. Northern blot analysis of poly(A)+ RNA from the tumour tissues revealed a constitutive expression of granulocyte (G), macrophage (M), and granulocyte-macrophage (GM) CSF genes in all tumours. Immunoassay specific for these CSFs revealed that G- and M-CSF immunoreactivity was detected in all CM and GM-CSF protein in two out of three CM. The plasma CSF levels also increased before operation and decreased to normal or near-normal range after operation. In contrast, tumour cell CM obtained from two lung cancer patients without leucocytosis neither stimulated haematopoietic colony formation nor contained immunoreactive CSFs. These results indicated that the neutrophilia found in the three patients was probably caused by constitutive production of multiple CSFs by lung cancer cells.


P-glycoprotein expression was demonstrated in two human intestinal adenocarcinoma cell-lines  by immunoprecipitation of a 170-180 kDa protein with monoclonal antibody JSB-1. Both HCT-8 and T84 formed functional epithelial cell layers of high transepithelial electrical resistance (greater than 700 omega.cm2) when grown on permeable matrices. These epithelial layers demonstrated vectorial secretion . These vectorial vinblastine secretions were sensitive to inhibition by verapamil. Passive transepithelial vinblastine permeation was limited by the presence of intercellular (tight) junctions, as demonstrated by the high transepithelial electrical resistance, and verapamil increased this passive vinblastine permeation concomitant with a reduction in the electrical resistance. Cellular vinblastine loading was significantly greater from the basal side, and this was also susceptible to inhibition by basal verapamil. The demonstration of vectorial transport of vinblastine in human intestinal colonic adenocarcinoma cell layers is direct evidence in favour of the hypothesis that the function of mdr1 in epithelial from the gastrointestinal tract is to promote detoxification by a process of epithelial secretion. This study also highlights that cellular vinblastine accumulation depends not only upon P-glycoprotein function, but also upon differential apparent membrane permeabilities and the presence of intercellular (tight) junctions that may restrict drug permeation and cellular accumulation to apical or basal membrane domains.


SR 4233  is the lead compound of the benzotriazene-di-N oxides which are selectively toxic to tumour cells under hypoxic conditions. However much higher concentrations given to rats caused bone marrow toxicity and necrosis of the low oxygen Zone 3 part of the liver. In the following effects of SR 4233 on hepatocytes under hypoxic vs aerobic conditions have been compared. (1) SR 4233 did not affect hepatocyte viability (as determined by plasma membrane disruption) or glutathione levels under aerobic conditions. SR 4233 however induced cyanide-resistant respiration, an indicator of redox cycling mediated oxidative stress and became cytotoxic if hepatocyte catalase or glutathione reductase was inactivated. Glutathione oxidation occurred well before cytotoxicity ensued. Addition of ascorbate markedly enhanced SR 4233 cytotoxicity to these compromised hepatocytes. (2) In contrast, SR 4233 was highly toxic to hypoxic hepatocytes. Addition of ascorbate to enhance SR 4233 reduction also caused a marked increase in hepatocyte toxicity and an SR 4233 radical was detected with ESR spectroscopy. SR 4233 cellular reduction and toxicity was prevented with fructose or inhibitors of NADPH:cytochrome P-450 reductase. Inactivation of catalase or glutathione reductase had no effect on SR 4233 toxicity and hepatocyte GSH was not oxidised indicating oxidative stress did not occur during hypoxic SR 4233 hepatocyte toxicity. (3) The lack of SR 4233 cytotoxicity under aerobic conditions could probably be attributed to the detoxification of the SR 4233 radical by mitochondrial oxidation as SR 4233, but not its metabolite SR 4317 markedly increased state III and IV mitochondrial respiration in the presence of NADH. The increased respiration was inhibited by the respiratory inhibitors KCN and antimycin A but not by rotenone. Furthermore SR 4233 cytotoxicity under aerobic conditions was markedly increased by partially inhibiting hepatocytes respiration with cyanide but not rotenone.


The prognostic value of cellular DNA content in melanoma metastases was investigated by flow cytometric analysis of fresh or paraffin-embedded tumour blocks from 95 consecutive patients referred to the Helsinki University Central Hospital Melanoma Team. Thirty-three per cent of the tumours were DNA diploid and 67% DNA aneuploid. S-phase fractions were lower in DNA diploid than in DNA aneuploid tumours (10.7% and 17.6%). Tumour ploidy and S-phase fraction were shown by multivariate Cox model analysis to be independent prognostic variables and major determinants of survival after first recurrence. Surprisingly, patients with DNA aneuploid tumours and with tumours with low SPF survived significantly longer than those with DNA diploid or high SPF tumours. This exceptional finding of favourable prognosis for DNA aneuploid tumours was more prominent among patients receiving intensive systemic therapy and among patients with stage IV disease, probably indicating a tendency for DNA aneuploid tumours to have higher sensitivity to systemic therapy.


Observation of Coincident arrhythmias is not uncommon but the co-existence of idiopathic verapamil sensitive left ventricular tachycardia (ILVT) with other arrhythmias is very rare. We hereby presented a 30 year old male patient with a history of frequent episodes of palpitations and sustained narrow complex tachycardia. During electrophysiologic study two arrhythmias, one with narrow complexes which was shown to be typical atrioventricular nodal re-entrant tachycardia and the other with wide QRS complexes and right bundle branch block and left axis morphology, compatible with ILVT, were inducible. Radiofrequency catheter ablation of both arrhythmias was done at two consecutive sessions. The patient has remained asymptomatic without antiarrhythmic therapy for the past six months. Idiopathic sustained ventricular tachycardia (VT) accounts for 10-20% of patients with sustained monomorphic VT , but coeWe hereby presented a 30 year old male patient with three years history of frequent episodes of palpitation. He had no structural heart disease. His previous medical records revealed several ECGs during arrhythmia all showing regular narrow complex tachycardia with a rate of 180beat/min. There was no documented wide complex tachycardia. During initial EP study a narrow complex tachycardia was reproducibly induced  which waearliest PP recording site  were introduced via left femoral vein and positioned at RV apex (and right ventricular outflow tract), His bundle and high right atrium. A 7F steerable decapolar catheter (2-5-2 mm) was introduced via left femoral artery  and positioned in LV on interventricular septum for recording intracardiac signals during sinus rhythm and tachycardia. This catheter was used as a guide for localization of earliest purkinje potential (PP) recording site by ablation catheter. A 7F ablation catheter  was introduced via right femoral artery and positioned in LV on septum for mapping and subsequent RF ablation. The ILVT was terminated by mechanical pressure at ing site . RF enerThis case describes the coincidence of AVNRT and ILVT. Occurrence of AVNRT in combination with idiopathic VT has been previously described. In patients with RVOT tachycardia the incidence of AVNRT has been reported to be as high as 15% . Our finearliest PP during tachycardia should be targeted.Wagshal AB, et al.  also repThis case represents the co-incidence of ILVT and AVNRT and underscores the importance of searching for dual AV node physiology and AVNRT in patients with ILVT.


The outcome of salvage treatment was reviewed in 55 patients relapsing during or after their primary chemotherapy for advanced malignant germ cell tumours. Fifty-two patients had been given cisplatin-based chemotherapy as their primary treatment, whereas three patients had received carboplatin-based chemotherapy. The median time to relapse was 2 months (range: 0-96 months) from discontinuation of the primary treatment. Two patients underwent radical surgery only, and one patient had radiotherapy to a brain metastasis as his only curatively intended salvage treatment. Six patients did not receive any treatment for their recurrent malignancy  except for purely palliative measures. The disease-free survival for the total group was 27% at 5 years. Complete response to primary treatment lasting for > or = 6 months was the only parameter which significantly predicted a favourable outcome .


A study involving the measurement of glutathione S-transferase activities and isoenzyme distributions in human ovarian tumours has been carried out. These tumours have been obtained either at initial debulking surgery, prior to cytotoxic chemotherapy, or at second look laparotomy following chemotherapy. The response rates of these two groups to chemotherapy differ markedly, with patients who have relapsed following initial chemotherapy showing a reduction in response rates to subsequent chemotherapy. Analysis of these data show no statistically significant differences between the glutathione S-transferase activity or isoenzyme distribution in these two groups of patients. Significant differences were observed in the glutathione-S-transferase activities (GST) between tumours and normal ovaries. GST activities in pre-chemotherapy tumours  and post-chemotherapy tumours  where significantly higher than the GST activity in normal ovaries (n = 15). One feature was the expression of the basic isoenzyme which is expressed more in normal ovaries than in tumours. No differences in these parameters were observed in normal peritoneal tissue taken from patients before or after chemotherapy. These data do not support the hypothesis that changes in glutathione S-transferase enzyme activity or isoenzyme expression are major determinants of response to chemotherapy in ovarian tumours.


ECG in sinus rhythm with ventricular preexcitation and changing QRS morphology was seen that was initially interpreted  as the multiple accessory pathway from elsewhere. .The following mechanisms are potentially involved in the electrogenesis of changing QRS morphology in WPW syndrome: 1) multiple accessory pathways ; 3) ventElectrophysiologic study showed short PR (75 ms) interval with wide QRS (152 ms) and negative HV (-12 ms) interval. No change in delta wave polarity was observed during HRA and CS pacing. In full preexcitation, no breakthrough was seen in the CS. During incremental ventricular pacing, atrial breakthrough site is initially recorded on the HRA catheter and then changed to distal pole of CS catheter with progressive decrease in pacing cycle length. During ventricular pacing at cycle length of 500 ms (S1), earliest atrial activity is recorded on HRA catheter.multiple APs because only right-sided AP had bidirectional conduction and no distal CS breakthrough was seen simultaneous with changing QRS morphology. The possibility of aberrant conduction is excluded by presence of negative HV interval in the beats with differing QRS morphology. No sinus cycle length variation before and after the beats with different morphologies are against the occurrence of functional LBBB. The prematurity of ventricular electrogram in His recording catheter with variable HV (H-electrogram is recorded after V-electrogram in second beat and before V-electrogram in third beat) and fixed V-RB intervals  are compatible with ventricular fusion of preexcited sinus impulse with ectopic ventricular impulse originating from parahissian area (explaining LBBB and inferior axis morphology of the beats with changing QRS) but not from the His bundle or RBB itself  (Changing QRS could not be explained by presence of r fusion).


Objective To study the microbial etiology of tubo-ovarian abscess (TOA).                Methods We recruited 11 women in Nairobi, Kenya who failed antibiotic therapy alone and requiredsurgical drainage of a presumptive TOA. Pus from the nine abscesses and two pyosalpinges were collected andcultured for aerobic, facultative and anaerobic microorganisms.Results Eleven women suspected of having a TOA were hospitalized and treated for a median of 6 days (range3–14 days) prior to surgical drainage of the abscess. Nine (82%) specimens were culture positive. Aerobeswere present in all nine specimens. Seven of the nine positive cultures (78%) were polymicrobial and five ofthe polymicrobial cultures contained both anaerobes and aerobes. Anaerobic Gram-negative bacilli  and Streptococcus sp.  were themost common microorganisms isolated. Neisseria gonorrhoeae and Chlamydia trachomatis were not isolated byculture or detected by polymerase chain reaction.Conclusions In Kenya, persistent TOAs are associated with endogenous flora similar to that normally foundin the gastrointestinal tract.


A 25 year old man underwent electrophysiology study for recurrent symptomatic paroxysmal palpitations. There were no documented episodes of supraventricular tachycardia on surface ECG.  Delta waves were absent on baseline ECG.A decapolar catheter was placed in the coronary sinus with the distal and proximal pair of electrodes configured as CS 1-2 and CS 9-10 respectively. Quadripolar catheters were positioned in the high right atrium, His-bundle region and right ventricular apex. During straight atrial pacing with a drive-cycle length of 340ms, the following arrhythmia was induced . The corThe surface ECG revealed a narrow complex supraventricular tachycardia with alternating R-R intervals at 310ms and 360ms . The int On close inspection of the intracardiac electrogram, two different alternating A-H intervals were observed, which presented as alternating R-R intervals on the surface ECG.The A-H intervals were 110ms and 160ms consecutively. We deduced that the antegrade limb of this macro-reentrant circuit involved both the fast and slow AV nodal pathways alternating with each other.Radiofrequency ablation of the left lateral pathway via a retrograde aortic approach was performed. The tachycardia was successfully terminated. Further electrophysiology studies confirmed the presence of dual AV nodal pathways. There were no further inducible arrhythmias even with isoproterenol infusion. The slow pathway was thus not ablated.The mechanism for this supraventricular tachycardia with alternating R-R intervals involved two alternating antegrade limbs  with different conduction times and a common retrograde concealed left sided accessory pathway. This electrophysiological mechanism had been postulated previously based on appearance of the surface ECG without intracardiac recordings .


The dentate gyrus is well known for its mossy fiber projection to the hippocampal field 3 (CA3) and its extensive associational and commissural connections. The dentate gyrus, on the other hand, has only few projections to the CA1 and the subiculum, and none have clearly been shown to extrahippocampal target regions.Using anterograde and retrograde tracer techniques in the Madagascan lesser hedgehog tenrec  it was shown in this study that the dentate hilar region gave rise to a faint, but distinct, bilateral projection to the most rostromedial portion of the olfactory tubercle, particularly its molecular layer. Unlike the CA1 and the subiculum the dentate gyrus did not project to the accumbens nucleus. A control injection into the medial septum-diagonal band complex also retrogradely labeled cells in the dentate hilus, but these neurons were found immediately adjacent to the heavily labeled CA3, while the tracer injections into the rostromedial tubercle did not reveal any labeling in CA3.The dentate hilar neurons projecting to the olfactory tubercle cannot be considered displaced cells of CA3 but represent true dentato-tubercular projection neurons. This projection supplements the subiculo-tubercular projection. Both terminal fields overlap among one another as well as with the fiber terminations arising in the anteromedial frontal cortex. The rostromedial olfactory tubercle might represent a distinct ventral striatal target area worth investigating in studies of the parallel processing of cortico-limbic information in tenrec as well as in cat and monkey. The dentate gyrus (Dt) may be known best for its input from the entorhinal cortex and its mossy fiber projection to the CA3 -3. In adEchinops telfairi, Et) will demonstrate some true dentate hilar cells projecting to a circumscribed region in the olfactory tubercle. The projection may represent an additional parallel pathway transferring cortico-limbic information to the ventral striatum [Continuing our attempts to elucidate the hippocampal and parahippocampal circuits in mammals with a poorly differentiated brain -29 the pstriatum -32.Among the experiments with tracer injections into the hippocampus Table  two caseThe latter two cases revealed the well-known hippocampo-striatal projection pattern to the nucleus accumbens (Acb) and the olfactory tubercle (Tu) . No hippIn Et01-21B the course of fibers towards Tu appeared to pass exclusively within the molecular layer of the hippocampal continuation  but getThe present data show for the first time a projection from the dentate gyrus to the rostromedial olfactory tubercle. Its cells of orgin, located in the dentate hilus, cannot be considered displaced neurons of CA3 but represent true dentato-tubercular projection neurons. The projection is assumed to be present in other mammals, too. In the tenrec the dentato-tubercular projection overlaps with the subiculo-tubercular projection and the fiber terminations arising in the anteromedial frontal cortex. The circumscribed ventral striatal target area appears worth investigating in studies of the parallel processing of cortico-limbic information in tenrec as well as in cat and monkey.Echinops telfairi, Et) is a member of the tenrecomorpha classically considered an insectivoran suborder [The lesser hedgehog tenrec . Thesuborder  built upsuborder ,81.Eleven tenrecs weighing between 80 and 140 gm were anesthetized with tribromoethanol  and injected with tracer into the hippocampus (n = 7), the ventral striatum (n = 4) and the septum (n = 1) following the German laws on protection of animals . Wheat germ agglutinin conjugated to horseradish peroxidase  and biotinylated dextran amine  were used as tracer substances. Most cases have been described previously with regard to other connections ,28,82. TThe brains were soaked overnight in a 30% phosphate-buffered sucrose solution, embedded in an albumin-gelatine mixture and cut on a freezing microtome at 40 μm in the frontal plane. The WGA-HRP staining was usually done on two out of four sections according to the standard tetramethylbenzidine technique  and a moBright-field illumination and polarized light were used for the analysis. Images were captured on an Ilford 50 negative film or a Fujichrome 64T positive film (using a Zeiss axiophot microscope), scanned (Nikon Coolscan 5000) and transported into Adobe Photoshop (v. 7.0) and Corel Draw (v. 11.0). The sharpness, contrast and brightness were adjusted to reflect the appearance of the labeling seen through the microscope. For the sake of clarity the most disturbing crystalline artefacts were removed, particularly in darkfield images. In some darkfield micrographs the light reflecting embedding medium was also removed. Furthermore to facilitate the analysis all micrographs are shown with the injection site on the left.Acb – nucleus accumbensBDA – biotinylated dextran amineCA1/3 – subfields of cornu ammoniscma – commissura anteriorDgMS – diagonal band-medial septum complexDt – dentate areaDtGr – dentate granule cell layerDtHi – dentate hilar regionDtMo – dentate molecular layerEt – Echinops telfairiHCt – hippocampal continuationSbi – subiculumTu – olfactory tubercleWGA-HRP – wheat germ agglutinin conjugated to horseradish peroxidase


A cytofluorometric assay allowing the measurement of thymidine phosphorylation in single cells had been established . This assay enables us to correlate intracellular thymidine kinase (TK) activity with the DNA content of single cells. Enzyme activity levels from neuroblastoma cells and normal fibroblasts derived from the same patient were determined. Using this cytofluorometric assay in a mixture of both cell types the neoplastic cells could be distinguished from the normal fibroblasts because of their higher TK level. A human lymphoblastoid cell line was compared with the cell line KG-1, derived from an acute myelogenous leukaemia, in the same way. The increased enzyme activity enabled us to detect KG-1 cells in a mixture with an 10,000-fold excess of Epstein Barr virus transformed lymphocytes.


Vasoactive drugs have a variety of effects upon splanchnic and hepatic haemodynamics which may alter tumour blood flow and potentiate the delivery of a chemotherapeutic drug to hepatic tumour. We have investigated the effects of vasopressin infusion on hepatic tumour blood flow in an experimental model of liver tumour. Hepatic tumour was induced by the intraportal inoculation of HSN sarcoma cells. Hepatic and splanchnic blood flow was determined using a dual reference microsphere technique before and after an intravenous infusion of vasopressin at a dose of 0.1 mU kg-1 min-1 for 10 min. There was a significant increase in systemic arterial blood pressure associated with a rise in portal venous inflow  and a significant fall in hepatic arterial flow (P less than 0.05). The tumour: liver blood flow ratio was significantly increased by vasopressin infusion (P less than 0.02). Vasopressin infusion decreases hepatic arterial flow and increases tumour blood flow which may potentiate the delivery of a regionally delivered chemotherapeutic drug to hepatic tumour.


The variation in survival of women with clinically similar breast cancers may lead to difficulty in clinical management so it is important to recognise factors which indicate the prognosis. Immunoperoxidase staining patterns of primary breast tumours using monoclonal antibody NCRC-11 have been shown to relate to overall survival  but the results have not been reproducible in other centres. In this study paraffin sections of 483 primary breast cancers were stained with NCRC-11 and 3E1.2 using an immunoperoxidase system. The tumour staining patterns were compared with overall survival using life tables and tested for relative prognostic significance by Cox's multivariate analysis. NCRC-11 related to survival in all 483 cases  but both antibodies achieved maximum prognostic significance in lymph node negative patients  in whom no other factor was more significant. Immunoperoxidase staining patterns produced by monoclonal antibodies NCRC-11 and 3E1.2 are important prognostic factors in breast cancer.


Central airway obstruction is a common complication of lung cancer. The microdebrider is a new device available for treatment of central airway obstruction.We report a case a 59-yr-old male with T3N2M1 non-small cell lung cancer with malignant distal left mainstem obstruction treated successfully with a novel elongated rotating tip microdebrider via rigid bronchoscopy with sufficient length to reach distal bronchial lesions.The microdebrider is an excellent addition to the spectrum of interventions available for the management of central airway obstruction with advantages including accuracy and immediate removal of debris without a need for separate suctioning or limitation in oxygenation. The microdebrider is a recent addition to the armeratarium available to the interventional bronchoscopist for central airway occlusion, following the successful application of laryngeal microdebriders for over a decade -3. HowevA 53-yr-old male with T3N2M1 non-small cell left upper lobe lung cancer with bilateral brain metastases diagnosed one-month prior was referred for management of dyspnea and hemoptysis. The patient had noted a dramatic increase in dyspnea over the prior 3 days. Past medical history was unremarkable and he had a negative smoking history.Physical exam revealed reduced breath sounds in the left lower zone. Chest x-ray demonstrated a left hilar mass with loss of volume and post obstructive pneumonia in the left lower lobe Figure . Axial C® (APC) was utilized to devitalize the tissue. The devitalized tissue was then debulked using the Straightshot® microdebrider with rotating tip  declare that they have no competing interests.


Sodium polyanethol sulphonate (SPS) when incorporated into rat erythrocyte lysate (REL) containing semi-solid agar medium at 1 mg ml-1. markedly enhanced colony formation by a number of anchorage-independent cell lines. REL usually needed to be included for the expression of SPS induced potentiation as in its absence SPS was generally cytotoxic. Studies suggested that SPS reduced the lag prior to colony initiation resulting in the earlier appearance of colonies and in a higher cloning efficiency. The effectiveness of SPS in potentiating colony formation by responsive cell lines was markedly influenced by the species of serum and to a lesser extent by differences between individual batches. Enhancement by SPS was greater with poorer foetal calf serum (FCS) batches than with better. This effect may have been partly due to SPS interfering with the action of a growth inhibiting serum component, possibly a lipoprotein. Studies in which delipidated FCS was substituted for normal FCS suggested that SPS was also able to compensate for the lack of a growth-promoting lipid component. Binding studies showed that initially 125I-SPS bound equally well at 4 degrees C and 37 degrees C with continued labelling occurring only at 37 degrees C. Autoradiography of cells labelled at 37 degrees C for 24 h revealed the presence of intracytoplasmic 125I-SPS.


In the article “Natural Variability and the Influence of Concurrent Control Values on the Detection and Interpretation of Low-Dose or Weak Endocrine Toxicities,” p-dioxin (TCDD)  . Second,There are many reasons why males from different rat strains obtained from different breeding facilities using different animal feed and housing conditions might differ in DSP. We thus disagree with the interpretation by


Eighty-one patients with small cell lung cancer (SCLC) with a survival of more than 2 years after start of chemotherapy were studied. Twenty-six of the 28 patients who died of relapsed SCLC had in fact relapsed before two years and of the 55 who had not then only two (4%) relapsed subsequently. It is stressed that with such observations treatment related factors should be taken in account. Second tumours were observed in ten patients, nine proven malignant. Of the eight patients with non-small cell lung cancer three had residual disease after initial chemotherapy. In our patient group after a 2 year disease-free interval the risk of developing non-small cell lung cancer seems higher than a subsequent relapse of SCLC.


Using immunohistochemistry and the monoclonal antibody C219 we have investigated P-glycoprotein expression in 26 locally advanced breast cancers. Twenty four patients had received four cycles of chemotherapy  prior to mastectomy; two received tamoxifen. Twelve tumours exhibited an objective response to the chemotherapy. A background pattern of isolated weakly positive (1+) stromal staining (myofibroblast) was observed in seven tumours, two of which had been treated by tamoxifen alone. Two of the tumours treated by induction chemotherapy showed positive staining (1+) within a very small number of isolated tumour cells (maximum of three) and macrophages. The significance of this staining is not clear although C219 may simply be cross reacting with myosin. We have failed to demonstrate a clear clinical utility for C219 in breast cancer, particularly regarding the identification of patients in whom MDR chemotherapy be avoided once metastases develop.


Phospholipids from malignant, benign and noninvolved human breast tissues were extracted by chloroform-methanol (2:1) and analysed by 31P MR spectroscopy at 202.4 MHz. Thirteen phospholipids were identified as constituents of the profiles obtained among the 55 tissue specimens analysed. Observed patterns in phospholipid tissues profiles were distinct, allowing qualitative characterisation of the three tissue groups. Multivariate analysis of lysophosphatidylcholine (LPC) and an uncharacterised phospholipid were shown to be independently significant in predicting benign tissue histology as either fibrocystic disease or fibroadenoma in 92% of cases. Univariate analysis of relative mole-percentage of phosphorus concentrations of individual phospholipids using the Scheffé comparison procedure revealed that in malignant tissues, phosphatidylethanolamine was significantly elevated compared to benign (+ 32%) and noninvolved tissues (+ 22%). Phosphatidylinositol (+ 33%) and phosphatidylcholine plasmalogen (PC plas) (+ 25%) were increased in malignant compared to benign and LPC was decreased (-44%) in malignant compared to noninvolved. LPC was significantly depressed (-39%) in benign tissue compared to normal. Phospholipid indices computed to further characterise the three tissue groups showed PC plas/PC elevated in malignant tissue compared to benign and PE plas/PE depressed in malignant tissue compared to noninvolved. These findings support previous investigations reporting that the alkyl-phospholipid analogues of phosphatidylcholine are released by malignant tissues and that levels of ethanolamine are elevated in malignant tissues. Indices describing the choline-containing phospholipids showed that these lipids are depressed significantly in malignant tissue relative to healthy tissue.


Epidermal growth factor receptors were measured using a radioligand binding assay in membrane preparations from 67 cancer and 25 non-malignant tissues. The binding characteristics of EGFr were similar in tumour and normal breast membranes. The concentrations were significantly higher in non-malignant tissue than in cancer. EGFr concentrations were directly correlated with steroid receptors in non-malignant tissue, whereas in cancer an inverse correlation between EGFr and steroid receptors was found.


There is paucity of literature describing complex elbow trauma in the pediatric population. We described a case of an uncommon pediatric elbow injury comprised of lateral condyle fracture associated with posterolateral dislocation of elbow.A 12-year-old boy sustained a direct elbow trauma and presented with Milch type II lateral condyle fracture associated with posterolateral dislocation of elbow. Elbow dislocation was managed by closed reduction. The elbow stability was assessed under general anaesthesia, followed by open K-wiring for the lateral condylar fracture fixation. The patient had an uneventful recovery with an excellent outcome at 39 months follow-up.Complex pediatric elbow injuries are quite unusual to encounter, the management of such fractures can be technically demanding. Concomitant elbow dislocation should be managed by closed reduction followed by open reduction and internal fixation (K-wires or cannulated screws) of the lateral condyle fracture. Traumatic elbow dislocation is a rare injury in children constituting 3–6% of all elbow injuries . It moreThere is limited evidence available in the literature describing complex elbow fracture dislocation in the pediatric population -7. This A 12-year-old boy presented with a grossly swollen and deformed left elbow after sustaining a fall off skateboard and directly landed on the elbow. There was no neurovascular deficit in the extremity. The anteroposterior and lateral radiographs showed posterolateral dislocation of the left elbow in association with Milch type II lateral condyle fracture  declare that they have no competing interests.Each author has equally contributed. HS collected the data and written up the manuscript, RA helped in scrutiny of the paper and obtaining illustrations. GRT had the idea and granted permission to use his patient data for preparing the manuscript.The pre-publication history for this paper can be accessed here:


Seventy-four patients previously treated in our department for germ cell tumour of the testis underwent a series of tests to determine the frequency of long-term therapeutic complications. All had received cisplatin-based chemotherapy as part of their treatment. There was a significant deterioration in renal function throughout the group. Eighteen (24%) had supine blood pressure greater than systolic 140 mmHg or diastolic 90 mmHg after treatment but hypertension did not correlate with renal impairment. Raynaud's phenomenon was common after chemotherapy (26/74) as was persistent sensory neuropathy (23/74). Although 34% had testosterone levels below the normal range, only six patients had a low free testosterone index with one testis still in situ; 18 patients have fathered children after chemotherapy. Approximately half of the patients completed a psychosexual questionnaire and some 30% of them admitted to sexual problems which they attributed to their treatment. Long-term sequelae of cisplatin-based chemotherapy for testicular malignancy are frequent and persistent, and follow-up of these patients should include prospective measurement of changes in blood pressure.


However, multiple rounds of amplification are required when assaying very small amounts of starting RNA. Moreover, certain cRNA-DNA mismatches are more stable than the analogous cDNA-DNA mismatches and this might increase non-specific hybridization. We sought to determine whether a recently developed linear isothermal amplification method (ribo-SPIA) that produces single stranded cDNA would offer advantages over traditional IVT-based methods for microarray-based analyses of transcript expression.The most widely used amplification method for microarray analysis of gene expression uses T7 RNA polymerase-driven A single round of ribo-SPIA amplification produced sufficient sscDNA for hybridizations when as little as 5 ng of starting total RNA was used. Comparisons of probe set signal intensities obtained from replicate amplifications showed consistently high correlations (r = 0.99). We compared gene expression in two different human RNA samples using ribo-SPIA. Compared with one round IVT, ribo-SPIA had a larger dynamic range and correlated better with quantitative PCR results even though we used 1000-fold less starting RNA. The improved dynamic range was associated with decreases in hybridization to mismatch control probes.The use of amplified sscDNA may offer substantial advantages over IVT-based amplification methods, especially when very limited amounts of starting RNA are available. The use of sscDNA targets instead of cRNA targets appears to improve hybridization specificity. PCR on [IVT] ,14, but on [IVT] .A new single primer, isothermal linear amplification method (ribo-SPIA) has been specifically developed for amplification of very small samples for use on DNA microarrays ,17. WithWe used the ribo-SPIA method to amplify several different total RNA samples. RNA was obtained from a pool of human tissues , mouse liver (Experiment 2), a second pool of human cells , and K562 human erythroleukemia cells . The amount of starting total RNA ranged from 5–100 ng and yields were in the range of ~6–12 μg of sscDNA . sscDNAs were fragmented in preparation for hybridization resulting in fragments of ~50–200 bp. These results are similar to those previously obtained using this method ,17.Each of the experiments included replicate amplifications . We hybridized each replicate to a separate microarray and calculated intensities for each probe set. Fig. 2 [mean K562 intensity/mean sUHR intensity]) and average signal intensity . M and A values obtained using sscDNA are shown in Fig. 7-fold higher in sUHR than K562 cell RNA (M < -7), but no differences of this magnitude were identified using cRNA. The number of probe sets associated with greater than 2-fold differences in expression (|M| > 1) was 1518 for sscDNA and 1043 for cRNA. 51% of the genes with >2-fold differences in expression on sscDNA arrays were not detected as >2-fold on cRNA arrays, but only 25% of the genes that were >2-fold different on cRNAs were not detected as >2-fold different on sscDNA arrays  primers for the synthesis of first strand cDNA, so this explanation is less likely.The new observation that sscDNA gave a wider range of relative expression (M) values despite lower average intensity (A) values could be explained by improved hybridization specificity under the conditions used in this study. This is plausible because the binding energy for DNA-DNA interactions is more sensitive to base pair mismatching than the binding energy for DNA-RNA interactions ,27. To lWe wished to compare how measurements made using amplified sscDNA and microarrays compared with measurements made using other approaches. We began by comparing results obtained using sscDNA and cRNA microarray hybridizations for all 12,625 probe sets on the arrays. Since the sscDNA and cRNA methods would be expected to introduce different systematic biases, we were not surprised that direct correlations between signal intensities obtained with the two different methods showed show relatively poor agreement . The finding indicates that it will not be useful to directly compare one array hybridized with sscDNA to another one hybridized with cRNA.We next compared differential gene expression measures  determined using sscDNA with those determined using cRNA. There was a clear correlation /(sUHR copy number), that could be directly compared to M values from arrays. 37 putative duplicate probe sets from 17 genes probe sets were hand-curated to confirm that they would correspond to the predicted qPCR product. In two cases probe sets were found to be misidentified in the GeneChip annotation and were removed from the analysis. In the remaining cases of duplication, the qPCR and microarray values were averaged across the duplicates. The final set of 106 curated genes and the associated data can be found at .Next we generated another set of expression measurements that could be used as a basis for comparison for the sscDNA and cRNA array results. qPCR is typically used as "gold standard" to confirm putative differentially expressed genes detected with microarrays. Since we saw a subset of genes for which expression differed between sscDNA and cRNA targets, we next assessed if either method tracked more closely to qPCR. We chose qPCR primers and probe sets from a large group of >1000 sets that have been developed for various studies. From these, primers and probes for four subsets of genes were selected for qPCR. The first set included all genes with >4 fold difference in expression between K562 and sUHR samples as determined using the sscDNA method, the cRNA method, or both methods (53 primer/probe sets). The second set included all other genes in which the two methods disagreed by more than 2-fold (29 primer probe sets). The third set consisted of a group of 33 empirically-derived 'housekeeping genes.' These were all genes that were nearly equally expressed (|M| < 0.1) in K562 and sUHR samples according to both the sscDNA and cRNA methods and gave strong signals (A > 5 for both methods). The fourth set included 8 housekeeping genes that had been previously validated as controls for qPCR in other experiments. We determined the gene copy number for each qPCR primer and probe set and then calculated a measure of relative expression, M = logThere were clear correlations between qPCR M values and array M values obtained using sscDNA Fig.  or cRNA We examined the suitability of a new isothermal linear amplification method for application to Affymetrix GeneChip microarrays. We performed a series of tests using starting amounts of RNA ranging from 5 to 100 ng for amplification yield and reproducibility. The amplification reactions consistently produced sufficient sscDNA for multiple array hybridizations. Pairwise comparison of technical replicates hybridized to microarrays by regression analysis showed excellent consistency. When we used sscDNA to analyze differential gene expression between two samples, we found a larger dynamic range than that obtained with cRNA hybridizations. The improved performance appears to be related to increased sscDNA hybridization specificity. The data obtained using this new method also more closely matched the results from qRT-PCR than data obtained using standard IVT reactions, even though the amount of starting RNA used was 1000-fold less. This new amplification method is a useful alternative approach for preparing targets that is especially well-suited for experiments involving small amounts of starting material.Clontech Human Universal Reference Pool total RNA (cUHR), derived by pooling RNA from a variety of human tissues, was purchased from BD Biosciences and used in Experiment 1. Mouse total liver RNA was isolated by standard methods from C57/BL6 mice according to procedures approved by the UCSF Committee on Animal Research and used in Experiment 2. For Experiment 3, we used Stratagene Human Universal Reference Pool and K562 erythroleukemia total RNAs from the same batches used in a previous study . All samsscDNA samples were prepared using the NuGEN Technologies Ovation RNA amplification and Biotin Labeling system (Version 1.0) according to the manufacturer's directions from the indicated amount of starting RNA (5–100 ng). All reactions were performed in 0.2 ml strip PCR tubes in an MJ GeneWorks PTC-100 thermocycler using recommended programs. Since the seal for PCR tubes and caps tends to deteriorate with repeated use, we replaced the caps for each tube before each resealing step in the protocol. Following amplification, sscDNA product was purified using QIAquick PCR purification kits (Qiagen). Samples were fragmented and end labeled with biotin. After stopping, each reaction was concentrated in a Microcon YM-3 column to a final volume of ~20 μl. The concentrated material was purified using a Centri-Sep 100 spin column (Princeton Separations). Negative control reactions were prepared by replacing input RNA with the appropriate volume of RNase free water.. K562 and sUHR microarray data were also analyzed using Microarray Suite 4.0 in order to calculate PM-MM values for each transcript probe set. Probe level analyses were performed using the BioConductor [All samples were placed in standard Affymetrix hybridization buffer. The sample denaturation time for the sscDNA samples was reduced from 5 to 2 minutes and hybridization time increased from 16 to 20 hours as recommended by NuGEN Technologies. cUHR gene expression was assayed using Affymetrix Human Genome U133A GeneChip arrays (Experiment 1). Mouse liver RNA was assayed using Murine Genome Mu6500A arrays (Experiment 2). K562 and sUHR RNAs were assayed using Human Genome U95Av2 arrays (Experiment 3). One template independent sample was also analyzed using a Human Genome U95Av2 array (Experiment 4). Arrays were stained with phycoerythrin-streptavidin according to the manufacturer's instructions. Metrics for all sample hybridizations including scaling factors, mean background intensities, and percent present calls have been provided  PCR was used to measure the expression of selected genes in sUHR and K562 cells. Gene-specific primers for multiplex real time RT-PCR were designed for each gene of interest using "Primer Express" software (Perkin-Elmer) and purchased from Biosearch Technologies. Sequence data for all oligonucleotides primers has been provided (see Additional File CSB conceived of the study, participated in the design and coordination, and wrote the manuscript. CG participated in the design of the study and both CG and KH performed the microarray experiments. GMD carried out the real time PCR studies. JYHY performed the statistical analyses. DJE participated in the study design and analysis and revised the manuscript. All authors read and approved the final manuscript.Hybridization Metrics is a .txt file suitable for opening in Microsoft Excel containing information about each individual hybridization.Click here for fileTaqman Primers provides sequence information for oligonucleotides used for qRT-PCR.Click here for file


The effects of shock waves generated by an experimental Siemens lithotripter in combination with cytostatic drugs or cytokines on several bladder cancer cell lines were examined in vitro. Proliferation after treatment was determined with the 3-4,5-dimethylthiazol-2,5 diphenyl tetrazolium bromide assay. Dose enhancement ratios were calculated for each drug and each shock wave application mode in order to characterise the sensitising effect of shock wave pretreatment. The influence of the time between shock wave and drug treatment as well as the effects of different sequences of shock wave and drug treatment or concomitant treatment were assessed for selected combinations of cell lines and drugs. It was found that shock wave treatment could render certain cell lines more susceptible to subsequent cis-platinum, mitomycin C or actinomycin D incubation. Cell lines sensitive to tumour necrosis factor alpha or interferon alpha were further sensitised to these cytokines by shock wave pretreatment. The enhanced sensitivity to cis-platinum and actinomycin D decreased rapidly during the first hours after shock wave treatment. The antiproliferative effect was most pronounced after concomitant shock wave and drug treatment. The sensitisation to interferon alpha diminishes more slowly after shock wave exposure. From the results presented in this study it is concluded that transient shock wave-induced permeabilisation of cell membrane not only enhances drug efficiency, but also causes damage to cell organelles and alterations in cellular metabolism.


Thrombolysis with recombinant tissue plasminogen activator (rtPA) is an established treatment in acute stroke. To prevent rethrombosis after rtPA therapy, secondary anticoagulation with heparin is commonly performed. However, the recommended time-point and extent of heparin treatment vary and are not well investigated.We report a 61-year-old man who developed an acute global aphasia and right-sided hemiparesis. Cranial CT was normal and systemic thrombolytic therapy with tPA was started 120 minutes after symptom onset. Low-dose subcutaneous heparin treatment was initiated 24 hours later. Transthoracic echocardiography (TTE) 12 hours after admission showed slightly reduced left ventricular ejection fraction (LVEF) but was otherwise normal. 48 hours later the patient suddenly deteriorated with clinical signs of dyspnea and tachycardia. TTE revelead a large left ventricular apical thrombus as well as a reduction of LVEF to 20 %. Serial further TTE investigations demonstrated a complete resolution of the thrombus and normalisation of LVEF within two days.Our case demonstrates an intracardiac thrombus formation following rtPA treatment of acute stroke, probably caused by secondary hypercoagulability. Rethrombosis or new thrombus formation might be an underestimated complication of rtPA therapy and potentially explain cases of secondary stroke progression. Recombinant tissue plasminogen activator (rtPA) is an approved treatment option for acute ischemic stroke within three hours of symptoms onset. It is well known that reperfusion of ischaemic brain tissue, when performed in a timely manner, improves clinical outcome ,2. HowevWe present a patient with acute ischaemic stroke who developed a transient left ventricular apical thrombus after systemic thrombolysis.A 61-year-old man was admitted to the Emergency Department immediately after he experienced global aphasia and right-sided hemiparesis. The clinical diagnosis of acute cerebral ischaemia in the middle cerebral artery (MCA) territory was made by a neurologist. A cranial CT-scan showed no early signs of cerebral infarct and no intracranial bleeding. Atrial fibrillation was recognised on the initial ECG and an cerebral infarct of cardiac embolic origin was assumed. The patient had no history of stroke but suffered from coronary heart disease and sustained an anterior cardiac wall infarction in the past. Further known vascular risk factors were hypertension and smoking. Even so, the patient took no medication at the time of admission.Thrombolysis with 70 mg rt-PA (0.9 mg/kg) was started two hours after the onset of symptoms. The neurological status during and after rt-PA treatment remained unchanged. Transthoracic echocardiography (TTE) performed 12 hours after initiation of thrombolytic therapy revealed a moderately reduced left ventricular systolic function (ejection fraction: 41%), left ventricular regional dyssynergy, and no intracardiac thrombus.The D-Dimer antigen plasma concentration as a marker of coagulation activation was raised 24 hours after rt-PA treatment and showed a tendency to normalisation during the following days.Extracranial Duplexsonography revealed moderate arteriosclerotic plaque formation in the carotid bulb on both sides but no hemodynamic stenosis of the carotid and vertebral arteries. Using transcranial duplexsonography a severe stenosis was found in the proximal segment of the left middle cerebral artery. No diabetes or hyperlipidaemia were diagnosed. Cerebral MRI, twenty-four hours after lysis revealed a cortical infarct with a clinically asymptomatic haemorrhagic transformation in the left-sided MCA territory. Antithrombotic treatment with subcutaneous low-dose heparin  was initiated 24 hours after thrombolysis. The fluid balance was continuously monitored and kept net 0 ml/24 hours to avoid fluid overload.rd day after admission, the patient developed acute dyspnea, tachypnea, tachycardia, and an elevated blood pressure. Acute myocardial infarction was excluded on the basis of ECG and blood tests . A CT-scan detected no pulmonary embolism, but massive bilateral pleural effusion. TTE revealed an apical thrombus measuring 2.0 × 2.0 cm in the left ventricle (figure On the 3e figure  with lefSystemic rt-PA thrombolysis in acute stroke has been implemented into daily clinical practice during the last decade. The treatment within three hours of stroke appears to be effective in reducing the neurological deficit ,7. HowevSome authors have reported a significantly higher incidence of parenchymal haematomas if thrombolysis was immediately followed by intravenous or subcutaneous heparin administration . Howeverrt-PA has a short biological half-life of 8–12 minutes but alters the physiological balance between coagulation and anticoagulation for a longer time period . After dOtherwise, clinical studies of fibrinolytic therapy in myocardial infarction show, that early heparin treatment starting immediately after thrombolysis significantly decreases the risk of cardiac vessel reocclusion . The capIn our patient, treated with low dose heparin, an apical thrombus developed within 72 hours after thrombolytic therapy, strongly suggesting a causal relationship of secondary hypercoagulability and thrombus formation. The patient's history and risk profile suggests an increased risk for cardiac events . We postCoronary angiography was not performed, considering the cerebral state of the patient and the observation that repeated ECGs were normal and cardiac enzymes not elevated. The clinical status stabilised spontaneously and the cardiac thrombus resolved within two days without initiation of a specific therapy, further supporting the hypothesis of a temporary status of hypercoagulation after thrombolysis.rd day and not directy after stroke which makes the diagnosis of neurogenic stunned myocardium further unlikely [Another hypothesis for the temporary cardiac detoriation in our patient could be an acute neurogenic stunned myocardium. This phenomenon is described as sudden, reversible left ventricular dysfunction with abnormal left ventricular wall motion and reduced ejection fraction. Levels of creatine kinase MB and troponin may be elevated and ECG alterations as depression or elevation of the ST segment or T wave inversion can be observed. However, acute neurogenic myocardial stunning has so far only been reported after subarachnoid hemorrhage -20 and iunlikely ,20,21.In summary, our case demonstrates an intracardiac thrombus formation following rtPA treatment of acute stroke, probably caused by secondary hypercoagulability. Rethrombosis or new thrombus formation might be an underestimated complication of rtPA therapy and potentially explain cases of secondary stroke progression. Early systemic anticoagulation with heparin might reduce the risk of rethrombosis but also increase the risk of a bleeding complication. Systematic studies concerning the incidence of thrombus formation after rtPA therapy and the effects of different post-thrombolysis anticoagulation strategies are required to assess the clinical relevance of the discussed secondary hypercoagulability. Closed echocardiographic monitoring in stroke patients treated with systemic thrombolysis might be useful for early detection of the described potential cardiac complications especially because repeated measurement of ECG and cardiac enzymes alone might fail.


Moisture content in commercially available milk powder was investigated using near infrared (NIR) diffuse reflectance spectroscopy with an Indian low-cost dispersive NIR spectrophotometer. Different packets of milk powder of the same batch were procured from the market. Forty-five samples with moisture range 4–10% were prepared in the laboratory. Spectra of the samples were collected in the wavelength region 800–2500 nm. Moisture values of all the samples were simultaneously determined by Karl Fischer (KF) titration. These KF values were used as reference for developing calibration model using partial least squares regression (PLSR) method. The calibration and validation statistics are


The mechanism of tumour necrosis photosensitised by liposome-delivered Zn(II) phthalocyanine (Zn-Pc) has been studied in mice bearing a transplanted MS-2 fibrosarcoma. Ultrastructural analyses of tumour specimens obtained at different times after red light-irradiation  indicate an early (3 h) photodamage of malignant cells especially at the level of the mitochondria and rough endoplasmic reticulum. The cellular damage becomes more evident between 6 h and 15 h after photodynamic therapy. On the other hand, the capillaries supplying the tumour tissue are modified at a much slower rate and appear to be severely damaged only after 15 h from irradiation, when the whole tissue becomes necrotic. Occasionally, mildly damaged capillaries are observed even at 72 h after irradiation. These findings support the hypothesis that low density lipoproteins (LDL) play a major role in the delivery of Zn-Pc to the tumour tissue; the photosensitiser is released specifically to malignant cells as a consequence of a receptor-mediated endocytosis of LDL.


Mediastinal pancreatic pseudocyst is a rare complication of acute or chronic pancreatitis.This case report describes the management of a difficult case of pancreatic pseudocyst with a mediastinal extension in a patient having chronic pancreatitis. Different management strategies were used until complete resolution of this complex pseudocyst occurred using open surgical cystogastrostomy.Despite the availablity of different minimally invasive techniques to treat pancreatic pseudocysts, management of complex mediastinal pseudocyst may still require open surgical drainage procedures. Mediastinal pancreatic pseudocyst is a rare complication of acute or chronic pancreatitis. Since its first description in 1951, approximately 50 cases have been reported in the world literature. We report a case of mediastinal pseudocyst associated with alcoholic pancreatitis successfully treated with cystogastrostomy.A 22-year-old female patient with a known history of chronic alcohol abuse was referred to hepato-biliary unit with recurrent upper abdominal related to chronic pancreatitis. Prior to this she has had multiple hospital admissions in previous one year with similar symptoms. In the past, ultrasonography had excluded gallstones and a CT scan 12 months ago had shown normal-looking pancreas. Her most notable symptomatic episode was 4 months ago when she was admitted with left lateral chest pain and dyspnoea. A CT-scan although excluded a pulmonary embolism but incidentally revealed a left gastric artery pseudoaneurysm  declare that they have no competing interests.All the authors have been involved in literature search, writing and final reviewing of this manuscript.


The renal function was evaluated with 131I-Hippuran clearance in 171 patients with malignant germ cell tumours. Assessments were performed before treatment and at three fixed times afterwards within 5 years. The patients were treated with surgery only (20 patients), infra-diaphragmatic radiotherapy only (median midplane dose 36 Gy) (48 patients), cisplatin-based chemotherapy  plus surgery (64 patients), cisplatin-based chemotherapy  with or without surgery (23 patients) or cisplatin-based chemotherapy  plus infra-diaphragmatic radiotherapy (16 patients). No renal impairment was observed for patients treated with surgery only. In patients who received radiotherapy no change of the renal function occurred during the first year post-treatment. Three to five years after treatment discontinuation a statistically significant reduction within the normal range was observed in patients who were greater than 40 years at the time of irradiation. Cisplatin-based chemotherapy led to a statistically significant irreversible renal impairment for all the three groups. The greatest reduction was seen in patients who received the highest total cisplatin dose or who were treated with irradiation in addition to chemotherapy. The clinical significance of the observed nephrotoxicity is still unknown.


The successful treatment of paroxysmal supraventricular tachycardia with adenosine 5` triphosphate (ATP) was initially reported by Somlo  in 1955.Non-invasive diagnosis of dual AV node physiology (DAVNP) in patients with AV nodal reentrant tachycardia by administration of ATP or adenosine has been reported by different authors ,5. ThougIn patients with palpitations of unclear etiology, ATP test identifies those who are likely to have AVNRT or AVRT  with a high positive predictive value . ATP/ade


Left ventricular free wall rupture after myocardial infarction has a high mortality. Suturing techniques of repair may be technically difficult and require cardiopulmonary bypass. We report a case of left ventricular rupture in a 47 year old man managed off pump employing a sutureless technique with Gelatine-Resorcin-Formalin glue and bovine pericardial patches. Left ventricular free wall rupture post myocardial infarction has a high mortality and therefore, rarely presents to the cardiac surgeon. Conventional approaches to this condition include ventricular repair with teflon buttressed sutures using cardiopulmonary bypass (CPB). Coronary artery bypass grafting may be performed concomitantly. Some authors have suggested sutureless techniques with or without cardiopulmonary bypass for this condition. We report a case of off-pump repair with bovine pericardial patches using Gelatine-Resorcin-Formalin (GRF) glue.A 47 year old man presented to the Emergency department with central chest pain. Lateral myocardial infarction (MI) was confirmed by typical changes in leads I, AVL and V6 and raised troponin T levels. The patient was not thombolysed. He was haemodynamically stable initially and a coronary angiogram was contemplated. However, before this could be performed, he developed further chest pain and became haemodynamically unstable with tachycardia, hypotension and collapse (at 48 hours). An urgent trans- thoracic echocardiogram (TTE) showed a pericardial effusion. Left ventricular (LV) free wall rupture was suspected. Contrast-enhanced computed tomography (CT) of the chest was performed which confirmed this, as well as a large pericardial effusion figure . He was Myocardial infarction complicated by left ventricular free wall rupture and pericardial tamponade occurs in 2–4% of cases and is associated with a high mortality . AutopsyTraditional left ventricular free wall rupture repair techniques such as a series of interrupted pledgeted sutures, reinforced linear teflon strips and circular patch repair often involve tying sutures though friable necrotic muscle . HeparinMariani et al  describeThe traditional approach is to proceed with infarctectomy and replacement with a prosthetic patch under CPB . Prior tBasically, there are two types of glue: synthetic (most commonly cyanoacrylates) and biological . Degradation of the cyanoacrylates can lead to tissue toxicity. The shorter-chain compounds (butyl) tend to have a higher degree of tissue toxicity than the longer-chain derivatives (octyl). These adhesives work by polmerising after which the adhesive may become brittle and is subject to fracturing. This is especially true in areas of high tension. These limitations of synthetic glues lead to the development of biological glues. The GRF glue should be used as a tissue reinforcer, the two-component fibrin sealer is preferable when haemostatic action must be accompanied with mechanical barrier and the cryoprecipitate glue can be used when haemostatic action is the only requirement.It is not known whether the benefit of CABG is worth the inherent risk involved in delaying definitive surgical therapy by performing coronary angiogram. In a review of literature, only 9 of the 87 long term survivors of free wall rupture repair had CABG as part of their surgical intervention [On the basis of our case report and the current literature we can say that left ventricular free wall rupture after myocardial infarction can potentially be repaired without the use of extra-corporeal circulation using the sutureless technique with glue and pericardial patch. Gluing is an alternative to suturing in such a desperate situation.


Fifty-two patients with epithelial ovarian cancer were treated with yttrium-90-labelled monoclonal antibody HMFG1 administered intraperitoneally following conventional surgery and chemotherapy as part of an extended phase I-II trial. The treatment was well tolerated and the only significant toxicity observed was reversible myelosuppression as previously described. Following conventional surgery and chemotherapy, 21 out of the 52 patients had no evidence of residual disease and were regarded as receiving treatment in an adjuvant setting. To date, two of these patients have died of their disease . This extended phase I-II study suggests that patients with advanced ovarian cancer who achieve a complete remission following conventional therapy may benefit from further treatment with intraperitoneal radioactive monoclonal antibody.


Recent studies have demonstrated a consistent inverse relationship between oestrogen receptor (ER) and epidermal growth factor receptor (EGFR) levels in female human breast cancer. Serial cross-section studies have suggested that separate populations of ER+/EGFR- and ER-/EGFR+ cancer cells exist in tumours deemed by immunocytochemical assay (ICA) to be positive for both. We have developed a dual ICA that is able to stain for both ER and EGFR on a single 5 microns frozen section sample of breast tissue. Twenty-two samples of female human breast cancer tissue that exhibited positivity for ER and EGFR by ER-ICA using the H222 monoclonal antibody and EGFR-ICA using the EGFR1 monoclonal antibody underwent the dual ICA. There was a significant correlation in receptor positivity between the single and dual assays for both ER  and EGFR . Individual cancer cells exhibited one of three staining patterns: nuclear staining only (ER+/EGFR-), membrane-associated and cytoplasmic staining only (ER-/EGFR+) or no staining (ER-/EGFR-). No cancer cells exhibited both nuclear and membrane/cytoplasmic staining. This is the first description of a simultaneous dual immunocytochemical assay system for ER and EGFR in clinical breast cancer specimens. The results suggest that ER and EGFR expression are mutually exclusive within an individual breast cancer cell in vivo with separate populations of ER+/EGFR- cells, ER-/EGFR+ cells and ER-/EGFR- cells coexisting.


Incisional sustained tachycardias are frequent in patients who have undergone a surgical repair of interatrial defect. A 43-year-old woman with drug refractory, highly symptomatic, persistent atrial tachycardia in the last year, was referred to our unit for catheter ablation. The patient had undergone a cardiac operation for repairing interatrial secundum ostium type defect with a patch five years before. A previous radiofrequency ablation procedure had been performed for common atrial flutter. We describe a case of incisional  atrial tachycardia ablation guided by the new EnSite NavX system equipped with a new electroanatomic mapping system. Incisional sustained tachycardias are frequent in patients who have undergone surgical repair of  an interatrial defects ,2. The mA 43-year-old woman with drug refractory, highly symptomatic, persistent atrial tachycardia in the last year, was referred to our unit for catheter ablation. Five years ago the patient was undergone  a cardiac operation for repairing an interatrial secundum ostium type defect. ECG showed an atrial tachycardia whose features resembled a common atrial flutter. Echocardiography images showed no  significant atrial dilation, no residual interatrial shunt and a normal ventricular systolic function.A quadripolar Josephson diagnostic catheter was inserted into the coronary sinus via left subclavian vein as a reference. A bipolar Cournard diagnostic catheter was inserted into the right ventricular apex. A  20-pole steerable catheter was placed around the tricuspid annulus. An 8 mm tip catheter (Boston Scientific EP Technologies) was used for mapping and ablation. The electrophysiological findings showed a stable atrial tachycardia of 230 ms cycle length which was firstly misunderstood as a common atrial flutter (counter clockwise) showing a caudocranial septal activation and craniocaudal activation along the right lateral atrial wall. The ablation catheter was therefore positioned on the cavo-tricuspid (CT) isthmus, considering this region crucial for the macroreentry circuit. But overdrive pacing at 210 ms cycle length from CT isthmus did not show any concealed entrainment. So we were inclined to believe that it was an incisional tachycardia and therefore we performed a reconstruction of the three-dimensional geometry of the right atrium  using the EnSite NavX system.An activation-voltage map was obtained moving the ablation catheter point by point inside the right atrium while the patient was still in stable atrial tachycardia. The voltage map showed a large no-signal area  on interatrial septum corresponding to a large patch used for repairing an interatrial defect . A largeRF application  in continuous way (70 Watts 55 °C) was then attempted along this isthmus drawing back the ablation catheter from the scar area to the tricuspid valve annulus, trying to connect the two areas of anatomic obstacles. We moved the ablation catheter along this line obtaining local electrogram disappearance or  voltage reduction more than 70%. EnSite NavX mapping system allowed us to move ablation catheter in  the right atrium without using fluoroscopy  during the procedure.After four RF applications along the isthmus between the patch on the atrial septum and the tricuspid valve annulus, atrial tachycardia was not yet interrupted though no significant local electrogram was still detectable along the ablation line we had performed. In order to verify ablation line continuity we performed an electroanatomic and activation remapping of the isthmus region which showed a gap on the ablation line  near theThe patient was followed up monthly after the procedure. Each month history, 12-lead ECG and 24 hour ECG Holter were analyzed. Ten months after the procedure though the patient had not taken any anti-arrhythmic drugs, there was no recurrence of palpitation or documented atrial tachycardia.The case shows that EnSite NavX system provides an excellent electroanatomic and activation map of any cardiac chamber and reliable monitoring of the ablation catheter. This system let us quickly made a diagnosis of incisional tachycardia with an anatomic circuit of macroreentry different from that of a common atrial flutter allowing us to identify a slow conduction isthmus in a place that was not the CT isthmus, though the electrocardiography feature of the atrial tachycardia was similar to a common atrial flutter. The Ensite NavX system rapidly identified a gap in the linear lesion we had performed, and allowed us to terminate the atrial tachycardia. NavX significantly reduces fluoroscopy time during ablation procedures and also allows to successfully perform difficult procedures with complex substrates. NavX supports detection of anatomic isthmus variations, particularly deep pouches or recesses ,14.


Echocardiography plays an integral role in the diagnosis of congestive heart failure including measurement of left heart pressure as well as mechanical dyssynchrony.In this report we describe novel therapeutic uses of echo pulsed wave Doppler in atrioventricular pacemaker optimization in patients who had either not derived significant symptomatic benefit post biventricular pacemaker implantation or deteriorated after deriving initial benefit. In these patients atrioventricular optimization showed novel findings and improved cardiac output and symptoms.In 3 patients with Cheyne Stokes pattern of respiration echo Doppler showed worsening of mitral regurgitation during hyperpneac phase in one patient, marked E and A fusion in another patient and exaggerated ventricular interdependence in a third patient thus highlighting mechanisms of adverse effects of Cheyne Stokes respiration in patients with heart failure. All 3 patients required a very short atrioventricular delay programming for best cardiac output. In one patient with recurrent congestive heart failure post cardiac resynchronization, mitral inflow pulse wave Doppler showed no A wave until a sensed atrioventricular delay of 190 ms was reached and showed progressive improvement in mitral inflow pattern until an atrioventricular delay of 290 ms. In 2 patients atrioventricular delay as short as 50 ms was required to allow E and A separation and prevent diastolic mitral regurgitation. All patients developed marked improvement in congestive heart failure symptoms post echo-guided biv pacemaker optimization.These findings highlight the value of echo-guided pacemaker optimization in symptomatic patients post cardiac resynchronization treatment. Echocardiography has become the gold standard for non invasive assessment of diastolic function . BesidesA 51 year old African American obese male with a history of non-ischemic dilated cardiomyopathy improved CHF symptoms from NYHA class III to class II after biv pacemaker implantation. Patient developed recurrent CHF with NYHA class III symptoms and an LVEF of 20% 8 months post CRT. On interrogation there was normal sinus rhythm and pacemaker was programmed in the DDD mode, AV delay of 160 ms and a VV delay of 0 ms. Mitral inflow PW Doppler and respirogram revealed cyclic variation of heart rate from 56 to 94 beats per minute during Cheyne Stokes respiratory cycle. Post apneac hyperpnea was associated with tachycardia Figure  and hypoA 76 year old Caucasian female developed increasing shortness of breath and NYHA class III symptoms for 8 weeks. Patient had a history of RV pacemaker implantation for a complete heart block followed by development of CHF class II-III. RV pacemaker was upgraded to a biv pacemaker that led to improvement in symptoms for about 7 months. A-pacing at 60 bpm and paced and sensed AVD of 170 and 150 ms respectively and LV pre-excitation of 20 ms were programmed. Moderate to severe mitral regurgitation (MR) with onset in diastole was seen at these settings Figure  and 3B. A 72 year old African American female with ischemic cardiomyopthy and prior left cerebrovascular accident and right hemiparesis presented with CHF and NYHA class III symptoms 1 year after biV pacing. LVEF was 25%, there was mild MR and peak pulmonary artery systolic pressure was 50 mm Hg. Cheyne Stokes respiratory pattern was seen on respirogram. Hyperpnea was associated with decreased RV-RA gradient during each inspiration Figure  and 6B. A 73 Caucasian male was referred for bi-V pacemaker optimization 7 months post CRT for ischemic cardiomyopathy. Symptoms of CHF improved initially, however worsening shortness of breath and fatigue occurred 5 months later to NYHA class III symptoms. BNP was 1040 pg/ml. The device was programmed in the DDD mode, with sensed AV delay of 150 ms and paced AV delay of 160 ms. PW Doppler showed absent mitral inflow A waves at baseline pacemaker settings despite presence of sinus rhythm Figure . AV delaAn 83-year-old Caucasian male with a history of ischemic cardiomyopathy, CHF and NYHA class II symptoms underwent biv pacemaker implantation. Sensed AVD was 100 ms and RV-LV offset was 20 ms. Routine pre-discharge optimization revealed late diastolic MR Figure  at an AVA 67 year old Caucasian female underwent biv pacemaker implantation for non ischemic dilated cardiomyopathy, severe mechanical dyssynchrony, left bundle branch block, NYHA class III and LVEF of 35%. She was referred for persistent shortness of breath upon exertion post CRT. Programmed AVD of 130 ms showed marked E and A fusion Figure  and diasOur findings describe the utility of AV pacemaker optimization using PW echocardiography in optimizing diastolic mitral inflow in patients who remain symptomatic post CRT. Our report also highlights the mechanisms of adverse effects of Cheyne Stokes pattern of respiration on cardiac performance.Other investigators have reported the incremental improvement in cardiac function and functional class by echocardiographic guided pacemaker optimization in patients who undergo CRT -3. StudiShortening of AV delay has been shown to reduce diastolic MR in patients with right sided pacemaker , and cauCentral sleep apnea is common among patients with CHF, being present in 30–40% of patients . It contOurs is not a consecutive series of patients, rather a collection of patients in whom echo Doppler during AV optimization allowed novel observations that provide insight into the mechanism of failure of CRT and adverse effects of Cheyne Stokes respiration. A systemic study is needed to prospectively evaluate prevalence of Cheyne Stokes respiration in symptomatic patients post CRT and effect of AVD optimization in this group.We report 3 separate mechanisms whereby Cheyne Stokes respiration can contribute to worsening of heart failure symptoms in patients with CHF post CRT. These include compromised diastolic filling during hyperpneac phase of respiration, increase in diastolic mitral regurgitation during hyperpneac phase and development of a constrictive physiology with enhanced ventricular interdependence. A short AVD helped improve diastolic filling, reduce diastolic MR and reduce ventricular interdependence in all 3 patients. We also report the use of short AVD in abolishing diastolic MR in patients without Cheyne Stokes respiration as well as improvement in diastolic filling by an extraordinarily long AVD in a patient with heart failure post CRT. Our findings extend the use of echo Doppler in evaluation of patients who remain symptomatic post CRT as well as in determining optimal AVD.AVD = Atrioventricular DelayCHF = Congestive Heart FailureCRT = Cardiac Resynchronization TreatmentCPAP = Continuous Positive Airway PressureLVEF = Left Ventricular Ejection FractionLV = Left Ventricle (Ventricular)MR = Mitral RegurgitationNYHA = New York Heart AssociationPW = Pulsed waveRA = Right AtriumRV = Right Ventricle (Ventricular)VTI = Velocity Times IntegralThe author(s) declare that they have no competing interests.TZN conceptualized the design of the study, imaging methodology and interpreted data during acquisition. AMR analyzed echocardiographic data and helped to draft the manuscript. All authors read and approved the final manuscript.


Twenty seven patients with malignant lymphoma of lymphocytic type/B-chronic lymphocytic leukaemia  diagnosed from lymph node and splenectomy specimens were studied histologically and immunologically. Lymph node biopsies showed a diffuse effacement of normal architecture by small round lymphocytes usually with scattered proliferation centres (PC). A1 spleens showed white pulp with or without red pulp involvement, sometimes with tumour nodules present. PC-like cells or PC were only found in the white pulp or tumour nodules. Studies of 13 specimens using the ABC immunoperoxidase technique on frozen sections with a large panel of monoclonal antibodies showed that although a part of the monoclonal B cell neoplasm, the proliferation centres or splenic white pulp have a different phenotype from the surrounding cells. Some of these phenotypic changes are similar to those reported with in vitro induction of "maturation" of ML, L/B-CLL cells. The implications for normal B-cell development are discussed. In contrast to reported peripheral blood findings, T cells, predominantly of T helper phenotype in lymph nodes, were present but usually not numerous.


Natural and lymphokine activated killer cells (NK and LAK) are believed to play an important role in the control of tumour progression and metastasis. Their specific receptors on tumours cells are still unknown. Several studies suggest that these cells recognise and eliminate abnormal cells with deleted or reduced expression of MHC class I molecules. Previous reports suggest that interferons (IFN), by increasing MHC class I expression on target cells, induce resistance to killing by NK cells. We investigated the role of MHC molecule expression by two human breast cancer cell lines T47D and ZR75-1 in their susceptibility to NK and LAK cells. These two cell lines spontaneously express low levels of HLA class I antigens but no HLA class II molecules. After IFN-gamma treatment they both overexpressed MHC class I and de novo expressed class II molecules as detected by flow cytometry, quantified by a radioimmunoassay and analysed by two-dimensional gel electrophoresis. Opposed to untreated cells these IFN-gamma treated cells were resistant to NK and LAK lysis. Furthermore, preincubation of IFN-gamma treated breast cancer cells with F(ab')2 fragments of monoclonal antibodies to HLA class I and HLA class II molecules was unable to restore lysis. In contrast, several complete monoclonal antibodies including anti-HLA class I and HLA class II induced the lysis of target cells whether or not they had been treated by IFN-gamma. The therapeutic use of monoclonal antibodies directed against antigens expressed on tumour cells (ADCC) in conjunction with interferon therapy should be discussed in lymphokine-based strategies for treatment of cancer patients.


A genetically reshaped human IgG1 monoclonal antibody (Hu2PLAP) with anti-tumour specificity, was radiolabelled with Indium-111 by chelation with a new macrocyclic compound (DOTA) which allows the production of stable radioimmunoconjugates for in vivo application. This was used to image seven patients with malignant disease, of whom two had been previously exposed to mouse monoclonal antibodies and had developed human anti-mouse antibodies (HAMA). Successful tumour localisation was seen in the four patients with active disease and antigen positive tumours. No patient showed any antibody responses against Hu2PLAP, but three out of six patients tested showed an immune response against the macrocycle DOTA. Reshaped human monoclonal antibodies with anti-tumour specificity may facilitate repeated administrations of radioactive antibodies, thus allowing new possibilities, both in the diagnosis and treatment of cancer.


Conditioned media from 14 short term fibroblast cell lines were mitogenic for human breast cancer cells with different steroid receptor profiles in serum-free culture. Fibroblast-conditioned medium stimulated tritiated thymidine incorporation in short term culture and growth in a longer proliferation study as measured by the MTT colorimetric assay. Conditioned media from benign and malignant epithelial cells were non-stimulatory for breast cancer cells but that derived from endothelial cells showed similar stimulation to fibroblasts. Partial purification of fibroblast-conditioned medium identified a peptide with a molecular weight of approximately 8 kDa that showed no affinity for heparin and was mitogenic for MCF-7 breast cancer cells.


Cells with "hand mirror" morphology have not, to the best of our knowledge, been described in a primary effusion sample. This paper describes a case of T-cell lymphoma with eosinophilia in a patient with suspected peritoneal carcinomatosis. Rarely, a T-cell lymphoproliferative process may mimic primary peritoneal carcinomatosis, clinically suggested by a presentation in CT imaging of omental caking with bilateral massive loculated effusions in a patient without lymphadenopathy or splenomegaly.A 60 year old caucasian male presented with vague abdominal discomfort and increasing abdominal girth. Computed tomography showed a two centimeter thick omental cake and a small loculated effusion. The clinical presentation and imaging findings were most consistent with peritoneal carcinomatosis. Cytologic evaluation of the effusion was undertaken for diagnostic study.Rapid intraprocedural interpretation of the effusion sample showed a monomorphic population of cells with "hand-mirror" cell morphology exhibiting cytoplasmic extensions (uropodia) with 3–5 course dark cytoplasmic granules and a rim of vacuolated cytoplasm capping the opposing "mirror head" side. These cells were seen within a background of mature eosinophils. Flow cytometric evaluation of the ascites fluid demonstrated an atypical T-cell population with the following immunophenotype: CD2-, CD3+, CD4-, CD5-, CD7-, CD8+, CD56+. T-cell receptor (TCR) gene rearrangement was positive for clonal TCR-gamma gene rearrangement, supporting the diagnosis of a T-lymphoprolifereative disorder.A T-cell lymphoproliferative process may present with "hand mirror" morphology in an effusion sample. These cells may show polar cytoplasmic vacuolization and 3–5 course granules within the "handle" of these unique cells. Cytoplasm shows peripheral constriction around the nucleus. Cells with "hand mirror" morphology have not, to the best of our knowledge, been described in a primary effusion fluids . The celWhen a suspected lymphoproliferative process is evaluated during an immediate FNA interpretation specimen adequacy evaluation is critical. This can facilitate optimal sample handling for immunohistochemistry, flow cytometry, molecular analysis or chemical/enzymatic evaluation .The present case describes a loculated eosinophilic peritoneal effusion with hand-mirror cells (HMC) in association with peritoneal caking. Clinical suspicion based on CT imaging included peritoneal carcinomatosis. This ascites fluid also showed an increased relative proportion of unremarkable eosinophils. Eosinophilic effusions (defined as exceeding 5% eosinophils) have been associated with many etiologies including trauma, infection, peritoneal dialysis and malignancy . Both thThis 60 year old Caucasian male presented to his primary care physician with a chief complaint of vague abdominal pain and increasing abdominal girth of 3 months duration. His past medical history was essentially unremarkable.Peripheral blood showed mildly elevated WBC and platelet counts with mild relative lymphocytopenia and normal eosinophil counts. Carcinoembryonic antigen, Alpha-feto protein, CA15-3 and CA125 were all within normal ranges. Calcium was consistently mildly decreased during the course of hospital stay with slightly elevated prothrombin times.Initial imaging of abdomen by computed tomography (CT) demonstrated large volume ascites, fatty changes of liver, and omental-peritoneal "caking" measuring up to 2.0 cm in thickness Figure: . There wUltrasound guided aspiration of ascites fluid was obtained for routine cytology using a 22 gauge needle. Following the initial ultrasound guided paracentesis a CT guided Fine needle aspirate samples of omental cake was performed using an 18 gauge core biopsy. Initial aspirate samples were evaluated onsite for specimen adequacy by the cytopathologist. Additional samples were retained for ThinPrep, cell block, core Bx and microbiologic culture. The patient tolerated the procedure without difficulty. Rapid interpretation documented a sample adequate for cytomorphologic evaluation, special studies and microbiologic culture.Cytologic interpretation of the FNA for both the left flank omental mass and ascites specimen showed an atypical lymphoid population with prominent hand-mirror cell morphology and frequent eosinophils. This was felt to be strongly suspicious for a lymphoproliferative process, possibly T-Cell derived Figure: . The nucFlow cytometric and Immunohistochemistry revealed an atypical CD56+ T-Cell population suggestive of a T-Cell lymphoproliferative process. PCR analysis for TCR gamma gene rearrangement revealed two monoclonal peaks with TCR gamma primers, corresponding to V-gamma 11 gene rearrangement and V-gamma [This was a case of a 60 year old man with loculated peritoneal eosinophilic effusion and omental caking who was diagnosed with T-cell lymphoma by FNA. Cytologically distinct hand-mirror forms comprised the majority of the cells in the effusion. The finding of hand mirror cells in a peritoneal effusion has not, to the best of our knowledge, been described in the current literature. The patient declined treatments, entered hospice care and died within three months of the diagnosis.The term hand-mirror cell differentiation is descriptive. The cell is characterized by an asymmetrical cytoplasmic elongation extending out from one pole of the nucleus (uropod). Both the nucleus and the elongated cytoplasmic "handle" together give the appearance of a hand-mirror. The hand-mirror cell morphology had in the past been classified as a distinct morphologic variant of acute lymphoblastic leukemia ; howeverHand-mirror cells have also been reported with many diagnostic entities including cutaneous natural killer cell lymphomas , AnaplasThe distinct morphology most likely represents an aspect of immune responsivity or challenge involving the incorporation of immune complexes. AlthougThe CT findings in this case were associated with peritoneal involvement by lymphomatosis however the CT observation of omental "caking" has been identified as an unreliable means to distinguish peritoneal carcinomatosis or tuberculous peritonitis from lymphomatosis. Ascites without loculation or septation and diffuse distribution of enlarged lymph nodes may however help to differentiate peritoneal lymphomatosis from other eitiologies .This case also illustrates advantages of on-site specimen adequacy evaluation which proved essential in obtaining samples for appropriate special studies.Immunophenotypic, flow cytometric and molecular studies in this case documented a T-Cell lymphoma. It also provided an opportunity to offer a rapid interpretation which in this case did not support the provisional diagnosis of peritoneal carcinomatosis which had been based on CT observation of omental caking. The preponderance of hand mirror cells and the presence of an eosionophilia in the effusion were consistent with the final diagnosis.


Serial frozen sections from eleven patients with malignant breast tumours and five patients with benign disease were studied by indirect immunoperoxidase using a panel of mouse monoclonal antibodies to human leucocyte antigens. More infiltrating leucocytes were seen in tumour sections than those of benign conditions. A considerable proportion of the infiltrating cells were T cells, and more of these were of the suppressor/cytotoxic subset than the helper/inducer subset. The T cells were apparently not all activated as indicated by lower levels of staining with anti HLA-DR than anti-leucocyte antibody. Diffuse staining was sometimes seen with HLA-DR and T cell subset antibodies. Tumour cells did not stain or were only very weakly positive with anti HLA-A, B, C.