Adipocyte-derived collagen VI affects early mammary tumor progression in vivo, demonstrating a critical interaction in the tumor/stroma microenvironment

The interactions of transformed cells with the surrounding stromal cells are of importance for tumor progression and metastasis. The relevance of adipocyte-derived factors to breast cancer cell survival and growth is well established. However, it remains unknown which specific adipocyte-derived factors are most critical in this process. Collagen VI is abundantly expressed in adipocytes. Collagen(-/-) mice in the background of the mouse mammary tumor virus/polyoma virus middle T oncogene (MMTV-PyMT) mammary cancer model demonstrate dramatically reduced rates of early hyperplasia and primary tumor growth. Collagen VI promotes its growth-stimulatory and pro-survival effects in part by signaling through the NG2/chondroitin sulfate proteoglycan receptor expressed on the surface of malignant ductal epithelial cells to sequentially activate Akt and beta-catenin and stabilize cyclin D1. Levels of the carboxyterminal domain of collagen VIalpha3, a proteolytic product of the full-length molecule, are dramatically upregulated in murine and human breast cancer lesions. The same fragment exerts potent growth-stimulatory effects on MCF-7 cells in vitro. Therefore, adipocytes play a vital role in defining the ECM environment for normal and tumor-derived ductal epithelial cells and contribute significantly to tumor growth at early stages through secretion and processing of collagen VI.     

P33Transfusion of Incompatible Units to a Patient with Anti-U: a Case Report

Introduction  U (MNS5) is a high frequency Ag with the U- phenotype blood found mainly in people of Black African origin. Anti-U has been reported to cause immediate and delayed haemolytic transfusion reactions (HTRs) as well as haemolytic disease of the fetus and newborn (HDFN). The selection of U - RBC is recommended for patients with anti-U. We present a case of a patient with anti-U who received serologically incompatible transfusions uneventfully.
Case report  A 31-year-old black male was admitted to hospital after injury to his right arm. The antibody (Ab) screen was positive and samples were referred to our laboratory for investigation. He grouped as B, D+, U- and anti-U was identified. Due to the severity of the injury the patient underwent emergency surgery without waiting for the Ab ID results. His Hb was 68 g L⁻¹. 5 units of unmatched group O RBC were transfused. During this episode the patient was not given any IVIg cover and there were no clinical or laboratory indications of a transfusion reaction beyond the positive DAT. The patient received four further U- RBC transfusions and was treated with parenteral iron.
Results   

     

Anemia during and at discharge from intensive care: the impact of restrictive blood transfusion practice

Objective To document the prevalence of anemia among patients admitted to intensive care (ICU) and, among survivors, at ICU discharge when restrictive transfusion practice was used.Design This was an observational cohort study.Setting Ten of the 26 general ICUs in Scotland.Patients and participants One thousand twenty-three sequential ICU admissions over 100 days, representing 44% of all ICU admissions in Scotland during the study period, studied daily from admission to discharge or death in the ICU.Interventions None.Measurements and results The median transfusion trigger used, in the absence of bleeding, was 78 g/l (interquartile range 73–84); <2% of transfusion triggers were above the upper limit of the national transfusion trigger guideline (100 g/l). Overall, 25% of admissions had a hemoglobin concentration <90 g/l at ICU admission. Seven hundred sixty-six patients admitted survived to ICU discharge. Among these, the prevalence of anemia (male <130 g/l; female <115 g/l) at ICU discharge was 87.0 (95% CI: 83.6 to 89.9)% for males and 79.6 (74.8 to 83.7)% for females. Of the male survivors 24.1 (20.3 to 28.3)% and of the female 27.9 (23.4 to 33.2)% had a hemoglobin <90 g/l at ICU discharge. The prevalence was similar for patients with and without pre-existing ischemic heart disease. Logistic regression found independent associations between having a hemoglobin concentration <90 g/l at ICU discharge and the first measured hemoglobin in ICU, the presence of acute renal failure and thrombocytopenia during ICU stay.Conclusions Anemia is highly prevalent in ICUs that use restrictive transfusion triggers. The impact of anemia on functional recovery after intensive care requires investigation.Electronic Supplementary Material Supplementary material is available in the online version of this article at The authors represent the Audit of Transfusion in Intensive Care in Scotland (ATICS) study group. The ATICS Study Group was made up of the organizing committee: T.S. Walsh (Chairman), M. Garrioch, C. Maciver (study coordinator), F. McArdle (study coordinator), J. Kinsella, R. Lee (statistician), G. Fletcher, D.B. McClelland, R. Green, A. Todd and F. MacKirdy and of the participating units: Aberdeen Royal Infirmary (S.A. Stott, J.L. Scott, M.G.K. Strachan); Borders General Hospital (T. Cripps, S. Hogg, D. Hedderly, L. Hume, J. Playfair, I. Gourlay); Glasgow Royal Infirmary (J. Kinsella, M.G. Booth, T. McMillan); Ninewells Hospital Dundee (S.L. Crofts, I. Mellor, S.J. Cole); Royal Alexandra Hospital Paisley (G. Fletcher, K. McIlravey); Royal Infirmary, Edinburgh (T.S. Walsh, F. McArdle, S.J. Dodds); Southern General Hospital, Glasgow (M. Garrioch, J. Sandbach, B. McMillan); St Johns Hospital, Livingston (M. Hughes, M. MacRury, L.M.M. Morrison); Western General Hospital, Edinburgh (C. Wallis, C.G. Battison, C. Hardcastle, E.D. Fox); Western Infirmary, Glasgow (A.R. Binning, M. Pollock, S. Kelly); Scottish National Blood Transfusion Service (D.B. McClelland, R.H.A. Green, A.M.M. Todd, I. McKechnie, C.R. Maciver); Scottish Intensive Care Society Audit Group (F. MacKirdy); Clinical Audit Resource Centre, Western General Hospital, Edinburgh (M.L. Hughes); Medical Statistics Unit, Edinburgh University (R.J. Lee).     

Systemic armed oncolytic and immunologic therapy for cancer with JX-594, a targeted poxvirus expressing GM-CSF.

Targeted oncolytic viruses and immunostimulatory therapeutics are being developed as novel cancer treatment platforms. These approaches can be combined through the expression of immunostimulatory cytokines from targeted viruses, including adenoviruses and herpesviruses. Although intratumoral injection of such viruses has been associated with tumor growth inhibition, eradication of distant metastases was not reported. The major limitations for this approach to date have been (1) inefficient intravenous virus delivery to tumors and (2) the lack of predictive, immunocompetent preclinical models. To overcome these hurdles, we developed JX-594, a targeted, thymidine kinase(-) vaccinia virus expressing human GM-CSF (hGM-CSF), for intravenous (i.v.) delivery. We evaluated two immunocompetent liver tumor models: a rabbit model with reproducible, time-dependent metastases to the lungs and a carcinogen-induced rat liver cancer model. Intravenous JX-594 was well tolerated and had highly significant efficacy, including complete responses, against intrahepatic primary tumors in both models. In addition, whereas lung metastases developed in all control rabbits, none of the i.v. JX-594-treated rabbits developed detectable metastases. Tumor-specific virus replication and gene expression, systemically detectable levels of hGM-CSF, and tumor-infiltrating CTLs were also demonstrated. JX-594 holds promise as an i.v.-delivered, targeted virotherapeutic. These two tumor models hold promise for the optimization of this approach.     

Evaluation of a novel mouse model of intracisternal strychnine-induced trigeminal allodynia

Purpose

Intractable neuropathic dynamic allodynia remains one of the major symptoms of human trigeminal neuropathy and is commonly accepted to be the most excruciatingly painful condition known to humankind. At present, a validated animal model of this disorder is necessary for efficient and effective development of novel drug treatments. Intracisternal strychnine in rats has been shown to result in localized trigeminal dynamic allodynia, thus representing a possible model of trigeminal neuralgia. The purpose of this study was to validate a mouse model of trigeminal glycinergic inhibitory dysfunction using established positive (carbamazepine epoxide) and negative (morphine) controls.

Methods

The actions of conventional first-line treatment (carbamazepine epoxide [CBZe]) and clinically ineffective morphine were tested for trigeminal dynamic mechanical allodynia produced by intracisternal strychnine. In mice under halothane anesthesia, we injected either strychnine (0.3 μg), strychnine with CBZe (4 ng), or artificial cerebrospinal fluid (aCSF) intracisternally (i.c.). In a separate set of experiments, subcutaneous morphine (3 mg·kg^?1 sc) was injected with intracisternal strychnine. Dynamic mechanical allodynia was induced by stroking the fur with polyethylene (PE-10) tubing. The response of each mouse was rated to determine its allodynia score, and scores of each group were compared. In addition, in a separate dichotomous disequilibrium study, pairs of mice were injected with strychnine/saline, strychnine/strychnine-CBZe, or strychnine/strychnine-morphine. A blinded observer recorded which mouse of each pair had the greater global pain behaviour.

Results

Strychnine (i.c.) produced higher quantitative allodynia scores in the trigeminal distribution (mean 81.5%; 95% confidence interval [CI] 76.4 to 86.6) vs the aCSF group (mean 11.3%; 95% CI 8.1 to 14.4) (P < 0.0001). Carbamazepine epoxide (i.c.) completely abolished allodynia when co-injected with strychnine (mean 83.2%; 95% CI 78.1 to 88.4) vs strychnine alone (mean 3.2%; 95% CI ?0.9 to 7.2) (P < 0.0001). Morphine co-injected with strychnine did not result in reduced allodynia (mean 65.7%; 95% CI 42.0 to 89.4) compared with strychnine alone (mean 87.6%; 95% CI 77.6 to 97.6) (P = 0.16). In a further global allodynia assessment, strychnine (i.c.) produced greater allodynia than both aCSF and strychnine administered with CBZe (P = 0.03). Morphine (ip) administered with strychnine did not result in reduced global allodynia compared with strychnine administered alone (P = 1.0).

Conclusion

In this study, we have developed and validated a novel murine model of trigeminal dynamic allodynia induced by intracisternal strychnine. The use of mice to study trigeminal allodynia has many benefits, including access to a vast repository of transgenic mouse variants, ease of handling, low cost, and minimal variance of results. The present model may have utility in screening drug treatments for dynamic mechanical allodynia resulting from trigeminal neuropathies.     

Antimicrobial Susceptibility Patterns for Recent Clinical Isolates of Anaerobic Bacteria in South Korea

We determined the antimicrobial susceptibilities of 255 clinical isolates of anaerobic bacteria collected in 2007 and 2008 at a tertiary-care hospital in South Korea. Piperacillin-tazobactam, cefoxitin, imipenem, and meropenem were highly active β-lactam agents against most of the isolates tested. The rates of resistance of Bacteroides fragilis group organisms and anaerobic Gram-positive cocci to moxifloxacin were 11 to 18% and 0 to 27%, respectively.Anaerobic bacterial resistance trends may vary greatly, depending on regions or institutions (1). The Clinical and Laboratory Standards Institute (CLSI) does not recommend routine susceptibility testing of all clinical isolates of anaerobic bacteria, except for the management of patients with serious infections (4). A recent survey indicated that only a few laboratories in the United States performed antimicrobial susceptibility testing of anaerobic bacteria due to the complex techniques and predictable susceptibilities involved (5). However, regional susceptibility patterns are pivotal in the empirical treatment of infected patients because these patterns are related to clinical outcomes (13). Therefore, periodic monitoring of the regional or institutional resistance trends of clinically important anaerobe isolates is recommended (4). Our investigation of resistance trends of Bacteroides fragilis group organisms from South Korea has been taking place since 1989 (9, 15). However, few studies have focused on the susceptibilities of other anaerobes. Therefore, the aim of this study was to determine the recent antimicrobial resistance patterns of frequently isolated anaerobes at a tertiary-care hospital in South Korea.Anaerobes were isolated from blood, normally sterile body fluid, and abscess specimens, but Clostridium difficile was isolated from stool specimens of suspected C. difficile-associated disease patients at Severance Hospital in 2007 and 2008. The isolates were identified by either conventional methods (19) or the ATB 32A system (bioMérieux, Marcy l''Etoile, France). A total of 255 nonduplicated isolates were used in this study, including 63 of B. fragilis, 57 of other B. fragilis group species, 28 of Prevotella spp., 9 of other Gram-negative bacilli, 15 of Anaerococcus prevotii, 15 of Peptoniphilus asaccharolyticus, 15 of Finegoldia magna, 13 of Peptostreptococcus spp., 15 of C. perfringens, 12 of C. difficile, and 13 of other Gram-positive bacilli.Antimicrobial susceptibility testing was performed using the CLSI agar dilution method (4). The medium used was Brucella agar (Becton Dickinson, Cockeysville, MD) supplemented with 5 μg hemin and 1 μg vitamin K₁ per ml and 5% laked sheep blood. The antimicrobial powders used were piperacillin and tazobactam (Yuhan, Seoul, South Korea), cefoxitin (Merck Sharp & Dohme, West Point, PA), cefotetan (Daiichi Pharmaceutical, Tokyo, Japan), clindamycin (Korea Upjohn, Seoul, South Korea), imipenem, metronidazole (Choong Wae, Seoul, South Korea), chloramphenicol (Chong Kun Dang, Seoul, South Korea), meropenem (Sumitomo, Tokyo, Japan), moxifloxacin (Bayer Korea, Seoul, South Korea), and vancomycin (Eli Lilly & Co., Indianapolis, IN). For the combination of piperacillin and tazobactam, a constant tazobactam concentration of 4 μg/ml was added.An inoculum of 10⁵ CFU was applied with a Steers replicator (Craft Machine Inc., Woodline, PA), and the plates were incubated in an anaerobic chamber (Forma Scientific, Marietta, OH) for 48 h at 37°C. The MIC of each antimicrobial agent was defined as the concentration at which there was a marked reduction in growth, such as from confluent colonies to a haze, <10 tiny colonies, or 1 to 3 normal-sized colonies. B. fragilis ATCC 25285 and B. thetaiotaomicron ATCC 29741 were used as controls.β-Lactamase production by anaerobic Gram-negative bacilli, with the exception of B. fragilis group organisms, was determined by applying test organisms to the Cefinase disks and recording the results after 30 min (Becton Dickinson, Cockeysville, MD).Table ​Table11 shows the MICs of antimicrobial agents and the resistance rates of the anaerobes tested. Among the 255 isolates, B. fragilis group organisms were the most prevalent (47%). These organisms are more virulent and more resistant to antimicrobial agents than most other anaerobes (3). In this study, piperacillin-tazobactam, cefoxitin, imipenem, and meropenem were highly active against B. fragilis group organisms, with resistance rates of less than 7%. The rates of resistance to imipenem and piperacillin-tazobactam were 4% and 7%, respectively, for other B. fragilis group organisms. However, much higher resistance rates were observed for piperacillin (27 to 51%), cefotetan (14 to 68%), and clindamycin (33 to 86%). These values were similar to those observed in 1997 to 2004 in the same hospital: piperacillin, 33 to 49%; cefotetan, 14 to 60%; clindamycin, 51 to 76% (15). A higher prevalence of resistance, in particular to clindamycin, was observed than in the United States, i.e.,19 to 35% (17). CLSI added a recommendation to test susceptibility to moxifloxacin in 2004 and 2007. In this study, the moxifloxacin resistance rates were 11% for B. fragilis and 18% for other B. fragilis group organisms. These rates were slightly higher than the 7 to 9% reported in Taiwan (11) but lower than those in Greece (14) and the United States (16 to 75% and 26 to 55%, respectively) (17).

TABLE 1.

Antimicrobial activities against 255 anaerobic bacteria isolated in 2007 to 2008

Protein Induced by Vitamin K Antagonist-II (PIVKA-II) Is a Reliable Prognostic Factor in Small Hepatocellular Carcinoma

Background

Hepatocellular carcinoma (HCC) <2 cm in diameter has a favorable prognosis. Therefore surgical resection of small HCC is associated with good outcomes. However, the predisposing factors of prognosis following resection of HCC remain ill-defined. The aims of the present study were to identify the clinicopathologic characteristics and outcomes of patients with small HCC and analyze the predisposing factors for tumor recurrence after surgery.

Methods

We retrospectively reviewed 180 patients with small HCC who underwent hepatectomy between 2006 and 2010. Independent predictors of tumor recurrence were identified with Cox regression analysis.

Results

The 1-year, 3-year, and 5-year disease-free survival rates and overall survival rates were 83.7, 68.0, 65.3, and 98.9, 96.5, 92.7 %, respectively. Multivariate analysis reported that protein induced by the vitamin K antagonist-II (PIVKA-II) ≥200 mAU/mL, alkaline phosphatase (ALP) ≥80 IU/mL, and microvascular invasion were important predisposing factors for tumor recurrence. Elevated serum PIVKA-II level was associated with microvascular invasion in small HCC, which was a powerful predisposing factor.

Conclusions

Although small HCC is generally associated with a good prognosis, serum PIVKA-II level ≥200 mAU/mL, ALP ≥ 80 IU/L, and microvascular invasion were predisposing factors for tumor recurrence. These factors can be used to stratify patients with respect to recurrence after resection. Elevated PIVKA-II was closely associated with microvascular invasion in small HCC. These data emphasize the importance of PIVKA-II in small HCC.     

Cross‐link stabilization does not affect the response of collagen molecules,fibrils, or tendons to tensile overload

We investigated whether immature allysine‐derived cross‐links provide mechanically labile linkages by exploring the effects of immature cross‐link stabilization at three levels of collagen hierarchy: damaged fibril morphology, whole tendon mechanics, and molecular stability. Tendons from the tails of young adult steers were either treated with sodium borohydride (NaBH₄) to stabilize labile cross‐links, exposed only to the buffer used during stabilization treatment, or maintained as untreated controls. One‐half of each tendon was then subjected to five cycles of subrupture overload. Morphologic changes to collagen fibrils resulting from overload were investigated using scanning electron microscopy, and changes in the hydrothermal stability of collagen molecules were assessed using hydrothermal isometric tension testing. NaBH₄ cross‐link stabilization did not affect the response of tendon collagen to tensile overload at any of the three levels of hierarchy studied. Cross‐link stabilization did not prevent the characteristic overload‐induced mode of fibril damage that we term discrete plasticity. Similarly, stabilization did not alter the mechanical response of whole tendons to overload, and did not prevent an overload‐induced thermal destabilization of collagen molecules. Our results indicate that hydrothermally labile cross‐links may not be as mechanically labile as was previously thought. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31:1907–1913, 2013     

Congenital cerebrospinal fluid-containing intracranial abnormalities: a sonographic classification

Ultrasound is useful as a primary modality for diagnosis of complex nonhydrocephalic intracranial malformations. We present 10 cases of intracranial cerebrospinal fluid containing abnormalities that may be diagnosed by ultrasound. Congenital abnormalities presented include holoprosencephaly, hydranencephaly, agenesis of the corpus callosum with interhemispheric cyst, porencephaly, schizencephaly, and arachnoid cyst. Ultrasound may be used in the fetus or neonate in detecting and separating these abnormalities from hydrocephalus.