bluuebunny/crossref_metadata_2025 huggingface dataset:
Starving the Addiction: New Opportunities for Durable Suppression of AR Signaling in Prostate Cancer
Authors: Knudsen Karen E. , Scher Howard I.
Published: July 2009

Clinical data and models of human disease indicate that androgen receptor (AR) activity is essential for prostate cancer development, growth, and progression. The dependence of prostatic adenocarcinoma on AR signaling at all stages of disease has thereby been exploited in the treatment of disseminated tumors, for which ablation of AR function is the goal of first-line therapy. Although these strategies are initially effective, recurrent tumors arise with restored AR activity, and no durable treatment has yet been identified to combat this stage of disease. New insights into AR regulation and the mechanisms underlying resurgent AR activity have provided fertile ground for the development of novel strategies to more effectively inhibit receptor activity and prolong the transition to therapeutic failure.
Sequence Variant Discovery in DNA Repair Genes from Radiosensitive and Radiotolerant Prostate Brachytherapy Patients
Authors: Pugh Trevor J. , Keyes Mira , Barclay Lorena , Delaney Allen , Krzywinski Martin , Thomas Dallas , Novik Karen , Yang Cindy , Agranovich Alexander , McKenzie Michael , Morris W. Jim , Olive Peggy L. , Marra Marco A. , Moore Richard A.
Published: July 2009

Purpose: The presence of intrinsic radiosensitivity within prostate cancer patients may be an important factor contributing to development of radiation toxicity. We investigated whether variants in genes responsible for detecting and repairing DNA damage independently contribute to toxicity following prostate brachytherapy.

Experimental Design: Genomic DNA was extracted from blood samples of 41 prostate brachytherapy patients, 21 with high and 20 with low late toxicity scores. For each patient, 242 PCR amplicons were generated containing 173 exons of eight candidate genes: ATM, BRCA1, ERCC2, H2AFX, LIG4, MDC1, MRE11A, and RAD50. These amplicons were sequenced and all sequence variants were subjected to statistical analysis to identify those associated with late radiation toxicity.

Results: Across 41 patients, 239 sites differed from the human genome reference sequence; 170 of these corresponded to known polymorphisms. Sixty variants, 14 of them novel, affected protein coding regions and 43 of these were missense mutations. In our patient population, the high toxicity group was enriched for individuals with at least one LIG4 coding variant (P = 0.028). One synonymous variant in MDC1, rs28986317, was associated with increased radiosensitivity (P = 0.048). A missense variant in ATM, rs1800057, associated with increased prostate cancer risk, was found exclusively in two high toxicity patients but did not reach statistical significance for association with radiosensitivity (P = 0.488).

Conclusions: Our data revealed new germ-line sequence variants, indicating that existing sequence databases do not fully represent the full extent of sequence variation. Variants in three DNA repair genes were linked to increased radiosensitivity but require validation in larger populations.
Radiofrequency Ablation Combined with KS-IL2 Immunocytokine (EMD 273066) Results in an Enhanced Antitumor Effect against Murine Colon Adenocarcinoma
Authors: Johnson Erik E. , Yamane Brett H. , Buhtoiarov Ilia N. , Lum Hillary D. , Rakhmilevich Alexander L. , Mahvi David M. , Gillies Stephen D. , Sondel Paul M.
Published: July 2009

Purpose: Radiofrequency ablation (RFA) is a common treatment modality for surgically unresectable tumors. However, there is a high rate of both local and systemic recurrence.

Experimental Design: In this preclinical study, we sought to enhance the antitumor effect of RFA by combining it with huKS-IL2 immunocytokine [tumor-specific monoclonal antibody fused to interleukin-2 (IL2)] in mice bearing CT26-KS colon adenocarcinoma. Mice were treated with RFA, huKS-IL2 via intratumoral injection, or combination therapy.

Results: Treatment of mice bearing s.c. tumors with RFA and huKS-IL2 resulted in significantly greater tumor growth suppression and enhanced survival compared with mice treated with RFA or huKS-IL2 alone. When subtherapeutic regimens of RFA or huKS-IL2 were used, tumors progressed in all treated mice. In contrast, the combination of RFA and immunocytokine resulted in complete tumor resolution in 50% of mice. Treatment of a tumor with RFA and intratumoral huKS-IL2 also showed antitumor effects against a distant untreated tumor. Tumor-free mice after treatment with RFA and huKS-IL2 showed immunologic memory based on their ability to reject subsequent challenges of CT26-KS and the more aggressive parental CT26 tumors. Flow cytometry analysis of tumor-reactive T cells from mice with complete tumor resolution showed that treatment with RFA and huKS-IL2 resulted in a greater proportion of cytokine-producing CD4 T cells and CD8 T cells compared with mice treated with RFA or huKS-IL2 alone.

Conclusions: These results show that the addition of huKS-IL2 to RFA significantly enhances the antitumor response in this murine model, resulting in complete tumor resolution and induction of immunologic memory.
A First-in-Man Phase I and Pharmacokinetic Study on CHR-2797 (Tosedostat), an Inhibitor of M1 Aminopeptidases, in Patients with Advanced Solid Tumors
Authors: Reid Alison H.M. , Protheroe Andrew , Attard Gerhardt , Hayward Nikki , Vidal Laura , Spicer James , Shaw Heather M. , Bone Elizabeth A. , Carter Joanne , Hooftman Leon , Harris Adrian , De Bono Johann S.
Published: July 2009

Purpose: To determine the maximum tolerated dose, dose-limiting toxicity, pharmacokinetics, and preliminary therapeutic activity profile of CHR-2797 (tosedostat), a novel, orally bioavailable inhibitor of the M1 family of aminopeptidases with antiproliferative and antiangiogenic activity in vitro.

Experimental Design: A phase I study of accelerated titration design that escalated through nine doses (10-320 mg) in patients (Eastern Cooperative Oncology Group performance status, ≤2) with advanced solid tumors. CHR-2797 was administered once daily.

Results: Forty patients (median age, 60 years; range, 24-80 years; male, 27; female, 13) were treated in 12 cohorts with once daily doses (10-320 mg). Dose-limiting toxicities were thrombocytopenia, dizziness, and visual abnormalities in one patient, and anemia, blurred vision, and vomiting in a second patient at 320 mg, resulting in an inability to complete 28 days of study drug. The most commonly observed toxicities were fatigue, diarrhea, peripheral edema, nausea, dizziness, and constipation. One patient had a partial response (renal cell carcinoma) and four patients had stable disease for >6 months. CHR-2797 and its active metabolite, CHR-79888, show dose-proportional increases in plasma AUC and Cmax. The terminal half-life for CHR-2797 is ∼1 to 3.5 hours and between 6 and 11 hours for CHR-79888. Intracellular (packed blood cells) exposure to CHR-79888 is consistent with intracellular levels that proved to be efficacious in xenograft models.

Conclusion: CHR-2797 is well tolerated and can be safely administered at doses that result in intracellular levels of CHR-79888 that are associated with activity in preclinical models. The recommended dose for single agent therapy in solid tumors is 240 mg/d.
Valproic Acid at Therapeutic Plasma Levels May Increase 5-Azacytidine Efficacy in Higher Risk Myelodysplastic Syndromes
Authors: Voso Maria Teresa , Santini Valeria , Finelli Carlo , Musto Pellegrino , Pogliani Enrico , Angelucci Emanuele , Fioritoni Giuseppe , Alimena Giuliana , Maurillo Luca , Cortelezzi Agostino , Buccisano Francesco , Gobbi Marco , Borin Lorenza , Di Tucci Anna , Zini Gina , Petti Maria Concetta , Martinelli Giovanni , Fabiani Emiliano , Fazi Paola , Vignetti Marco , Piciocchi Alfonso , Liso Vincenzo , Amadori Sergio , Leone Giuseppe
Published: July 2009

Purpose: Epigenetic changes play a role and cooperate with genetic alterations in the pathogenesis of myelodysplastic syndromes (MDS). We conducted a phase II multicenter study on the combination of the DNA-methyltransferase inhibitor 5-azacytidine (5-AZA) and the histone deacetylase inhibitor valproic acid (VPA) in patients with higher risk MDS.

Experimental Design: We enrolled 62 patients with MDS (refractory anemia with excess blasts, 39 patients; refractory anemia with excess blasts in transformation, 19 patients; and chronic myelomanocytic leukemia (CMML), 4 patients) and an International Prognostic Scoring System (IPSS) rating of Intermediate-2 (42 patients) or high (20 patients). VPA was given to reach a plasma concentration of >50 μg/mL, then 5-AZA was added s.c. at 75 mg/m2 for 7 days in eight monthly cycles.

Results: The median overall survival was 14.4 months. At a median follow-up of 12 months (range, 0.7-21.0), the disease progressed in 20 patients, with 21% cumulative incidence of progression. Of 26 patients who completed eight cycles, 30.7% obtained complete or partial remission, 15.4% had a major hematologic improvement, whereas 38.5% showed stable disease. Drug-related toxicity was mild. Favorable prognostic factors for survival were IPSS Intermediate-2 and plasma VPA of ≥50 μg/mL (log rank = 0.013 and 0.007, respectively). Analysis of polymorphisms important for the metabolism of the drugs used in the trial showed that carriers of the CYP2C19*2 variant of cytochrome P450 required higher VPA doses to achieve the target VPA plasma concentration of 50 μg/mL on day 1 of 5-AZA treatment (P = 0.0021).

Conclusion: Our data show that the 5-AZA/VPA combination is active and safe in patients with MDS with a poor prognosis. Achievement of VPA therapeutic levels may indeed increase 5-AZA efficacy.
Interleukin-6 Regulates Androgen Synthesis in Prostate Cancer Cells
Authors: Chun Jae Yeon , Nadiminty Nagalakshmi , Dutt Smitha , Lou Wei , Yang Joy C. , Kung Hsing-Jien , Evans Christopher P. , Gao Allen C.
Published: July 2009

Purpose: The standard systemic treatment for prostate cancer patients is androgen deprivation therapy. Although serum testosterone concentrations were significantly reduced after androgen deprivation therapy, levels of intraprostatic androgens are reproducibly measured at concentrations sufficient to activate androgen receptor and stimulate tumor growth, suggesting that prostate cancer cells may survive androgen deprivation therapies by increasing intracrine androgen synthesis within the prostate. However, factors that regulate de novo intracrine androgen synthesis have not been identified. Interleukin-6 (IL-6) has been implicated in the modulation of androgen receptor activation and growth and differentiation in prostate cancer. In this study, we investigate whether IL-6 regulates intraprostatic androgen synthesis in prostate cancer cells.

Experimental Design: Quantitative reverse transcription-PCR and Western blotting were done to detect expression levels of steroidogenic enzymes. AKR1C3 promoter reporter was constructed and analyzed for IL-6–mediated AKR1C3 transcriptional activity. IL-6–mediated signaling was knocked down using small interfering RNA specific to IL-6 receptor and gp130, and the effect on AKR1C3 expression was examined. Intraprostatic androgen levels in prostate cancer cells in culture and in tumors were measured by an enzyme immunoassay (Testosterone EIA kit).

Results: We found that IL-6 increases the expression of genes encoding many steroidogenic enzymes, including HSD3B2 and AKR1C3, involved in androgen biosynthesis. Down-regulation of IL-6 receptor and gp130 expression using specific small interfering RNA abolished IL-6–mediated AKR1C3 expression, suggesting that IL-6 signaling is responsible for AKR1C3 expression. IL-6 increases AKR1C3 promoter activity, indicating that the increase in IL-6–mediated AKR1C3 expression is in part at the transcriptional level. Treatment of IL-6 increased testosterone level in LNCaP cells. The tumor testosterone levels were detected at 378 pg/g in tumors generated from IL-6–overexpressing LNCaP-IL6+ cells inoculated orthotopically into the prostates of castrated male nude mice.

Conclusions: These results suggest that IL-6 increases levels of intracrine androgens through enhanced expression of genes mediating androgen metabolism in prostate cancer cells.
Regulation of Colorectal Cancer Cell Apoptosis by the n-3 Polyunsaturated Fatty Acids Docosahexaenoic and Eicosapentaenoic
Authors: Giros Anna , Grzybowski Mike , Sohn Vanessa R. , Pons Elisenda , Fernandez-Morales Jessica , Xicola Rosa M. , Sethi Puja , Grzybowski Jessica , Goel Ajay , Boland C. Richard , Gassull Miquel A. , Llor Xavier
Published: July 2009

Several studies have suggested that the n-3 fatty acids Docosahexaenoic (DHA) and Eicosapentaenoic (EPA) have an important protective effect on colorectal cancer, and this could be at least partly due to their proapoptotic activity. It is unclear, however, how this phenomenon is triggered and what mechanisms are implicated. Here, we show that both DHA and EPA have an important proapoptotic effect on colorectal cancer cells with different molecular phenotypes but not in noncancerous cells. Apoptosis is caspase dependent, and both intrinsic and extrinsic pathways are implicated. The dimerization of Bax and Bak, the depolarization of the mitochondrial membrane, and the subsequent release of cytochrome c and Smac/Diablo to the cytosol evidence the activation of the intrinsic pathway. The implication of the extrinsic pathway is shown by the activation of caspase-8, along with the down-regulation of FLIP. The timing of caspase-8 activation, and the oligomerization of Bid with Bax, suggest a cross-talk with the intrinsic pathway. None of the death receptors that commonly initiate the extrinsic pathway: FAS, TNF-R1, and TRAIL-R2 are found to be responsible for triggering the apoptosis cascade induced by DHA and EPA. Neither PPARγ nor cyclooxygenase-2, two likely candidates to regulate this process, play a significant role. Our findings suggest that the down-regulation of two key regulatory elements of the extrinsic and intrinsic pathways, FLIP and XIAP, respectively, is determinant in the induction of apoptosis by DHA and EPA. These fatty acids could potentially be useful adjuvant anticancer agents in combination with other chemotherapeutic elements.
A Population-Based Case-Control Study of Marijuana Use and Head and Neck Squamous Cell Carcinoma
Authors: Liang Caihua , McClean Michael D. , Marsit Carmen , Christensen Brock , Peters Edward , Nelson Heather H. , Kelsey Karl T.
Published: July 2009

Cannabinoids, constituents of marijuana smoke, have been recognized to have potential antitumor properties. However, the epidemiologic evidence addressing the relationship between marijuana use and the induction of head and neck squamous cell carcinoma (HNSCC) is inconsistent and conflicting.

Cases (n = 434) were patients with incident HNSCC disease from nine medical facilities in the Greater Boston, MA area between December 1999 and December 2003. Controls (n = 547) were frequency matched to cases on age (±3 years), gender, and town of residence, randomly selected from Massachusetts town books. A questionnaire was adopted to collect information on lifetime marijuana use (decade-specific exposures) and associations evaluated using unconditional logistic regression.

After adjusting for potential confounders (including smoking and alcohol drinking), 10 to 20 years of marijuana use was associated with a significantly reduced risk of HNSCC [odds ratio (OR)10-<20 years versus never users, 0.38; 95% confidence interval (CI), 0.22-0.67]. Among marijuana users moderate weekly use was associated with reduced risk (OR0.5-<1.5 times versus <0.5 time, 0.52; 95% CI, 0.32-0.85). The magnitude of reduced risk was more pronounced for those who started use at an older age (OR15-<20 years versus never users, 0.53; 95% CI, 0.30-0.95; OR≥20 years versus never users, 0.39; 95% CI, 0.17-0.90; Ptrend < 0.001). These inverse associations did not depend on human papillomavirus 16 antibody status. However, for the subjects who have the same level of smoking or alcohol drinking, we observed attenuated risk of HNSCC among those who use marijuana compared with those who do not.

Our study suggests that moderate marijuana use is associated with reduced risk of HNSCC.
Flavones as Colorectal Cancer Chemopreventive Agents—Phenol-O-Methylation Enhances Efficacy
Authors: Cai Hong , Sale Stewart , Schmid Ralf , Britton Robert G. , Brown Karen , Steward William P. , Gescher Andreas J.
Published: July 2009

Flavonoids occur ubiquitously in plants, and some possess preclinical cancer chemopreventive activity. Little is known about molecular features that mediate chemopreventive efficacy of flavonoids. Here, three related flavones, apigenin (4′,5,7-trihydroxyflavone), tricin (4′,5,7-trihydroxy-3′,5′-dimethoxyflavone), and 3′,4′,5′,5,7-pentamethoxyflavone (PMF), were compared in terms of their effects on (a) adenoma development in ApcMin mice, a model of human gastrointestinal malignancies; (b) growth of APC10.1 mouse adenoma cells in vitro; and (c) prostaglandin E-2 generation in HCA-7 human-derived colorectal cancer cells in vitro. Life-long consumption of PMF with the diet at 0.2% reduced ApcMin mouse adenoma number and burden by 43% and 61%, respectively, whereas apigenin was inactive. Tricin has previously shown activity in this model. IC50 values for murine adenoma cell growth inhibition by PMF, tricin, and apigenin were 6, 13, and 18 μmol/L, respectively. In ApcMin mice that received flavones (0.2%) for 4 weeks, adenoma cell proliferation as reflected by Ki-67 staining was reduced by PMF and tricin, but not by apigenin. On incubation with HCA-7 cells for 6 hours, PMF reduced prostaglandin E-2 generation with an IC50 of 0.8 μmol/L, a fraction of the respective values reported for tricin or apigenin. In silico PMF docked into the cyclooxygenase active site with greater affinity than tricin or apigenin. The results suggest that the rank order of cancer chemopreventive efficacy in ApcMin mice is PMF > tricin > apigenin, supporting the notion that the presence of O-methyl in the flavone molecular scaffold promotes gastrointestinal cancer chemopreventive efficacy.
Effect of Nest Box Availability on the Circadian Activity Rhythm of Common Marmosets (Callithrix jacchus)
Authors: Gonçalves Fabiana B. , Belísio Aline S. , Azevedo Carolina V.M.
Published: July 2009

The effect of nest box availability on the circadian activity rhythm (CAR) was compared in a diurnal primate kept in two conditions: with and without nest box, in a light-dark (LD) cycle (12:12) and constant light (LL). Marmosets kept in LD without nest box showed a longer duration of the activity phase (α), smaller day-to-day variability in activity onset and offset and higher levels of activity, particularly towards the end of the activity time. In LL, all animals maintained with nest box expressed free-running CAR, but without nest box, 3 of the 4 individuals showed relative coordination with the outdoor LD cycle, possibly due to social synchronization with captive marmosets maintained outdoors. All animals showed longer α and higher levels of activity without nest box. Thus, the availability of a nest box is an important aspect in maintaining marmosets, influencing CAR expression in this diurnal primate in LD and LL conditions, possibly due to differences in light exposure that induces masking. Further studies are needed to evaluate the effects of the unavailability of a nest box on CAR in LL conditions.
Cervical Epithelial Cells from Chlamydia trachomatis-Infected Sites Coexpress Higher Levels of Chlamydial Heat Shock Proteins 60 and 10 in Infertile Women than in Fertile Women
Authors: Jha Rajneesh , Vardhan Harsh , Bas Sylvette , Salhan Sudha , Mittal Aruna
Published: July 2009

Background/Aims: The objective of the present study was to examine the possible relationship between the chlamydial heat shock proteins (cHSP) 60 and 10 expression and the damaging sequelae of a Chlamydia trachomatis infection, such as infertility. Methods: Seven fertile and 7 infertile female patients infected with C. trachomatis attending the gynecology outpatient department of Safdarjung hospital (New Delhi, India) were enrolled. The relative transcript levels and intracellular expression of cHSP60 and cHSP10 in cervical cells were assessed using real-time RT-PCR and flow cytometry, respectively. Results: Quantitative real-time RT-PCR analysis showed that transcript levels of both cHSP60 (p = 0.007) and cHSP10 (p = 0.0006) were higher in infertile women than in fertile women. Flow cytometric analysis showed significantly higher intracellular levels of cHSP60 (p = 0.0006) and cHSP10 (p = 0.0041) in fertile women infected with Chlamydia than in infertile women. However, the percentage of double-positive cells (both cHSP60- and cHSP10-expressing cells) were higher (p = 0.0006) in infertile women than in fertile women. Conclusion: Our results suggest that cHSP60 and cHSP10 have a different pattern of expression in infertile women compared to fertile women reflecting a probable difference in the metabolic state of Chlamydia with the presence of an abnormal cryptic form of C. trachomatis in infertile women.
The Chemotactic Activity of Native and Denatured Serum Albumin
Authors: Wilkinson P.C. , McKay I.C.
Published: July 2009

Native human serum albumin (HSA) is not chemotactic for neutrophil leucocytes. Certain treatments such as acidification or reduction-alkylation may cause HSA preparations to become moderately chemotactic, whereas, after other treatments such as heat-denaturation, very little increase in chemotactic activity is observed. The chemotactic activity which follows denaturation is generally confined to monomeric forms of HSA. Acquisition of such activity correlates well with physical measurements of conformational change in the HSA molecule inasmuch as the viscosity and surface activity of chemotactically active HSA preparations are greater than those of preparations without activity. A hypothesis is proposed whereby chemotactically active sites on protein molecules are associated with hydrophobic areas of the molecule and are therefore concealed in the native state, but may become exposed during unfolding so that denatured proteins acquire chemotactic activity.
Effect of Polyspecific Rabbit Anti-Immunoglobulin Antisera on the Response of Human Lymphocytes in Mixed Lymphocyte Culture
Authors: Fröland S.S. , Natvig J.B.
Published: July 2009

The effect of pretreatment of human peripheral lymphocytes with rabbit anti-F(ab′)2 on reactivity in mixed lymphocyte cultures (MLC) was studied in 19 experiments using a one-way system and in 10 experiments with two-way stimulation. In some ‘two-way cultures’, an additive effect of allogeneic lymphocytes and undiluted anti-F(ab′)2 was observed. In the one-way system and in the majority of the two-way MLC tests a marked potentiating effect of the two stimulants could be seen, the activity in most cases being much higher than the sum of the single effects. With neither system could inhibition be observed. In the one-way MLC system, the effect of diluting the antiserum was studied to see if any inhibitory effect could be discovered. At the highest dilution studied (1:10,000), however, there was still no inhibition. It is concluded that the present study does not give evidence of immunoglobulin receptors necessary for the MLC response. The possibility that the lymphocytes stimulated by anti-F(ab′)2 may be different from the cells engaged in the MLC response, is discussed.
Increase in the Sensitivity of the Rat to Histamine Following Infection with Nippostrongylus brasiliensis
Authors: Keller R. , Beeger I.
Published: July 2009

The present results show that following infection of rats with the nematode, Nippostrongylus brasiliensis, the toxicity of histamine is increased but the hyperglycemic response to histamine and epinephrine is unchanged. In rats pretreated with Haemophilus pertussis vaccine, on the contrary, the increase in toxicity to histamine is accompanied by a blockade of the hyperglycemic response to histamine and epinephrine. Further observations have shown that the hypersensitivity of nematode-infected rats to histamine is not diminished by pretreatment with Β-adrenergic blockers or cortisone together with L-epinephrine. Thus, blockade of the adrenergic function is not an important feature in the increase in histamine lethality of the nematode-infected rat.
Measurement of Antibody-Producing Capacity in Man
Authors: Lee A.K.Y. , Mackay I.R. , Rowley M.
Published: July 2009

Antibody-producing capacity in man was assessed in various diseases, including chronic bronchitis and infections, after immunization with monomeric flagellin from Salmonella adelaide. For each disease group studied, results were compared with those for controls matched for age and sex and derived from a hospital population with miscellaneous illnesses not affecting the immunological system. A previous study showed that the humoral immune response in disease was less than that in health; in the present study, patients with chronic bronchitis, infections, peptic ulcer, ischaemic heart disease, diabetes mellitus, chronic alcoholism and postgastrectomy states exemplified this general impairment of antibody-producing capacity in disease. Moreover, in chronic bronchitis, infections and multiple myeloma the response to flagellin was significantly more impaired than in other diseases. Explanations for this immunodepression associated with chronic bronchitis and infections were prolonged antigenic stimulation, antigenic competition, or effects of antibiotics; alternatively there may be predisposition to infection in patients with an impaired antibody-producing capacity.
The Role of two Retina Antigens in Production of Experimental Allergic Uveitis and its Suppression by Mycrobacteria
Authors: Wacker W.B. , Lipton M.M.
Published: July 2009

Of two tissue specific antigens demonstrated in guinea pig retina, only one was active in production of allergic uveitis in this species, even though antibodies produced to either antigen were demonstrably reactive with the animals own retina. Further, the frequency and severity of disease was a function of the amount of antigen used, as well as the quantitative relationships between mycobacteria and antigen in the immunizing emulsion. Nearly 100% of guinea pigs developed uveitis when the proper amounts of each component were in the inoculum, but a small increase of mycobacteria beyond an optimal amount was found to suppress the immunologic and pathologic reactions otherwise elicited by retina antigen.
Immunological Effects of Aspirin Anhydride, a Contaminant of Commercial Acetylsalicylic Acid Preparations
Authors: De Weck A.L.
Published: July 2009

Aspirin anhydride (ASAN) has been found to be a frequent contaminant of commercial acetylsalicylic acid (ASA) preparations. ASAN is a highly immunogenic substance susceptible to form protein conjugates in vitro, to induce the formation of anti-aspiryl antibodies and of contact hypersensitivity to ASAN in guinea pigs and rabbits. Commercial ASA samples varied greatly in their capacity to induce hypersensitivity to ASAN under standardized conditions in guinea pigs. As demonstrated with various types of aspiryl-protein conjugates, antibodies induced by ASAN and detected by passive cutaneous anaphylaxis are mostly of aspiryl specificity. Antibodies of similar specificity have been detected in patients ingesting aspirin. A few patients presenting a reliable history of generalized urticaria and acute angioneurotic edema after ingestion of aspirin showed skin hypersensitivity to aspirylpolylysine. The possible significance of contamination of acetylsalicylic acid preparations by aspirin anhydride in the development of untoward reactions to aspirin is discussed.
Aggregation of Platelets of Normal and Bordetella pertussis Vaccinated Mice by ATP, ADP, and Sodium Fluoride
Authors: O’Bryan Brenda S. , Fishel Ch.W.
Published: July 2009

The results of the present studies have shown that platelets obtained from B. pertussis vaccinated mice were more sensitive to the aggregative action of ATP, ADP, and sodium fluoride than were their normal counterparts. Aggregation initiated by any of these substances was suppressed by exogenous cAMP but was enhanced in the presence of NE. These data suggested a role of adenyl cyclase and adrenergic receptors in the mouse platelet aggregation with the exaggerated responsiveness of ‘pertussis’ components due to either hyperreactive α-type or blockade of the β-type receptors.
Precipitation Reactions of an IgA Myeloma Protein with Nitrophenylated Proteins
Authors: Patterson R. , Andersen B. , McAninch J. , Roberts Mary
Published: July 2009

The reactions of a human serum with an IgA monoclonal component which precipitates with dinitrophenyl (DNP) conjugated proteins are described. The IgA existed in polymeric and monomeric forms. Only the polymeric forms precipitated DNP-proteins, but binding of DNP proteins by the monomer was demonstrated by autoradiographic immunoelectrophoresis. The precipitating reactions of the IgA occurred with certain populations of DNP-proteins and were higher with more heavily haptenized preparations. The binding affinity of the strongly precipitating IgA serum for DNP-lysine was low and the precipitation reaction with DNP-proteins was not inhibited by DNP-lysine. The results indicated that precipitation involves the reaction of polymeric forms of IgA with DNP-protein configurations or closely associated DNP-DNP configurations in heavily haptenized conjugates.
Immunological Events Associated with Immunization by Sperm in Incomplete Freund’s Adjuvant
Authors: Tung K.S.K. , Unanue E.R. , Dixon F.J.
Published: July 2009

Guinea pigs were immunized with sperm antigen(s) in complete or incomplete Freund’s adjuvant. Those immunized with sperm in CFA developed orchids, made high titers of complement fixing antibody; their lymphocytes could transfer orchitis when inoculated into normal recipients and produced in vitro migration inhibitory factors. Those immunized with sperm in ICFA did not develop orchitis and made high titers of anaphylactic antibodies; their lymphocytes did not transfer orchitis into normal guinea pigs and did not inhibit migration of macrophages. Lymphocytes from guinea pigs immunized with CFA could transfer orchitis when inoculated into the testes of guinea pigs immunized with ICFA. Hence the lack of disease in guinea pigs immunized with ICFA is not because of a protection of their testes by non-phlogogenic antibodies.
Effect of Number of Haptens per Carrier Molecule on an Adoptive Secondary Anti-Hapten Response
Authors: Kontiainen S.
Published: July 2009

Primary antihapten response to a monovalent conjugate of NIP (4-hydroxy-3-iodo-5-nitrophenylacetic acid) and bovine serum albumin (BSA) tended to be rich in 7S and primary response to a polyvalent conjugate NIP34-BSA rich in 19S anti-NIP antibodies. Monovalent conjugate NIP1.1-BSA tended to boost cells primed with polyvalent conjugates of NIP to a secondary anti-NIP response rich in 7S and polyvalent conjugate, NIP34-BSA, to a response rich in 19S anti-NIP antibodies. Monovalent conjugate, NIP1.1-BSA, did not prime the cells detectably, although it induced a primary antihapten response.
Antibody-Induced Lysis of Nucleated Cells in Agar Gel
Authors: Juji T. , Kano K. , Milgrom F.
Published: July 2009

Cytolysis of nucleated cells incorporated into agar gel was studied using hetero- and isoantibodies. Evidence was presented that IgG and IgM antibodies may participate in these reactions. Studies on the lytic activity of rabbit anti-rat sera showed that antibodies inducing lysis of rat thymus cells are directed against species-specific cell surface antigens. Evidence was presented that some of these antigens are shared by nucleated cells and erythrocytes whereas other antigens are present on nucleated cells only. Tests with rabbit anti-sheep erythrocyte serum demonstrated that Forssman antibodies elicit lysis of nucleated cells originating from Forssman-positive species. It was also shown that infectious mononucleosis sera are capable of inducing lysis of sheep and bovine nucleated cells. In studying human renal transplantation sera, cytolysis of kidney cells was induced by sera obtained from recipients after removal of rejected grafts or pretransplantation sera of recipients who rejected hyperacutely their grafts. These results were consistent with those observed in previous studies by means of mixed agglutination.
In vivo Passive Sensitization of Normal Rabbit Leukocytes with Sera Demonstrating Homocytotropic Antibody Activity
Authors: Colwell E.J. , Ortaldo J.R. , Schoenbechler M.J. , Barbaro J.F.
Published: July 2009

Normal rabbit leukocytes were passively sensitized, as determined by in vitro antigen-induced histamine release, by the i.v. administration of homologous sera demonstrating homocytotropic antibody activity. There was a significant correlation between the magnitude of passive sentization and the PCA titers of the donor sera. Those parameters that affected skin sensitization also affected leukocyte sensitization. Maximal leukocyte sensitization was observed 3 days after serum administration and by the 10th day no significant histamine release was demonstrable.
Mechanism of Histamine Release by Toluidine Blue from Rat Mast Cells
Authors: Yamasaki H. , Komoto S.
Published: July 2009

Toluidine blue (TB) releases histamine from isolated rat mast cells suspended in buffered physiological solution (pH 7.0, 37 °C), with or without accompanying degranulation depending upon the concentration of drug used. Numerous vesicular protrusions, which are probably distinct from degranulation and do not occur with compound 48/80, appear over the whole cell surface at the concentrations in which histamine release is maximal; an indication of intracellular release of histamine. Histamine release by TB is completed in 5 sec and morphological changes of mast cells are accomplished within 15 sec at latest. These processes probably occur before the dye enters the cell, as judged by serial color microphotographs. The reactions may be triggered by the binding of the dye to the receptive sites on the cell membrane. Histamine release by TB depends upon the cellular energy-generating ability, as with other basic histamine liberators.
Neonatal Thymectomy and the Antibody Response to Sheep Erythrocytes in Mice
Authors: Lemmel E.-M. , Cooper M.D. , Good R.A.
Published: July 2009

Neonatal thymectomy of Swiss albino mice and C3H mice leads to deficiency of production of hemagglutinins when mice are immunized at age of 6 weeks with sheep erythrocytes. However, the antibody response of thymectomized mice seems to be dose dependent – with a higher dose of antigen higher titers of hemagglutinins were produced. With large enough antigen dose the primary response was only delayed and not reduced. The capability for a secondary response appears unimpaired in neonatally thymectomized animals. It is suggested that the thymus-dependent ‘antigen reactive’ cells are not obligatory participants in the immune response to sheep erythrocytes but rather act as ‘amplifiers’ of a thymus-independent humoral immune response.
Immunization of Neonatal Rabbits with Bovine Serum Albumin Associated with Allogeneic Thymus Cells
Authors: Winfield J.B. , Martin W.J. , Mage Rose G.
Published: July 2009

Bovine serum albumin (BSA) associated with thymus cells from a donor differing for 2 or 3 allotypic markers (a, b, A14–15) from recipient animals was immunogenic when administered to newborn rabbits. The same dose (15 μg)of antigen, as well as larger doses, administered in solution were usually tolerogenic. Recipients of thymus cells develop detectable levels of donor type immunoglobulin markers persisting for 8–10 weeks. The anti-BSA appeared to be exclusively of host origin; no relationship existed between the presence of the donor type immunoglobulin and the antibody response. Cellular chimeras in the peripheral blood were not demonstrated using sex chromosomes as markers.
Alteration of Immunological Reactivity in Chickens by the in ovo Administration of Antisera Against Central Lymphoid Tissue
Authors: Toben H.R. , St. Pierre R.L.
Published: July 2009

Antisera of high specificity have been produced against thymic and bursal lymphocytes in the chicken. By the in ovo administration of the antibodies directed against central lymphoid tissues, further evidence has been demonstrated for the functional dichotomy of the immune system in the chicken by the selective inhibition of immunologic function of thymus or bursa of Fabricius. A model system for human sex-linked agammaglobulinemia and thymic aplasia can be developed in the chicken by the selective inhibition of central lymphoid organs through the use of specific antilymphocytic globulin.
Myosin Vc, a Member of the Actin Motor Family Associated with Rab8, Is Involved in the Release of DV2 from HepG2 Cells
Authors: Xu Xiao-Feng , Chen Zong-Tao , Gao Na , Zhang Jun-Lei , An Jing
Published: July 2009

Objective: The pathogenesis of the dengue virus (DV) infection has not been well defined. We have reported that actin and Rab8 are involved in DV2 infection. Myosin Vc (Myo5c) is a novel member of the class V myosins and regulates the actin-mediated membrane trafficking associated with Rab8. In this study, the involvement of Myo5c in the release of DV2 was investigated in HpeG2 cells. Methods: Distributions of actin, Myo5c, DV2 and Rab8 were revealed by fluorescent staining. HepG2Myo5c-tail cells expressing a dominant-negative mutant of Myo5c were constructed by transfection and were assessed by Western blotting. The viral titers were detected by plaque assay, and the expression of Rab8 was analyzed by flow cytometry. Results: DV2 infection altered the distribution pattern of Myo5c, which might be associated with the depolymerization of actin, though colocalization rates of Myo5c with DV2 or actin were low. Furthermore, the release of DV2, but not the intracellular viral production, was reduced from HepG2Myo5c-tail cells. Moreover, Myo5c colocalized with Rab8 and an increase of Rab8 was associated with the decrease of the viral release caused by the Myo5c tail.Conclusions: Our data suggest that Myo5c associated with Rab8 is involved in the release of DV2 from HepG2 cells.
Time Course of Corneal Biomechanical Parameters after Laser in situ Keratomileusis
Authors: Kamiya Kazutaka , Shimizu Kimiya , Ohmoto Fumiko
Published: July 2009

Purpose: To assess the time course of corneal biomechanics after laser in situ keratomileusis (LASIK). Methods: We examined 36 eyes of 20 consecutive patients undergoing LASIK for low to moderate myopia. We quantitatively assessed the values of corneal biomechanics characterized by corneal hysteresis (CH) and corneal resistance factor (CRF) using an Ocular Response Analyzer™ (Reichert Ophthalmic Instruments) before and 1 week, 1, 3, and 6 months after surgery. We carried out this measurement 3 times, and the average value was used for statistical analysis. Results: The CH was 10.6 ± 1.7 (mean ± SD) mm Hg preoperatively, and 8.6 ± 1.2, 9.0 ± 1.7, 9.0 ± 1.4, and 8.9 ± 1.5 mm Hg 1 week, 1, 3, and 6 months postoperatively, respectively. The CRF was 10.0 ± 1.7 mm Hg preoperatively, and 7.3 ± 1.5, 7.6 ± 2.0, 7.8 ± 1.6, and 7.7 ± 1.6 mm Hg 1 week, 1, 3, and 6 months postoperatively, respectively. The variances of the data were statistically significant (p < 0.001 for both CH and CRF). Multiple comparisons demonstrated significant differences between measurements made before surgery and at all postoperative times (at 1 week and 1, 3, and 6 months; p < 0.001 for CH and CRF, Fisher’s least significant difference test). Conclusions: Over the study period, the largest changes in corneal biomechanical parameters occurred within 1 week after surgery, and these then became nearly stable. No progressive deterioration of the corneal biomechanics was observed at any time during the 6-month follow-up period.
Segmental Ischemia of the Liver – Microdialysis in a Novel Porcine Model
Authors: Winbladh A. , Sandström P. , Olsson H. , Svanvik J. , Gullstrand P.
Published: July 2009

Background: Segmental liver ischemia is often used in rodents to study ischemia and reperfusion injuries (IRI). There are no reports of protocols using segmental ischemia in porcine models. Microdialysis (MD) provides the opportunity to study local effects of IRI in vivo. Methods: Eight pigs received an MD catheter placed in liver segments IV and V, respectively. All circulation to segment IV was stopped for 80 min, and reperfusion was followed for 240 min. Results: During ischemia the levels of lactate, glycerol and glucose increased 3-fold (p < 0.001), 40-fold (p < 0.001) and 4-fold (p < 0.01), respectively, in the ischemic segment compared to the perfused segment, whereas the levels of pyruvate fell to a tenth of the preischemic level (p < 0.001). All values returned to baseline after reperfusion. Serum levels of aspartate aminotransferase increased (p < 0.05). Polymorphonuclear cells increased in both segments, although the density was significantly higher in segment IV. Conclusion: Clamping of one liver segment in pigs is a simple, stable and reproducible model to study IRI with minimal systemic effects. MD revealed no signs of anaerobic metabolism in the perfused segment but still there was an increase in the number of polymorphonuclear neutrophils in this segment, although it was lower than that in the ischemic segment.
Role of the Endocannabinoid System in Abdominal Obesity and the Implications for Cardiovascular Risk
Authors: Rosenson Robert S.
Published: July 2009

Several cardiometabolic factors present in obese and insulin-resistant individuals represent a continuum of increasing risk for the development of type 2 diabetes and cardiovascular disease. The importance of abdominal obesity as an independent risk factor is underscored by its association with adverse endocrine function. Recent evidence from animal and human studies has shown a role for the endocannabinoid system in maintaining energy balance and glucose and lipoprotein metabolism, with overactivity linked to aberrant glycemic and lipoprotein control, and a link to adiposity. Modulation of this system through endocannabinoid-receptor blockade has resulted in an improvement in a number of important risk factors in clinical trials, including visceral and subcutaneous abdominal adipose tissue, glucose tolerance, dyslipidemia and measures of inflammation. These findings may have significant implications for the management of patients at risk of developing cardiovascular and metabolic disease; however, the occurrence of psychiatric adverse events with rimonabant may preclude further development of centrally active endocannabinoid receptor antagonists for the treatment of cardiometabolic disorders. Future research is needed to explore the role of selective peripheral CB1 receptor antagonists in the treatment of patients at high cardiometabolic risk.
How Well Do the ADAS-cog and its Subscales Measure Cognitive Dysfunction in Alzheimer’s Disease?
Authors: Benge Jared F. , Balsis Steve , Geraci Lisa , Massman Paul J. , Doody Rachelle S.
Published: July 2009

Background/Aims: The Alzheimer’s Disease Assessment Scale-cognitive (ADAS-cog) is regularly used to assess cognitive dysfunction in Alzheimer’s disease (AD) clinical trials. Yet, little is known about how the instrument and its subscales measure cognition across the spectrum of AD. The current investigation used item response theory (IRT) analyses to assess the measurement properties of the ADAS-cog across the range of cognitive dysfunction in AD. Methods: We used IRT-based analyses to establish the relationship between cognitive dysfunction and the probability of obtaining observed scores on each subscale and the test as a whole. Data were obtained from 1,087 patients with AD and amnestic mild cognitive impairment. Results: Results showed that the ADAS-cog and its subscales provide maximum information at moderate levels of cognitive dysfunction. Raw score differences toward the lower and higher ends of the scale corresponded to large differences in cognitive dysfunction, whereas raw score differences toward the middle of the scale corresponded to smaller differences. Conclusions: The utility of the ADAS-cog and its subscales is optimal in the moderate range of cognitive dysfunction, but raw score differences in that region correspond to relatively small differences in cognitive dysfunction. Implications for tracking and staging dementia and for clinical trials are discussed.
Very Early Mobilisation and Complications in the First 3 Months after Stroke: Further Results from Phase II of A Very Early Rehabilitation Trial (AVERT)
Authors: Sorbello D. , Dewey H.M. , Churilov L. , Thrift A.G. , Collier J.M. , Donnan G. , Bernhardt J.
Published: July 2009

Background: Interventions that may reduce the number and severity of potentially harmful post-stroke complications are desirable. This study explored whether very early and frequent mobilisation (VEM) affected complication type (immobility/stroke related), number and severity. Methods: Secondary analysis from phase II, randomised controlled trial. Patients admitted within 24 h of stroke, whose physiological parameters fell within set limits, were randomised to either VEM, commencing <24 h, or standard care. Complications to 3 months were recorded by a blinded assessor and classified by a neurologist. Analysis was intention to treat. Results: Seventy-one patients were recruited (standard care 33; VEM 38).There were no significant group differences in the number, type or severity of complications by 3 months, and most patients (81.6%) experienced one or more complications. Falls were common, while depression was absent. The multivariate analysis showed older age (OR 1.10, 95% CI: 1.02–1.18, p = 0.009) and longer length of stay (OR 1.18, 95% CI: 1.06–1.32, p = 0.002) were associated with experiencing an immobility-related complication. Conclusion: Interventions that promote recovery and reduce complications may consequently reduce length of stay. The larger phase III trial currently underway may shed light on whether increasing mobilisation reduces complications after stroke.
Thiazolidinediones as Therapy for Endometriosis: A Case Series
Authors: Moravek Molly B. , Ward Elizabeth A. , Lebovic Dan I.
Published: July 2009

Background: Current medical therapies for endometriosis result in delayed conception and have not been shown to provide any fertile benefit subsequent to treatment. Thiazolidinediones (TZDs) do not impede conception and have been shown to reduce endometriotic lesions in animal models; however, no studies have been performed in humans. The aim of this study was to provide preliminary data about the effectiveness of a TZD in treating endometriosis-related pain. Methods: Case series of women with endometriosis recruited from the University of Michigan as part of an open-label prospective phase 2a clinical trial. Participants were given rosiglitazone, 4 mg daily, for 6 months. Subjective endometriosis symptoms were assessed using a modified Biberoglu and Behrman symptom severity scale and the McGill pain questionnaire. Results: Two of the 3 patients exhibited improvement in severity of symptoms and pain levels with a concurrent decrease in pain medication, while 1 experienced no change. Rosiglitazone was well tolerated by all patients. Conclusions: Combined with data gathered from studies in rats and nonhuman primates, the results from this study offer positive justification for using TZDs as a well-tolerated treatment for endometriosis that can address pain without impeding ovulation and without the need for add-back therapy.