Vaccination with human amniotic epithelial cells confer effective protection in a murine model of Colon adenocarcinoma.

As a prophylactic cancer vaccine, human amniotic membrane epithelial cells (hAECs) conferred effective protection in a murine model of colon cancer. The immunized mice mounted strong cross-protective CTL and antibody responses. Tumor burden was significantly reduced in tumor-bearing mice after immunization with hAECs. Placental cancer immunotherapy could be a promising approach for primary prevention of cancer. In spite of being the star of therapeutic strategies for cancer treatment, the results of immunotherapeutic approaches are still far from expectations. In this regard, primary prevention of cancer using prophylactic cancer vaccines has gained considerable attention. The immunologic similarities between cancer development and placentation have helped researchers to unravel molecular mechanisms responsible for carcinogenesis and to take advantage of stem cells from reproductive organs to elicit robust anti-cancer immune responses. Here, we showed that vaccination of mice with human amniotic membrane epithelial cells (hAECs) conferred effective protection against colon cancer and led to expansion of systemic and splenic cytotoxic T cell population and induction of cross-protective cytotoxic responses against tumor cells. Vaccinated mice mounted tumor-specific Th1 responses and produced cross-reactive antibodies against cell surface markers of cancer cells. Tumor burden was also significantly reduced in tumor-bearing mice immunized with hAECs. Our findings pave the way for potential future application of hAECs as an effective prophylactic cancer vaccine.

Regulation of luteinizing hormone receptor in hippocampal neurons following different long-lasting treatments of castrated adult rats.

The aim of this study was to investigate the effects of different Luteinizing hormone (LH) and steroid hormones levels on LH receptor (LHR) expression in the hippocampal cells. Rats (24 males and 24 females) were assigned to four groups: one control and three experimental [gonadectomy (GDX), gonadectomy + gonadotropin releasing hormone analogue (GDX+GnRHa) and GDX+GnRHa+estradiol (E2) or testosterone (T)] independently for each gender. All experimental rats were gonadectomized; then GnRHa was administrated to GDX+GnRHa group, and GnRHa plus steroid hormone to GDX+GnRHa+E2 or T group in both genders for four-month. LHR mRNA expression and its protein level in hippocampal cells were measured using QRT-PCR and Western blotting. Quantification of mRNA revealed a decrease in LHR transcripts level in GDX+GnRHa group of females. A significant change was observed between GDX groups and GDX+GnRHa+E2 or T versus GDX+GnRHa group in females. High levels of LH decreased significantly the immature isoform of LHR in GDX group compared to control group in both genders, but low LH concentrations in GDX+GnRHa group induced immature LHR isoform production only in females. Therefore increased LH concentration induces production of incomplete LHR transcripts in hippocampal cells and decreases immature LHR at the protein level. This implies that LH decreases the efficiency of translation through either producing non-functional LHR molecules or preventing their translation.

Biocatalytic synthesis of polymeric nanowires by micellar templates of ionic surfactants.

Micelle-templated polyguaiacol nanowires were successfully prepared via polymerization oxidation of guaiacol (o-methoxy phenol) by peroxidase enzyme in the presence of hydrogen peroxide at mild reaction conditions. The dimensions of the prepared nanowires were controlled by tuning the size and shape of the micelle structure via changing and controlling the type, chain length and molar concentrations of the ionic surfactant. The progress of the reaction and estimation of the size of soft micellar templates were followed by UV-Vis spectroscopy and dynamic light scattering (DLS). The resulting micelle encapsulated or purified polyguaiacol nanowires were characterized using transmission electron microscopy (TEM).

HCV genotype 3 is associated with a higher hepatocellular carcinoma incidence in patients with ongoing viral C cirrhosis.

Liver steatosis is a main histopathological feature of Hepatitis C (HCV) infection because of genotype 3. Steatosis and/or mechanisms underlying steatogenesis can contribute to hepatocarcinogenesis. The aim of this retrospective study was to assess the impact of infection with HCV genotype 3 on hepatocellular carcinoma (HCC) occurrence in patients with ongoing HCV cirrhosis. Three hundred and fifty-three consecutive patients (193 men, mean age 58 ± 13 years), with histologically proven HCV cirrhosis and persistent viral replication prospectively followed and screened for HCC between 1994 and 2007. Log-rank test and Cox model were used to compare the actuarial incidence of HCC between genotype subgroups. The patients infected with a genotype 3 (n = 25) as compared with those infected with other genotypes (n = 328) had a lower prothrombin activity [78 (interquartile range 60-85) vs 84 (71-195) %, P = 0.03] and higher rate of alcohol abuse (48%vs 29%, P = 0.046). During a median follow-up of 5.54 years [2.9-8.6], 11/25 patients (44%) and 87/328 patients (26%) with a genotype 3 and non-3 genotype, respectively, develop a HCC. HCC incidences were significantly different among the genotype subgroups (P = 0.001). The 5-year occurrence rate of HCC was 34% (95% CI, 1.3-6.3) and 17% (95% CI, 5.7-9.2) in genotype 3 and non-3 genotype groups, respectively (P = 0.002). In multivariate analysis, infection with a genotype 3 was independently associated with an increased risk of HCC occurrence [hazard ratio 3.54 (95% CI, 1.84-6.81), P = 0.0002], even after adjustment for prothrombin activity and alcohol abuse [3.58 (1.80-7.13); P = 0.003]. For patients with HCV cirrhosis and ongoing infection, infection with genotype 3 is independently associated with an increased risk of HCC development.

Superficial community-acquired skin infections: prevalence of bacteria and antibiotic susceptibility in France.

OBJECTIVE: Evaluation of the susceptibility to currently used antibiotics of bacteria, particularly S. aureus isolated from superficial community- acquired skin infection and to compare results with those from an earlier study. METHODS: Every dermatologist in community practice participating in the study was asked to include the first two patients consulting them for superficial cutaneous bacterial infection. Swab specimens collected from the skin infection were sent to a central laboratory. RESULTS: The dermatologist enrolled 390 patients in the study. The rate of positive culture was 49%, 259 bacterial strains were isolated. S. aureus was the major species (56.8% of all isolated strains). S. aureus was resistant to methicillin in 4%. All strains of S. aureus were susceptible to pristinamycin and mupirocin. CONCLUSIONS: The results of the two epidemiological studies of superficial community acquired skin infections with a comparable methodology at a 6-year interval demonstrated that the prevalence of CA-MRSA skin infection remained low in this setting.

[First-intent monotherapy: persistence rate of antihypertensive treatment is better with angiotensin receptor blockers. Ol'métrise Cardio survey results]. / Monothérapie de première intention: le traitement antihypertenseur est plus souvent poursuivi, seul ou combiné, quand il s'agit d'un ARA II. Enquête Ol'métrise Cardio.

OBJECTIVES: To describe, according to the type of first-line antihypertensive monotherapy (FLAM) the persistence rate and the cumulative probability of its potential given up to the advantage of another antihypertensive class (rotation) in the following 3 years. METHODS: retrospective survey (to avoid any "survey effect" undertaken by 551 French cardiologists, in treated hypertensive patients. The survey described the FLAM, patients characteristics when FLAM was introduced, when given up in case of rotation and at the inclusion visit, as well as the dates of introduction, rotation and visit. STATISTICAL ANALYSIS: Kaplan Meier's method, Cox model. RESULTS: One thousand nine hundred and fifty five patients (62 +/- 11 years, males 57%, diabetes 13%, coronary disease 11%, LVH 20%) were analysed. At the end of 2.1 +/- 0.7 years, 1407 patients (72%) continued the FLAM alone (N=738, 52%) or combined (N=669, 48%). Survival analysis shows i) that the probability of rotation is roughly constant over time whatever the FLAM ii) that the probability of rotation against time is different according to the FLAM (p<0.0001, log rank) and lesser with ARB. These results are confirmed by the multivariate analysis (Cox model adjusted on age, sex and comorbidities).

Denaturation of jack-bean urease by sodium n-dodecyl sulphate: a kinetic study below the critical micelle concentration.

Kinetics of urease denaturation by anionic surfactant (sodium n-dodecyl sulphate, SDS) at concentrations below the critical micelle concentration (CMC) is investigated spectrophotometrically at neutral pH and the corresponding two-phase kinetic parameters of the process are estimated from a three-state reversible process using a binomial exponential relation based on the relaxation time method as: Using a prepared computer program, the experimental data are properly fitted into a binomial exponential relation, considering a two-phase denaturation pathway including a kinetically stable folded intermediate formed at SDS concentration of 1.1 mM. Forward and backward rate constants are estimated as: k(1)=0.2141+/-4.5 x 10(-3), k(2)=5.173 x 10(-3)+/-8.3 x 10(-5), k(-1)=0.09432+/-3.6 x 10(-4) and k(-2)=2.079 x 10(-3)+/-5.6 x 10(-5)s(-1) for the proposed mechanism. The rate-limiting step as well as the reaction coordinates in the denaturation mechanism are established. The mechanism involves formation of a kinetically stable folded native like intermediate through the electrostatic interactions. The intermediate was found to be more stable even than the native form (by about 9 kJmol(-1)) and still hexamer, because no loss of amplitude was observed. Electrophoresis experiments on the native and surfactant/urease complexes indicated a higher mobility for the kinetically folded native like intermediate.

[Prevalence and mechanism of masked hypertension: the ol'mesures survey]. / Prevalence et mécanismes de l'hypertension masquée. Enquête OL'MESURES.

RATIONALE: Masked hypertension (MH) and uncontrolled hypertension (UCH) have both bad prognosis. The influence of measurement circumstances on MH prevalence and reproducibility are little known. OBJECTIVE: To evaluate the prevalence and reproducibility of MH after excluding confusing factors [method and time of blood pressure (BP) measurement, antihypertensive treatment] by a standardization procedure. METHODS: 2189 hypertensive patients (61+/- 12 years, men 57%) having been treated in monotherapy by an angiotensin II receptor inhibitor for at least 8 weeks Were evaluated in a French multicenter prospective observational survey. Three BP successive office measurements were performed by the GPs during 2 visits (V) at similar times 13 +/- 9 days apart (BP: V1 149 +/- 19 / 85 +/- 11 mmHg, V2 145 +/- 19/83 +/- 11 mmHg) and home BP self-measurements (HBPM) were performed morning and evening for 3 consecutive days (HBPM morning + evening : n=18 +/- 1; 142 +/- 16/81 +/- 9 mmHg) and at the time of the visit (daytime HBPM: n=9 +/- 1; 140 +/- 16/80 +/- 10 mmHg) by the patients (Omron-705CP). RESULTS: [table: see text]. CONCLUSION: the observed MH prevalence is similar to previous published studies and is independent of: treatment, BP measurement methods, measurements frequency and HBPM time but it depends on office BP values. Consequently, its reproducibility is directly dependent of the quality of office BP measurements.

Postconditioning is protective in renal reperfusion injury only in male rats. A gender difference study.

PURPOSE: We investigated the impact of sex on the protective effect of postconditioning (POC), a series of brief ischemia-reperfusion (IR) cycles at the reperfusion onset, as a recently described novel approach to attenuate renal IR injury. In this study, the left renal pedicles of uni-nephrectomized male and female rats were clamped for 45 minutes followed by 24 hours of reperfusion as IR groups. Uni-nephrectomized, sham-operated male and female rats served as control groups. Ischemic postconditioning was performed using 4 cycles of 10 seconds of IR of renal pedicle at the end of the ischemia. Twenty-four hours later, BUN (blood urea nitrogen), plasma creatinine (Cr), and renal histological changes, as well as kidney levels of MDA (malondialdehyde) and SOD (superoxide dismutase) as oxidative stress markers were evaluated to detect the protective effect of POC against IR injury in rats. RESULTS: Induction of IR resulted in significant reduction in renal function, demonstrated by increase in plasma Cr and BUN, histological changes and oxidative stress in both genders. Application of POC afforded significant protection against these injuries in male rats, namely decreased levels of BUN and Cr, histological improvements and less oxidative damages. However, there were no significant differences in the above-mentioned parameters in female rats. CONCLUSION: While POC is shown to be beneficial against renal IR injury in male rats, it did not show any protective effect in female rats.

Microbiological assay for the analysis of certain macrolides in pharmaceutical dosage forms.

Clarithromycin (CLA) and roxithromycin (ROX) are macrolide antibiotics with an expanded spectrum of activity that are commercially available as tablets. A microbiological assay, applying the cylinder-plate method and using a strain of Micrococcus luteus ATCC 9341 as test organism, has been used and validated for the quantification of two macrolide drugs; CLA and ROX in pure and pharmaceutical formulations. The validation of the proposed method was carried out for linearity, precision, accuracy and specificity. The linear dynamic ranges were from 0.1 to 0.5µg/mL for both compounds. Logarithmic calibration curve was obtained for each macrolide (r>0.989) with statistically equal slopes varying from 3.275 to 4.038, and a percentage relative standard deviation in the range of 0.24-0.92%. Moreover, the method was applied successfully for the assay of the studied drugs in pharmaceutical tablet dosage forms. Recovery from standard addition experiments in commercial products was 94.71-96.91% regarding clarithromycin and 93.94-98.12% regarding roxithromycin, with a precision (%RSD) 1.32-2.11%. Accordingly, this microbiological assay can be used for routine quality control analysis of titled drugs in tablet formulations.

Sphingosine, oleylamine and stearylamine inhibit both CD11a/CD18-dependent and -independent homotypic aggregation: demonstration by cytofluorimetry.

An CD11a/CD18-dependent homotypic aggregation pathway is induced by triggering CD45 molecules on human thymocytes. By contrast, a CD11a/CD18-independent homotypic aggregation process is induced by triggering the CD99 molecule (E2, MIC2 gene product) expressed at the surface of either Jurkat T cells or human thymocytes. A new quantitative method based on FACS analysis of aggregated cells was used and allowed to show that both types of aggregation (CD11a/CD18-dependent and CD11a/CD18-independent) were inhibited with sphingosine, oleylamine or stearylamine. These three compounds had no effect on the expression of CD99, CD45, CD11a, CD18 or other [correction of others] known integrins expressed at the surface of the cells studied.

Effect of Mn2+, Co2+, Ni2+, and Cu2+ on horseradish peroxidase: activation, inhibition, and denaturation studies.

The effects of transition metal ions (M2+) such as Mn2+, Co2+, Ni2+, and Cu2+ on the functional and structural stabilities of horseradish peroxidase (HRP) were investigated with respect to reversible chemical denaturation, Michaelis-Menten kinetics, chemical modification and time-dependent catalytic activity. Conformational Gibbs free energy (deltaGo(H2O)) as a structural stability criterion and transition concentrations of metal ions ([M2+] 1/2) were estimated using a two-state chemical denaturation model. Activation and inhibitory concentration ranges for each metal ion were specified by the steady-state enzyme kinetics. Results of a pH-profile method confirmed by chemical modification indicate that a histidine residue interacts in the activation concentration range, whereas carboxylic residues (Asp and Glu) contribute to interaction in the inhibitory concentration range. Incubation of the enzyme with the metal ion at activation concentration leads to long-term functional stability of peroxidase. Thus, such metal ions as potent effectors induced the enhancement of conformational and functional stabilities of horseradish peroxidase.

[The acute respiratory distress syndrome in malaria: is it always the prerogative of Plasmodium falciparum?]. / Le syndrome de détresse respiratoire aiguë au cours du paludisme: est-il toujours l'apanage du Plasmodium falciparum?

Severe malaria causes nearly one million deaths annually in endemic areas and is a public health priority worldwide. Severity associated with the occurrence of acute respiratory distress syndrome (ARDS) is a well-known complication of infection with Plasmodium falciparum and can reach 25% of infected adults. However, ARDS is less often described with other Plasmodium species. We report the case of a young Moroccan soldier who died in an array of ARDS related to malaria of Plasmodium ovale 7 months after his return from an endemic country.

Monoclonal antibodies against ROR1 induce apoptosis of chronic lymphocytic leukemia (CLL) cells.

ROR1 is a receptor tyrosine kinase (RTK) recently identified to be overexpressed at the gene and protein levels in chronic lymphocytic leukemia (CLL). Monoclonal antibodies (MAbs) against RTKs have been successfully applied for therapy of solid tumors. We generated five MAbs against the Ig (n = 1), cysteine-rich (CRD) (n = 2) and kringle (KNG) (n = 2) domains, respectively, of the extracellular part of ROR1. All CLL patients (n = 20) expressed ROR1 on the surface of the leukemic cells. A significantly higher frequency of ROR1 expression was found in patients with progressive versus non-progressive disease, and in those with unmutated versus mutated IgVH genes. All five MAbs alone induced apoptosis in the absence of complement or added effector cells (Annexin-V and MTT, as well as cleavage of poly-(ADP ribose)-polymerase, caspase-8 and caspase-9) of CLL cells but not of normal B cells. Most effective were MAbs against CRD and KNG, significantly superior to rituximab (P < 0.005). Cross-linking of anti-ROR1 MAbs using the F(ab')(2) fragments of anti-Fc antibodies significantly augmented apoptosis. Two of the MAbs induced complement-dependent cytotoxicity (CDC) similar to that of rituximab and one anti-ROR1 MAb (KNG) (IgG1) showed killing activity by antibody-dependent cellular cytotoxicity. The identified ROR1 epitopes may provide a basis for generating human ROR1 MAbs for therapy.

Cereblon is a direct protein target for immunomodulatory and antiproliferative activities of lenalidomide and pomalidomide.

Thalidomide and the immunomodulatory drug, lenalidomide, are therapeutically active in hematological malignancies. The ubiquitously expressed E3 ligase protein cereblon (CRBN) has been identified as the primary teratogenic target of thalidomide. Our studies demonstrate that thalidomide, lenalidomide and another immunomodulatory drug, pomalidomide, bound endogenous CRBN and recombinant CRBN-DNA damage binding protein-1 (DDB1) complexes. CRBN mediated antiproliferative activities of lenalidomide and pomalidomide in myeloma cells, as well as lenalidomide- and pomalidomide-induced cytokine production in T cells. Lenalidomide and pomalidomide inhibited autoubiquitination of CRBN in HEK293T cells expressing thalidomide-binding competent wild-type CRBN, but not thalidomide-binding defective CRBN(YW/AA). Overexpression of CRBN wild-type protein, but not CRBN(YW/AA) mutant protein, in KMS12 myeloma cells, amplified pomalidomide-mediated reductions in c-myc and IRF4 expression and increases in p21(WAF-1) expression. Long-term selection for lenalidomide resistance in H929 myeloma cell lines was accompanied by a reduction in CRBN, while in DF15R myeloma cells resistant to both pomalidomide and lenalidomide, CRBN protein was undetectable. Our biophysical, biochemical and gene silencing studies show that CRBN is a proximate, therapeutically important molecular target of lenalidomide and pomalidomide.

Bone mineral density assessed by dual-energy X-ray absorptiometry in patients with viral or alcoholic compensated cirrhosis. A prospective study.

BACKGROUND/AIM: Cirrhosis is considered as a risk factor for osteoporosis whose prevalence is poorly known. The aim was to assess prospectively bone mineral density (BMD) in patients with alcoholic or viral compensated cirrhosis. METHODS: From 2006 to 2008, patients with viral or alcoholic compensated cirrhosis had BMD assessment by dual-energy X-ray absorptiometry. The prevalence of osteopenia (-2.5SD