This complement activation pathway was attributed to the structural similarities between the EO monomers of poloxamine and a region of D-mannose [144]. The brush-like conformation minimizes the MBL and ficolin binding to PEG backbone and consequently reduces the complement activation via the lectin pathway [145]. Thus, the conformation and the mobility of surface projected PEO chains of poloxamine on nanoparticles are paramount to modulate the complement Inhibitors,research,lifescience,medical activation pathway [146]. 2.5. “Long Circulation” Revealed PEG-and poloxamine-coated nanocarriers have been demonstrated to undergo immunoglobulin, fibronectin, and apolipoprotein association [14, 29, 33, 118, 122–124, 147] as

well as Inhibitors,research,lifescience,medical C3 opsonisation that mediates the biorecognition by macrophages through specific complement receptors (CR1 and CR3, CD11b/CD18) [18]. However, these systems possess long-lasting profiles in blood [148]. The prolonged circulation in the bloodstream is due to the steric hindrance of the surface polymers [134] that prevents the macrophage approach [124]. Furthermore, the C3b adsorbed on the polymer corona of the particle surface can be proteolytically degraded to fragments that by assembling

with Inhibitors,research,lifescience,medical other cofactors inhibit the recognition by the macrophage receptors [149]. The factor C3bn of the complement adsorbed on PEG-coated liposomes may also bind CR1 receptor Inhibitors,research,lifescience,medical associated with the erythrocytes membrane, which can also explain the prolonged circulation time of PEGylated liposomes [150]. The steric shielding effect conveyed by polymer coating on long circulation properties of stealth nanocarriers was demonstrated by Moghimi using poloxamine-908-coated particles. These particles, incubated with Inhibitors,research,lifescience,medical serum obtained from a poloxamine-908 preinjected

animal, showed a higher protein adsorption as compared to particles incubated with serum obtained from animals that were not preexposed to poloxamine. The protein-coated nanoparticles showed similar pharmacokinetic profiles when administered to animals never exposed to poloxamine. This evidence reinforces the explanation that the improved circulation time of Parvulin stealth nanoparticles is not solely ascribable to reduced protein adsorption on particle surface [151] which surely takes place for sterically stabilized nanocarriers. Improved circulation time can be mainly attributable to the prohibited biorecognition of the adsorbed opsonic proteins by the macrophages. 2.6. Nanocarrier Coating with Hydrophilic Polymers: Physical and Chemical Strategies Sterically Pexidartinib price protective polymer can be physically or chemically conjugated to the nanocarrier surface. Physically conjugation involves the hydrophobic adsorption of polymer fragments on the particle surface while the chemical conjugation is obtained by chemical reaction of polymers with surface functions to yield covalent bonds.

These methods will probably help in the further development of personalized preventive interventions. Selected abbreviations and acronyms AF attributable fraction ER exposure rate NNT number needed to treat OR odds ratio
In 2002 we contributed

an article to Dialogues in Clinical Neuroscience which discussed the neuroprotective actions of estrogens.1 In this review we build on the understanding Inhibitors,research,lifescience,medical we had at that point, and will discuss the importance of the accumulating data that point to the complexities of estrogen action. Clearly, many factors, including the type of estrogen used, the dose and route of administration, and the age and previous hormonal and health status Inhibitors,research,lifescience,medical of the women being treated, must be taken into consideration when designing clinical studies and when interpreting results. Women usually undergo the menopausal transition when they are about 51 years of age. This dramatic physiological change, which may be abrupt or may occur over a period of a few years, is marked by a decline in ovarian hormone

secretion and cessation of reproductive fertility. The postmenopausal period is often associated with vaginal dryness, urinary symptoms, osteoporosis, and a panel of neurological manifestations Inhibitors,research,lifescience,medical such as hot flushes, and greater instances of sleep disturbance, emotional instability, cerebrovascular stroke, and Alzheimer’s disease. Over the past century the average life expectancy in the United States has increased to over 80 years, while the age of menopause has remained fixed Inhibitors,research,lifescience,medical at age 51. Thus,

today, more than ever before, a greater number of women and a larger proportion of women are destined to spend over three decades of their lives in a hypoestrogenic state. This makes it imperative that we understand the intricacies of estrogen action, when it is protective, and Inhibitors,research,lifescience,medical when it increases risk. Estrogens and progestins Estradiol and progesterone belong to a family of steroid hormones with complex actions. Estradiol-17β (E2), the predominant and crotamiton most biologically active estrogen, is an 18 carbon (C-18) steroid with an aromatic A-ring. It is synthesized mainly by the ovary; however, other organs and tissues, including adipose tissue, the brain (neurons, astrocytes, and microglia), cells of the immune system, and bone, are thought to produce it as well. Progesterone is a C-21 steroid hormone, which is not only an active hormone in and of itself, but is also a precursor to estrogens. In addition to the estrogens and progestins produced in human tissues, a wide variety of estrogenic and learn more progestogenic compounds are synthesized in other species or are pharmacologically manufactured through pathways that have been developed by researchers and have been used widely by the pharmaceutical industry.

g. A-EA-2005. Seven anti-FMDV bovine post-vaccinal sera were used in the study. Two were against the two existing vaccine strains, A-KEN-05-1980 and A-ETH-06-2000 raised in Kenya and Ethiopia [21], respectively, by administering the commercially prepared vaccine. The animals vaccinated with A-KEN-05-1980 were bled on 21 day following vaccination. The animals vaccinated with A-ETH-06-2000 received a boost on 21-day post-vaccination and bled one week later. ABT 888 The rest five bvs were raised in cattle

against one existing vaccine strain (A-ERI-1998) and four candidate vaccine strains (A-EA-1981, A-EA-1984, A-EA-2005 and A-EA-2007) following the method previously described [23]. The candidate vaccine strains were selected taking into account the genotypes currently circulating in the region. For each Modulators antigen, sera from four or five animals were pooled for use in the neutralisation test. The homologous neutralising antibody titres of each pooled serum

are presented in Table 1a. The 2D-VNT test was carried out using the pooled post-vaccination bovine sera according to Rweyemamu and colleagues. [24]. Antibody titres were calculated from regression data as the log10 reciprocal antibody dilution required for 50% neutralisation of 100 tissue culture infective units of virus (log10SN50/100 TCID50). The antigenic relationship of viruses is given by the ratio: ‘r1’ = neutralising antibody titre against the heterologous virus/neutralising antibody titre against the homologous virus.

buy NVP-BEZ235 The significance of differences between ‘r1-values’ obtained by the polyclonal antiserum was evaluated according to standard criteria [25]. The sequences of the entire capsid coding region (P1) of the viruses were generated. RNA extraction from the cell culture grown viruses, reverse transcription (RT), polymerase chain reaction (PCR) to amplify the P1 region, sequencing, sequence analysis and assembling, and alignment were performed as described previously [26]. MEGA 5 [27] was used to determine nucleotide and aa variations. The aa variability of the capsid coding region of the type A viruses were determined as described by Valdar [28]. The aligned, complete P1 nucleotide sequences were used to determine the most suitable nucleotide substitution model using jModelTest Sitaxentan [29] and MEGA [27] resulting in the selection of a General time reversal (GTR) model with a combination of gamma distribution and proportion of invariant sites (GTR + G + I). Then, Bayesian analysis was performed using the BEAST software package v1.5.4 [30]. In BEAUti v1.5.4, the ages of the viruses were defined by the date of sample collection and the analysis used GTR + G + I model to describe rate heterogeneity among sites. Variations in substitution rate among branches were evaluated by comparing four different clocks in BEAST. The maximum clade credibility (MCC) phylogenetic tree was inferred using the Bayesian Markov Chain Monte Carlo (MCMC) method.

These potential confounding factors make detection of anticipation in BP disorder difficult. The hypothesis that anticipation in BP disorder reflects causative expanding trinucleotide CTG repeat sequences has generated genomic searches for such sequences,138-141 using the repeat expansion detection Inhibitors,research,lifescience,medical method.142 Increased lengths of CTG repeats were thus noted in BP disorders, especially among those with familial

disease. However, not all studies have reported this difference,143 and no report shows transmission of an expanding repeat within BP families, the definitive evidence. Furthermore, greater than 90% of the expanded CTG repeats detected by the repeat expansion detection method142 are from two apparently nonpathogenic unstable CTG repeats on 17q and 18q21.144 The hypothesis that unstable trinucleotide repeats represent BP susceptibility

factors Inhibitors,research,lifescience,medical deserves continued study. Selected abbreviations and acronyms APM affected pedigree member ASP affected sibling pair BP bipolar disorder BP I bipolar disorder I – manic and depressive episodes BP II bipolar disorder II – hypomank and depressive episodes IBD identical-by-descent LOD logarithm of the odds of linkage MZ Inhibitors,research,lifescience,medical monozygotic UP unipolar disorder
ALK inhibitor gabapentin Inhibitors,research,lifescience,medical enjoys a wide spectrum of psychopharmacological and neuropharmacological indications. Curiously, we found only a single article on the efficacy of gabapentin for treating neuroleptic-induced akathisia [Pfeffer et al. 2005]. This is counterintuitive on theoretical and clinical grounds. Theoretically, gabapentin enhances the activity Inhibitors,research,lifescience,medical of gamma-aminobutyric acid (GABA), an inhibitory neurotransmitter that would be predicted to suppress the abnormal

involuntary movements of akathisia. Clinically, gabapentin carries US Food and Drug Administration approval for restless legs syndrome (RLS), a neurological to disorder. RLS and neuroleptic-induced akathisia are not identical conditions, but they are probably related, so one wonders why the efficacy of gabapentin has not been more thoroughly investigated in the latter disorder. The purpose of the present article is to investigate the efficacy of gabapentin for treating neuroleptic-induced akathisia in a private-practice setting. The patients’ anonymity was carefully protected and the study was performed with informed consent and pursuant to all guidelines for study with human subjects as required by the institutions with which the authors are affiliated.

Consequently, differences between StreptInCor and the M protein sequences do not affect opsonization of the target strain, indicating that StreptInCor have broad capacity of coverage against the diverse M-types around the world. Previously we showed

that StreptIncor can be recognized by several HLA class II molecules, making it a candidate vaccine with broad capacity of coverage. The binding prediction of the C-terminal Palbociclib order amino acid sequences of the M1, M5, M6, M12 and M87 proteins with different HLA class II molecules shows that the possibility of recognition/processing of M proteins and peptides in the pockets (P1, P4, P6 and P9) of different HLA class II molecules agree with previous human studies from our group [26]. Libraries Another important data present here is that the anti-StreptInCor opsonizing and neutralizing antibodies did not induce cross-reactivity with human valve protein extracts, indicating the absence of cross-reactive antibodies. These results agrees with previous studies with HLA class II transgenic mice, in which no cross reactivity against heart-tissue derived proteins and

no tissue lesions were observed in several organs up to one year post-vaccination [29]. The present work reinforces the safety of and strong immune response triggered by the StreptInCor mice vaccination. Productions of antibodies that opsonize and neutralize a broad range of S. pyogenes GPCR Compound Library mw strains indicate

the potential of StreptInCor to prevent streptococcal infections without causing deleterious reactions. The authors declare that there is no conflict of interest. StreptInCor intellectual properties are in the names of Luiza Guilherme and Jorge Kalil. This work was supported by grants from “Fundação de Amparo à Pesquisa do Estado de Sao Paulo (FAPESP)” and “Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)”. Karine De Amicis’s benefits were supported by “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)”. not “
“Global molecular analyses are exploited to enhance our understanding of novel vaccination strategies. High-throughput technologies, including microarray analyses and RNA deep sequencing, allow genome-wide profiling of gene expression within different study groups. Similarly, targeted assays enable study of the expression of a dedicated number of genes [e.g. dual colour reverse transcription multiplex ligation-dependent probe amplification (dcRT-MLPA) assay], cell-expressed molecules (e.g. flow cytometry) or secreted molecules (multiplex assays). Expectations of data output from these analyses in vaccine trials are high, and it is hoped that through the systematic analysis of biomarkers using modern bioassays, predictive biomarkers, which can be used as (surrogate) markers of clinical endpoints or of adverse events, can be identified.

17 Thus, PPI deficits have also been found in obsessive-compulsive disorder (OCD),Tourette’s syndrome, Huntington’s disease, panic disorder,19 and manic patients with bipolar disorder.20 These disorders are all characterized by PPI deficits and abnormalities of gating in sensory, motor, or cognitive domains.

It should also be noted, however, that deficient PPI is not found in several other psychiatric disorders.17 Antipsychotic effects on PPI in animals PPI models in rodents The cross-species nature of startle and PPI enables the use of animal models of induced deficits that are extremely similar to the gating deficits seen Inhibitors,research,lifescience,medical in schizophrenia. Beginning with the initial Inhibitors,research,lifescience,medical demonstrations of the ability of dopamine agonists to disrupt PPI in rats, the rodent PPI models have evolved into at least four distinct models.21 These models have PPI measures in common, but are differentiated by the manipulations Inhibitors,research,lifescience,medical used to disrupt PPI: (i) psychostimulant dopamine agonists; (ii) hallucinogenic serotonin agonists; (iii) psychotomimetic N-methyl-D-aspartate (NMDA) receptor antagonists; and (iv) developmental manipulations, such as isolation rearing or neonatal lesions of the ventral hippocampus.

Inhibitors,research,lifescience,medical The first three models are based on changes induced by acutely administered psychotomimetic drugs. While pharmacological approaches that alter PPI help identify relevant neural substrates, they do not assess environmental or developmental contributions to PPI deficits. In contrast, the fourth PPI model is based on the loss of PPI in adult rats subsequent to social isolation during

development.22 Although this isolation rearing model has proven to be of value in Selleck GSK1349572 testing antipsychotic treatments,23 only the dopamine and Inhibitors,research,lifescience,medical NMDA models are particularly relevant for the present discussion. The dopamine PPI mafosfamide model As reviewed in detail elsewhere,21 PPI disruptions that mimic those seen in schizophrenia were first produced in animals by the administration of direct or indirect dopamine agonists, such as apomorphine or d-amphetaminc.24 ‘Ihc original dopamine model focused primarily on testing the ability of antipsychotic drugs to block the PPIdisruptive effects of apomorphine in rats.25 In brief, these effects of apomorphine in rats are reliably prevented by virtually all antipsychotics that have appreciable affinity for dopamine D2 receptors. There is an excellent correlation between the clinical potency of an antipsychotic and its ability to block the PPI -disruptive effects of the dopamine agonist apomorphine in rats.

In addition, up to 200 patients diagnosed as extreme risk but whose iliofemoral anatomy precludes placement of an 18-Fr sheath will undergo either an axillary or direct aortic approach described below. The CoreValve US Pivotal Trial includes 790 high-risk patients deemed to have an estimated 30-day mortality

of between 10% and 15% due to the presence Inhibitors,research,lifescience,medical of comorbidities. Patients are assigned in 1:1 fashion to either TAVR or to sAVR. The primary endpoint, 1-year all-cause mortality, will assess the noninferiority of TAVR with sAVR. Up to 20% of patients can be treated using a noniliofemoral approach. Patients with significant residual coronary artery disease are excluded as coronary artery bypass surgery is allowed at the time of sAVR. An important aspect of these studies is the inclusion of patients who are treated with an alternative noniliofemoral access route. In patients with a minimal lumen iliofemoral diameter of <6.0 mm Inhibitors,research,lifescience,medical in a noncalcified vessel and <7.0 mm in a calcified vessel, those with aneurysmal dilatation of the abdominal aorta or with prior surgical or percutaneous aneurysm repair will be treated using either the subclavian (axillary) or direct aortic approaches.26-30 The ADVANCE Registry was a prospective, multicenter, observational study in 1,015 patients undergoing TAVR with CoreValve

Inhibitors,research,lifescience,medical in Europe. CoreValve implantation was performed in 996 patients. In a preliminary report of this registry, the primary endpoint, a composite of major adverse cardiac and cerebrovascular

events at 30 days, Inhibitors,research,lifescience,medical occurred in 8.3% of patients, with a 30-day all-cause mortality rate of 4.3%.31 Intermediate-Risk Patients With the noninferiority of TAVR demonstrated in patients at high-risk for sAVR, there is general interest in expanding the clinical trial portfolio to include lower-risk patients (Figure 1). An STS-PROM >4 comprises the highest 25% risk of patients currently undergoing sAVR, and an STS-PROM >3 LBH589 research buy identifies the highest Inhibitors,research,lifescience,medical 33% risk.17 Two studies have been designed to address this population of intermediate-risk patients. Figure 1. Spectrum of surgical risk in patients with aortic stenosis. The PARTNER II Cohort A Trial is a noninferiority study of up to 2,000 patients with severe, symptomatic aortic valve stenosis who have an elevated risk for traditional open-heart surgery (STS-PROM ≥4).17 Patients without coronary artery disease will be randomly assigned to TAVR (SAPIEN XT) or sAVR.17 Patients with all coronary artery disease will be randomly assigned to TAVR (SAPIEN XT), percutaneous coronary intervention or sAVR, and coronary artery bypass graft surgery. Those undergoing TAVR will be treated with either a transfemoral or transapical approach. The primary endpoint to be evaluated is a composite of death and major stroke at 2 years, with secondary endpoints that include valve performance and quality-of-life indicators.

No associations were found with dementia or AD. It is important to note that this analysis was based upon self-reports of health habits rather than clinical assessment of health status.

Exercise and other physical activity interventions have been shown to improve cognition in older adults. In a randomized trial, Hassmen et al262 found that participants randomly assigned to an exercise group (regular walking, three times a week for 3 months) exhibited Inhibitors,research,lifescience,medical significantly better performance than controls on complex cognitive tasks following the intervention. Most recently, there has been an increased focus on the role of cognitive activity and social engagement in maintaining good cognitive function with age. Investigators of the Victoria Longitudinal Study mTOR inhibitor examined the hypothesis that maintaining intellectual engagement through participation in everyday activities buffers individuals against cognitive decline in later life.263 In a longitudinal study, they examined the relationships among changes in lifestyle variables and cognition. Decreases in intellectually Inhibitors,research,lifescience,medical related activities were associated with decline in cognitive functioning. However, as the investigators point out, while their findings suggest, that intellectually engaging activities buffer against cognitive decline, an alternative Inhibitors,research,lifescience,medical explanation is that the pursuit

of intellectually active Inhibitors,research,lifescience,medical lives may be confounded with educational level and socioeconomic status, such that individuals pursuing such activities throughout their life span continue to do so until cognitive decline in old age limits these activities. Still other investigators have suggested that, social engagement, defined as the maintenance of

many social connections and a high level of participation in social activities, guards against cognitive decline in elderly persons. Bassuk et al264 examined the relationship between a global social disengagement scale, Inhibitors,research,lifescience,medical which included information on presence of a spouse, monthly visual contact with three or more relatives or friends, yearly nonvisual contact, Rolziracetam with relatives or friends, attendance at religious services, group membership, and regular social activities, and cognitive performance as assessed by the Short, Portable Mental Status Questionnaire. These investigators found that individuals with minimal social ties were at increased risk for cognitive decline, and suggested that social disengagement may be a risk factor for cognitive impairment among elderly persons. As with intellectual activities, it is difficult, to know whether lower levels of social engagement reflect rather than precipitate cognitive decline. Further studies are required to more fully address these issues. Current issues Many of the same concerns that impact, our interpretation of clinical trials in AD, also limit our interpretation of similar approaches in nondemented populations.

“Sporadic Late-onset Nemaline

Myopathy” (SLONM) Clinical Manifestations: SLONM affects both sexes equally at ages ranging from 20 to 50. The most common clinical syndrome is one of proximal limb weakness of subacute onset and progression, sometimes severe and disabling. The neck extensors may be affected, resulting in the dropped head syndrome. Dysphagia and respiratory failure may appear. Tendon reflexes are usually absent. Fasciculation is not often seen but may be. Sometimes Inhibitors,research,lifescience,medical there is evidence of a second muscle disorder, which may be dermatomyositis or polymyositis. Progressive external ophthalmoplegia was reported in two patients (5, 6). The association with HIV was first Inhibitors,research,lifescience,medical described by Dalakas and associates (7). It is not clear how often patients with nemaline disease are HIV-positive. Laboratory Findings: Diagnostic studies are usually indicative of myopathy but sometimes show evidence of denervation. By definition, the muscle biopsy must show the deposits, which are seen as dark red. At the Mayo Clinic, Chahin et al. (2) examined 3-μm-thick frozen sections stained trichromatically or immunostained

for α-actinin or myotilin. Electron microscopy in 12 cases identified the rods in all and revealed additional structural abnormalities. CK values were normal or low. Seven of their 14 patients had monoclonal gammopathy and were followed for 1 to 5 years; five died Inhibitors,research,lifescience,medical of respiratory failure. Five patients without monoclonal gammopathy were followed for 4 to 23 years and none died of the disease. The presence of gammopathy therefore may be ominous. HIV was excluded in 3 of 6 patients and the other 3 were deemed to have no HIV risk factors. In other Inhibitors,research,lifescience,medical reports, a patient with monoclonal gammopathy had features of both nemaline disease and “trabecular” or “lobulated” muscle fibers as well as biclonal gammopathy (8). Another patient was being treated for dermatomyositis when muscle biopsy

showed nemaline rods (9). Therapy: Immunosuppressive Inhibitors,research,lifescience,medical therapy with melphalan, intravenous immunoglobulins (IVIG) or both may be helpful (2, 10-12) . Prednisone is sometimes effective (13, 14) but was uniformly ineffective in the Mayo report (2). buy SB203580 Autologous stem therapy Suplatast tosilate has also brought benefit (15, 16). Long term immunosuppression with rituximab may be considered. Physical therapy is also indicated to maintain gait and general strength. HIV and Motor Neuron Diseases In 1985, the fourth year of the emerging AIDS epidemic, Hoffman et al. (17) described a 26 year old man with both upper and lower motor neuron signs. He was still alive a year later. Many feared there would be an epidemic of viral ALS. However, that fear was never realized. By mid-2002, there had been reports of 19 patients with motor neuron disorders, with no evidence that HIV infection increases the likelihood of developing ALS. 13 of the 19 clearly had a disorder that was unlike ALS in one major way, the rapidity of progression.

8, containing 4% (w/v) SDS, 10% (w/v) glycerol, 5% (v/v) 2-mercaptoethanol and 0.002% (w/v) bromophenol blue] and then boiled for 5 min. SDS-polyacrylamide gel electrophoresis (SDS-PAGE, 10%) and subsequent gel staining with coomassie blue were used for detection of protein expression. The fusion protein was purified from IPTG-induced bacteria in denaturing conditions via a standard nickel resin purification protocol (Qiagen, Valencia, CA). In-gel digestion with trypsin and protein identification via nano-liquid chromatography–linear ion trap quadrupole mass spectrometry (Nano-LC–LTQ-MS) analysis (Thermo Electron Corp., Waltham, MA) were performed following the protocols described previously

[24]. After IPTG induction, E. coli harboring the expression vector with inserted FomA gene [E. Sotrastaurin nmr coli BL21(DE3) FomA] were spread on a sterilized surface and irradiated with UV at total

energy of 7000 J/m2 by an UV cross-linker (Spectronics, Westbury, NY). The viability of UV-irradiated E. coli was determined by observing the growth of bacterial colonies on LB agar plates. For immunization, female ICR (Institute of Cancer Research) mice (3–6 weeks old; Harlan, Indianapolis, IN) were intranasally immunized by inoculating 25 μl of UV-irradiated E. coli BL21(DE3) FomA (108 CFU) into the nasal cavity of each mouse for 9 weeks at a 3-week interval. The second and third inoculations were administered ATM inhibitor in the same manner as the first immunization. Mice immunized with an UV-irradiated E. coli harboring expression vector for green fluorescence protein (GFP) [E. coli BL21(DE3) GFP] (108 CFU) served as a control group. The concentrations of purified recombinant FomA and GFP were determined

by a Bradford crotamiton assay (Bio-Rad, Hercules, CA). The sample (25 μg) was electrophoresed in a 10% (w/v) SDS-PAGE and electrophoretically transferred onto a polyvinylidene fluoride (PVDF) membrane (Millipore, Billerica, MA) for 90 min at a current of 75 V. The membrane was pre-incubated in Tris-buffered saline [with 0.1% (v/v) Tween 20] containing 5% (w/v) skim milk, and then incubated at 4 °C overnight with serum (1:1000 dilution) obtained from mice immunized with UV-irradiated E. coli BL21(DE3) FomA or GFP for 9 weeks. Bound antibodies (IgG) were detected with anti-mouse horseradish peroxidase (HRP)-conjugated IgG (1:5000 dilution, Promega, Madison, WI). The peroxidase activity was developed with a western lighting chemiluminescence kit (PerkinElmer, inhibitors Boston, MA). To induce gum swelling and abscesses, the immunized mice were inoculated with live bacteria as previously described [25]. Briefly, an aliquot of 100 μl of live F. nucleatum (4 × 108 CFU/2 ml in PBS), P. gingivalis (103 CFU/1 ml in PBS) or F. nucleatum plus P. gingivalis (4 × 108 CFU plus 103 CFU/3 ml in PBS) were suspended in 100 μl of PBS, and then inoculated into the oral cavities of immunized mice everyday for 3 days.