INTRODUCTION: In hepatitis C virus (HCV)-related mixed cryoglobulinemia (MCG), the nonenveloped HCV core protein (HCV-Cp) is a constituent of the characteristic cold-precipitating immune complexes (ICs). A possible correlation between HCV-Cp, virologic, laboratory, and clinical parameters in both untreated MCG patients and those undergoing specific treatment was explored. METHODS: HCV-Cp was quantified by a fully automated immune assay. Correlations between HCV-Cp and HCV RNA, cryocrit, and virus genotype (gt) were investigated in 102 chronically HCV-infected MCG patients. RESULTS: HCV-Cp concentrations strongly correlated with HCV RNA levels in baseline samples. An average ratio of 1,425 IU and 12,850 IU HCV RNA per picogram HCV-Cp was estimated in HCV gt-1 and gt-2 patients, respectively. This equation allowed us to estimate that, on average, HCV-Cp was associated with the viral genome in only 3.4% of the former and in 35% of the latter group of patients. The direct relation between HCV-Cp and the cryocrit level suggests that the protein directly influences the amount of cryoprecipitate. Although the therapy with rituximab (RTX) as a single agent resulted in the enhancement of HCV-Cp levels, in patients treated with RTX in combination with a specific antiviral therapy (pegylated interferon-α plus ribavirin), the prompt and effective clearance of HCV-Cp was documented. CONCLUSIONS: Our data provide evidence that HCV-Cp has a direct effect on the cold-precipitation process in a virus genotype-dependence in HCV-related MCG patients.

Background: Several  studies  have  shown  a  genetic  role  in  the  pathogenesis  of   many  childhood  psychiatric  disorders.  The  most  common  childhood  psychiatric   disorder  is  the  attention  deficit  hyperactivity  disorder  (ADHD)  and  some  reports   showed  the  co-­occurance  in  ADHD  children  and  Autism  Spectrum  Disorders  (ASD).   Many  investigators  focused  their  attention  on  polymorphisms  affecting  gene  regions   coding  for  dopamine  receptors.  In  this  study  we  evaluate  the  association  of  three   single-­nucleotide  polymorphisms  (SNPs)  with  clinically  significant  level  of  autistic   symptoms  among  children  with  ADHD. Methods: We  enrolled  150  children  who   were  divided  into  four  groups:  children  with  ADHD,  children  with  ASD,  children  with   co-­occurance  of  ADHD/ASD,  and  control  subjects.  We  investigated  rs265975  C/T   (174862195C>T)  for  dopamine  receptor  D1  gene,  rs1076560  C/A  (113283688C>A)   and  rs1079597  G/A  (113296286C>T)  for  dopamine  receptor  D2  gene  utilizing   previous  DNA  extraction  and  amplification,  restriction  enzymes  that  recognized  one   of  two  allelic  variants. Results: Our  results  demonstrated  that  homozygosis  T/T  for   rs265975  had  a  lower  frequency  in  ADHD  patients  compared  to  other  groups,whereas  small  differences  were  seen  in  heterozygosis  C/T.  Both  heterozygosis  C/A   for  rs1076560  and  heterozygosis  G/A  for  rs1079597  showed  higher  frequency  in   ASD  group  with  respect  to  control  children  and  ADHD  patients,  whereas  in   ADHD/ASD  group  a  ratio  3:1  vs  unaffected  people  was  seen.  The  same  trend,  but   with  slight  differences,  was  observed  in  homozygosis  A/A  for  rs1076560  and   rs1079597. Conclusions: These  preliminary  data  pointing  to  differences  between   ADHD/ASD  and  other  groups  must  be  confirmed  and  encourage  us  to  enlarge  our   study  populations.

In seeking more specific biomarkers of the cystic fibrosis (CF) lung inflammatory disease that would be sensitive to antibiotic therapy, we sought to evaluate the gene expression profiles of neutrophils in CF patients before treatment in comparison with non-CF healthy individuals and after antibiotic treatment. Genes involved in neutrophil-mediated inflammation, i.e. chemotaxis, respiratory burst, apoptosis, and granule exocytosis, were the targets of this study. Microarray analysis was carried out in blood and airway neutrophils from CF patients and in control subjects. A fold change (log) threshold of 1.4 and a cut-off of p,0.05 were utilized to identify significant genes. Community networks and principal component analysis were used to distinguish the groups of controls, pre- and post-therapy patients. Control subjects and CF patients before therapy were readily separated, whereas a clear distinction between patients before and after antibiotic therapy was not possible. Blood neutrophils before therapy presented 269 genes down-regulated and 56 up-regulated as compared with control subjects. Comparison between the same patients before and after therapy showed instead 44 genes downregulated and 72 up-regulated. Three genes appeared to be sensitive to therapy and returned to ‘‘healthy’’ condition: phorbol-12-myristate-13-acetate-induced protein 1 (PMAIP1), hydrogen voltage-gated channel 1 (HVCN1), and b-arrestin 1 (ARRB1). The up-regulation of these genes after therapy were confirmed by real time PCR. In airway neutrophils, 1029 genes were differentially expressed post- vs pre-therapy. Of these, 30 genes were up-regulated and 75 down-regulated following antibiotic treatment. However, biological plausibility determined that only down-regulated genes belonged to the gene classes studied for blood neutrophils. Finally, it was observed that commonly expressed genes showed a greater variability in airway neutrophils than that found in blood neutrophils, both before and after therapy. These results indicate more specific targets for future interventions in CF patients involving respiratory burst, apoptosis, and granule exocytosis.

Background: Cystic  fibrosis  (CF)  lung  disease  is  characterized  by  massive   extravasation  of  neutrophils  into  the  airways  that  undergo  apoptosis  and  thereby  do   not  clear  respiratory  infections.  The  surrogate  end-­points  that  describe  this  process   and  the  effect  of  antibiotic  therapy,  such  as  spirometry,  are  not  sensitive  and  non-­   specific.  We  sought  to  evaluate  the  gene  expression  profile  of  circulating  neutrophils   in  CF  patients  before  and  after  antibiotic  treatment. Methods: Microarray  analysis   (28,869  genes,  Affymetrix  GeneChip  Gene  1.0  ST  Array  System)  was  performed  in   blood  neutrophils  from  10  CF  patients  before  and  after  treatment  for  clinical   exacerbation  with  antibiotics  and  7  healthy  control  subjects. Results: Blood   neutrophils  before  therapy  presented  293  down-­regulated  genes  and  57  up-­ regulated  genes  as  compared  with  control  subjects  (considering  as  cut-­off  P  <  0.05   by  ANOVA).  Comparison  between  the  same  patients  before  and  after  therapy  (with   the  same  cut-­off  by  paired  t  test)  showed  instead  that  1,422  genes  were  down-­ regulated  and  282  up-­regulated  following  antibiotic  treatment.  Interestingly,  three   genes  appeared  to  be  sensitive  to  therapy  and  returned  to  “healthy”  condition:   phorbol-­12-­myristate-­13-­acetate-­induced  protein  1  (PMAIP1),  hydrogen  voltage-­ gated  channel  1  (HVCN1),  and  dom-­3  homolog  Z  (DOM3Z).  The  up-­regulation  of   these  genes  after  therapy  were  confirmed  by  RT -­PCR  in  blood  neutrophils  (n=5)  and   in  sputum  neutrophils  obtained  from  the  same  patients  (n=7). Conclusions: These   results  suggest  the  feasibility  of  investigating  novel  biomarkers  of  therapeutic   efficacy  by  a  global  gene-­wide  platform  and  indicate  more  specific  targets  for  future   interventions  involving  respiratory  burst  and  apoptosis.

Current evidence indicates that periodontal disease is frequently due to inappropriate levels of gingival granulocyte functions. Reason of this failure may be the toxic effects of a number of local or systemic exogenous factors, capable of spreading through the gingival crevice environment, and strongly conditioning the granulocyte activities. The wide list includes bacteria and granulotoxic products, hedonistic drugs (mainly tobacco), and chemotherapeutic agents (especially antimicrobials used for preventing or reducing the accumulation of dental plaque). Almost always, their presence induces a time- and/or dose-dependent toxicity.

BACKGROUND: It is not known whether antibiotic therapy for lung disease in cystic fibrosis (CF) has an influence on circulating polymorphonuclear neutrophil (PMN) function and apoptosis. PATIENTS AND METHODS: Blood PMNs were obtained from 14 CF patients before and after antibiotic treatment for an acute exacerbation, and from 10 healthy controls. PMNs were evaluated for production of reactive oxygen species (ROS) by spectrophotometry, of cytokines in the conditioned medium by ELISA, and apoptotic response by cytofluorimetry. RESULTS: ROS and interleukin (IL)-8 were produced at higher levels by CF PMNs pre-therapy than control PMNs under basal conditions. IL-8 levels further increased after therapy. Early apoptotic response was higher in CF PMNs pre-therapy than in control PMNs, and this pattern did not change after antibiotic treatment. CONCLUSIONS: Circulating PMNs are primed in CF acute patients. Further studies are needed to consider PMN-produced IL-8 as a biomarker to evaluate response to antibiotic therapy in CF patients.

OBJECTIVE: Single-nucleotide polymorphisms (SNP) in the interleukin 28B (IL-28B) gene region are strongly predictive of the response of infected patients to antiviral therapy for hepatitis C virus (HCV). We sought to determine the prevalence of SNP IL-28B rs12979860 C/C and non-C/C (C/T plus T/T) genotypes in HCV-related cryoglobulinemic vasculitis (CV), as compared with HCV-positive patients without CV. We also searched for their association with peculiar clinical manifestations of CV and potential influence on the complete response (virological, molecular, and immunological) to the therapy. METHODS: The study cohort comprised 159 and 172 HCV-infected patients with and without CV, respectively, prospectively followed starting from 1990. SNP rs12979860 genotyping was performed by Taq-Man allelic discrimination. In 106 patients (66.6%) with CV, the profile of circulating B cell clonalities was determined as well. All patients with CV were treated with pegylated interferon-α/ribavirin-based antiviral therapy. RESULTS: The T/T IL-28B genotype was more common in patients with CV than in those without (17% vs 8.1%, p = 0.02). In patients with CV, compared with non-C/C variants, the IL-28B C/C genotype was associated with a higher rate of complete response (52.6% vs 39.2%, p = 0.13), whereas a treatment response of 61.4% was demonstrated when solely virological response was considered (p = 0.008). A higher frequency of expanded B cell clonalities in the circulation (84.2% vs 55.9%; p = 0.005), kidney involvement (21% vs 2.9%; p = 0.003), and B cell non-Hodgkin lymphoma (17.5% vs 6.8%; p = 0.048), were also observed. CONCLUSION: In HCV-positive patients with CV, the IL-28B C/C genotype is distinguished biologically by a higher frequency of restriction of B cell response and clinically by a higher risk of cryoglobulinemic nephropathy and B cell malignancies, while acting as an independent predictor of a sustained virological response to antiviral therapy. In addition, we found that IL-28B T/T variant was more prevalent in patients with CV than in those without.

OBJECTIVES: p120ctn is a component of the catenin family. To date, there have only been two studies examining expression levels of p120ctn in oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: Paraffined specimens of 113 OSCCs and 12 of normal mucosa were examined by immunohistochemistry. Frozen samples of 20 OSCCs and 5 of normal mucosa were examined by Western blot (WB). Results were correlated with clinicopathological parameters. Five cell lines were examined by immunofluorescence, immunocytochemistry, and WB to show immunoreactivity and cellular localization of p120ctn. RESULTS: Altered p120ctn expression was observed in 109/113 cases of OSCC. Heterogenous cytoplasmic/nuclear expression was associated with loss of membranous distribution (88/113 cases). Complete loss of expression was noted in 21/113 cases. Increased cytoplasmic expression was evident in all positive cases, without significant correlation among p120ctn staining/pattern and grading/stage. Reduction/absence of p120ctn expression was related to poor prognosis (P < .05). CONCLUSION: p120ctn delocalization/loss of expression could be an independent prognostic marker in OSCC.

This study illustrates the use and efficacy of a combination of pegylated interferon-alpha (Peg-IFN-alpha) and ribavirin (RBV), with or without rituximab (RTX), in hepatitis C virus (HCV)-related mixed cryoglobulinemia (MC). Twenty-two patients with HCV-related MC received Peg-IFN-alpha (2a: 180 mug or 2b: 1.5 mug/kg) weekly plus RBV (1000 or 1200 mg) daily for 48 weeks, and RTX (375 mg/m(2)) once a week for 1 month followed by two 5-monthly infusions (termed PIRR). Fifteen additional patients received Peg-IFN-alpha/RBV with the same modalities as the PIRR schedule. Complete response was achieved in 54.5% (12/22) and in 33.3% (5/15) of patients who received PIRR and Peg-IFN-alpha/RBV, respectively (P < .05). Clearance of HCV RNA and conversion of B-cell populations from oligoclonal to polyclonal in liver, bone marrow, and peripheral blood was maintained for up to 3 years in 10 of 12 (83.3%) and in 2 of 5 (40%) patients receiving PIRR and Peg-IFN-alpha/RBV, respectively (P < .01). Cryoproteins in 22.7% (5/22) of patients with PIRR and in 33.3% (5/15) with Peg-IFN-alpha/RBV persisted despite sustained HCV RNA clearance. No response occurred in remaining 5 patients of both groups. PIRR therapy is well tolerated and more effective than Peg-IFN-alpha/RBV combination in HCV-related MC. Its effect may last for more than 3 years.

Survivin (SVV) is a protein that belongs to the inhibitor of apoptosis proteins (IAP) family and is involved in the G2/M phase progression of the cell cycle as a spindle-associated molecule. The biological features of this protein are well documented and its activity appears to be involved in mitochondria-dependent and -independent antiapoptotic pathways. Overexpression of SVV at the transcriptional and translational level has been associated with cancer, a multifactorial disorder in which the occurrence of a -31G to C polymorphism in the promoter region may significantly contribute to the development of this pathology. To verify this hypothesis, the occurrence of a single nucleotide polymorphism (SNP) in cis-acting cell cycle-dependent elements (CDEs) and in cell cycle homology regions (CHRs) of the survivin TATA-less promoter was investigated. A total of 23 oral squamous cell carcinoma (OSCC) cell lines and normal epithelium-derived normal human epidermal keratinocyte (NHEK) cell lines were analyzed by RFLP and direct DNA sequencing of their promoter region. Furthermore, survivin expression at the transcriptional and translational levels was evaluated in these cells by RT-PCR and Western blotting, respectively. The findings indicate that the presence of a G or C allele is not directly correlated to survivin expression, at the mRNA or at the protein level, at least in the OSCC lines analyzed in this study.