Identifying the subproteome of kinetically stable proteins via diagonal 2D SDS/PAGE

Most proteins are in equilibrium with partially and globally unfolded conformations. In contrast, kinetically stable proteins (KSPs) are trapped by an energy barrier in a specific state, unable to transiently sample other conformations. Among many potential roles, it appears that kinetic stability (KS) is a feature used by nature to allow proteins to maintain activity under harsh conditions and to preserve the structure of proteins that are prone to misfolding. The biological and pathological significance of KS remains poorly understood because of the lack of simple experimental methods to identify this property and its infrequent occurrence in proteins. Based on our previous correlation between KS and a protein's resistance to the denaturing detergent SDS, we show here the application of a diagonal 2D (D2D) SDS/PAGE assay to identify KSPs in complex mixtures. We applied this method to the lysate of Escherichia coli and upon proteomics analysis have identified 50 nonredundant proteins that were SDS-resistant (i.e., kinetically stable). Structural and functional analyses of a subset (44) of these proteins with known 3D structure revealed some potential structural and functional biases toward and against KS. This simple D2D SDS/PAGE assay will allow the widespread investigation of KS, including the proteomics-level identification of KSPs in different systems, potentially leading to a better understanding of the biological and pathological significance of this intriguing property of proteins.     

胸腔镜内乳淋巴结清扫:一种针对乳腺癌内乳淋巴结的分期技术(英文)

Objective  

The aim of our study was to investigate the feasibility and safety of thoracoscopic internal mammary lymphadenectomy as a method to refine and thereby improve nodal staging in breast cancer.     

Laparoscopic hysterectomy in the treatment of endometrial cancer: NCI experience

BackgroundThe standard treatment for women with endometrial cancer is total abdominal hysterectomy and pelvic lymphadenectomy for surgical staging. Total laparoscopic radical hysterectomy (TLH) is an alternative approach providing surgical and patient related advantages to laparoscopy.MethodsTwenty female patients with early stage endometrial cancer were operated upon by TLH and pelvic lymphadenectomy, aiming to assess the safety and efficacy of TLH.ResultsThe mean operative time was 296.8 min conversion to laparotomy was done in one patient due to bleeding from the uterine vessels. The mean blood loss was 517.5 cc. The uterus was removed transvaginally in 18 patients (90%) and via a small Pfannenstiel incision in two patients (10%). The mean number of pelvic lymph nodes retrieval was 21.2. Postoperative bleeding occurred in one patient (5%) which necessitated exploration. One patient (5%) suffered a pulmonary embolism. Four patients (20%) developed pyrexia, and one patient (5%) suffered from a chest infection. One patient (5%) had wound infection. The mean hospital stay was 4.5 days (range 3–10).ConclusionTLH with pelvic lymphadenectomy is a safe and effective approach in the treatment of early endometrial carcinoma.     

Immunoadjuvant capacity of flagellin and mannosamine-coated poly(anhydride) nanoparticles in oral vaccination

Bioadhesive poly(anhydride) nanoparticles coated with mannose (M-NP) or Salmonella Enteritidis derived flagellin (F-NP) were designed to be applied in oral vaccination strategies using ovalbumin (OVA) as antigen model. Nanoparticles formulations (OVA-M-NP, OVA-F-NP and control OVA-NP) were characterized and evaluated in BALB/c mice. OVA-M-NP and OVA-F-NP displayed a size of about 300–400 nm and were efficiently coated with the respective ligand, Systemic and mucosal immune responses reported after S.C. and oral administration, indicated that a single dose of OVA-M-NP and OVA-F-NP, elicited higher and balanced systemic specific antibody responses [IgG1 (Th2-response) and IgG2a (Th1-response)] compared to non-coated ones. In addition, oral immunization using OVA-M-NP or OVAF-NP was able to elicit a higher levels of intestinal secretory IgA compared to S.C. In summary, oral immunization by bioadhesive mannosylated or flagellin nanoparticles demonstrated strong long lasting systemic and mucosal immune responses than the respective non-conjugated vectors.     

In vivo and in vitro adjuvant activities of the B subunit of Type IIb heat-labile enterotoxin (LT-IIb-B5) from Escherichia coli

The pentameric B subunit of the Escherichia coli LT-IIb enterotoxin (LT-IIb-B₅) activates TLR2 signaling in macrophages. Herein we demonstrate that LT-IIb-B₅, in contrast to a TLR2-nonbinding point mutant, induces functional activation of bone marrow-derived dendritic cells and stimulates CD4⁺ T cell proliferation, activities which suggested that LT-IIb-B₅ might function as an adjuvant in vivo. Indeed, in an intranasal mouse immunization model, LT-IIb-B₅ augmented specific mucosal and serum antibody responses to a co-administered immunogen, at levels which were almost comparable to those induced by intact LT-IIb holotoxin, a potent but toxic adjuvant. Therefore, LT-IIb-B₅ displays useful adjuvant properties which, combined with lack of enterotoxicity and relative stability against degradation, may find application in mucosal vaccines.     

Induction Docetaxel and Cisplatin Followed by Weekly Docetaxel and Cisplatin with Concurrent Radiotherapy in Locally Advanced Stage III Non Small Cell Lung Cancer (LA-NSCLC)-A Phase II Study

Purpose: The objective of this phase II study was to document the activity and to evaluate the toxicity of docetaxel and cisplatin as induction chemotherapy followed by concurrent docetaxel and cisplatin with thoracic radiation in locally advanced stage III non small cell lung cancer. Patients and Methods: Twenty-seven patients with stage III locally advanced non-small cell lung cancer received induction chemotherapy with two cycles of docetaxel 75mg/m2 and cisplatin 75mg/m2 D1 every 3 weeks. Patients without disease progress after induction chemotherapy were assigned to concurrent chemoradiotherapy 20mg/m2 docetaxel&25mg/m2 cisplatin administrated on day 1 every week for 6 weeks along with concurrent radiotherapy at a dose of 60Gy in 30 fractions (2 Gy/fraction and 5 fractions per week). The primary endpoint was to determine the overall response rate (ORR), the secondary endpoint was to evaluate time to progression (TTP) and safety profile. Results: After induction chemotherapy, the overall response rate (ORR) was 44.4%, 23 patients without disease progress were assigned to concurrent treatment with an overall response rate of 65%. Median survival time was 17 months, time to progression was 11.5 months and the one-year survival was 58%. Neutropenia was the most common toxicity during induction therapy (26% expressed grade 3-4) whereas esophagitis was the most common toxicity during concurrent phase (17.3% expressed grade 3-4); toxicities were manageable. Conclusion: Induction chemotherapy by docetaxel and cisplatin followed by weekly docetaxel and cisplatin with concurrent thoracic radiation therapy is feasible and tolerable. These results warrant further large randomized studies to document and confirm the effectiveness of this regimen. Key Words: Lung cancer , Docetaxel , Cisplatin , Concurrent chemoradiotherapy.     

Negative ANA-IIF in SLE patients: what is beyond?

Clinical Rheumatology - The antinuclear antibody (ANA) test has high sensitivity in diagnosing and classifying systemic lupus erythematosus (SLE). To describe the immunological pattern of SLE...     

Does cigarette smoking affect the diagnostic reliability of hemoglobin α2δ2 (HbA2)?

Quantitation of hemoglobin alpha 2 delta 2 (HbA2) is a basic and confirmatory test in diagnosing the carrier state of beta-thalassemia. The present study was designed to investigate the effect of cigarette smoking on the diagnostic reliability of HbA2. A total of 2,867 (654 smokers and 2,213 never smokers) male subjects were involved in the present study. The subjects were categorized into three groups according to their laboratory findings: beta-thalassemia minor, iron deficient, and normal groups. Complete blood count (CBC) parameters and HbA2 levels were compared between smokers and never smokers of each group according to the independent-samples t-test using the SPSS program, significance results were reported at P<0.05. The results showed a significant increase in red blood cell (RBC) mass (RBC count and hematocrit [Hct]) and Hb concentration in smokers of all groups; however, no significant differences were reported in the HbA2 level between smokers and never smokers in all groups. It was concluded that cigarette smoking does not affect the diagnostic reliability of the HbA2 test.     

T4 sympathectomy for palmar hyperhidrosis: looking for the right operation

Most surgeons still perform T2 or T2-3 sympathectomy for palmar hyperhidrosis (PH), but both of these treatments can cause severe side effects. Some recent articles advocating T4 sympathectomy have obtained satisfactory results. The aim of this study was to compare the outcomes of 3 different levels of sympathectomy. Between July 2003 and July 2006, we treated 60 patients (20 men and 40 women, mean age 26 years) who suffer from palmar hyperhidrosis by endoscopic thoracic sympathectomy (ETS). Patients were divided into 3 groups according to the level of sympathectomy: ETS2, ETS3, and ETS4 (20 patients in each group). Data were collected by review of medical charts, outpatient clinic notes, and telephone interviews. Patients were asked whether they considered their symptoms to be "cured" or "unchanged." The degree of hand dryness was assessed subjectively. Postoperative complications (wound infection, chest pain, and Horner's syndrome) were assessed. Any occurrence of gustatory sweating, rhinitis, presence and region of reflex compensatory sweating, and recurrence was noted. Patient satisfaction was assessed. Treatment success at follow-up was 90% for the ETS2, 95% for ETS3 patients, and 100% for the ETS4 patients. In the ETS2 and ETS3 groups, a higher rate of overdryness of limbs was observed (35% and 20%, respectively). The compensatory sweating was mild in the ETS4 group, whereas moderate-to-severe reflex sweating was more common in the ETS2 and ETS3 groups. About 40% of ETS2 groups and 25% of ETS3 group patients were unsatisfied with their operation. All patients of the ETS4 group were satisfied with the outcome. In conclusion, ETS4 sympathectomy is an effective method for treating PH and it decreases the rate of compensatory hydrosis (CH).