Recombinant Secreted Antigens from Mycoplasma hyopneumoniae Delivered as a Cocktail Vaccine Enhance the Immune Response of Mice

Mycoplasma hyopneumoniae is the etiological agent of porcine enzootic pneumonia (EP), which is a respiratory disease responsible for huge economic losses in the pig industry worldwide. The commercially available vaccines provide only partial protection and are expensive. Thus, the development of alternatives for the prophylaxis of EP is critical for improving pig health. The use of multiple antigens in the same immunization may represent a promising alternative. In the present study, seven secreted proteins of M. hyopneumoniae were cloned, expressed in Escherichia coli, and evaluated for antigenicity using serum from naturally and experimentally infected pigs. In addition, the immunogenicity of the seven recombinant proteins delivered individually or in protein cocktail vaccines was evaluated in mice. In Western blot assays and enzyme-linked immunosorbent assays, most of the recombinant proteins evaluated were recognized by convalescent-phase serum from the animals, indicating that they are expressed during the infectious process. The recombinant proteins were also immunogenic, and most induced a mixed IgG1/IgG2a humoral immune response. The use of these proteins in a cocktail vaccine formulation enhanced the immune response compared to their use as antigens delivered individually, providing evidence of the efficacy of the multiple-antigen administration strategy for the induction of an immune response against M. hyopneumoniae.     

Older patients with low Charlson score and high-risk prostate cancer benefit from radical prostatectomy

Introduction

The aim of the study was to identify the appropriate level of Charlson comorbidity index (CCI) in older patients (>70 years) with high-risk prostate cancer (PCa) to achieve survival benefit following radical prostatectomy (RP).

Methods

We retrospectively analyzed 1008 older patients (>70 years) who underwent RP with pelvic lymph node dissection for high-risk prostate cancer (preoperative prostate-specific antigen >20 ng/mL or clinical stage ≥T2c or Gleason ≥8) from 14 tertiary institutions between 1988 and 2014. The study population was further grouped into CCI < 2 and ≥2 for analysis. Survival rate for each group was estimated with Kaplan–Meier method and competitive risk Fine-Gray regression to estimate the best explanatory multivariable model. Area under the curve (AUC) and Akaike information criterion were used to identify ideal ‘Cut off’ for CCI.

Results

The clinical and cancer characteristics were similar between the two groups. Comparison of the survival analysis using the Kaplan–Meier curve between two groups for non-cancer death and survival estimations for 5 and 10 years shows significant worst outcomes for patients with CCI ≥ 2. In multivariate model to decide the appropriate CCI cut-off point, we found CCI 2 has better AUC and p value in log rank test.

Conclusion

Older patients with fewer comorbidities harboring high-risk PCa appears to benefit from RP. Sicker patients are more likely to die due to non-prostate cancer-related causes and are less likely to benefit from RP.

     

Immune responses of a chimaeric protein vaccine containing Mycoplasma hyopneumoniae antigens and LTB against experimental M. hyopneumoniae infection in pigs

A recombinant chimaeric protein containing three Mycoplasma hyopneumoniae antigens (C-terminal portion of P97, heat shock protein P42, and NrdF) fused to an adjuvant, the B subunit of heat-labile enterotoxin of Escherichia coli (LTB), was used to immunize pigs against enzootic pneumonia. The systemic and local immune responses, as well as the efficacy of the chimaeric protein in inducing protection against experimental M. hyopneumoniae infection were evaluated. In total, 60 male piglets, purchased from a M. hyopneumoniae-free herd, at 4 weeks of age were randomly allocated to six different experimental groups of 10 animals each: recombinant chimaeric protein by intramuscular (IM) (1) or intranasal (IN) (2) administration, commercial bacterin by IM administration (3), and the adjuvant LTB by IM (4, control group A) or IN (5, control group B) administration. All groups were immunized at 24 and 38 days of age and challenged at 52 days of age. The sixth group that was not challenged was used as the negative control (IN [n = 5] or IM [n = 5] administration of the LTB adjuvant). Compared with the non-challenged group, administration of the chimaeric protein induced significant (P < 0.05) IgG and IgA responses against all individual antigens present in the chimaera, but it could not confer a significant protection against M. hyopneumoniae infection in pigs. This lack of effectiveness points towards the need for further studies to improve the efficacy of this subunit-based vaccine approach.     

Cycloalkyl indole-2-carboxylates as useful tools for mapping the "north-eastern" region of the glycine binding site associated with the NMDA receptor

A novel series of indole-2-carboxylate analogues of GV 150526 (1) in which the terminal phenyl ring belonging to the side chain present in the position C-3 has been replaced with a bridged cycloalkyl group was synthesized and evaluated for its pharmacological profile. Modelling studies on this class of novel glycine antagonist allowed us to identify an asymmetric lipophilic pocket present in the "North-Eastern" region of the pharmacophoric model of the glycine binding site associated to the NMDA receptor. Among the derivatives prepared, 3-[2-(1-adamantylaminocarbonyl)ethenyl]-4,6-dichloroindole-2 -carboxylic acid 6b and 3-[2-(norbornylaminocarbonyl)ethenyl]-4,6-dichloroindole-2-c arboxylic acid 6l were found to be antagonists acting at the strychnine-insensitive glycine binding site, showing nanomolar affinity for the glycine binding site (Ki = 63 and 19 nM, respectively), coupled with high glutamate receptor selectivity (IC50 > 10(-5) M at the NMDA, AMPA, KA binding sites) and high in vivo potency after systemic administration by inhibition of convulsion induced by NMDA in mice.     

Percutaneous ultrasound-guided treatment of shoulder tendon calcifications: Clinical and radiological follow-up at 6 months

IntroductionCalcific tendinitis of the shoulder is a common condition characterized by chronic pain and/or very painful acute episodes. Different treatments are used during painful flare-up, but they are often ineffective. US-guided percutaneous needle aspiration/lavage is proving to be an effective means for eliminating these calcifications.Materials and methodsWe treated 123 consecutive patients (mean age 48 years) with calcific tendinitis of the shoulder. Fifty-five patients had persistent symptoms requiring 2 or more treatments with lavage and intrabursal steroid infiltration. Before and after treatment, US studies were done independently by 2 radiologists with experience in musculoskeletal ultrasound. Results were concordant in over 90% of the cases. Constant Shoulder Scores were calculated before and 6 months after treatment. At 6 months, MRI was performed to identify impingement and/or bursitis.ResultsPost-treatment Constant scores were significantly improved in all 68 patients treated once (Group 1: mean scores 28.6 vs. 81.4) and in 52 of the 55 treated twice or more (Group 2: mean scores 34.1 vs. 71.1) (p < 0.0001 in both cases). Pretreatment Constant scores were similar in patients with and without shoulder impingement on MRI (31.2 vs. 30.9, respectively), but after treatment the impingement group’s scores were significantly higher (82.2 vs. 73.3, respectively; p < 0.001).ConclusionsUS-guided percutaneous needle aspiration/lavage is an effective and economic treatment for calcific tendinitis of the shoulder. Pretreatment MRI should be done to check for impingement since it is often associated with an incomplete response to the first treatment.     

Hyperechoic pseudotumors in segment IV of the liver.

Hyperechoic pseudotumors usually are considered "spared areas" in a fatty liver; they frequently are detected at the fourth hepatic segment close to the portal vein. Over a 3 year period, we observed 14 patients with a hyperechoic pseudotumor in otherwise normal livers; all of these lesions resembled a hyperechoic pseudotumor of the fourth segment with respect to site and morphology. In all cases echographic findings did not significantly change during the follow-up period ranging from 4 to 12 months. Computed tomographic examination was normal in two cases, whereas in the remaining 12 cases the hyperechoic lesion was appreciable as a hypodense area on both direct and dynamic scans; the contrast enhancement was never typical for a malignancy of a hemangioma. Six patients also underwent a color Doppler and power Doppler examination, which never demonstrated intralesional or perilesional abnormalities in the vascular signals. Three patients, who underwent surgery for adenocarcinoma of the large bowel, had intraoperative sonography and sonographically guided biopsy; a hepatic steatosis was diagnosed at histologic examination in all cases. The uniqueness of the cases presented here lies on the finding of focal steatosis at a site where, according to the most credible hypothesis, intracellular deposition of triglycerides is less likely because of possible variation of the regional portal circulation. From a practical point of view it should be emphasized that, in addition to the more frequent hypoechoic pseudolesions, hyperechoic pseudonodular images just anterior to the portal vein can be observed in normal livers; in our experience these lesions should be interpreted as a focal steatosis in an atypical site.