Lack of protective efficacy in buffaloes vaccinated with Fasciola gigantica leucine aminopeptidase and peroxiredoxin recombinant proteins

Gene coding for leucine aminopeptidase (LAP), a metalloprotease, was identified in the tropical liver fluke, Fasciola gigantica; that on sequence analysis showed a close homology (98.6%) with leucine aminopeptidase of the temperate liver fluke, Fasciola hepatica. The recombinant leucine aminopeptidase protein was expressed in Escherichia coli. F. gigantica peroxiredoxin, a hydrogen peroxide scavenger and an immunomodulating protein, was also cloned and expressed in E. coli. A vaccination trial in buffaloes was conducted with these two recombinant proteins, with 150 and 300 μg of leucine aminopeptidase and a cocktail of 150 μg each of recombinant leucine aminopeptidase and peroxiredoxin in three groups, respectively. Both Th1- and Th2-associated humoral immune responses were elicited to immunization with these antigens. A challenge study with 400 metacercariae did not show a significant protection in terms of reduction in the worm burden (8.4%) or anti-fecundity/embryonation effect in the immunized groups, as to the non-immunized control animals. Our observations in this buffalo vaccination trial are contrary to the earlier promise shown by leucine aminopeptidase of F. hepatica as a leading candidate vaccine molecule. Identification of leucine aminopeptidase gene and evaluation of the protein for its protective efficacy in buffaloes is the first scientific report on this protein in F. gigantica.     

Safety and results of cryoablation in patients <5 years old and/or <15 kilograms

Current recommendations discourage elective radiofrequency ablation in patients <5 years old and/or weighing <15 kg, primarily because of the greater complication rate. To describe the current use, complications, and immediate outcomes of cryoablation in this patient population, a multicenter retrospective review of all patients <5 years old and/or weighing <15 kg who were treated with cryoablation for arrhythmia was performed. Eleven centers contributed data for 68 procedures on 61 patients. Of those, 34% were elective and 24% (n = 16) were both cryoablation and radiofrequency ablation. The median age and weight at ablation was 3.5 years (range 8 days to 9.9 years) and 15.2 kg (range 2.3 to 23), respectively. Congenital heart disease was present in 23% of the patients. The immediate success rate of cryoablation alone was 74%. No major complications occurred with cryoablation only; however, 2 of the 16 patients who underwent cryoablation and radiofrequency ablation had major complications. Of the 50 patients receiving cryoablation, 8 (16%) had variable degrees of transient atrioventricular block. The recurrence rate was 20% after cryoablation and 30% after cryoablation plus radiofrequency ablation. In conclusion, cryoablation appears to have a high safety profile in these patients. Compared to older and larger patients, the efficacy of cryoablation in this small, young population was lower and the recurrence rates were higher. Cryoablation's effect on the coronary arteries has not been fully elucidated and requires additional research.     

Significant association of combination of OCT4, NANOG,and SOX2 gene polymorphisms in susceptibility and response to treatment in North Indian breast cancer patients

Purpose

Dysregulations of regulatory genes in embryonic stem cells (ESCs) gene polymorphisms may lead to breast cancer cell growth, differentiation, and tumor metastasis.

Methods

Polymorphisms in OCT4 (rs3130932), NANOG (rs11055786), LIN28 (rs4274112), and SOX2 (rs11915160) genes were evaluated for susceptibility in 297 breast cancer females and 273 healthy controls from north Indian population. Response to neo-adjuvant chemotherapy was followed in 128 locally advanced breast cancer patients along with clinicopathological features. Genotyping was done using TaqMan allelic discrimination assays. Statistical analysis was performed using SPSS and multifactor dimensionality reduction (MDR).

Results

For OCT4 gene polymorphism, protective effect of genotypes AC [P _corr = 0.031, OR = 0.63 (0.44–0.91)] and AC+CC [P _corr = 0.031, OR = 0.68 (0.48–0.95)] was seen in patients. However, no association of NANOG, LIN28, and SOX2 gene polymorphisms was found with overall breast cancer susceptibility. Further, significant association of AG+GG genotype [P _corr = 0.021, OR = 6.08 (1.83–20.15)] and G allele [P _corr = 0.021, OR = 3.07 (1.21–7.77)] of rs4274112 polymorphism was seen with positive lymph node. For OCT4, significant association of allele C was seen with patients having negative hormone receptor [P _corr = 0.021, OR = 0.51 (0.29–0.90)], but no association of any of the studied polymorphisms individually was found with response to NACT. On MDR analysis, we found combination of SNPs SOX2 rs11915160, OCT4 rs3130932, and NANOG rs11055786 to be the best interaction model for predicting breast cancer risk [p for permutation test <10^?3, OR = 2.04 (1.43–2.910] and response to NACT [p for permutation test = 0.005, OR = 2.09 (1.24–3.52)].

Conclusion

Combination of genetic variants of ESCs gene may have a profound effect in breast cancer risk and response to NACT.     

Cartilage Support for Fascia Graft in Type I Tympanoplasty

To evaluate the success rate, in terms of anatomical and functional results, in our technique of cartilage support for fascia graft in type I tympanoplasty. Retrospective study of tragal cartilage support for fascia graft in tympanoplasty for large central perforation in 748 patients was carried at an academic institution during January 2004 to March 2012. Patients’ age ranged from 11 to 65 years. 325 (43.4 %) male and 423 (56.6 %) female patients were operated and mean post-operative follow up was of 24 months (range 6–48 months). The inclusion criteria were large central or subtotal perforation, anterior quadrant perforation, anterior tympanosclerotic patch with perforation and revision myringoplasty. Small central perforation, posterior perforations, traumatic perforations and Ossiculplasty were excluded from this study. In this technique, a piece of tragal cartilage carved in semi lunar shape is inserted medial to anterior remnant of tympanic membrane. Temporalis fascia graft is sandwiched between cartilage and anterior remnant of tympanic membrane. Post-operative closure of perforation was noted. Pure-tone average pre and post-operative air-bone gap in dB at 250, 500, 1000, 2000, and 4,000 Hz were compared. The overall success rate of our technique was 98.3 % in terms of graft uptake and within 13.35 ± 5.22 dB of air bone gap closure in terms of hearing improvement. This study reveals that cartilage support for fascia graft in type I tympanoplasty is a highly reliable technique and gives significant improvement in graft take-up and hearing status.     

Etiopathological Study of 100 Patients of Hoarseness of Voice: In a Rural Based Hospital

Hoarseness of voice is one of the commonest symptoms in otolaryngological practice and it indicates diseases ranging from totally benign condition to the most malignant condition. This is a study to know the etiology, predisposing factor, and clinical profile of patients having hoarseness of voice. The study was carried out in the department of ENT, Mahatma Gandhi Institute of Medical Sciences, Sewagram. Patients coming to our OPD were selected. All the patients then underwent detailed history and routine investigations. Stroboscopic evaluation was done to reach to a diagnosis. A total of 100 patients having hoarseness of voice with male to female ratio 1.50:1 were analysed. Age group varied from 12 to 82 years. Largest group comprised of labourer (33 %) followed by house wives (30 %) and vocal paralysis was found in 33 % of the cases. Upper respiratory tract infection (24 %) and smoking (33 %) were found to be the common predisposing factors. Functional disorders were found in 14 % of the cases.     

Optimization, in vitro-in vivo evaluation, and short-term tolerability of novel levofloxacin-loaded PLGA nanoparticle formulation

A novel poly(lactic-co-glycolic acid) (PLGA)-based nanoformulation of levofloxacin was developed for multidrug-resistant tuberculosis with the purpose of achieving sustained release in plasma. After lyophilization of levofloxacin-loaded nanoparticles, the average size, charge, and polydispersity index were 268 ± 18 nm, -10.2 ± 1.5 mV, and 0.15 ± 0.03, respectively. The maximum drug encapsulation efficiency and loading capacity were 36.9 ± 6.1% (w/w) and 7.2 ± 1.2 mg/100 mg nanopowder, respectively. Biphasic extended-release profile was produced in vitro. Scanning electron microscopy and Fourier transform infrared studies showed spherical shape of drug-loaded nanoparticles and no drug-polymer interactions were observed. After single oral administration in mice, levofloxacin-loaded PLGA nanoparticles produced sustained release of levofloxacin for 4 days in plasma against 24 h for free levofloxacin. Levofloxacin was detected in organs (lung, liver, and spleen) for up to 4-6 days in case of levofloxacin-loaded nanoparticles, whereas free levofloxacin was cleared within 24 h. This novel formulation did not show any significant adverse effects on body weight and clinical signs in mice. No treatment-related changes were found in hematological and biochemical parameters and on histopathological evaluation. These results indicate the feasibility of development of an orally efficacious safe formulation of levofloxacin with sustained-release properties.     

Cellular quantitative structure-activity relationship (Cell-QSAR): conceptual dissection of receptor binding and intracellular disposition in antifilarial activities of Selwood antimycins

We present the cellular quantitative structure-activity relationship (cell-QSAR) concept that adapts ligand-based and receptor-based 3D-QSAR methods for use with cell-level activities. The unknown intracellular drug disposition is accounted for by the disposition function (DF), a model-based, nonlinear function of a drug's lipophilicity, acidity, and other properties. We conceptually combined the DF with our multispecies, multimode version of the frequently used ligand-based comparative molecular field analysis (CoMFA) method, forming a single correlation function for fitting the cell-level activities. The resulting cell-QSAR model was applied to the Selwood data on filaricidal activities of antimycin analogues. Their molecules are flexible, ionize under physiologic conditions, form different intramolecular H-bonds for neutral and ionized species, and cross several membranes to reach unknown receptors. The calibrated cell-QSAR model is significantly more predictive than other models lacking the disposition part and provides valuable structure optimization clues by factorizing the cell-level activity of each compound into the contributions of the receptor binding and disposition.