Cancer management presents multifarious problems. Triple negative breast cancer (TNBC) is associated with inaccurate prognosis and limited chemotherapeutic options. Betulinic acid (BA) prevents angiogenesis and causes apoptosis of TNBC cells. NIH recommends BA for rapid access in cancer chemotherapy because of its cell-specific toxicity. BA however faces major challenges in therapeutic practices due to its limited solubility and cellular entree. We report lactoferrin (Lf) attached BA nanoparticles (Lf-BAnp) for rapid delivery in triple negative breast (MDA-MB-231) and laryngeal (HEp-2) cancer cell types. Lf association was confirmed by SDS-PAGE and FT-IR analysis. Average hydrodynamic size of Lf-BAnp was 147.7 ± 6.20 nm with ζ potential of -28.51 ± 3.52 mV. BA entrapment efficiency was 75.38 ± 2.70% and the release mechanism followed non-fickian pattern. Impact of Lf-BAnp on cell cycle and cytotoxicity of triple negative breast cancer and its metastatic site laryngeal cancer cell lines were analyzed. Lf-BAnp demonstrated strong anti-proliferative and cytotoxic effects, along with increased sub-G1 population and reduced number of cells in G1 and G2/M phases of the cell cycle, confirming reduced cell proliferation and significant cell death. Speedy intracellular entry of Lf-BAnp occurred within 30 min. Lf-BAnp design was explored for the first time as safer chemotherapeutic arsenals against complex TNBC conditions.
Introduction: Laryngectomy patients undergo voice rehabilitation that requires implantation of trachea-oesophagal speech valves (TESV). Usually, laryngeal cancer patients require insertion of these devices post-operatively to im-prove their quality of life. Implantation of TESV dates back to 1979 by pioneering work of Blom and Singer. There are cases of aspiration of TESV wearer reported, and obstruction of the TESV causes leakage through the valve and is suggested as a main reason for replacement of the device. The dysfunctional failure may be caused by microbial colonization on the valve or physical malfunction and requires immediate replacement is desirable. The aim of this study is to identify the microbial community members of selected TESVs using both culture-independent techniques (Next-generation sequencing) to analyse the microbiota, including unculturable species, and routine microbiology techniques (culture-dependent method) and to obtain representative isolates that can form the basis for experiments to enable increased understanding of the community. Methods: Biofilms were harvested from 16 explanted speech valves from patients visiting the ENT clinic in Freeman Hospital, Newcastle, UK. Routine microbiology techniques (culture-dependent method) including ChromeID® plates and Matrix-Assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) Mass Spectrometry were used for identification of TESV microbiome. Sequencing of the samples was performed at MR DNA (www.mrdnalab.com, USA) on a MiSeq following the manufacturer’s guidelines in order to determine the bacteria and candida composition in the biofilm community. Results: The most frequently isolated fungal species was C. albicans, which was cultured from 11 out of 16 TESVs (79%), followed by five TESVs with C. tropicalis (36%), three TESVs had C. glabrata (21%) and only one TESV contained S. cerevisiae (7%). Interestingly no biofilm communities contained more than two fungal species and 2 TESVs (12%) possessed only bacterial species. There were only 16 species of bacteria cultured and identified by MALDI-TOF MS. This was far lower than the 91 species that were detected by NGS. Species from the genus Lactobacillus were found in 10 of 16 TESVs (63%), the highest frequency of any bacterial genus isolated from TESVs followed by S. aureus found in eight TESVs of 16. S. epidermidis was identified in two TESVs (13%), Streptococcus spp., K. oxytoca and O. anthropi were both identified in five different TESVs, while the gut bacterium E. faecium was found in four TESVs. Only one TESV contained E. coli. Conclusion: TESV biofilm composition was dominated by Candida spp. and occasionally contained other types of eukaryote such as Saccharomycetes. It was not uncommon for more than one Candida species to be present. The biofilms also harboured a mixture of bacteria, with lactic acid producers (Lactobacillus sp. and Streptococcus sp.) normally accompanying Candida sp. in the biofilm.
The association of human papillomavirus (HPV) with juvenile laryngeal papillomatosis has been well documented. We report two cases of juvenile laryngeal papillomatosis and correlated these cases with presence of HPV, p53 and c-erbB-2 proteins. The first case was a one-year-old male patient and the second a six-year-old female patient. Formalin-fixed paraffin-embedded biopsy specimens were tested for the presence of HPV genome by the technique of in situ hybridisation using wide spectrum and type specific biotinylated probes while the immunohistochemical expression of p53 (D07, 1:50) and c-erbB-2 (DAKO A0485, 1:300) proteins were evaluated with commercially available antibodies. Histologically the tumours in both cases showed papillary configuration of squamous papilloma. The first case detected HPV type 6, HPV type 11 and p53 protein expression while the second case showed only HPV type 6. Both cases of HPV showed positive signals confined to the nuclei in the superficial squamous epithelium. The first case showed p53 positivity seen from the basal region up to one third of the epithelium of laryngeal papillomas and the subsequent recent repeat biopsy showed the positivity of p53 had extended throughout the upper layers of the epithelium. Expression of c-erbB-2 protein was not detected in both cases. These findings were similar as in other studies where follow-up of the cases was recommended since they tend to recur.
A 57-year-old male chronic smoker with underlying diabetes mellitus presented with dysphonia associated with cough, dysphagia and reduced effort tolerance of 3 months' duration. Videoendoscope finding revealed bilateral polypoidal and erythematous true and false vocal fold with small glottic airway. The patient was initially treated as having tuberculous laryngitis and started on antituberculous drug. However, no improvement was observed. CT of the neck showed erosion of thyroid cartilage, which points to laryngeal carcinoma as a differential diagnosis. However, the erosion was more diffuse and appeared systemic in origin. The diagnosis of laryngeal perichondritis was made when the histopathological examination revealed features of inflammation, and the tracheal aspirate isolated Pseudomonas aeruginosa The patient made a good recovery following treatment with oral ciprofloxacin.