Functional Outcomes Following Total Laryngectomy and Pharyngolaryngectomy: A 20-Year Single Center Study.

BACKGROUND: Laryngeal cancer accounts for 1% of all cancers in men and 0.3% of all cancers in women. Pharyngolaryngectomy (TPL) and total laryngectomy (TL) are central surgical techniques in the management of advanced laryngeal malignancies but are associated with significant morbidity. In addition, optimal reconstruction following TPL remains an area of active research. METHODS: Here, we compared speech and swallowing outcomes following circumferential and partial pharyngeal resection alongside total laryngectomy in patients with laryngeal and hypolaryngeal tumors. We performed a systemic analysis of patient demographics, tumor characteristics, treatment modality, and pharyngeal reconstruction technique following TPL and TL, leveraging data collected over a 20-year period at a large tertiary referral center. RESULTS: Analyzing 155 patients the results show circumferential pharyngeal defects and prior radiotherapy have a significant impact on surgical complications. CONCLUSION: Pharyngeal resection carries a substantial risk of incurring impaired speech and swallowing in patients. Moreover, our results support poorer functional outcomes with more radical pharyngeal resections and show a clear trend toward worse swallowing outcomes in salvage surgery.

Rapid identification of highly potent human anti-GPCR antagonist monoclonal antibodies.

Complex cellular targets such as G protein-coupled receptors (GPCRs), ion channels, and other multi-transmembrane proteins represent a significant challenge for therapeutic antibody discovery, primarily because of poor stability of the target protein upon extraction from cell membranes. To assess whether a limited set of membrane-bound antigen formats could be exploited to identify functional antibodies directed against such targets, we selected a GPCR of therapeutic relevance (CCR1) and identified target binders using an in vitro yeast-based antibody discovery platform (AdimabTM) to expedite hit identification. Initially, we compared two different biotinylated antigen formats overexpressing human CCR1 in a 'scouting' approach using a subset of the antibody library. Binders were isolated using streptavidin-coated beads, expressed as yeast supernatants, and screened using a high-throughput binding assay and flow cytometry on appropriate cell lines. The most suitable antigen was then selected to isolate target binders using the full library diversity. This approach identified a combined total of 183 mAbs with diverse heavy chain sequences. A subset of clones exhibited high potencies in primary cell chemotaxis assays, with IC50 values in the low nM/high pM range. To assess the feasibility of any further affinity enhancement, full-length hCCR1 protein was purified, complementary-determining region diversified libraries were constructed from a high and lower affinity mAb, and improved binders were isolated by fluorescence-activated cell sorting selections. A significant affinity enhancement was observed for the lower affinity parental mAb, but not the high affinity mAb. These data exemplify a methodology to generate potent human mAbs for challenging targets rapidly using whole cells as antigen and define a route to the identification of affinity-matured variants if required.

An unusual cause of community-acquired pneumonia.

We present a case of fatal community-acquired pneumonia (CAP) due to Acinetobacter baumannii, which is rarely reported in the northeastern United States. Previously reported cases originate from tropical and subtropical climates, and infection tends to have an aggressive course with a poor outcome. Appropriate antimicrobial therapy is crucial; however, the associated systemic inflammatory response may overwhelm host defenses, especially in patients with certain co-morbidities.

Long-term functional outcomes in surgically treated patients with oropharyngeal cancer.

OBJECTIVES/HYPOTHESIS: As survival rates in oropharyngeal cancer improve, long-term functional outcomes are increasingly important to understand. We report long-term functional outcomes in a cohort of surviving patients with oropharyngeal squamous cell carcinoma treated with primary surgery ± radiotherapy. STUDY DESIGN: Cross-sectional study. METHODS: Patients undergoing primary surgery for oropharyngeal cancer in Oxford, United Kingdom, between 2000 and 2010 were identified. The University of Washington Quality-of- Life and MD Anderson Dysphagia Inventory questionnaires were sent to all patients. Multivariate analysis was performed to determine the relationship between clinical factors and swallowing outcomes. RESULTS: Twenty percent of patients required gastrostomy-tube placement (mean feed duration, 114 days). On multivariate analysis, increased age, advanced T stage, and an open surgical approach were associated with significantly reduced quality-of-life scores. CONCLUSIONS: Mean functional scores were comparable to previously published series of patients treated with primary surgery. Gastrostomy insertion rate was lower than in many previously published studies. Furthermore, specific variables have been identified that are associated with adverse functional outcome.

Increased expression of G protein-coupled receptor kinases in cystic fibrosis lung.

A reduction in airway beta-adrenoceptor density has been reported in cystic fibrosis lung but the mechanism underlying this defect remains unclear. In this study, we have investigated whether the decrease in beta2-adrenoceptor associates with altered G protein-coupled receptor kinase (GRK) levels. We assessed GRK activity by rhodopsin phosphorylation, and beta2-adrenoceptor and GRK at the mRNA and protein levels by Northern and Western blotting in peripheral lung samples from normal donors and patients with cystic fibrosis. GRK activity was significantly increased in peripheral cystic fibrosis lung with parallel increases in GRK2/5 mRNAs and protein expression. Functionally, isoproterenol-stimulated adenylyl cyclase activity was also diminished by 65% in cystic fibrosis lung homogenates. These data suggest that the increase in GRK activity may be one of the mechanisms underlying alterations in the coupling between beta2-adrenoceptor and adenylyl cyclase via G-protein and may thus contribute to the downregulation of beta2-adrenoceptor in cystic fibrosis lung.

vanG element insertions within a conserved chromosomal site conferring vancomycin resistance to Streptococcus agalactiae and Streptococcus anginosus.

Three vancomycin-resistant streptococcal strains carrying vanG elements (two invasive Streptococcus agalactiae isolates [GBS-NY and GBS-NM, both serotype II and multilocus sequence type 22] and one Streptococcus anginosus [Sa]) were examined. The 45,585-bp elements found within Sa and GBS-NY were nearly identical (together designated vanG-1) and shared near-identity over an ~15-kb overlap with a previously described vanG element from Enterococcus faecalis. Unexpectedly, vanG-1 shared much less homology with the 49,321-bp vanG-2 element from GBS-NM, with widely different levels (50% to 99%) of sequence identity shared among 44 related open reading frames. Immediately adjacent to both vanG-1 and vanG-2 were 44,670-bp and 44,680-bp integrative conjugative element (ICE)-like sequences, designated ICE-r, that were nearly identical in the two group B streptococcal (GBS) strains. The dual vanG and ICE-r elements from both GBS strains were inserted at the same position, between bases 1328 and 1329, within the identical RNA methyltransferase (rumA) genes. A GenBank search revealed that although most GBS strains contained insertions within this specific site, only sequence type 22 (ST22) GBS strains contained highly related ICE-r derivatives. The vanG-1 element in Sa was also inserted within this position corresponding to its rumA homolog adjacent to an ICE-r derivative. vanG-1 insertions were previously reported within the same relative position in the E. faecalis rumA homolog. An ICE-r sequence perfectly conserved with respect to its counterpart in GBS-NY was apparent within the same site of the rumA homolog of a Streptococcus dysgalactiae subsp. equisimilis strain. Additionally, homologous vanG-like elements within the conserved rumA target site were evident in Roseburia intestinalis. Importance: These three streptococcal strains represent the first known vancomycin-resistant strains of their species. The collective observations made from these strains reveal a specific hot spot for insertional elements that is conserved between streptococci and different Gram-positive species. The two GBS strains potentially represent a GBS lineage that is predisposed to insertion of vanG elements.

Identification of a Novel and Selective Series of Itk Inhibitors via a Template-Hopping Strategy.

Inhibition of Itk potentially constitutes a novel, nonsteroidal treatment for asthma and other T-cell mediated diseases. In-house kinase cross-screening resulted in the identification of an aminopyrazole-based series of Itk inhibitors. Initial work on this series highlighted selectivity issues with several other kinases, particularly AurA and AurB. A template-hopping strategy was used to identify a series of aminobenzothiazole Itk inhibitors, which utilized an inherently more selective hinge binding motif. Crystallography and modeling were used to rationalize the observed selectivity. Initial exploration of the SAR around this series identified potent Itk inhibitors in both enzyme and cellular assays.