Vitamin D3 correlates inversely with systemic dendritic cell numbers and bone erosion in chronic rhinosinusitis with nasal polyps and allergic fungal rhinosinusitis

Vitamin D₃ (VD₃) is a steroid hormone that regulates bone health and numerous aspects of immune function and may play a role in respiratory health. We hypothesized that T helper type 2 (Th2) disorders, chronic rhinosinusitis with nasal polyps (CRSwNP) and allergic fungal rhinosinusitis (AFRS) would have VD₃ deficiencies, resulting in increased mature dendritic cells (DCs) and bone erosion. We conducted a retrospective study examining VD₃ levels in patients with AFRS (n = 14), CRSwNP (n = 9), chronic rhinosinusitis without nasal polyps (CRSsNP) (n = 20) and cerebrospinal fluid leak repair (non‐diseased controls) (n = 14) at time of surgery. Circulating immune cell levels were determined by immunostaining and flow cytometric analysis. Plasma VD₃ and immune regulatory factors (granulocyte–macrophage colony‐stimulating factor and prostaglandin E₂) were measured by enzyme‐linked immunosorbent assay. It was observed that CRSwNP and AFRS demonstrated increased circulating DCs, while chronic rhinosinusitis without nasal polyps displayed increased circulating macrophages. CRSwNP and AFRS were to found to have insufficient levels of VD₃ which correlated inversely with circulating numbers of mature DCs, DC regulatory factors and bone erosion. CRSsNP displayed no change in circulating DC numbers or VD₃ status compared to control, but did display increased numbers of circulating macrophages that was independent of VD₃ status. Lastly, VD₃ deficiency was associated with more severe bone erosion. Taken together, these results suggest support a role for VD₃ as a key player in the immunopathology of CRSwNP and AFRS.     

Analgesic requirements and pain experience after caesarean section under neuraxial anesthesia in women with preeclampsia

Objective: We determined pain experience and analgesic usage in women with preeclampsia (PE) after caesarean section (CS). Method: We conducted a one-year retrospective case (PE), control (healthy pregnancy HP) study in women undergoing CS. Results: Sixty-two women were included. Cases received more intrathecal bupivacaine (mean difference 0.4 mg) and in the first six hours postoperatively received (mean ± SD, % or median (interquartile range, IQR)), less oxycodone (11.5 ± 3.9 mg versus 14.3 ± 5.1 mg, p < 0.031), less often received parecoxib (43% versus 100%, p < 0.001), and reported less maximum pain scores (0 (0,5) versus 4 (3,6), p < 0.005). Conclusion: Women with PE received less analgesia and experienced less pain compared to controls.     

The role of genetic breast cancer susceptibility variants as prognostic factors

Recent genome-wide association studies identified 11 single nucleotide polymorphisms (SNPs) associated with breast cancer (BC) risk. We investigated these and 62 other SNPs for their prognostic relevance. Confirmed BC risk SNPs rs17468277 (CASP8), rs1982073 (TGFB1), rs2981582 (FGFR2), rs13281615 (8q24), rs3817198 (LSP1), rs889312 (MAP3K1), rs3803662 (TOX3), rs13387042 (2q35), rs4973768 (SLC4A7), rs6504950 (COX11) and rs10941679 (5p12) were genotyped for 25 853 BC patients with the available follow-up; 62 other SNPs, which have been suggested as BC risk SNPs by a GWAS or as candidate SNPs from individual studies, were genotyped for replication purposes in subsets of these patients. Cox proportional hazard models were used to test the association of these SNPs with overall survival (OS) and BC-specific survival (BCS). For the confirmed loci, we performed an accessory analysis of publicly available gene expression data and the prognosis in a different patient group. One of the 11 SNPs, rs3803662 (TOX3) and none of the 62 candidate/GWAS SNPs were associated with OS and/or BCS at P<0.01. The genotypic-specific survival for rs3803662 suggested a recessive mode of action [hazard ratio (HR) of rare homozygous carriers=1.21; 95% CI: 1.09-1.35, P=0.0002 and HR=1.29; 95% CI: 1.12-1.47, P=0.0003 for OS and BCS, respectively]. This association was seen similarly in all analyzed tumor subgroups defined by nodal status, tumor size, grade and estrogen receptor. Breast tumor expression of these genes was not associated with prognosis. With the exception of rs3803662 (TOX3), there was no evidence that any of the SNPs associated with BC susceptibility were associated with the BC survival. Survival may be influenced by a distinct set of germline variants from those influencing susceptibility.     

Characterisation of the human GFRalpha-3 locus and investigation of the gene in Hirschsprung disease

BACKGROUND—The GDNF family receptor alpha (GFRα) proteins are extracellular cell surface bound molecules that act as adapters in binding of the GDNF family of soluble neurotrophic factors to the RET receptor. These molecules are essential for development of many neural crest derived cell types and the kidney. Mutations in RET and in two members of the GDNF ligand family are associated with Hirschsprung disease (HSCR), a congenital absence of the enteric ganglia. Members of the GFRα family are also candidates for HSCR mutations. One such gene is GFRα-3, which is expressed in the peripheral nervous system and developing nerves.
OBJECTIVE—We have characterised the structure of the human GFRα-3 locus and investigated the gene for sequence variants in a panel of HSCR patients.
METHODS—Long range PCR or subcloning of PAC clones was used to investigate GFRα-3 intron-exon boundaries. A combination of single strand conformation polymorphism (SSCP) analysis and direct sequencing was used to investigate GFRα-3 sequence variants.
RESULTS—GFRα-3 spans eight coding exons and has a gene structure and organisation similar to that of GFRα-1. We identified three polymorphic variants in GFRα-3 in a normal control population, a subset of which also occurred in HSCR patients. We did not detect any sequence variants within the coding sequence of GFRα-3. We found a base substitution in the 5' UTR of GFRα-3, 15 base pairs upstream of the translation start site. A second substitution was identified in intron 4 (IVS4-30G>A) between the splice branch site and the splice acceptor site. The final variant was a 2 base pair insertion within the splice donor consensus sequence of exon 7 (IVS7+4ins GG).
CONCLUSIONS—We did not detect any correlation between variants of GFRα-3 and the HSCR phenotype. Our data suggest that mutations of this gene are not a cause of HSCR.


Keywords: GFRα-3; Hirschsprung disease; RET     

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Background: Homeostatic maintenance and repair of the bladder urothelium has been attributed to proliferation of keratin 5‐expressing basal cells (K5‐BC) with subsequent differentiation into superficial cells. Recent evidence, however, suggests that the intermediate cell layer harbors a population of progenitor cells. We use label‐retaining cell (LRC) methodology in conjunction with a clinically relevant model of uropathogenic Escherichia coli (UPEC)‐induced injury to characterize urothelial ontogeny during development and in response to diffuse urothelial injury. Results: In the developing urothelium, proliferating cells were dispersed throughout the K5‐BC and intermediate cells layers, becoming progressively concentrated in the K5‐BC layer with age. When 5‐bromo‐2‐deoxyuridine (BrdU) was administered during urothelial development, LRCs in the adult were found within the K5‐BC, intermediate, and superficial cell layers, the location dependent upon time of labeling. UPEC inoculation resulted in loss of the superficial cell layer followed by robust proliferation of K5‐BCs and intermediate cells. LRCs within the K5‐BC and intermediate cell layers proliferated in response to injury. Conclusions: Urothelial development and regeneration following injury relies on proliferation of K5‐BC and intermediate cells. The existence and proliferation of LRCs within both the K5‐BC and intermediate cell layers suggests the presence of two populations of urothelial progenitor cells. Developmental Dynamics 243::988–998, 2014. © 2014 Wiley Periodicals, Inc.     

RET Signaling in Endocrine Tumors: Delving Deeper into Molecular Mechanisms

The rearranged during transfection (RET) proto-oncogene encodes a receptor tyrosine kinase that is implicated in the development of endocrine tumors of the thyroid and adrenal glands. In humans, activating RET mutations are found in the inherited cancer syndrome multiple endocrine neoplasia 2 and in sporadic medullary and papillary thyroid carcinomas. The specific type and location of RET mutations are strongly correlated with the disease phenotype and have both diagnostic and prognostic value. Recent advances in the molecular characterization of the RET receptor and its mutants have begun to define the mechanisms underlying the transforming ability of the different RET mutant forms. This information has revealed key functional features of these mutant proteins that distinguish the different clinically recognized mutations and provide clues as to the functional origins of the phenotypes associated with specific RET mutations. The elucidation of molecular mechanisms involved in RET-mediated transformation is a key step in the development of much needed therapeutics that target RET’s oncogenic properties. Recent advances have begun to provide a deeper understanding of the receptor’s function, and dysfunction, in human tumors that may guide this process.     

Cross-reactive CD8+ T cell epitopes identified in US adolescent minorities

Vaccines designed to bring forth CD8+ T cell responses in different racial and ethnic groups will require inclusion of T cell epitopes presented by various MHC class I molecules. This study was designed to identify new CD8+ T cell epitopes in HIV-infected African American and Hispanic youth as well as to determine the frequency of responses to both novel and previously described HIV-1 epitopes in a cohort of racially and ethnically diverse individuals. We found 8 MHC class I-restricted CD8+ T cell epitopes that had not been previously described, another 8 epitopes that were restricted by class I alleles not previously associated with these epitopes, and 8 additional epitopes that have been described previously. In a larger cohort, we demonstrated that 11 (69%) of these 16 newly described immunogens were recognized by individuals of different race or ethnicity. Most HIV-1-specific CD8+ T cell epitopes identified were either novel or restricted by alternative MHC class I alleles. Frequent recognition of several of these CTL epitopes in persons of diverse racial backgrounds bodes well for the development of a broadly reactive HIV-1 vaccine.