pH sensing in skin tumors: Methods to study the involvement of GPCRs,acid-sensing ion channels and transient receptor potential vanilloid channels

Solid tumors exhibit an inversed pH gradient with increased intracellular pH (pH_i) and decreased extracellular pH (pH_e). This inside-out pH gradient is generated via sodium/hydrogen antiporter 1, vacuolar-type H + ATPases, monocarboxylate transporters, (bi)carbonate (co)transporters and carboanhydrases. Our knowledge on how pH_e-signals are sensed and what the respective receptors induce inside cells is scarce. Some pH-sensitive receptors (GPR4, GPR65/TDAG8, GPR68/OGR1, GPR132/G2A, possibly GPR31 and GPR151) and ion channels (acid-sensing ion channels ASICs, transient receptor potential vanilloid receptors TRPVs) transduce signals inside cells. As little is known on the expression and function of these pH sensors, we used immunostainings to study tissue samples from common and rare skin cancers. Our current and future work is directed towards investigating the impact of all the pH-sensing receptors in different skin tumors using cell culture techniques with selective knockdown/knockout (siRNA/CRISPR-Cas9). To study cell migration and proliferation, novel impedance-based wound healing assays have been developed and are used. The field of pH sensing in tumors and wounds holds great promise for the development of pH-targeting therapies, either against pH regulators or sensors to inhibit cell proliferation and migration.     

Horseshoe Le Fort I osteotomy in combination with endosteal implants — a median-term follow-up study

Fourteen patients with Class VI resorption of the maxilla were treated with horseshoe Le Fort I osteotomy. In 11 cases, the procedure was followed by the placement of endosteal implants. In six patients, simultaneous placement of implants was carried out, while in five patients this was done in a second procedure. Ten patients wore their implant-supported dentures. In one patient, 5/8 implants were lost due to nonintegration. Three patients lost one implant each. The total number of implants placed was 76, and the survival rate of the implants was 88.1%. In the one-step procedure (n=42), the survival rate was 84.8%; in the two-step procedure (n=34), 92.3%. In comparison of the one-step to the two-step procedure, there was no statistically significant difference (P>0.11) between the amount of marginal peri-implant bone loss and the condition of the peri-implant soft tissues as measured 2 years after implantation.     

Voxel-based morphometry studies of personality: issue of statistical model specification--effect of nuisance covariates

There are an increasing number of studies on the localization of personality using voxel-based morphometry. Due to the complex analytic challenge in volumetric studies, the specification and treatment of the nuisance covariate (such as age, gender, and global measures) is currently not consistent. Here, we present a study in which we conducted voxel-based morphometry with Five-Factor Model (FFM) of personality traits (extraversion, neuroticism, openness to experience, agreeableness, and conscientiousness) that aimed to test the influence of NC specification in the determination of the results. In this study, 62 healthy subjects underwent MRI investigation and completed a German version of the FFM personality questionnaire. Voxel-based morphometry was used to investigate the correlation between the FFM personality traits and subtle brain structure. Different NC combinations were used during the model specification. Significant clusters were found only under the condition of some of the NC combinations but not under the others. In addition, we use the structure equation modeling (automated specification search from AMOS) to narrow down the possible choices of NC combinations according to a set of goodness-of-fit indices to identify well-fitted statistic models. As a final step, theoretical implications of the results are discussed, before accepting the selected model.     

Temporal changes in the causes of aortic stenosis: a surgical pathologic study of 646 cases

Among 646 patients with pure aortic stenosis who underwent valve replacement at our institution between 1981 and 1985, the three most frequent causes were calcification of congenitally bicuspid aortic valves (38%), degenerative (senile) calcification of tricuspid aortic valves (33%), and postinflammatory (presumably rheumatic) calcification and fibrosis (24%). Among the 324 patients younger than 70 years of age, calcified bicuspid valves were observed in 50%. In contrast, among 322 patients 70 years of age or older, degenerative calcification accounted for 48% of the stenotic aortic valves. During the 5 years of the study, the relative frequency of postinflammatory disease decreased from 30% to 18%, and that of bicuspid valves decreased from 37% to 33%. In contrast, the relative frequency of degenerative calcification increased from 30% to 46%. Consequently, degenerative (senile) calcification is currently the most common cause of aortic stenosis among patients undergoing valve replacement at our institution. This finding may be related to changes in life expectancy in the general population, alterations in patient referral practices, and an increased willingness of surgeons to operate on older patients. Regardless of cause, the observed temporal changes in etiologic factors for aortic stenosis may indicate a potential source of increasing health-care costs among the elderly population.     

Phosphorylation state–dependent modulation of spinal glycine receptors alleviates inflammatory pain

Diminished inhibitory neurotransmission in the superficial dorsal horn of the spinal cord is thought to contribute to chronic pain. In inflammatory pain, reductions in synaptic inhibition occur partially through prostaglandin E₂- (PGE₂-) and PKA-dependent phosphorylation of a specific subtype of glycine receptors (GlyRs) that contain α3 subunits. Here, we demonstrated that 2,6-di-tert-butylphenol (2,6-DTBP), a nonanesthetic propofol derivative, reverses inflammation-mediated disinhibition through a specific interaction with heteromeric αβGlyRs containing phosphorylated α3 subunits. We expressed mutant GlyRs in HEK293T cells, and electrophysiological analyses of these receptors showed that 2,6-DTBP interacted with a conserved phenylalanine residue in the membrane-associated stretch between transmembrane regions 3 and 4 of the GlyR α3 subunit. In native murine spinal cord tissue, 2,6-DTBP modulated synaptic, presumably αβ heteromeric, GlyRs only after priming with PGE₂. This observation is consistent with results obtained from molecular modeling of the α-β subunit interface and suggests that in α3βGlyRs, the binding site is accessible to 2,6-DTBP only after PKA-dependent phosphorylation. In murine models of inflammatory pain, 2,6-DTBP reduced inflammatory hyperalgesia in an α3GlyR-dependent manner. Together, our data thus establish that selective potentiation of GlyR function is a promising strategy against chronic inflammatory pain and that, to our knowledge, 2,6-DTBP has a unique pharmacological profile that favors an interaction with GlyRs that have been primed by peripheral inflammation.