Effect of oxybutynin and reboxetine on obstructive sleep apnea: a randomized, placebo-controlled, double-blind, crossover trial.

STUDY OBJECTIVES: Although recent investigations combining noradrenergic and antimuscarinic drugs have shown promising short-term results to treat obstructive sleep apnea (OSA), the mid-term effect and optimal dosage remain uncertain. The present study aimed to evaluate the effect of 1 week of 5 mg oxybutynin and 6 mg reboxetine (oxy-reb) on OSA versus placebo. METHODS: We performed a randomized, placebo-controlled, double-blind, crossover trial comparing the effect of 1 week of oxy-reb versus 1 week of placebo on OSA severity. At-home polysomnography was performed at baseline and after each week of intervention. RESULTS: Fifteen participants (male 66.7%) aged 59 [44-62] years (median [interquartile range]) with a mean body mass index of 33.1 ± 6.6 kg/m2 were included. No significant difference in apnea-hypopnea index (AHI) was observed between conditions (estimated marginal means [95% confidence interval] at baseline: 39.7 [28.5-55.3]; oxy-reb: 34.5 [22.7-52.3]; placebo: 37.9 [27.1-52.9]; p = 0.652), but oxy-reb improved average oxygen desaturation (p = 0.016) and hypoxic burden (p = 0.011) and lowered sleep efficiency (p = 0.019) and rapid eye movement sleep (p = 0.002). Moreover, participants reported reduced sleep quality during the week of oxy-reb compared to the week of placebo (4.7 [3.5; 5.9] vs. 6.5 [5.5; 7.5] on a 0-10 visual analogic scale, respectively; p = 0.001). No significant differences in sleepiness, vigilance, and fatigue were observed. No serious adverse events occurred. CONCLUSIONS: Administration of oxybutynin 5 mg and reboxetine 6 mg did not improve OSA severity assessed by AHI, but did alter sleep architecture and sleep quality. Reduced average oxygen desaturation and hypoxic burden were also observed. CLINICAL TRIAL: ClinicalTrials.gov, https://clinicaltrials.gov, NCT04394143.

Finding biomarkers is getting easier.

Single biomarkers are rarely accurate. Even suites of biomarkers can give conflicting results. Ideally potent combinations of variables are isolated which accurately identify specific analytes and their level of toxicity. The search for such combinations can be done by reducing the thousands of candidate variables to the small number necessary for treatment classification. When the key variables are recognized by machine learning (ML) the results are quite surprising, given the apparent failure of other searching methods to produce good diagnostics. Proteins seem especially useful for portable field tests of a variety of adverse conditions. This review shows how ML, in particular artificial neural networks, can find potent biomarkers embedded in any type of expression data, mainly proteins in this article. A computer does multiple iterations to produce sets of proteins which systematically identify (to near 100% accuracy) the treatment classes of interest. Whether these proteins are useful in actual diagnoses is tested by presenting the computer model with unknown classes. Finding the biomarkers is getting easier but there still must be confirmation, by multivariable statistics and with field studies.

Protein expression profiling.

Protein expression profiling is defined in general as identifying the proteins expressed in a particular tissue, under a specified set of conditions and at a particular time, usually compared to expression in reference samples. This information is useful in drug discovery and diagnosis as well as in understanding response mechanisms at the protein level. We may identify all the proteins responding to a particular stimulus and select those whose expression changes most. Or we may isolate significant protein variables and then identify them. These definitive sets of proteins (protein expression signatures; PES) are specific to diseases, toxicants, physical stresses, and to degrees of stress severity. Here we describe a method, based on machine learning, for isolating the sets of proteins, before identifying them by name, which classify accurately the treatment classes in a study. The principle in this chapter is that if proteins associated with known classes of interest can be used to identify unknown classes then the proteins are definitive for diagnosis.The proteins in each class, including controls, are converted to digital data and serve as input to artificial neural network (ANN) models. Multiple two-dimensional electrophoresis (2DE) gel patterns are included in each treatment class. A training subset of digitized individual, not composite, gel images is used to construct an ANN model which is then applied to a test set of images. Successful classification of the unknown (test) data confirms that the variables included in the model are indeed significant in discrimination among the classes. In the study described here the misclassifications were 5% or less using the ANN models. The ANN method seems to be a useful complement to image analysis, described in Chapter "Troubleshooting Image Analysis in 2DE". The reduction in protein variables permits multivariable statistics such as cluster and discriminant analyses.

Phylogenetic analysis of the Neks reveals early diversification of ciliary-cell cycle kinases.

BACKGROUND: NIMA-related kinases (Neks) have been studied in diverse eukaryotes, including the fungus Aspergillus and the ciliate Tetrahymena. In the former, a single Nek plays an essential role in cell cycle regulation; in the latter, which has more than 30 Neks in its genome, multiple Neks regulate ciliary length. Mammalian genomes encode an intermediate number of Neks, several of which are reported to play roles in cell cycle regulation and/or localize to centrosomes. Previously, we reported that organisms with cilia typically have more Neks than organisms without cilia, but were unable to establish the evolutionary history of the gene family. METHODOLOGY/PRINCIPLE FINDINGS: We have performed a large-scale analysis of the Nek family using Bayesian techniques, including tests of alternate topologies. We find that the Nek family had already expanded in the last common ancestor of eukaryotes, a ciliated cell which likely expressed at least five Neks. We suggest that Neks played an important role in the common ancestor in regulating cilia, centrioles, and centrosomes with respect to mitotic entry, and that this role continues today in organisms with cilia. Organisms that lack cilia generally show a reduction in the number of Nek clades represented, sometimes associated with lineage specific expansion of a single clade, as has occurred in the plants. CONCLUSION/SIGNIFICANCE: This is the first rigorous phylogenetic analysis of a kinase family across a broad array of phyla. Our findings provide a coherent framework for the study of Neks and their roles in coordinating cilia and cell cycle progression.

Loss of Bardet Biedl syndrome proteins causes defects in peripheral sensory innervation and function.

Reception and interpretation of environmental stimuli is critical for the survival of all organisms. Here, we show that the ablation of BBS1 and BBS4, two genes mutated in Bardet-Biedl syndrome and that encode proteins that localize near the centrioles of sensory neurons, leads to alterations of s.c. sensory innervation and trafficking of the thermosensory channel TRPV1 and the mechanosensory channel STOML3, with concomitant defects in peripheral thermosensation and mechanosensation. The thermosensory phenotype is recapitulated in Caenorhabditis elegans, because BBS mutants manifest deficient thermosensory responses at both physiological and nociceptive temperatures and defective trafficking of OSM-9, a polymodal sensory channel protein and a functional homolog of TRPV1 or TRPV4. Our findings suggest a hitherto unrecognized, but essential, role for mammalian basal body proteins in the acquisition of mechano- and thermosensory stimuli and highlight potentially clinical features of ciliopathies in humans.

Chlorfenapyr and mallard ducks: overview, study design, macroscopic effects, and analytical chemistry.

The first commercial pesticide derived from a class of compounds known as halogenated pyrroles was registered for use in the United States in 2001. Chlorfenapyr degrades slowly in soil, sediment, and water and is highly toxic to birds. Information on biochemical or histological endpoints in birds is lacking; therefore, a two-year study was conducted to provide information needed to develop diagnostic criteria for chlorfenapyr toxicosis. In the first year, male mallard ducks were fed concentrations of 0, 2, 5, or 10 ppm technical chlorfenapyr or 5 ppm of a formulated product in their diet during a 10-week chronic exposure study. Survival, body weight, feed consumption (removal), behavior, and molt progression were monitored. Feed and liver were analyzed for chlorfenapyr and two metabolites. Five of 10 ducks in the 10-ppm group died, and neurotoxic effects were observed in the 5- and 10-ppm groups. Feed removal increased for ducks receiving chlorfenapyr and body weights of 5- and 10-ppm ducks were reduced. Loss of body fat, muscle atrophy, and bile retention were suggestive of metabolic disruption or a decreased ability to digest and absorb nutrients. Liver and kidney weights and liver and kidney weight/body weight ratios exhibited a positive response to concentrations of chlorfenapyr in the diet. Emaciation and elevated organ weight/body weight ratios are candidates for a suite of indicators of chronic chlorfenapyr exposure. Liver is the preferred tissue for chemical confirmation of exposure.

A NIMA-related kinase, Cnk2p, regulates both flagellar length and cell size in Chlamydomonas.

The cycle of ciliogenesis and ciliary disassembly is coordinated with cell division. In the unicellular alga Chlamydomonas, the two flagella are maintained at constant and equal length during interphase, and are reabsorbed prior to mitosis. We report that the NIMA-related kinase, Cnk2p, is an axonemal protein that affects flagellar length via effects on disassembly rate and also plays a role in the cellular assessment of size prior to committing to mitosis. This is the second NIMA-related kinase shown to affect ciliary function and cell cycle progression in Chlamydomonas. We speculate that members of the NIMA family have evolved nuanced roles to coordinate cilia/cell cycle regulation.

Identification and sequence analysis of six new members of the NIMA-related kinase family in Chlamydomonas.

The NIMA kinases are an evolutionarily conserved protein family with enigmatic roles in the regulation of mitosis. We report six new members of this family in Chlamydomonas, in addition to the previously identified NIMA-related kinase, Fa2p. Chlamydomonas NIMA-related kinases (CNKs) 1-6 were sequenced from subclones generated by RT-PCR using information from EST libraries and the recently sequenced Chlamydomonas genome. Phylogenetic and bioinformatic approaches were used to determine the relationships of the six new members with known members of the NIMA-related kinase family. Although humans express at least eleven NIMA-related kinases, the eukaryotic microbes that have been studied to date express only one or two members of the family. Thus, the discovery that Chlamydomonas expresses a total of at least seven NIMA-related kinases is intriguing. Our analyses suggest that members of this family may play roles in the assembly and function of cilia.

Protein expression signatures: an application of proteomics.

The methods of proteomics, the study of the protein complement of the genome, are applicable to environmental testing. Sets of proteins specific to different stressors can be isolated using computer imaging software. Individual proteins can be identified by mass spectrometry. The Protein Expression Signatures (PES) obtained have potential in diagnosing adverse factors in the environment. The challenge is to demonstrate their feasibility in complex environments. We have shown that PES for three endocrine disrupting compounds in trout (Onchorhynchus mykiss), can be detected in mixed sewage effluent. Other studies support these results. As protein databases expand, identification becomes routine, and capture molecules specific to each protein are developed, the possibility of simple field tests for multiple stressors becomes real.

Zinc concentration effect at the organismal, cellular and subcellular levels in the eastern oyster.

The purpose of this research was to demonstrate a concentration effect of zinc exposure at organismal, cellular and sub-cellular levels in the eastern oyster and to find associated protein expression signatures (PES) for each concentration of zinc. Oysters were exposed to six concentrations of zinc for 48 h in a controlled environment. At the organismal level, fecal material was observed as a measure of physiological health during metal exposures. At the cellular level, lysosomal destabilization was measured using hemolymph. This cellular response was significant only at the highest concentration, when the fecal index was lowest. Protein responses were monitored in the oyster following exposure to zinc. Gill tissue was excised and homogenized, and then analyzed using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and digital image analysis. Protein expression signatures were found to be specific to each concentration. The protein responses were linked to the other biological parameters measured, each of which followed a concentration gradient of zinc.

The role of chemotherapy and radiation in organ-preservation strategies for muscle-invasive bladder cancer.

Radical cystectomy with pelvic lymph node dissection has been accepted as the standard treatment for muscle-invasive bladder cancer. Radiation therapy and chemotherapy are increasingly being implemented in bladder-preservation protocols to provide an alternative treatment to cystectomy. We review experience with radiation and chemotherapy in treating bladder cancer and their use in bladder-preservation protocols. Multimodality organ-sparing treatment strategies offer overall survival rates comparable to radical cystectomy and pelvic lymph node dissection in selected cases. However, bladder-preservation techniques risk local recurrence of potentially aggressive tumors whose long-term effect on cancer-specific survival has not been fully characterized. No improvement in quality of life has clearly been demonstrated with bladder-preservation regimens. Bladder-preservation protocols are costly and require precise coordination of multiple specialists as well as strict, life-long patient compliance. Bladder-preservation protocols should only be performed at tertiary care centers with experience in their administration and be limited to patients desiring an alternative cystectomy or who are not surgical candidates.

DNA damage, histological changes and DNA repair in larval Japanese medaka (Oryzias latipes) exposed to ultraviolet-B radiation.

Cyclobutane dimer formation, photorepair capability and histological damage were compared among four differently pigmented strains of larval Japanese medaka (Oryzias latipes) to determine whether pigmentation modifies the level of UV-B radiation (290-320 nm) inducible damage in these fish. One-day post-hatch medaka were exposed to one of several UV-B fluence rates with or without photoreactivating light for 5 days for 7 h per day. Their DNA was extracted for analysis by ELISA for cyclobutane pyrimidine dimers or the larvae were processed for histological examination. At the higher UV-B fluence rates tested, wild-type melanophore-containing medaka formed significantly more dimers than at least one of the other strains tested. Wild-type medaka also showed significantly less photorepair capability than the white melanophore-lacking medaka. The wild-type larvae had significantly more necrosis than the orange-red melanophore-lacking larvae at the lower UV-B fluence rate tested and at the higher fluence rate used, the wild-type medaka also exhibited significantly more necrosis than the white melanophore-lacking larvae. Of the 19 medaka observed with cellular hyperplasia, six were wild-type. These six individual larvae showed the greatest degree of cellular hyperplasia. Cellular hyperplasia appeared to be greatest at the lowest UV-B fluence rate used. The presence of melanophores in the wild-type medaka may have contributed to an increased level of tissue damage in this strain when compared to the other strains.