Sperm associated antigen 7 is activated by T3 during Xenopus tropicalis metamorphosis via a thyroid hormone response element within the first intron.

Thyroid hormone (T3) affects many diverse physiological processes such as metabolism, organogenesis, and growth. The two highly related frog species, diploid Xenopus tropicalis and pseudo tetraploid Xenopus laevis, have been used as models for analyzing the effects of T3 during vertebrate development. T3 regulates T3-inducible gene transcription through T3 receptor (TR)-binding to T3-response elements (TREs). We have previously identified sperm associated antigen 7 (spag7) as a candidate T3 target gene that is potentially involved in adult stem cell development and/or proliferation during intestinal metamorphosis. To investigate whether T3 regulates spag7 directly at the transcriptional level via TR, we first conducted qRT-PCR to analyze its expression during natural and T3-induced metamorphosis and found that spag7 was up-regulated during natural metamorphosis in the intestine, tail, brain and hindlimb, peaking at the climax of metamorphosis in all those organs, and upon T3 treatment of premetamorphic tadpoles. Next, we demonstrated that an intronic TRE in spag7, first identified through bioinformatic analysis, could bind to TR in vitro and in vivo during metamorphosis. A dual luciferase assay utilizing a reconstituted frog oocyte transcription system showed that the TRE could mediate promoter activation by liganded TR. These results indicate that spag7 expression is directly regulated by T3 through the TRE in the first intron during metamorphosis, implicating a role for spag7 early during T3-regulated tissue remodeling and resorption.

Furosemide Exposure and Prevention of Bronchopulmonary Dysplasia in Premature Infants.

OBJECTIVE: To evaluate the association between furosemide exposure and risk of bronchopulmonary dysplasia (BPD). STUDY DESIGN: This retrospective cohort study included infants (2004-2015) born at 23-29 weeks gestational age and 501-1249 g birth weight. We compared the demographic and clinical characteristics of infants exposed and not exposed to furosemide between postnatal day 7 and 36 weeks postmenstrual age. We examined the association between furosemide exposure and 2 outcomes: BPD and BPD or death. We performed multivariable probit regression models that included demographic and clinical variables in addition to 2 instrumental variables: furosemide exposure by discharge year, and furosemide exposure by site. RESULTS: Of 37 693 included infants, 19 235 (51%) were exposed to furosemide; these infants were more premature and had higher respiratory support. Of 33 760 infants who survived to BPD evaluation, 15 954 (47%) had BPD. An increase in the proportion of furosemide exposure days by 10 percentage points was associated with a decrease in both the incidence of BPD (4.6 percentage points; P = .001), and BPD or death (3.7 percentage points; P = .01). CONCLUSIONS: More days of furosemide exposure between postnatal day 7 and 36 weeks was associated with decreased risk of BPD and a combined outcome of BPD or death.

Preparation of pyrite concentrate powder from the Thackaringa mine for quantitative phase analysis using X-ray diffraction.

The quantitative phase analysis using X-ray diffraction of pyrite ore concentrate samples extracted from the Thackaringa mine is problematic due to poor particle statistics, microabsorption and preferred orientation. The influence of sample preparation on these issues has been evaluated, with ball milling of the powder found most suitable for accurate and precise quantitative phase analysis. The milling duration and other aspects of sample preparation have been explored, resulting in accurate phase reflection intensities when particle sizes are below 5 µm. Quantitative phase analysis on those samples yielded precise phase fractions with standard deviations below 0.3 wt%. Some discrepancy between the elemental composition obtained using X-ray powder diffraction data and that determined using wavelength-dispersive X-ray fluorescence was found, and is thought to arise from unaccounted for crystalline phase substitution and the possible presence of an amorphous phase. This study provides a methodology for the precise and accurate quantitative phase analysis of X-ray powder diffraction data of pyrite ore concentrate from the Thackaringa mine and a discussion of the limitations of the method. The optimization process reveals the importance of confirming reproducibility on new samples, with as much prior knowledge as possible.

Highly Efficient and Marker-free Genome Editing of Human Pluripotent Stem Cells by CRISPR-Cas9 RNP and AAV6 Donor-Mediated Homologous Recombination.

Genome editing of human pluripotent stem cells (hPSCs) provides powerful opportunities for in vitro disease modeling, drug discovery, and personalized stem cell-based therapeutics. Currently, only small edits can be engineered with high frequency, while larger modifications suffer from low efficiency and a resultant need for selection markers. Here, we describe marker-free genome editing in hPSCs using Cas9 ribonucleoproteins (RNPs) in combination with AAV6-mediated DNA repair template delivery. We report highly efficient and bi-allelic integration frequencies across multiple loci and hPSC lines, achieving mono-allelic editing frequencies of up to 94% at the HBB locus. Using this method, we show robust bi-allelic correction of homozygous sickle cell mutations in a patient-derived induced PSC (iPSC) line. Thus, this strategy shows significant utility for generating hPSCs with large gene integrations and/or single-nucleotide changes at high frequency and without the need for introducing selection genes, enhancing the applicability of hPSC editing for research and translational uses.

Ear pain and its treatment in hypobaric chamber training in the Japan Air Self-Defense Force.

BACKGROUND: We have documented that ear pain is the most prevalent physiologic incident during hypobaric chamber training in the Japan Air Self-Defense Force. Ear pain may increase also in flight in the future because it is closely related to allergic rhinitis. Therefore, it is very important to know the characteristic of ear pain and the efficacy of its treatment. METHODS: The incidence of ear pain was calculated in each training profile from 1990 to 1998. Type III chamber flight records were further analyzed for the characteristics of ear pain: relationship with a trainee occupational category, time of occurrence of ear pain, and efficacy of treatment. RESULTS: Of 17,935 exposures, 740 trainees (4.1%) had ear pain. Of 7,047 trainees, 429 (6.1%) complained of ear pain especially in Type III, totaling 625 times. Fighter pilots and cargo pilots complained of ear pain one twelfth and one third the number of times, respectively, compared with passengers. Of the 625 episodes, 616 (98.6%) occurred during descent. Three kinds of treatment were administered until the pain was relieved in the following order: Valsalva maneuver, Politzer bag, and decompression. The efficacy rates were 35.8, 92.3, and 83.9%, respectively. Only 5 trainees (0.07%) could not complete training due to ear pain despite treatment. CONCLUSION: The combination treatment of Valsalva maneuver, Politzer bag, and decompression is very effective for relieving ear pain encountered during hypobaric chamber training. A health specialist needs to understand ear pain and its treatment in hypobaric environment such as aircraft.

Pilot cerebral oxygen status during air-to-air combat maneuvering.

BACKGROUND: Successful monitoring of in-flight cerebral oxygen status (COS; cerebral hemoglobin concentration changes and oxygenation changes under dynamic flight conditions) was recently achieved using near-infrared spectroscopy (NIRS). In this study, we examined the effects of air-to-air combat maneuvering on COS. METHOD: Six F-15 fighter pilots performed 2-vs.-1 air-to-air combat one to three times in each of eight sorties. We took continuous measurements of the pilots' in-flight COS using a commercial NIRS system. We measured the direct effects of G-forces on COS as evidenced by relative concentrations of oxy- and deoxy-hemoglobin. RESULTS: With respect to the G-levels reached during air combat maneuvering (Gz range of -0.4 to + 9.5), oxyhemoglobin concentration (O2Hb) and tissue oxygenation index (TOI, the ratio of oxygenated to total tissue hemoglobin) decreased with increasing G-forces during aerial combat maneuver (ACM). Maximum changes in relative O2Hb ranged from -4.2 to -26 micromol x L(-1). Subjects' experience as measured by total fighter time was an independent determinant of the magnitude of decrease in relative oxygenation. CONCLUSIONS: 1. Pilots' COS declined with dynamic G-forces experienced under aerial combat conditions. 2. Fighter pilots with more flying hours maintained a higher cerebral oxygen level at the same level of G-forces than pilots with less flying time. 3. NIRS technology in the form of the NIRO-300G has matured for continuous monitoring of in-flight cerebral oxygen status under vigorous field conditions.

Cerebral infarction in a 24-year-old pilot.

Ischemic stroke is a rare event in young adults. We report on a 24-yr-old pilot with cerebral infarction of undetermined etiology, temporally associated with chain smoking. The patient exhibited dysphasia, stupor (confused consciousness), and right facial-nerve palsy. Computed-tomography revealed a low-density area in the left insular cortex. Cerebroangiography showed severe stenosis in a branch of the left middle cerebral artery. After admission, the patient made a rapid and uneventful recovery within 72 h. MRI showed an area of hyperintensity on T2-weighted images 2 mo after the attack. Based on the hyperintense area on FLAIR (fluid attenuated inversion recovery sequence) images obtained in MRI performed 10 mo after the attack, we diagnosed a cerebral infarction. In the Japan Air Self-Defense Force, cerebral infarction is an aeromedically disqualifying condition. However, in the evaluation 2 mo after the attack, differentiation from reversible ischemic neurological deficit was difficult. We discuss the criteria used for diagnosis and the risk factors for cerebral infarction in young adults, as well as the aeromedical disposition of young pilots.

Physiological incidents during 39 years of hypobaric chamber training in Japan.

BACKGROUND: Hypobaric chamber training for military aircrew is very important for flight safety. Since we began hypobaric training in our laboratory in 1960, some trainees have suffered physiological incidents. This study will characterize the physiological incidents during hypobaric chamber training at the Japan Air Self-Defense Force (JASDF). METHODS: All available training records from 1960-1998 were reviewed and the frequency of physiological incidents counted and analyzed. RESULTS: There were 29,677 trainees and 58,454 exposures. Overall frequency of physiological incidents was 6.3%. Physiological incidents included ear pain, paranasal sinus pain, abdominal pain, hypoxia, hyperventilation, joint pain, and toothache. Decompression sickness (DCS-I, simple joint pain only) was rare. In cases of DCS-I, joint pain was easily relieved with controlled descent. During the last three decades, overall prevalence of physiological incidents has gradually increased from 5.3 to approximately 6.1% before 1991, to 6.8-9.9% after 1991. However, prevalence rate showed no change through out the period when ear pain was factored out. The increase in prevalence was entirely due to an increased frequency of ear pain: 3.6 to approximately 4.6% before 1991, and 5.4 to approximately 7.2% after 1991. CONCLUSIONS: DCS has not been a problem in the JASDF hypobaric chamber training experience. The majority of physiological incidents during hypobaric chamber training in JASDF have been ear pain, a minor but frequent obstacle to hypobaric training. The exact cause of the observed increase in frequency of Eustachian tube dysfunction currently remains unclear.

D2-class dopamine receptor inhibition of NMDA currents in prefrontal cortical neurons is platelet-derived growth factor receptor-dependent.

NMDA receptor function is modulated by both G-protein-coupled receptors and receptor tyrosine kinases. In acutely isolated rat hippocampal neurons, direct activation of the platelet-derived growth factor (PDGF) receptor or transactivation of the PDGF receptor by D4 dopamine receptors inhibits NMDA-evoked currents in a phospholipase C (PLC)-dependent manner. We have investigated further the ability of D2-class dopamine receptors to modulate NMDA-evoked currents in isolated rat prefrontal cortex (PFC). We have demonstrated that, similar to isolated hippocampal neurons, the application of PDGF-BB or quinpirole to isolated PFC neurons induces a slow-onset and long-lasting inhibition of NMDA-evoked currents. However, in contrast to hippocampal neurons, the inhibition of NMDA-evoked currents by quinpirole in PFC neurons is dependent upon D2/3, rather than D4, dopamine receptors. In PFC slices, application of both PDGF-BB and quinpirole induced a phosphorylation of the PDGF receptor at the PLCgamma binding and activation site, Tyr1021. The PDGF receptor kinase inhibitor, tyrphostin A9, and the D2/3 dopamine receptor antagonist, raclopride, inhibited quinpirole-induced Tyr1021 phosphorylation. These finding suggest that quinpirole treatment inhibits NMDAR signaling via PDGF receptor transactivation in both the hippocampus and the PFC, and that the effects of quinpirole in these regions are mediated by D4 and D2/3 dopamine receptors, respectively.

Allergic rhinitis and ear pain in flight.

BACKGROUND: Ear pain is the most common physiologic incident in hypobaric chamber training in the Japan Air Self-Defense Force, and the incidence of ear pain has been gradually increasing. There has been a concomitant increase in the incidence of allergic rhinitis in Japan. We hypothesized that the increased incidence of ear pain may be due to the increased incidence of eustachian tube dysfunction associated with allergic rhinitis. OBJECTIVE: To explore whether there is an association between allergic rhinitis and ear pain experienced in hypobaric chamber training. METHODS: We examined 9 years of training records and noted whether there was a seasonal influence on the occurrence of ear pain in 7047 trainees. In addition, we studied the prevalence of allergic rhinitis among trainees during a representative training year using a questionnaire method, noting association between allergic rhinitis and the occurrence of ear pain. RESULTS: Comprehensive review of training records showed 429 (6.1%) of 7047 trainees complained of ear pain. Ear pain occurred more often in spring than the other 3 seasons. The questionnaire data showed 202 (23.0%) of 878 trainees had allergic rhinitis. Trainees with allergic rhinitis complained of ear pain more often in the spring than trainees without allergic rhinitis. CONCLUSIONS: Active allergic rhinitis is closely associated with the occurrence of ear pain in hypobaric chamber training. A similar phenomenon might occur in an actual flight. Special attention should be paid to not only common cold but also allergic rhinitis in both hypobaric chamber training and actual flights.