Adaptive thermogenesis is essential for survival, and therefore is tightly regulated by a central neural circuit. Here, we show that microRNA (miR)-33 in the brain is indispensable for adaptive ...thermogenesis. Cold stress increases miR-33 levels in the hypothalamus and miR-33
mice are unable to maintain body temperature in cold environments due to reduced sympathetic nerve activity and impaired brown adipose tissue (BAT) thermogenesis. Analysis of miR-33
dopamine-β-hydroxylase (DBH)-Cre mice indicates the importance of miR-33 in Dbh-positive cells. Mechanistically, miR-33 deficiency upregulates gamma-aminobutyric acid (GABA)
receptor subunit genes such as Gabrb2 and Gabra4. Knock-down of these genes in Dbh-positive neurons rescues the impaired cold-induced thermogenesis in miR-33
DBH-Cre mice. Conversely, increased gene dosage of miR-33 in mice enhances thermogenesis. Thus, miR-33 in the brain contributes to maintenance of BAT thermogenesis and whole-body metabolism via enhanced sympathetic nerve tone through suppressing GABAergic inhibitory neurotransmission. This miR-33-mediated neural mechanism may serve as a physiological adaptive defense mechanism for several stresses including cold stress.
Purpose
4-Borono-2-
18
Ffluoro-
l
-phenylalanine (
18
FFBPA) synthesized with
18
FF
2
, produced using the
18
O(p, n)
18
F reaction, has been reported for increasing radioactivity. However, a ...dedicated system and complex procedure is required to reuse the costly
18
OO
2
gas; also, the use of
18
FF
2
as a labeling agent reduces the labeling rate and radiochemical purity. We developed a stable and practical method for
18
FFBPA synthesis by combining
18
FF
2
, produced using a
18
OO
2
single-use system, and a
18
FCH
3
COOF labeling agent.
Methods
The produced
18
FF
2
was optimized, and then
18
FFBPA was synthesized. For passivation of the target box, 0.5% F
2
was pre-irradiated in argon. Gaseous products were discarded; the target box was filled with
18
OO
2
gas, and then irradiated (first irradiation). Then, the
18
OO
2
gas was discarded, 0.05–0.08% F
2
in argon was fed into the target box, and it was again irradiated (second irradiation). The
18
FF
2
obtained after this was passed through a CH
3
COONa column, converting it into the
18
FCH
3
COOF labeling agent, which was then used for
18
FFBPA synthesis.
Results
The mean amount of as-obtained
18
FF
2
was 55.0 ± 3.3 GBq and that of as-obtained
18
FCH
3
COOF was 21.6 ± 1.4 GBq after the bombardment. The radioactivity and the radiochemical yield based on
18
FF
2
of
18
FFBPA were 4.72 ± 0.34 GBq and 12.2 ± 0.1%, respectively. The radiochemical purity and molar activity were 99.3 ± 0.1% and 231 ± 22 GBq/mmol, respectively.
Conclusion
We developed a method for
18
FFBPA production, which is more stable and practical compared with the method using
18
OO
2
gas-recycling and
18
FF
2
labeling agent.
Background Micro RNA (miR)-33 targets cholesterol transporter ATP -binding cassette protein A1 and other antiatherogenic targets and contributes to atherogenic progression. Its inhibition or deletion ...is known to result in the amelioration of atherosclerosis in mice. However, mice lack the other member of the miR-33 family, miR-33b, which exists in humans and other large mammals. Thus, precise evaluation and comparison of the responsibilities of these 2 miRs during the progression of atherosclerosis has not been reported, although they are essential. Methods and Results In this study, we performed a comprehensive analysis of the difference between the function of miR-33a and miR-33b using genetically modified mice. We generated 4 strains with or without miR-33a and miR-33b. Comparison between mice with only miR-33a (wild-type mice) and mice with only miR-33b (miR-33a
/miR-33b
) revealed the dominant expression of miR-33b in the liver. To evaluate the whole body atherogenic potency of miR-33a and miR-33b, we developed apolipoprotein E-deficient/miR-33a
/miR-33b
mice and apolipoprotein E-deficient/miR-33a
/miR-33b
mice. With a high-fat and high-cholesterol diet, the apolipoprotein E-deficient/miR-33a
/miR-33b
mice developed increased atherosclerotic plaque versus apolipoprotein E-deficient/miR-33a
/miR-33b
mice, in line with the predominant expression of miR-33b in the liver and worsened serum cholesterol profile. By contrast, a bone marrow transplantation study showed no significant difference, which was consistent with the relevant expression levels of miR-33a and miR-33b in bone marrow cells. Conclusions The miR-33 family exhibits differences in distribution and regulation and particularly in the progression of atherosclerosis; miR-33b would be more potent than miR-33a.
Display omitted
•KUS121 was developed to selectively inhibit the adenosine triphosphatase activity of valosin-containing protein without affecting other cellular functions of valosin-containing ...protein.•KUS121 preserved adenosine triphosphate levels, reduced endoplasmic reticulum stress, and suppressed cell death in H9C2 rat cardiomyoblast cells, treated with tunicamycin or hydrogen peroxide, or cultured in glucose-free medium.•In murine ischemia and reperfusion injury models, KUS121 treatment after reperfusion attenuated the infarcted size and preserves cardiac function by maintaining adenosine triphosphate levels and reducing ER stress.•In porcine ischemia and reperfusion injury models, intracoronary administration of KUS121 also attenuated the infarcted area in a dose-dependent manner.•These results indicated that KUS121 is a promising novel therapeutic agent for myocardial infarction.
No effective treatment is yet available to reduce infarct size and improve clinical outcomes after acute myocardial infarction by enhancing early reperfusion therapy using primary percutaneous coronary intervention. The study showed that Kyoto University Substance 121 (KUS121) reduced endoplasmic reticulum stress, maintained adenosine triphosphate levels, and ameliorated the infarct size in a murine cardiac ischemia and reperfusion injury model. The study confirmed the cardioprotective effect of KUS121 in a porcine ischemia and reperfusion injury model. These findings confirmed that KUS121 is a promising novel therapeutic agent for myocardial infarction in conjunction with primary percutaneous coronary intervention.
The Hippo signaling pathway is involved in the pathophysiology of various cardiovascular diseases. Yes‐associated protein (YAP) and transcriptional enhancer activator domain (TEAD) transcriptional ...factors, the main transcriptional complex of the Hippo pathway, were recently identified as modulators of phenotypic switching of vascular smooth muscle cells (VSMCs). However, the intrinsic regulator of YAP/TEAD‐mediated gene expressions involved in vascular pathophysiology remains to be elucidated. Here, we identified Homeobox A4 (HOXA4) as a potent repressor of YAP/TEAD transcriptional activity using lentiviral shRNA screen. Mechanistically, HOXA4 interacts with TEADs and attenuates YAP/TEAD‐mediated transcription by competing with YAP for TEAD binding. We also clarified that the expression of HOXA4 is relatively abundant in the vasculature, especially in VSMCs. In vitro experiments in human VSMCs showed HOXA4 maintains the differentiation state of VSMCs via inhibition of YAP/TEAD‐induced phenotypic switching. We generated Hoxa4‐deficient mice and confirmed the downregulation of smooth muscle‐specific contractile genes and the exacerbation of vascular remodeling after carotid artery ligation in vivo. Our results demonstrate that HOXA4 is a repressor of VSMC phenotypic switching by inhibiting YAP/TEAD‐mediated transcription.
Synopsis
HOXA4 suppresses vascular smooth muscle cell phenotypic switching and vascular remodeling by inhibiting YAP/TEAD‐mediated transcription.
Unbiased shRNA screen identified HOXA4 as a repressor of YAP/TEAD transcriptional activity.
HOXA4 competes with YAP for binding to TEADs to preserve VSMC differentiation.
Hoxa4‐deficient mice showed exacerbated neointima formation after carotid artery ligation.
HOXA4 suppresses vascular smooth muscle cell phenotypic switching and vascular remodeling by inhibiting YAP/TEAD‐mediated transcription.
4-Borono-2-
Ffluoro-l-phenylalanine (
FFBPA) synthesized with
FF
, produced using the
O(p, n)
F reaction, has been reported for increasing radioactivity. However, a dedicated system and complex ...procedure is required to reuse the costly
OO
gas; also, the use of
FF
as a labeling agent reduces the labeling rate and radiochemical purity. We developed a stable and practical method for
FFBPA synthesis by combining
FF
, produced using a
OO
single-use system, and a
FCH
COOF labeling agent.
The produced
FF
was optimized, and then
FFBPA was synthesized. For passivation of the target box, 0.5% F
was pre-irradiated in argon. Gaseous products were discarded; the target box was filled with
OO
gas, and then irradiated (first irradiation). Then, the
OO
gas was discarded, 0.05-0.08% F
in argon was fed into the target box, and it was again irradiated (second irradiation). The
FF
obtained after this was passed through a CH
COONa column, converting it into the
FCH
COOF labeling agent, which was then used for
FFBPA synthesis.
The mean amount of as-obtained
FF
was 55.0 ± 3.3 GBq and that of as-obtained
FCH
COOF was 21.6 ± 1.4 GBq after the bombardment. The radioactivity and the radiochemical yield based on
FF
of
FFBPA were 4.72 ± 0.34 GBq and 12.2 ± 0.1%, respectively. The radiochemical purity and molar activity were 99.3 ± 0.1% and 231 ± 22 GBq/mmol, respectively.
We developed a method for
FFBPA production, which is more stable and practical compared with the method using
OO
gas-recycling and
FF
labeling agent.
The effects of sodium nitroprusside (SNP) on cerebral blood flow and glucose metabolism were investigated by the microinfusion of SNP into rat cerebral cortex. A significant enhancement in glucose ...metabolism, as measured using 14Cdeoxyglucose (DG), was observed throughout widespread areas of the cerebral cortex within 1 h of microinjection of 50 nmol/microl SNP. Using a kinetic analysis, the increase in glucose metabolism was found to be due to an increase in the phosphorylation of 14CDG in the brain. On the other hand, regional cerebral blood flow, as measured using 14Ciodoantypirine, was not significantly altered by the SNP infusion. No significant cell death was detected by 2,3,5-triphenyltetrazolium chloride (TTC) staining 1 h after the SNP infusion. The uncoupling of flow and metabolism was almost completely prevented by pretreatment with an NMDA antagonist, MK-801. However, pretreatment with MK-801 did not prevent the SNP-induced neural cell death detected 6 h after the SNP infusion. These results suggest that the SNP-induced uncoupling of flow and metabolism was not directly related to neural cell death in the cerebral cortex.
Nonalcoholic steatohepatitis (NASH) can lead to cirrhosis and hepatocellular carcinoma in their advanced stages; however, there are currently no approved therapies. Here, we show that microRNA ...(miR)-33b in hepatocytes is critical for the development of NASH. miR-33b is located in the intron of sterol regulatory element-binding transcription factor 1 and is abundantly expressed in humans, but absent in rodents. miR-33b knock-in (KI) mice, which have a miR-33b sequence in the same intron of sterol regulatory element-binding transcription factor 1 as humans and express miR-33b similar to humans, exhibit NASH under high-fat diet feeding. This condition is ameliorated by hepatocyte-specific miR-33b deficiency but unaffected by macrophage-specific miR-33b deficiency. Anti-miR-33b oligonucleotide improves the phenotype of NASH in miR-33b KI mice fed a Gubra Amylin NASH diet, which induces miR-33b and worsens NASH more than a high-fat diet. Anti-miR-33b treatment reduces hepatic free cholesterol and triglyceride accumulation through up-regulation of the lipid metabolism-related target genes. Furthermore, it decreases the expression of fibrosis marker genes in cultured hepatic stellate cells. Thus, inhibition of miR-33b using nucleic acid medicine is a promising treatment for NASH.
AIM:To assess the diagnostic ability of endoscopic ultrasonography(EUS)for evaluating causes of dista biliary strictures shown on endoscopic retrograde chol angiopancreatography(ERCP)or magnetic ...resonance cholangiopancreatography(MRCP),even without iden tifiable mass on computed tomography(CT). METHODS:The diagnostic ability of EUS was retro spectively analyzed and compared with that of routine cytology(RC)and tumor markers in 34 patients with distal biliary strictures detected by ERCP or MRCP a Dokkyo Medi...
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