Development of multifunctional pyrrole-imidazole polyamides that increase hepatocyte growth factor and suppress transforming growth factor-ß1.

PURPOSE: The DNA recognition peptide compounds pyrrole-imidazole (PI) polyamides bind to the minor groove and can block the binding of transcription factors to target sequences. To develop more PI polyamides as potential treatments for fibrotic diseases, including chronic renal failure, we developed multifunctional PI polyamides that increase hepatocyte growth factor (HGF) and decrease transforming growth factor (TGF)-ß1. METHODS: We designed seven PI polyamides (HGF-1 to HGF-7) that bind to the chicken ovalbumin upstream promoter transcription factor-1 (COUP-TF1) binding site of the HGF promoter sequence. We selected PI polyamides that increase HGF and suppress TGF-ß1 in human dermal fibroblasts (HDFs). FINDINGS: Gel shift assays showed that HGF-2 and HGF-4 bound the appropriate dsDNAs. HGF-2 and HGF-4 significantly inhibited the TGF-ß1 mRNA expression in HDFs stimulated by phorbol 12-myristate 13-acetate. HGF-2 and HGF-4 significantly inhibited the TGF-ß1 protein expression in HDFs with siRNA targeting HGF, indicating that HGF-2 and HGF-4 directly inhibited the expression of TGF-ß1. CONCLUSION: The designed and synthetic HGF PI polyamides targeting the HGF promoter, which increased the expression of HGF and suppressed the expression of TGF-ß, will be a potential practical medicine for fibrotic diseases, including progressive renal diseases.

Hypolipidemic and Anti-Inflammatory Effects of Curcuma longa-Derived Bisacurone in High-Fat Diet-Fed Mice.

Turmeric (Curcuma longa) contains various compounds that potentially improve health. Bisacurone is a turmeric-derived compound but has been less studied compared to other compounds, such as curcumin. In this study, we aimed to evaluate the anti-inflammatory and lipid-lowering effects of bisacurone in high-fat diet (HFD)-fed mice. Mice were fed HFD to induce lipidemia and orally administered bisacurone daily for two weeks. Bisacurone reduced liver weight, serum cholesterol and triglyceride levels, and blood viscosity in mice. Splenocytes from bisacurone-treated mice produced lower levels of the pro-inflammatory cytokines IL-6 and TNF-α upon stimulation with a toll-like receptor (TLR) 4 ligand, lipopolysaccharide (LPS), and TLR1/2 ligand, Pam3CSK4, than those from untreated mice. Bisacurone also inhibited LPS-induced IL-6 and TNF-α production in the murine macrophage cell line, RAW264.7. Western blot analysis revealed that bisacurone inhibited the phosphorylation of IKKα/ß and NF-κB p65 subunit, but not of the mitogen-activated protein kinases, p38 kinase and p42/44 kinases, and c-Jun N-terminal kinase in the cells. Collectively, these results suggest that bisacurone has the potential to reduce serum lipid levels and blood viscosity in mice with high-fat diet-induced lipidemia and modulate inflammation via inhibition of NF-κB-mediated pathways.

Research activities contributing to evidence-based practice implementation in Japanese rehabilitation professionals.

[Purpose] This study aimed to determine whether certain research activities improve the attitude of rehabilitation professionals towards evidence-based practice and its implementation in Japan. [Participants and Methods] We included physical, occupational, and speech therapists currently working in clinical settings. We employed hierarchical multiple regression analyses to assess the attitude of rehabilitation professionals towards evidence-based practice and research activities. Scores of the five dimensions of the Health Sciences-Evidence Based Practice questionnaire were considered the dependent variables. The five dimensions were as follows: Dimension 1, attitude towards evidence-based practice; Dimensions 2-4, evidence-based practice implementation; and Dimension 5, work environment related to evidence-based practice barriers-facilitators. The four sociodemographic variables (gender, academic degree, clinical experience, and the number of therapists at work) were initially included, following which self-reported research achievements were supplemented as independent variables (the number of case studies, literature reviews, cross-sectional studies, and longitudinal studies). [Results] We analyzed data from 167 participants. In addition to sociodemographic variables, the research achievements that statistically increased F-values of the modeling were case study achievements in Dimensions 2-3, cross-sectional study achievements in Dimensions 2 and 4, and longitudinal study achievements in Dimension 5. [Conclusion] Case studies and cross-sectional studies could improve evidence-based practice implementation among rehabilitation professionals in Japan.

A study on the diagnostic support system of the repetitive brain concussion based on the reconstruction analysis of the accident.

In this paper the effectiveness of the support system which predicts the risk of the repetitive brain concussion is studied biomechanically. In the risk prediction system, the accident that caused the concussion is reconstructed by analyzing the game video via multibody dynamics and the resulting brain injury is calculated in detail by the finite element method. In order to calculate the aggravation of the brain injury by the repeated brain concussion, the following two methods are examined. In the first method, the material properties of the part of the brain damaged by the1st impact are changed in the simulation of the 2nd impact. In the second method, each brain damage caused by the repeated impacts is accumulated. The system was applied to the real-life accidents that occurred during Judo and American football games. As a result of the simulations, the aggravation of the brain damage due to repetitive concussion was determined numerically in terms of the maximum strain of the brain and the brain damage rate of the whole brain. The biomechanical process of the collision accidents and the resulting brain damage were reconstructed based on the video and the results are effective to prevent the future repeated concussion accidents. J. Med. Invest. 70 : 213-220, February, 2023.

Gut-Microbiota Dysbiosis in Stroke-Prone Spontaneously Hypertensive Rats with Diet-Induced Steatohepatitis.

Metabolic-dysfunction-associated fatty-liver disease (MAFLD) is the principal worldwide cause of liver disease. Individuals with nonalcoholic steatohepatitis (NASH) have a higher prevalence of small-intestinal bacterial overgrowth (SIBO). We examined gut-microbiota isolated from 12-week-old stroke-prone spontaneously hypertensive-5 rats (SHRSP5) fed on a normal diet (ND) or a high-fat- and high-cholesterol-containing diet (HFCD) and clarified the differences between their gut-microbiota. We observed that the Firmicute/Bacteroidetes (F/B) ratio in both the small intestines and the feces of the SHRSP5 rats fed HFCD increased compared to that of the SHRSP5 rats fed ND. Notably, the quantities of the 16S rRNA genes in small intestines of the SHRSP5 rats fed HFCD were significantly lower than those of the SHRSP5 rats fed ND. As in SIBO syndrome, the SHRSP5 rats fed HFCD presented with diarrhea and body-weight loss with abnormal types of bacteria in the small intestine, although the number of bacteria in the small intestine did not increase. The microbiota of the feces in the SHRSP5 rats fed HFCD was different from those in the SHRP5 rats fed ND. In conclusion, there is an association between MAFLD and gut-microbiota alteration. Gut-microbiota alteration may be a therapeutic target for MAFLD.

Synthesis and evaluation of a deltic guanidinium analogue of a cyclic RGD peptide.

A guanidine group is abundantly found in natural products and drugs. Guanidine has the highest basicity among many common functional groups in nature. Because of its high basicity, it generally exists as a protonated guanidinium and functions as a cationic hydrogen bond donor. Finding an appropriate bioisostere of guanidinium is challenging because of its high basicity and unique trigonal planar shape. In this study, we explored the possibility of "deltic guanidinium" as a bioisostere of guanidinium using a cyclic arginine-glycine-aspartic acid (RGD) peptide as a parent compound. We synthesized c(deltic RGDyK), in which a guanidinium group of an arginine residue in c(RGDyK) is replaced with deltic guanidinium. A target binding assay, biodistribution study, and metabolic stability assay were conducted with c(deltic RGDyK) and its radioiodinated variant. The deltic guanidinium analog peptides exhibited similar biological properties to the parent peptides and improved in vivo stability, indicating that deltic guanidinium could work as a unique bioisostere of guanidinium.

Possible solutions to enhance evidence-based practice proposed by rehabilitation professionals in Japan: a Delphi study.

[Purpose] We aimed to identify possible solutions to enhance evidence-based practice (EBP) in rehabilitation professionals in Japan. [Participants and Methods] A three-round Delphi method was undertaken among a cohort of clinical therapists (328 physical therapists, 55 occupational therapists, and 6 speech therapists). In the first round, the participants listed possible solutions for promoting EBP, other than 12 solutions presented in a previous study; subsequently, a new list was created. In the second round, a newly-created list of solutions was presented, and the participants responded on a 5-point Likert scale on how much they agreed with the solutions promoting EBP in Japanese rehabilitation professionals. In the third round, the distribution of responses obtained in the second round was presented, and participant's agreement was again assessed on a 5-point Likert scale. [Results] Across the three rounds, data were collected from 33.7% to 47.0% of all eligible participants. After the first round, 17 possible solutions were developed, and a list of 29 solutions was used in the second round. After the third round, 10 solutions reached the predetermined criteria for consensus. [Conclusion] In this study, ten possible solutions to promote EBP were proposed by the Japanese rehabilitation professionals.

Relationship between tissue characteristics and mechanical properties of coronary plaques: a comparison between integrated backscatter intravascular ultrasound (IVUS) and speckle-tracking IVUS.

High-risk coronary plaques have certain morphological characteristics. Thus, comprehensive assessment is needed for the risk stratification of plaques in patients with coronary artery disease. Integrated backscatter intravascular ultrasound (IB-IVUS) has been used successfully used to evaluate the tissue characteristics of coronary plaques; however, the mechanical properties of plaques have been rarely assessed. Therefore, we developed Speckle-tracking IVUS (ST-IVUS) to evaluate the mechanical properties of coronary plaque. This study aimed to evaluate the relation between the tissue characteristics of coronary plaques using IB-IVUS and their mechanical properties using ST-IVUS. We evaluated 95 non-targeted plaques in 95 patients undergoing elective percutaneous coronary intervention to the left anterior descending artery. We set regions of interest (ROIs) in the cross-sectional images of coronary plaques where we divided 120 degree plaques into four quadrants (every 30 degrees), with the center at the area of maximum atheroma thickness. We measured relative calcification area (%CA, relative fibrous area (%FI) and relative lipid pool area (%LP) in a total of 380 ROIs. In ST-IVUS analysis, we measured strain in the circumferential direction of the lumen area (LA strain: %), the external elastic membrane area strain (EEM strain: %), and strain in the radial direction (radial strain: %). On global cross-sectional area IB-IVUS analysis, the %CA was 1.2 ± 1.2%; the %FI was 49.0 ± 15.9%, and the %LP was 49.7 ± 16.5%. In ST-IVUS analysis, the LA strain was 0.67 ± 0.43%; the EEM strain was 0.49 ± 0.33%, and the radial strain was 2.02 ± 1.66%. On regional analysis, the %LP was not associated with the LA strain (r = - 0.002 p = 0.97), the EEM strain (r = - 0.05 p = 0.35), or with the radial strain (r = - 0.04 p = 0.45). These trends were seen between the %FI and the LA strain (r = 0.02 p = 0.74), the %FI and the EEM strain (r = 0.05 p = 0.35), and the %FI and the radial strain (r = 0.04 p = 0.50). A significant correlation was only observed between the %CA and the LA strain (r = - 0.15 p = 0.0038). Our findings indicate that the associations between mechanical properties and tissue characteristics lacked statistical significance, more often than not, and that it is necessary to evaluate the mechanical properties as well as plaque characteristics for risk stratification of coronary plaques.

Impact of directional coronary atherectomy followed by drug-coated balloon strategy to avoid the complex stenting for bifurcation lesions.

Although the simple single stenting rather than complex double stenting is recommended on percutaneous coronary intervention (PCI) for bifurcation lesions, double stenting cannot always be avoided. We investigated the impact of directional coronary atherectomy (DCA), followed by drug-coated balloon (DCB) treatment to reduce the number of stents and avoid complex stenting in PCI for bifurcation lesions and short-term patency. DCA treatment without stents was attempted for 27 bifurcation lesions in 25 patients, of those, 26 bifurcation lesions in 24 patients were successfully treated and 3-month follow-up angiography and optical coherence tomography (OCT) were performed. Sixteen lesions (59.3%) were related to left main trunk distal bifurcations, and 7 (25.9%) were true bifurcation lesions. Among the true bifurcation lesions, 4 lesions (57.1%) needed 1 stent, and the other 3 lesions (42.9%) needed no stents. Among the non-true bifurcation lesions, 1 lesion (5.0%) needed bailout stent and other lesions (95.0%) needed no stents. According to DCA followed by DCB treatment, the angiographic mean diameter stenosis improved from 65.5 ± 15.0% to 7.8 ± 9.8%, and the mean plaque area in intravascular ultrasound improved from 80.4 ± 10.5% to 39.0 ± 11.5%, respectively. Angiographic and OCT late lumen loss values were 0.2 ± 0.6 mm and 1.4 ± 1.9 mm, respectively. No patient had in-hospital major adverse cardiac events (MACE) and 3-month MACE. In conclusion, compared with standard provisional side branch stenting strategy, DCA followed by DCB treatment might reduce the number of stents, avoid complex stenting for major bifurcation lesions and provide good short-term outcomes.

TWIST1 transcriptionally upregulates complement 3 in glomerular mesangial cells from spontaneously hypertensive rats.

We have shown that complement 3 (C3) is upregulated in cardiovascular and renal organs, which induces the synthetic phenotype and exaggerates the growth of mesenchymal cells from spontaneously hypertensive rats (SHRs). However, the mechanisms of the upregulation of C3 have remained unclear. In the present study, we investigated the role of TWIST1, a transcription factor that regulates mesodermal embryogenesis, in the upregulation of C3 in glomerular mesangial cells (GMCs) from SHRs and Wistar-Kyoto (WKY) rats. Immunocytochemical staining and western blot analysis showed that the expression of TWIST1 in GMCs from SHRs was higher than that in GMCs from WKY rats in vivo and in vitro. Real-time PCR analysis showed increases in the expression of Twist1 mRNA with attenuated expression of miR-151-3p in GMCs from SHRs compared to that in cells from WKY rats. Chromatin immunoprecipitation assays showed increases in TWIST1 binding to the C3 promoter in GMCs from SHRs compared to that in cells from WKY rats. Transfection of Twist1 cDNA by a lentiviral vector increased the expression of C3 mRNA in GMCs from WKY rats. TWIST1 siRNA significantly decreased the mRNA expression of C3 and osteopontin in GMCs from SHRs. These results indicate that the increases in TWIST1 expression, attenuation of miR-151-3p, and strong binding of TWIST1 upregulate C3 gene expression in GMCs from SHRs. The enhanced TWIST1-C3 system induces the synthetic phenotype of mesenchymal tissue that may be associated with cardiovascular and renal remodeling in hypertension.

Work productivity in the office and at home during the COVID-19 pandemic: A cross-sectional analysis of office workers in Japan.

The coronavirus disease 2019 (COVID-19) pandemic has drastically changed work styles and environments. Given the coexistence of work in the office and work from home (WFH) in the future, studies are needed to identify ways to increase productivity when working in both places. We conducted a questionnaire survey and environment measurements of 916 workers in 22 offices across 2 weeks in November-December 2020 in Japan. While average workdays at the offices decreased from 4.9 to 3.9 days/week, those at homes increased from 0.1 to 1.1 days/week due to COVID-19, indicating an increase in the relative importance of WFH. Compared to the office, the satisfaction rate was lower for lighting, spatial, and information technology (IT) environments, but higher for thermal, air, and sound environments at home. Although it was easier to concentrate on work and to refresh at home, workers experienced challenges associated with business communication from home. Meanwhile, in the office, satisfaction with COVID-19 countermeasures was significantly associated with work productivity. Furthermore, lower PM2.5 concentration was associated with greater satisfaction with COVID-19 countermeasures, indicating that reducing PM2.5 may increase satisfaction with COVID-19 countermeasures and work productivity. We expect these findings will help improve work productivity in the New Normal era.

[Development of Photocatalysts-Coated Polyethylene Terephthalate Film for Degradation and Elimination of Chemotherapy Drugs].

Prolonged exposure to anticancer drugs can lead to health damage in medical professionals as well as in patients and their families, and can be a serious issue. The risk of occupational acute exposure to anticancer drugs in medical professionals has largely been reduced following the establishment of guidelines for safe handling of hazardous drugs. However, the problem of pollution associated with residual anticancer drugs attached to the surface of floors and walls in the prescription laboratory, treatment room and the lavatory remains unsolved. We have recently developed a polyethylene terephthalate(PET) film coated with photocatalysts such as titanium dioxide and tungsten oxide. In the present study, the ability of the PET film to degrade anticancer drugs was tested by dripping anticancer drug solution on to the surface of the film followed by irradiation with LED light. The contents of several anticancer drugs, including 5-fluorouracil, cyclophosphamide, and cisplatin, were greatly reduced after 24 hr irradiation. Therefore, the present photocatalysts-coated PET film may become a useful tool to reduce the risk for exposure to hazardous agents including anticancer drugs.

Forecasting model of activities of the city-level for management of CO2 emissions applicable to various cities.

CO2 reduction has become one of the most critical issues globally, and considering sustainable development, many countries are implementing and reviewing CO2 reduction policies. To examine the effect of the CO2 reduction policies, a forecasting model that considers the relationship between variables such as population, building area, industries, vehicle use, and the environment is required. Moreover, this model should also be applicable to various cities to support effective policymaking. In this study, we develop a model that can predict CO2 emissions from the relationship between the variables using System Dynamics, a method to model cities to represent one system composed of various variables. To expand the applicability of the model to various cities in Japan, the proposed model assigns statistical data as input data that can be obtained in any city and standardizes the system structure and variables of the model. In this study, we selected three cities, namely Fukuoka, Kashiwa, and Kumano, which had different populations and industrial characteristics. The calculation accuracy error of CO2 emissions for the three cities was found to be less than 6%. In addition, through the parameter study, it was confirmed that the proposed model can be used to examine the sectors that require CO2 reduction policies, along with the optimal application period. This study aims to provide an effective model that can help in CO2 forecasting and thus in environmental and sustainable development policymaking. Our approach to the CO2 forecasting model facilitates calculating effective CO2 reductions in various cities.

Increased expression of acyl-CoA oxidase 2 in the kidney with plasma phytanic acid and altered gut microbiota in spontaneously hypertensive rats.

We performed a DNA microarray analysis of the renal medulla and cortex from spontaneously hypertensive rats (SHRs), stroke-prone SHRs (SHRSPs), and Wistar-Kyoto (WKY) rats to identify pivotal molecules in the kidney associated with the onset of hypertension and found increased expression of acyl-CoA oxidase 2 (Acox2) mRNA. Real-time polymerase chain reaction revealed that Acox2 mRNA expression in the renal medulla and cortex of SHRs and SHRSPs was increased in comparison to WKY rats. These findings indicate that increased renal ACOX2 (an enzyme that induces the ß-oxidation of fatty acids) is associated with the onset of hypertension. Immunostaining of ACOX2 in the distal tubules from SHRs was stronger than that in the distal tubules from WKY rats. Western blot analysis showed increased expression of ACOX2 protein in renal medulla from SHRs. Regarding the overexpression of ACOX2, plasma levels of phytanic acid in SHRs were significantly higher than those in WKY rats. There were no differences in other short-chain fatty acids. Plasma phytanic acid was affected by the gut microbiota through the conversion from phytol by yeast in the intestinal tract. We compared the gut microbiota profile in three strains of 5-week-old rats by the terminal-restriction fragment length polymorphism method. The gut microbiota profile and ratio of Firmicutes/Bacteroides differed between SHRs and WKY rats. These findings suggest that the increased expression of ACOX2 in the kidney along with increases in plasma phytanic acid and the altered gut microbiota may be involved in the oxidation in the kidney and the pathogenesis of hypertension.

Identification of Clock Genes Related to Hypertension in Kidney From Spontaneously Hypertensive Rats.

BACKGROUND: There is a diurnal variation in the blood pressure fluctuation of hypertension, and blood pressure fluctuation abnormality is considered to be an independent risk factor for organ damage including cardiovascular complications. In the current study, we tried to identify molecules responsible for blood pressure circadian rhythm formation under the control of the kidney biological clock in hypertension. METHODS: DNA microarray analysis was performed in kidneys from 5-week-old spontaneously hypertensive rats (SHRs)/Izm, stroke-prone SHR rats (SHRSP)/Izm, and Wistar Kyoto (WKY)/Izm rats. To detect variation, mouse tubular epithelial cells (TCMK-1) were stimulated with dexamethasone. We performed immunostaining and western blot analysis in the renal medulla of kidney from 5-week-old WKY rats and SHRs. RESULTS: We extracted 1,032 genes with E-box, a binding sequence for BMAL1 and CLOCK using a Gene Set Enrichment Analysis. In a microarray analysis, we identified 12 genes increased as more than 2-fold in the kidneys of SHRs and SHRSP in comparison to WKY rats. In a periodic regression analysis, phosphoribosyl pyrophosphate amidotransferase (Ppat) and fragile X mental retardation, autosomal homolog 1 (Fxr1) showed circadian rhythm. Immunocytochemistry revealed PPAT-positivity in nuclei and cytoplasm in the tubules, and FXR1-positivity in the cytoplasm of TCMK-1. In 5-week-old WKY rat and SHR kidneys, PPAT was localized in the nucleus and cytoplasm of the proximal and distal tubules, and FXR1 was localized to the cytoplasm of the proximal and distal tubules. CONCLUSIONS: PPAT and FXR1 are pivotal molecules in the control of blood pressure circadian rhythm by the kidney in hypertension.

Nitric Oxide Enters Quorum Sensing via the H-NOX Signaling Pathway in Vibrio parahaemolyticus.

Nitric oxide (NO) plays a major role in the regulation of mammalian biological functions. In recent years, NO has also been implicated in bacterial life cycles, including in the regulation of biofilm formation, and the metabolism of the bacterial second messenger signaling molecule cyclic-di-GMP. In a previous study, we reported the discovery of an NO-responsive quorum sensing (QS) circuit in Vibrio harveyi. Here, we characterize the homologous QS pathway in Vibrio parahaemolyticus. Spectroscopic analysis shows V. parahaemolyticus H-NOX is an NO sensory protein that binds NO in 5/6-coordinated mixed manner. Further, we demonstrate that through ligation to H-NOX, NO inhibits the autophosphorylation activity of an H-NOX-associated histidine kinase (HqsK; H-NOX-associated quorum sensing kinase) that transfers phosphate to the Hpt (histidine-containing phosphotransfer protein) protein LuxU. Indeed, among the three Hpt proteins encoded by V. parahaemolyticus, HqsK transfers phosphate only to the QS-associated phosphotransfer protein LuxU. Finally, we show that NO promotes transcription of the master quorum sensing regulatory gene opaR at low cell density.

Impact of neointimal tissue characterization and heterogeneity of bare-metal stents and drug-eluting stents on the time course after stent implantation evaluated by integrated backscatter intravascular ultrasound.

Pathological studies have suggested the different process of in-stent restenosis (ISR) of bare-metal stents (BMS) and drug-eluting stents (DES). Here, we evaluated the components of neointimal tissue using integrated backscatter intravascular ultrasound (IB-IVUS) and focused on the time course after stent implantation and tissue signal distribution. We evaluated 125 lesions of 125 patients who underwent target lesion revascularization for ISR (BMS: n = 73, DES: n = 52). Volume analysis of a 4-mm length centered on a minimum lumen area in every 1-mm cross-sectional area was performed. For IB-IVUS analysis, color-coded maps were constructed from the default setting based on the integrated backscatter (IB) values (middle-IB value, green: fibrous and low-IB value, blue: lipid pool). For the neointimal tissue volume, we evaluated the ratios of the green (%G) and blue (%B) areas. Tissue signal distribution (TD) was also obtained from the default setting based on IB values in each pixel of IB-IVUS imaging. We compared values of neointimal tissues measured by IB-IVUS between the DES and BMS and time course. The observed period was longer after BMS implantation than after DES implantation (BMS: 2545 days, DES: 1233 days, p < 0.001). Overall, %G and %B were similar between the BMS and DES groups (%G: 55% and 51%, respectively, p = 0.10; %B: 36% and 38%, respectively, p = 0.51); however, TD was significantly higher in the DES group than in the BMS group (1091 vs. 1367, p < 0.001). TD in the DES group remained high during the follow-up periods. However, TD in the BMS group was low in the early phase and significantly increased over time (r = 0.56, p < 0.001). When analyzing the ISR within 2 years after stent implantation, the BMS was distinguished with a sensitivity of 66% and a specificity of 90% (cut-off value: TD = 1135, area under the curve 0.83, 95% confidence interval 0.74-0.92). TD could differentiate neointimal tissue after BMS implantation in the early phase. TD can be a useful index in the observation of neoatherosclerosis.

Lipoprotein Lipase Deficiency Arising in Type V Dyslipidemia.

A 40-year-old Japanese man presented with child-onset hypertriglyceridemia recently complicated by diabetes mellitus. The patient's diabetes mellitus was maintained, but he had persistent insulin resistance. The patient also had persistent severe hypertriglyceridemia (1,224-4,104 mg/dL), despite the administration of bezafibrate and ezetimibe. Type V dyslipidemia was revealed by agarose gel electrophoresis and the refrigerator test, and a significantly reduced post-heparin lipoprotein lipase mass of 26 ng/mL was confirmed. Genetic testing confirmed two heterozygous LPL variants, p.Tyr88X and p.Gly215Glu in trans; thus, the patient was diagnosed with lipoprotein lipase deficiency. Lipoprotein lipase deficiency typically arises in type I dyslipidemia, but is latent in type V dyslipidemia.

The adrenal gland circadian clock exhibits a distinct phase advance in spontaneously hypertensive rats.

The circadian clock influences a multitude of cellular and biological processes, including blood pressure control. Spontaneously hypertensive rats (SHR) exhibit aberrant circadian rhythms affecting cardiovascular parameters, and they also have abnormal clock gene expression profiles in several organs. Given the important role of the adrenal gland in orchestrating circadian oscillations, we investigated the adrenal gland circadian clock in SHR and control Wistar-Kyoto rats maintained under a 12-hour light-dark cycle. Adrenal glands, livers, and serum samples were collected every 4 h and mRNA was extracted for analysis of clock gene expression. Serum levels of corticosterone and aldosterone were also analyzed. Overall, the circadian profiles of Bmal1, Per2, Per3, Cry1, RevErba, Revervb, and Dbp gene expression in SHR adrenal glands were phase-advanced relative to controls. The expression profile of StAR (a representative gene under circadian control in the adrenal gland), as well as the circadian rhythms of serum concentrations of corticosteroid and aldosterone were also phase advanced. E4bp4 gene expression was significantly higher during the dark period, yet the expression of its transcriptional activator, Rora, was significantly lower throughout the 24 h period in SHR adrenal glands than in controls. This paradoxical high E4bp4 gene expression was, however, not observed in the liver. In addition, Per1, Per2, Per3, Reverba, and Reverbb mRNA tended to be lower in SHR adrenal glands than in controls. Thus, we conclude that SHR possess an abnormal adrenal circadian clock, which may affect the transcriptional regulation of clock-controlled genes, and steroid hormone secretion by the adrenal gland.

Involvement of complement 3 in the salt-sensitive hypertension by activation of renal renin-angiotensin system in spontaneously hypertensive rats.

We previously showed that complement 3 (C3) is highly expressed in mesenchymal tissues in spontaneously hypertensive rats (SHR). We targeted C3 gene by zinc-finger nuclease (ZFN) gene-editing technology and investigated blood pressure and phenotype in SHR. Blood pressure was measured by tail-cuff and telemetry methods. Histology and expression of liver X receptor α (LXRα), renin, Krüppel-like factor 5 (KLF5), and E-cadherin were evaluated in kidneys. Mesangial cells (MCs) were removed from glomeruli from three strains, and we evaluated the phenotype in vitro. SHR showed the salt-sensitive hypertension that was abolished in C3 knockout (KO) SHR. Proliferation of MCs from SHR was higher than that from Wistar-Kyoto (WKY) rats and showed a synthetic phenotype. Renal injury scores were higher in SHR than in WKY rats and C3 KO SHR. Expression of E-cadherin was lower, and expression of renin was higher in the nephrotubulus from SHR than WKY rats and C3 KO SHR. Expression of C3 α-chain protein and α-smooth muscle actin protein was significantly higher in renal medulla from SHR than from WKY rats. Expression of angiotensinogen, LXRα, renin, and KLF5 mRNA was increased in kidney from SHR compared with C3 KO SHR. Intrarenal angiotensin II levels were significantly higher in kidney from SHR than WKY rats and C3 KO SHR. Urinary epinephrine and norepinephrine excretions were significantly higher in SHR than in WKY rats and C3 KO SHR. These findings showed that increased C3 induces salt-sensitive hypertension with increases in urinary catecholamine excretion and intrarenal activation of the renin-angiotensin system by the dedifferentiation of mesenchymal tissues in kidney from SHR.