Evaluation of Local Vascular Perfusion in the Lower Extremities on Intravoxel Incoherent Motion Imaging before and after Endovascular Therapy.

PURPOSE: To evaluate improvement in local vascular perfusion of the lower limbs on intravoxel incoherent motion (IVIM) imaging after endovascular therapy (EVT). MATERIALS AND METHODS: IVIM imaging was performed on 20 lower limbs of 16 patients with lower extremity arterial diseases before and after EVT. To estimate IVIM, diffusion-weighted lower-limb axial images (number of slices = 25 and slice thickness = 3.5 mm) were acquired using different b values (0, 300, and 1000 s/mm2). IVIM imaging with the simplified IVIM techniques was performed. The perfusion-related coefficient (D* [10-3 mm2/s]), perfusion fraction (f [%]), and D*f product (10-3 mm2/s %) were calculated before and 2-3 days after EVT. The ankle brachial index (ABI), mean D* (10-3 mm2/s), mean f (%), and mean D*f product (10-3 mm2/s %) before and after EVT were compared. RESULTS: Successful revascularization was achieved in all cases. After EVT, the mean ABI significantly increased from 0.59 ± 0.19 to 0.87 ± 0.15 (p < 0.001, paired t test). The mean D* (10-3 mm2/s) (22.08 ± 3.26 versus 24.87 ± 2.65, p = 0.005, paired t test), and D*f product (10-3 mm2/s%) (551.03 ± 79.02 versus 634.55 ± 76.96, p = 0.002, paired t-test) of the lower limbs significantly increased after EVT, whereas f (%) (25.00 ± 1.28 versus 25.52 ± 1.61, p = 0.261, paired t-test) did not significantly increased after EVT. CONCLUSION: D* (10-3 mm2/s) and D*f product (10-3 mm2/s %) on IVIM imaging could evaluate improvement in local vascular perfusion of the lower limbs after EVT. LEVEL OF EVIDENCE: Level 4, Case Series.

Recent advances in delivering RNA-based therapeutics to mitochondria.

INTRODUCTION: After the emergence of lipid nanoparticles (LNP) containing therapeutic mRNA as vaccines for use against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the clinical usefulness of nucleic acid-encapsulated LNPs is now a fact. In addition to the nucleus and cytoplasm, mitochondria, which have their own genome, are a site where nucleic acids function in the cell. Gene therapies targeting mitochondria are expected to pave the way for the next generation of therapies. AREAS COVERED: Methods for delivering nucleic acids to mitochondria are needed in order to realize such innovative therapies. However, only a few reports on delivery systems targeting mitochondria have appeared. In this review, we summarize the current state of research on RNA-based therapeutics targeted to mitochondria, with emphasis on mitochondrial RNA delivery therapies and on therapies that involve the use of mitochondrial genome editing devices. EXPERT OPINION: We hope that this review article will focus our attention to this area of research, stimulate more interest in this field of research, and lead to the development of mitochondria-targeted nucleic acid medicine. It has the potential to become a major weapon against urgent and unknown diseases, including SARS-CoV-2 infections.

The use of a MITO-Porter to deliver exogenous therapeutic RNA to a mitochondrial disease's cell with a A1555G mutation in the mitochondrial 12S rRNA gene results in an increase in mitochondrial respiratory activity.

We report on validating a mitochondrial gene therapeutic strategy using fibroblasts derived from patients with an A1555G point mutation in mitochondrial DNA coding 12S ribosomal RNA (rRNA (12S)). Wild-type rRNA (12S) as a therapeutic RNA was encapsulated in a mitochondrial targeting liposome, a MITO-Porter (rRNA-MITO-Porter), and an attempt was made to deliver the MITO-Porter to mitochondria of the diseased cells. It was confirmed that the rRNA-MITO-Porter treatment significantly decreased the ratio of the mutant rRNA content. Moreover, it was shown that the mitochondrial respiratory activities of the diseased cells were improved as the result of the mitochondrial transfection of the rRNA-MITO-Porter.

Validation of Gene Therapy for Mutant Mitochondria by Delivering Mitochondrial RNA Using a MITO-Porter.

Here, we report on validating a mitochondrial gene therapy by delivering nucleic acids to mitochondria of diseased cells by a MITO-Porter, a liposome-based carrier for mitochondrial delivery. We used cells derived from a patient with a mitochondrial disease with a G625A heteroplasmic mutation in the tRNAPhe of the mitochondrial DNA (mtDNA). It has been reported that some mitochondrial gene diseases are caused by heteroplasmic mutations, in which both mutated and wild-type (WT) genes are present, and the accumulation of pathological mutations leads to serious, intractable, multi-organ diseases. Therefore, the decrease of the mutated gene rate is considered to be a useful gene therapy strategy. To accomplish this, wild-type mitochondrial pre-tRNAPhe (pre-WT-tRNAPhe), prepared by in vitro transcription, was encapsulated in the MITO-Porter. The pre-WT-tRNAPhe encapsulated in the MITO-Porter was transfected into diseased mitochondrial cells, and the resulting mutant levels were examined by an amplification refractory mutation system (ARMS)-quantitative PCR. The mutation rate of tRNAPhe was decreased, and this therapeutic effect was sustained even on the 8th day after transfection. Furthermore, mitochondrial respiratory activity of the disease cells was increased after the transfection of therapeutic pre-WT-tRNAPhe. These results support the conclusion that the mitochondrial delivery of therapeutic nucleic acids represents a viable strategy for mitochondrial gene therapy.

Development of a nanoparticle that releases nucleic acids in response to a mitochondrial environment.

The delivery of nucleic acids targeting mutant mtDNA represent a potential strategy for addressing a variety of mitochondria-related diseases. We previously developed a MITO-Porter, a nano carrier that is capable of delivering nanoparticles of nucleic acids to mitochondria of human cells. Here, we report on an investigation of a series of nanoparticles formed with various poly cationic peptides that can release nucleic acids in response to a mitochondrial environment. A significant relationship was found between the number of and the location of arginine and histidine residues in the peptide sequence and the release of nucleic acids in a mitochondrial environment.

The value of CT findings for prognostic prediction of spontaneous superior mesenteric artery dissection.

BACKGROUND: Spontaneous superior mesenteric artery (SMA) dissection is rare cause of acute abdomen. Time-dependent change of SMA dissection has not been established. PURPOSE: To determine Sakamoto classification (SC) type of acute and chronic SMA dissection (aSMAD and cSMAD) to predict the treatment methods and outcome. MATERIAL AND METHODS: From April 2003 to March 2017, unenhanced and contrast-enhanced CT were used to diagnose acute symptomatic or chronic asymptomatic SMA dissection in 25 consecutive patients without aortic dissection. Correlations between SCs and treatment methods and outcomes were investigated. RESULTS: All 13 patients with aSMAD initially received conservative treatment. Initial SCs in aSMAD were type I = 1, type III = 9, and type IV = 3. Three of nine initial type III and two of three initial type IV changed to type I at follow-up. One of nine type III changed to type II at follow-up. Ohers did not change. One with initial type III required vascular repair, so the final SC was not available. Three patients required bowel resection. In cSMAD of 12 patients, the initial/final SC were type I and IV in ten and two patients, respectively, without change during follow-up. cSMAD was significantly older than aSMAD. The initial length of dissection of aSMAD was longer than in the cSMAD group. In aSMAD, the final length of dissection was significantly shorter than in the initial computed tomography scan. CONCLUSION: Initial SC differed significantly between aSMAD and cSMAD. Initial SC types in aSMAD were type III and IV mainly, and changed during the observation period. In cSMAD, SC types were I and IV without change.

Nonfatal air embolism complicating percutaneous CT-guided lung biopsy and VATS marking: Four cases from a single institution.

Systemic air emboli occur as a rare complication of percutaneous needle biopsy of the lung and video-assisted thoracoscopic surgery (VATS) marking. Here we present four cases of systemic air emboli from single institution and the imaging findings and embolism' kinetics using contrast-enhanced media during VATS color marking with indocyanine green. We suggest that early detection using routine whole-lung CT is required for asymptomatic patients with abnormal air. If abnormal air is found, we should keep the patient to the appropriate posture in order to prevent moving the air until it dissipates. Early detection of abnormal air can prevent severe complications.

The value of adding diffusion-weighted images for tumor detection and preoperative staging in renal pelvic carcinoma for the reader's experience.

PURPOSE: This retrospective study aimed to assess the value of adding diffusion-weighted magnetic resonance imaging (DWI) or gadolinium-enhanced fat-suppressed T1WI (CEI) to T2-weighted imaging (T2WI) for preoperative T categorization in renal pelvic carcinoma by the reader's experience using surgical specimens as the reference standard. METHODS: Two radiologists (Reader 1; 3 years, 2; 13 years) reviewed 49 cases with urothelial carcinoma who underwent magnetic resonance imaging examination before surgery, independently, using three image sets: T2WI alone, T2WI plus DWI, and T2WI plus CEI for tumor detection and T categorization. The differences in the apparent diffusion coefficient values between tumors and renal parenchyma, histopathologic grade were analyzed. RESULTS: T2WI plus CEI or DWI had high detection rates (93.4%) compared to T2WI alone. When discriminating T3a/T3b, for Reader 1, the use of T2WI plus DWI (88.0%) and T2WI plus CEI (92.0%) was significantly more accurate than T2WI alone (73%), with AUCs of 0.86, 0.86 and 0.77, respectively. For Reader 2, the accuracies were high on all image sets, with AUCs of 0.87-0.95, and the mean ADC of the tumors was significantly lower than that of the normal renal parenchyma. In addition, the mean ADC values of high-grade tumors were significantly lower than that of low-grade tumors. CONCLUSIONS: DWI and CEI could be more helpful than T2WI alone for preoperative T categorization by less-experienced reader and DWI could be used for preoperative T categorization and for predicting the histopathologic grade of renal pelvic carcinoma.

Chronic orofacial pain in dental patients: retrospective investigation over 12 years.

Orofacial pain is often difficult to diagnose and treat. However, there have been few reports on the clinical observation of dental patients with orofacial pain. We retrospectively investigated the characteristics of 221 dental patients who had suffered from persistent orofacial pain. Data were collected from the outpatient medical records in our clinic over the past 12 years. More than half of the patients (53.8%) had suffered with pain for more than 6 months from pain onset until the first visit to our clinic. The main diagnoses were neuropathic pain (30.3%), myofascial pain (23.5%), psychogenic pain (20.4%), odontogenic toothache (17.2%), and others (7.7%) such as temporomandibular disorders and glossitis. The treatments included pharmacotherapy, splint therapy, and others such as nerve block, dental treatment, physiotherapy, and/or psychotherapy. Excluding the patients (52 of 221 initially enrolled patients) with unknown responses to treatment, 65.7% showed remission or a significant improvement in pain in response to treatment. Although only a small group of patients had odontogenic toothache, the rate of improvement was highest for this disorder. In conclusion, early consultation with a dentist is useful to prevent chronicity of odontogenic pain and to make a differential diagnosis in patients with orofacial pain.

C-reactive protein levels and risk of disabling dementia with and without stroke in Japanese: the Circulatory Risk in Communities Study (CIRCS).

OBJECTIVE: Studies have shown that elevated high-sensitivity C-reactive protein (hs-CRP) predicts stroke, which is a risk factor for dementia. It remains, however, unclear whether hs-CRP increases risk of dementia. METHODS: A prospective nested case-control study of Japanese 40-69 years of age was conducted using frozen serum samples collected from approximately 7531men and women who participated in cardiovascular risk surveys from 1984 to 1994 in one community and 1989-1995 in another community under the Circulatory Risk in Communities Study (CIRCS). Two control subjects per case were matched by sex, age, community, and year of serum storage. The hs-CRP was measured using a latex particle-enhanced immunonephelometric assay. RESULTS: Between 1999 and 2013, we identified 275 disabling dementia cases (96 cases with history of stroke and 179 without it). There was a positive association between hs-CRP levels and risk of dementia with history of stroke. No significant association was observed between hs-CRP levels and risk of dementia without history of stroke. After adjustment for hypertension, diabetes and other confounding variables, the positive association remained statistically significant. The multivariable odds ratios associated with 1-SD increment of log hs-CRP were 1.02 (0.87-1.20) for total dementia, 1.35 (1.02-1.79) for dementia with history of stroke, and 0.89 (0.72-1.10) for dementia without history of stroke. CONCLUSION: Elevated hs-CRP levels were associated with increased risk of disabling dementia in individuals with history of stroke but not in those without it.

Development of hypoxia-inducible factor (HIF)-1α inhibitors: effect of ortho-carborane substituents on HIF transcriptional activity under hypoxia.

A series of substituted ortho-carboranylphenoxyacetanilides were synthesized and evaluated for their ability to inhibit hypoxia-induced HIF-1 transcriptional activity using a cell-based reporter assay in HeLa cells expressing the HRE-dependent firefly luciferase reporter construct (HRE-Luc) and constitutively expressing CMV-driven Renilla luciferase reporter, and their ability to inhibit cell growth (GI(50)) using the MTT assay. Among the compounds synthesized, 1g and 1l showed significant inhibition of hypoxia-induced HIF-1 transcriptional activity (IC(50): 1.9 ± 0.4 and 1.4 ± 0.2 µM, respectively). Both compounds suppressed HIF-1α accumulation in a concentration-dependent manner. The porcine heart malate dehydrogenase (MDH) refolding assay revealed that compound 1l inhibited human Hsp60 chaperone activity (IC(50): 6.80 ± 0.25 µM) and this inhibition activity was higher than that of ETB (IC(50): 10.9 ± 0.63 µM).

Risk factors for sudden cardiac death among Japanese: the Circulatory Risk in Communities Study.

OBJECTIVE: There is little evidence concerning risk factors for sudden cardiac death (SCD) among Asians. PATIENTS AND METHODS: A prospective, nested, case-control study of Japanese patients aged between 30 and 84 years was undertaken using data collected from 26 870 participants in cardiovascular risk surveys conducted in four communities between 1975 and 2005. The incidence of SCD was ascertained by systematic surveillance, with 239 cases of SCD identified over this period. For each case of SCD, three control patients were selected, matched by age, sex, examination year, follow-up time, and community. RESULTS: Hypertension, diabetes mellitus, smoking, major ST-T abnormalities, left high amplitude R waves, and increased heart rate (≥77 beat/min) were all independently associated with a 1.5-3.2-fold increase in SCD risk, whereas no associations were observed for body mass index and hypercholesterolemia. The population-attributable fraction [95% confidence interval (CI)] was 23.0% (2.9-39.0) for hypertension, 15.3% (3.8-25.5) for current smoking, 14.5% (8.0-20.5) for major ST-T abnormalities, and 8.1% (2.2-13.7) for diabetes mellitus. The number of SCD risk factors (hypertension, diabetes, smoking, and ECG abnormalities) was positively associated with increased SCD risk. The odds ratio for increased SCD risk with three or more risk factors versus zero risk factors was 5.76 (95% CI 3.20-10.39). CONCLUSIONS: Among the Japanese population, hypertension, smoking, major ST-T abnormalities, left high amplitude R waves, and diabetes mellitus were associated with an increased incidence of SCD, whereas there were no associations of body mass index or hypercholesterolemia with SCD incidence.

Trends in sudden cardiac death and its risk factors in Japan from 1981 to 2005: the Circulatory Risk in Communities Study (CIRCS).

OBJECTIVE: There is little evidence whether sudden cardiac death (SCD) is increasing in Asia, although the incidence of coronary heart disease among urban middle-aged Japanese men has increased recently. We examined trends in the incidence of SCD and its risk factors in the Circulatory Risk in Communities Study. DESIGN AND SETTING: This was a population-based longitudinal study. Surveillance of men and women for SCD incidence and risk factors was conducted from 1981 to 2005. SUBJECTS: The surveyed population was all men and women aged 30-84 years who lived in three rural communities and one urban community in Japan. MAIN OUTCOME MEASURES: Trends in SCD incidence and its risk factors. RESULTS: Age-adjusted and sex-adjusted incidence of SCD decreased from 1981-1985 to 1991-1995, and plateaued thereafter. The annual incidence per 100 000 person-years was 76.0 in 1981-1985, 57.9 in 1986-1990, 39.3 in 1991-1995, 31.6 in 1996-2000 and 36.8 in 2001-2005. The prevalence of hypertension decreased from 1981-1985 to 1991-1995, and plateaued thereafter for men and women. The age-adjusted prevalence of current smoking for men decreased while that of diabetes mellitus increased for both sexes from 1981-1985 to 2001-2005. CONCLUSIONS: The incidence of SCD decreased from 1981 to 1995 but was unchanged from 1996 to 2005. Continuous surveillance is necessary to clarify future trends in SCD in Japan because of an increasing incidence of diabetes mellitus.

Boron-containing phenoxyacetanilide derivatives as hypoxia-inducible factor (HIF)-1alpha inhibitors.

A series of boron-containing phenoxyacetanilide derivatives 8a-f, 9a-f, 15, and 16 were synthesized as hypoxia-inducible factor (HIF)-1alpha inhibitors. Among the compounds synthesized, carboranylphenoxyacetanilide 16 (GN26361) was found to be a potent inhibitor against HIF-1alpha accumulation under hypoxic conditions and inhibited the hypoxia-induced HIF-1 transcriptional activity in HeLa cells (IC50=0.74 microM). Compound 16 suppressed hypoxia-induced HIF-1alpha accumulation and vascular endothelial growth factor mRNA expression in a concentration-dependent manner without affecting the expression of HIF-1alpha mRNA.