Impaired Glycosylation of Gastric Mucins Drives Gastric Tumorigenesis and Serves as a Novel Therapeutic Target.

BACKGROUND & AIMS: Gastric cancer is often accompanied by a loss of mucin 6 (MUC6), but its pathogenic role in gastric carcinogenesis remains unclear. METHODS: Muc6 knockout (Muc6-/-) mice and Muc6-dsRED mice were newly generated. Tff1Cre, Golph3-/-, R26-Golgi-mCherry, Hes1flox/flox, Cosmcflox/flox, and A4gnt-/- mice were also used. Histology, DNA and RNA, proteins, and sugar chains were analyzed by whole-exon DNA sequence, RNA sequence, immunohistochemistry, lectin-binding assays, and liquid chromatography-mass spectrometry analysis. Gastric organoids and cell lines were used for in vitro assays and xenograft experiments. RESULTS: Deletion of Muc6 in mice spontaneously causes pan-gastritis and invasive gastric cancers. Muc6-deficient tumor growth was dependent on mitogen-activated protein kinase activation, mediated by Golgi stress-induced up-regulation of Golgi phosphoprotein 3. Glycomic profiling revealed aberrant expression of mannose-rich N-linked glycans in gastric tumors, detected with banana lectin in association with lack of MUC6 expression. We identified a precursor of clusterin as a binding partner of mannose glycans. Mitogen-activated protein kinase activation, Golgi stress responses, and aberrant mannose expression are found in separate Cosmc- and A4gnt-deficient mouse models that lack normal O-glycosylation. Banana lectin-drug conjugates proved an effective treatment for mannose-rich murine and human gastric cancer. CONCLUSIONS: We propose that Golgi stress responses and aberrant glycans are important drivers of and promising new therapeutic targets for gastric cancer.

Denosumab-induced hypocalcemia in patients with solid tumors and renal dysfunction: a multicenter, retrospective, observational study.

BACKGROUND: Bone metastases are frequently observed in advanced cancer, and bone modifying agents are used to prevent or treat skeletal-related events. Zoledronic acid is contraindicated in patients with severe renal impairment (Ccr < 30 mL/min), but it is not completely known whether denosumab can be used in them. We aimed to determine the association between renal function and hypocalcemia development during denosumab treatment. METHODS: We included patients with solid cancer and bone metastases who started denosumab treatment between April 2017 and March 2019. They were classified into four groups based on creatinine clearance (Ccr; mL/min): normal (Ccr ≥ 80), mild (50 ≤ Ccr ˂80), moderate (30 ≤ Ccr ˂50), and severe (Ccr ˂30). Hypocalcemia was evaluated using the Common Terminology Criteria for Adverse Events (v5.0) based on the albumin-adjusted serum calcium levels; its incidence (stratified by renal function) and risk factors were investigated using a Chi-square test and logistic regression analysis. RESULTS: Of 524 patients (age: 69 ± 11 years; 303 men), 153 had a normal renal function and 222, 117, and 32 had mild, moderate, and severe renal dysfunction. The albumin-adjusted serum calcium level was higher than the measured (total) calcium level in most patients. The incidence of grade ≥ 1 hypocalcemia was 32.0% in the normal group and 37.4%, 29.9%, and 62.5% in the mild, moderate, and severe renal dysfunction groups, respectively. It was, therefore, higher in the severe renal dysfunction groups than in the normal group (P = 0.002). The incidence of grade ≥ 3 hypocalcemia did not differ significantly among the groups. Pre-treatment low serum calcium levels and severe renal dysfunction were risk factors for hypocalcemia. CONCLUSIONS: Evaluating denosumab-induced hypocalcemia required albumin adjustment, and its incidence was high among patients with severe renal dysfunction. Reduced serum calcium levels and severely impaired renal function were associated with an elevated hypocalcemia risk.

Clinical Comparison of Drug-Coated Balloon and Drug-Eluting Stent for Femoropopliteal Lesions in Chronic Limb-Threatening Ischemia With Wounds.

BACKGROUND: Endovascular therapy (EVT) with devices such as drug-coated balloons (DCBs) and drug-eluting stents (DESs) for atherosclerotic disease in the femoropopliteal (FP) artery has been established. However, EVT using drug-based devices for chronic limb-threatening ischemia (CLTI) remains challenging. The optimal device for FP lesions in patients with CLTI remains unknown. This study compared the clinical efficacy of DCB and DES in patients with CLTI and FP lesions.Methods and Results: This retrospective single-center study included 539 consecutive patients (562 lesions) treated with EVT between January 2018 and December 2022; 166 patients with CLTI and Rutherford Class 5 or 6 wounds underwent EVT with DCB or DES. Clinical outcomes were compared between 53 pairs after propensity score matching. There were no significant differences between the DCB and DES groups in the incidence of complete wound healing without death or major amputation (84.8% vs. 80.2%, respectively; P=0.99), primary patency (69.4% vs. 75.6%, respectively; P=0.65), and freedom from target lesion revascularization at 1 year (78.6% vs. 78.0%, respectively; P=0.92). Multivariate analysis showed that complete wound healing at 1 year is negatively associated with hemodialysis and Wound, Ischemia, and foot Infection Stage 4, but positively associated with Global Limb Anatomic Staging System FP Grade 3 or 4. CONCLUSIONS: No significant differences in clinical outcomes were found between DCB and DES for patients with CLTI and FP lesions.

Guideline-Directed Medical Therapy for Elderly Patients With Acute Myocardial Infarction Who Undergo Percutaneous Coronary Intervention　- Insights From a Retrospective Observational Study.

BACKGROUND: The efficacy of guideline-directed medical therapy (GDMT) in the elderly remains unclear. This study evaluated the impact of GDMT (aspirin or a P2Y12inhibitor, angiotensin-converting enzyme inhibitor or angiotensin receptor blocker, ß-blocker, and statin) at discharge on long-term mortality in elderly patients with acute myocardial infarction (AMI) who had undergone percutaneous coronary intervention (PCI). METHODS AND RESULTS: Of 2,547 consecutive patients with AMI undergoing PCI in 2009-2020, we retrospectively analyzed 573 patients aged ≥80 years. The median follow-up period was 1,140 days. GDMT was prescribed to 192 (33.5%) patients at discharge. Compared with patients without GDMT, those with GDMT were younger and had higher rates of ST-segment elevation myocardial infarction and left anterior descending artery culprit lesion, higher peak creatine phosphokinase concentration, and lower left ventricular ejection fraction (LVEF). After adjusting for confounders, GDMT was independently associated with a lower cardiovascular death rate (hazard ratio [HR] 0.35; 95% confidence interval [CI] 0.16-0.81), but not with all-cause mortality (HR 0.77; 95% CI 0.50-1.18). In the subgroup analysis, the favorable impact of GDMT on cardiovascular death was significant in patients aged 80-89 years, with LVEF <50%, or with an estimated glomerular filtration rate ≥30 mL/min/1.73 m2. CONCLUSIONS: GDMT in patients with AMI aged ≥80 years undergoing PCI was associated with a lower cardiovascular death rate but not all-cause mortality.

Basic Performance Evaluation of a Radiation Survey Meter That Uses a Plastic-Scintillation Sensor.

After the Fukushima nuclear power plant accident in 2011, many types of survey meters were used, including Geiger-Müller (GM) survey meters, which have long been used to measure ß-rays. Recently, however, a novel radiation survey meter that uses a plastic-scintillation sensor has been developed. Although manufacturers' catalog data are available for these survey meters, there have been no user reports on performance. In addition, the performance of commercial plastic-scintillation survey meters has not been evaluated. In this study, we experimentally compared the performance of a plastic-scintillation survey meter with that of a GM survey meter. The results show that the two instruments performed very similarly in most respects. The GM survey meter exhibited count losses when the radiation count rate was high, whereas the plastic-scintillation survey meter remained accurate under such circumstances, with almost no count loss at high radiation rates. For measurements at background rates (i.e., low counting rates), the counting rates of the plastic-scintillation and GM survey meters were similar. Therefore, an advantage of plastic-scintillation survey meters is that they are less affected by count loss than GM survey meters. We conclude that the plastic-scintillation survey meter is a useful ß-ray measuring/monitoring instrument.

Comparison of shielding effects of over-glasses-type and regular eyewear in terms of occupational eye dose reduction.

Given the new recommendations for occupational eye lens doses, various lead glasses have been used to reduce irradiation of interventional radiologists. However, the protection afforded by lead glasses over prescription glasses (thus over-glasses-type eyewear) has not been considered in detail. We used a phantom to compare the protective effects of such eyewear and regular eyewear of 0.07 mm lead-equivalent thickness. The shielding rates behind the eyewear and on the surface of the left eye of an anthropomorphic phantom were calculated. The left eye of the phantom was irradiated at various angles and the shielding effects were evaluated. We measured the radiation dose to the left side of the phantom using RPLDs attached to the left eye and to the surface/back of the left eyewear. Over-glasses-type eyewear afforded good protection against x-rays from the left and below; the average shielding rates on the surface of the left eye ranged from 0.70-0.72. In clinical settings, scattered radiation is incident on physicians' eyes from the left and below, and through any gap in lead glasses. Over-glasses-type eyewear afforded better protection than regular eyewear of the same lead-equivalent thickness at the irradiation angles of concern in clinical settings. Although clinical evaluation is needed, we suggest over-glasses-type Pb eyewear even for physicians who do not wear prescription glasses.

Examination and comparison of the RNA extraction methods using mouse serum.

Serum microRNAs (miRNAs) are considered useful as non-invasive biomarkers for different diseases. However, the optimal method for extracting RNAs from serum is currently unknown. In the present study, several RNA extraction kits were used to examine the optimal kit. RNAs were extracted from the serum of 8-week-old C57BL/6NJcl male mice following the protocol of each RNA extraction kit. The yield of the extracted RNA samples was calculated, and an Agilent Bioanalyzer was used to assess the electrophoretic patterns. An Agilent mouse miRNA microarray was utilized to confirm the expression patterns of the extracted RNA samples. The results revealed significant differences in RNA yields from the miRNeasy Serum/Plasma Advanced kit and mirVana™ PARIS™ RNA and Native Protein Purification Kit compared with almost all other samples. Further, two peaks were determined in the miRNeasy Serum/Plasma Advanced kit using a small RNAs kit of Agilent Bioanalyzer, including one at 20-40 nucleotides (nt) and another at ~40-100 nt, whereas the other reagents had a single peak. This revealed that the extracted RNAs may differ in composition based on the RNA extraction method. Some types of miRNAs were only detected with certain RNA extraction reagents. This suggested that different RNA extraction reagents may cause differences in the types of miRNAs detected. On the other hand, the miRNAs commonly expressed by the three RNA extraction reagents are highly correlated in expression levels.

Iatrogenic acute limb ischemia with complete traumatic rupture of the popliteal artery associated with total knee arthroplasty.

Acute limb ischemia (ALI) related to total knee arthroplasty (TKA) is rare. Most occlusions are caused by thrombus formation in the popliteal artery (PA). Currently such cases are revascularized using less invasive approaches such as endovascular therapy or Fogarty thrombectomy. We report a case of ALI in a 65-year-old woman with complete rupture of the PA due to a TKA procedure. She had resting pain and motor paralysis in her right lower extremity after TKA. Contrast-enhanced computed tomography showed occlusion of the right femoropopliteal artery. Subsequently, she was referred to our hospital with a diagnosis of ALI. Initially, a less invasive revascularization procedure was unsuccessfully attempted. Therefore, we performed an emergency distal bypass and succeeded in revascularization. Intraoperative examination revealed a complete rupture of the PA. Postoperatively, the patient exhibited no signs of myonephropathic metabolic syndrome. Although there was significant motor impairment, the affected limbs were successfully salvaged. ALI with complete rupture of the PA associated with TKA has not been reported previously. In cases of iatrogenic ALI after TKA, it would be essential to consider diagnostic and revascularization methods that account for the possibility of severe injury to the PA. Learning objective: Acute limb ischemia after total knee arthroplasty is a rare and life- and limb-threatening condition. The underlying pathological mechanism is often thrombus occlusion due to mechanical stimuli of the popliteal artery (PA). There are no established treatments for this condition, and less invasive approaches such as endovascular procedures and Fogarty thrombectomy are often used. However, in cases involving severe damage to the PA, bypass surgery may be necessary, and revascularization procedures should be considered accordingly.

Extracapsular Dissection With Narrow-Band Imaging Using the Transnasal Endoscopic Tri-port Approach for Extracranial Trigeminal Schwannoma: A Case Report.

The pterygopalatine fossa and infratemporal fossa are often approached through an external incision because of their deep facial location, but this can present problems such as facial scarring and deformity. In schwannoma surgery, intraneural dissection is a useful surgical technique for achieving gross total resection while preserving the capsule, including the nerves. For appropriate enucleation and preservation of the functional nerve, it is indispensable to distinguish between the pseudocapsule and the tumor capsule. This case report presents a case of endonasal surgical intervention for an extracranial trigeminal schwannoma employing the tri-port approach and narrow-band imaging.

[Temperature Dependence of Scintillation Survey Meter with and without Temperature Compensation Function].

PURPOSE: The objective of this study was to compare the temperature dependence of a scintillation survey meter with and without the temperature compensation function. Investigation of temperature dependence is important to make precise measurements in various environments. METHOD: The experiment was conducted using the NaI (Tl) scintillation survey meter with the temperature compensation function (TCS-1172) and the NaI (Tl) and CsI (Tl) scintillation survey meters without the temperature compensation function (TCS-171, PDR-111). In all, 1 cm dose equivalent rate (µSv/h) was measured by changing the room temperature from 10 to 40 degree Celsius. RESULT: The results showed that the scintillation survey meter with the temperature compensation function had almost no change in the measured values with changes in room temperature, whereas the 1 cm dose equivalent rate of the scintillation survey meter without the temperature compensation function changed by a maximum of -7.2 (%/10°C) as temperature increased. CONCLUSION: This study confirms that the scintillation survey meter with the temperature compensation function was less dependent on temperature, and stable measurement was possible. However, it was suggested that the scintillation survey meter without the temperature compensation function might cause a drop in the measured value as the temperature rises.

Changes in cognitive ability and serum microRNA levels during aging in mice.

Mild cognitive impairment (MCI) is an early stage that can result in dementia. MCI can be reversed, and diagnosis at an early stage is crucial to control the progression to dementia. Dementia is currently diagnosed based on interviews and screening tests; however, novel biomarkers must be identified to allow early MCI detection. Therefore, the present study aimed to identify novel biomarkers in the form of blood microRNAs (miRNAs/miRs) for the diagnosis of MCI or early dementia. Blood samples were collected from C57BL/6NJcl male mice at four time points, including 4-week-old (4W), 8-week-old (8W), 36-week-old (36W) and 58-week-old (58W), and serum was isolated. Body weight and blood total cholesterol levels were increased, and blood alkaline phosphatase was decreased with aging. The 8W mice exhibited the highest cognitive ability in the Morris water maze test, whereas the 58W mice demonstrated decreased cognitive ability. The serum RNA concentrations of the 4W, 8W, 36W and 58W mice demonstrated no significant differences. Furthermore, small RNA levels were detected in the serum of all mice. miRNA microarray analysis revealed a >1.5-fold increase in the serum expression of two miRNAs (miR-21a-5p and miR-92a-3p) and a >1.5-fold decrease in the serum expression of two other miRNAs (miR-6769b-5p and miR-709) in 58W mice compared with those in 8W mice. In the future, we aim to further analyze aged mice to discover novel MCI biomarkers.

Narrow-band imaging to enhance intraneural dissection in head and neck schwannoma surgery: a quantitative evaluation.

OBJECTIVE: The objective of this study was to assess the utility of narrow-band imaging (NBI) for improving intraneural dissection during gross total resection of head and neck schwannoma. Specifically, we aimed to quantitatively evaluate whether NBI can enhance the identification of pseudocapsule and true capsule within the tumor. METHODS: Nine schwannoma surgery cases conducted between February 2018 and October 2022 were retrospectively analyzed. The surgical procedures followed established principles with a specific focus on utilizing NBI to distinguish between the pseudocapsule and true capsule. Intraneural dissection was performed by searching for a tumor surface with a fascicle-free window, followed by longitudinal incision of the pseudocapsule. NBI was used to distinguish between the pseudocapsule and true capsule. Surgical views were captured under both white light (WL) illumination and NBI for further analysis. The brightness and contrast of the pseudocapsule and true capsule were quantitatively measured using ImageJ and were compared. RESULTS: Under NBI, the pseudocapsule consistently appeared greenish-gray, whereas the true capsule exhibited a white appearance. Quantitative analysis revealed a statistically significant difference (p < 0.0001) in brightness between the pseudocapsule (mean grayscale value 52.1, 95%CI; 46.4-75.3) and true tumor capsule (mean grayscale value 120.8, 95%CI; 155.7-109.0) under NBI. Conversely, there was no statistically significant difference in the brightness of these structures under WL (p = 0.2067). NBI also showed significantly higher contrast between the two structures than did WL (contrast 73.6, 95%CI; 53.1-89.5 vs. 30.9, 95%CI; 1.0-47.5, p = 0.0034). Further spectral analysis revealed that the most substantial difference in brightness between the pseudocapsule and the true tumor capsule was observed in the red spectrum, with a difference in brightness of -0.6 (95%CI; -16.8-14.8) under WL and 83.5 (95%CI; 50.3-100.0) under NBI (p < 0.0001). CONCLUSION: NBI proved to be a valuable tool for enhancing the identification of pseudocapsule and true capsule during intraneural dissection in head and neck schwannoma surgery. The improved contrast and membrane visibility offered by NBI might have the potential to reduce postoperative neurological deficits and improve surgical outcomes. Further research is warranted to validate our findings and explore the broader applications of NBI in schwannoma surgery.

Effect of backscatter radiation on the occupational eye-lens dose.

We quantified the level of backscatter radiation generated from physicians' heads using a phantom. We also evaluated the shielding rate of the protective eyewear and optimal placement of the eye-dedicated dosimeter (skin surface or behind the Pb-eyewear). We performed diagnostic X-rays of two head phantoms: Styrofoam (negligible backscatter radiation) and anthropomorphic (included backscatter radiation). Radiophotoluminescence glass dosimeters were used to measure the eye-lens dose, with or without 0.07-mm Pb-equivalent protective eyewear. We used tube voltages of 50, 65 and 80 kV because the scattered radiation has a lower mean energy than the primary X-ray beam. The backscatter radiation accounted for 17.3-22.3% of the eye-lens dose, with the percentage increasing with increasing tube voltage. Furthermore, the shielding rate of the protective eyewear was overestimated, and the eye-lens dose was underestimated when the eye-dedicated dosimeter was placed behind the protective eyewear. We quantified the backscatter radiation generated from physicians' heads. To account for the effect of backscatter radiation, an anthropomorphic, rather than Styrofoam, phantom should be used. Close contact of the dosimeter with the skin surface is essential for accurate evaluation of backscatter radiation from physician's own heads. To assess the eye-lens dose accurately, the dosimeter should be placed near the eye. If the dosimeter is placed behind the lens of the protective eyewear, we recommend using a backscatter radiation calibration factor of 1.2-1.3.

Gut Bacteria-derived Membrane Vesicles Induce Colonic Dysplasia by Inducing DNA Damage in Colon Epithelial Cells.

BACKGROUND & AIMS: Colorectal cancer (CRC) is the third most common cancer in the world. Gut microbiota has recently been implicated in the development of CRC. Actinomyces odontolyticus is one of the most abundant bacteria in the gut of patients with very early stages of CRC. A odontolyticus is an anaerobic bacterium existing principally in the oral cavity, similar to Fusobacterium nucleatum, which is known as a colon carcinogenic bacterium. Here we newly determined the biological functions of A odontolyticus on colonic oncogenesis. METHODS: We examined the induction of intracellular signaling by A odontolyticus in human colonic epithelial cells (CECs). DNA damage levels in CECs were confirmed using the human induced pluripotent stem cell-derived gut organoid model and mouse colon tissues in vivo. RESULTS: A odontolyticus secretes membrane vesicles (MVs), which induce nuclear factor kappa B signaling and also produce excessive reactive oxygen species (ROS) in colon epithelial cells. We found that A odontolyticus secretes lipoteichoic acid-rich MVs, promoting inflammatory signaling via TLR2. Simultaneously, those MVs are internalized into the colon epithelial cells, co-localize with the mitochondria, and cause mitochondrial dysfunction, resulting in excessive ROS production and DNA damage. Induction of excessive DNA damage in colonic cells by A odontolyticus-derived MVs was confirmed in the gut organoid model and also in mouse colon tissues. CONCLUSIONS: A odontolyticus secretes MVs, which cause chronic inflammation and ROS production in colonic epithelial cells, leading to the initiation of CRC.

Circulating T follicular helper 2 cells, T follicular regulatory cells and regulatory B cells are effective biomarkers for predicting the response to house dust mite sublingual immunotherapy in patients with allergic respiratory diseases.

The relationships between T follicular helper (Tfh) cells and antigen-specific immunoglobulins (sIgs) in patients with allergic respiratory diseases who are receiving antigen immunotherapy (AIT) have not been fully clarified. Therefore, we started to perform house dust mite sublingual immunotherapy (HDM-SLIT) for 20 patients with atopic asthma comorbid with allergic rhinitis (AA+AR) who were already receiving ordinary treatments including inhaled corticosteroid (ICS). We examined percentages of circulating T follicular helper (cTfh) and regulatory (cTfr) cells and percentages of circulating regulatory T (cTreg) and B (cBreg) cells by FACS and we examined levels of Der-p/f sIgs by ELISA. Based on the symptom score (asthma control questionnaire: ACQ) and medication score ((global initiative for asthma: GINA) treatment step score) in patients with AA, the patients were divided into responders and non-responders. The percentage of cTfh2 cells significantly decreased and the percentage of cTfh1 cells significantly increased within the first year. Der-p/f sIgEs decreased after a transient elevation at 3 months in both groups. Notably, the percentage of cTfh2 cells and the ratio of cTfh2/cBreg cells and Der-p/f sIgEs greatly decreased in responders from 6 months to 12 months. The percentages of cTfr and cTreg cells showed significant negative correlations with the percentage of cTfh2 cells. The percentage of IL-4+ cTfh cells were significantly decreased and the percentage of IFN-γ+ cTfh cells were increased before treatment to 24 months in 6 patients examined (4 responders and 2 non-responders). We performed multi plelogistic regression analysis based on these results, the ratios of cTfh2/cTfr cells and cTfh2/cBreg cells at the start of therapy were statistically effective biomarkers for predicting the response to HDM-SLIT in patients with AA+AR.