Bakuchiol targets mitochondrial proteins, prohibitins and voltage-dependent anion channels: New insights into developing antiviral agents.

We previously reported that bakuchiol, a phenolic isoprenoid anticancer compound, and its analogs exert anti-influenza activity. However, the proteins targeted by bakuchiol remain unclear. Here, we investigated the chemical structures responsible for the anti-influenza activity of bakuchiol and found that all functional groups and C6 chirality of bakuchiol were required for its anti-influenza activity. Based on these results, we synthesized a molecular probe containing a biotin tag bound to the C1 position of bakuchiol. With this probe, we performed a pulldown assay for Madin-Darby canine kidney cell lysates and purified the specific bakuchiol-binding proteins with SDS-PAGE. Using nanoLC-MS/MS analysis, we identified prohibitin (PHB) 2, voltage-dependent anion channel (VDAC) 1, and VDAC2 as binding proteins of bakuchiol. We confirmed the binding of bakuchiol to PHB1, PHB2, and VDAC2 in vitro using Western blot analysis. Immunofluorescence analysis showed that bakuchiol was bound to PHBs and VDAC2 in cells and colocalized in the mitochondria. The knockdown of PHBs or VDAC2 by transfection with specific siRNAs, along with bakuchiol cotreatment, led to significantly reduced influenza nucleoprotein expression levels and viral titers in the conditioned medium of virus-infected Madin-Darby canine kidney cells, compared to the levels observed with transfection or treatment alone. These findings indicate that reducing PHBs or VDAC2 protein, combined with bakuchiol treatment, additively suppressed the growth of influenza virus. Our findings indicate that bakuchiol exerts anti-influenza activity via a novel mechanism involving these mitochondrial proteins, providing new insight for developing anti-influenza agents.

Cardamonin decreases inflammatory mediator expression in IL-1ß-stimulated human periodontal ligament cells.

BACKGROUND: Cardamonin is classified as a natural chalcone, and has been reported to possess various bioactive effects. However, there have been limited attempts to utilize cardamonin in the treatment of periodontitis. This study aimed to investigate whether cardamonin has anti-inflammatory effects on human periodontal ligament cells (HPDLCs), which are a component cell of periodontal tissue. Specifically, the study seeks to determine whether cardamonin affects the expression of inflammatory mediators, such as cytokines and adhesion molecules, induced by interleukin-1ß (IL-1ß) in HPDLCs, as well as the signaling pathways activated by IL-1ß. METHODS: Cytokine and chemokine levels in supernatants of HPDLCs were measured by ELISA. Western blot analysis was used to measure protein expression and signal transduction pathway activation in HPDLCs. RESULTS: We found that IL-1ß-induced CC chemokine ligand (CCL)2, CCL5, CCL20, CXC-chemokine ligand (CXCL)10, and interleukin (IL)-6 production and intercellular adhesion molecule (ICAM)-1 and cyclooxygenase (COX)-2 expression in HPDLCs were suppressed by cardamonin treatment. We also found that cardamonin suppressed IL-1ß-activated nuclear factor (NF)-κB pathway, and the phosphorylation of signal transducer and activator of transcription (STAT)3. Furthermore, cardamonin treatment enhanced the expression of the antioxidant enzymes, heme oxygenase (HO)-1 and NAD(P)H dehydrogenase [quinone] 1 (NQO1), in HPDLCs. CONCLUSION: In this study, we found that cardamonin could suppress the production of inflammatory mediators in HPDLCs as well as the activation of several signaling pathways induced by IL-1ß treatment.

Cardamonin inhibits the expression of inflammatory mediators in TNF-α-stimulated human periodontal ligament cells.

OBJECTIVE: Periodontis is a chronic inflammatory disease induced by periodontopathogenic bacteria. The excessive immune response caused by persistent bacterial infection leads to alveolar bone resorption and ultimately tooth loss. Cardamonin is a biologically active substance that is found in the Zingiberaceae family, such as Alpinia zerumbet, and is classified as a natural chalcone. There have been no attempts to use cardamonin for the treatment of periodontitis, and no reports have examined the effects of cardamonin on periodontal tissue component cells. The aim of this study was to analyze effects of cardamonin on expression of inflammation mediators produced by TNFα-stimulated human periodontal ligament cells (HPDLCs), including its effects on signal transduction molecules. METHODS: Cytokine and chemokine levels were measured by ELISA. Protein expression in HPDLCs and activations of signal transduction pathway were determined by Western blotting. RESULTS: Our results indicate that cardamonin suppresses C-C motif chemokine ligand (CCL)2, CCL20, C-X-C motif chemokine ligand (CXCL)10, and interleukin (IL)-6 production and intercellular adhesion molecule (ICAM)-1 and cyclooxygenase (COX)-2 expression in TNF-α-stimulated HPDLCs. In addition, cardamonin induced the expression of the antioxidant enzyme, Heme Oxygenase (HO)-1, in HPDLCs. Furthermore, cardamonin suppressed TNF-α-stimulated c-Jun N-terminal kinase (JNK), nuclear factor (NF)-κB, and signal transducer and activator of transcription (STAT)3 signaling pathways in HPDLCs. CONCLUSION: We show that cardamonin reduces inflammatory mediator production by inhibiting the activation of several signaling pathways in this manuscript.

Transcriptome Analysis of PC12 Cells Reveals That trans-Banglene Upregulates RT1-CE1 and Downregulates abca1 in the Neurotrophic Pathway.

trans-Banglene and cis(c)-banglene possess neurotrophin-like activity in rat neurons. However, the molecular mechanisms underlying t-banglene-induced neurotrophic activity in rat and human neurons remain unclear. Here, we performed transcriptome analysis in PC12 cells, a rat adrenal gland pheochromocytoma cell line treated with t-banglene, using comprehensive RNA sequencing. The differentially expressed gene analysis of the sequencing data revealed that the expression of RT1 class I, locus CE1 (RT1-CE1) was upregulated, and that of ATP binding cassette subfamily A member 1 (abca1), myosin light chain 6, and hippocampus abundant transcript 1 was downregulated in t-banglene-treated PC12 cells, with statistically significant differences. We also confirmed the RT1-CE1 upregulation and abca1 downregulation in t-banglene-treated PC12 cells by real-time reverse transcription quantitative PCR. RT1-CEl is a major histocompatibility complex class I (MHCI) protein. ABCAl is a major cholesterol transporter that regulates efflux of intracellular cholesterol and phospholipids. Thus, our results suggest an exciting link between MHCI, cholesterol regulation, and neural development.

[A case of amebic colitis that developed after corticosteroid therapy in which perforation occurred during metronidazole treatment].

Although amebiasis is usually asymptomatic, fulminant amebic colitis is associated with a high mortality rate. Here, we report the case of a patient with amebic colitis in which bowel perforation occurred despite treatment with metronidazole. A man in his 70s underwent steroid pulse therapy to treat serious acute hepatitis A. After corticosteroid therapy, he developed acute abdomen because of amebic colitis. We immediately administered metronidazole and his symptoms improved. After completing treatment, he developed colon perforation. Amebic colitis can progress to bowel perforation even after administering the appropriate medication, so this abdominal symptom requires careful attention.

The effects of berteroin on inflammatory mediators and antioxidant enzymes expression in human periodontal ligament cells.

Berteroin is a bioactive substance classified as an isothiocyanate found in cruciferous vegetables such as cabbage, arugula, and salad leaves. In this study, we aimed to determine whether berteroin exerts anti-inflammatory effects on human periodontal ligament cells (HPDLCs), a resident cells of periodontal tissue. Berteroin suppressed interleukin (IL)-1ß or tumor necrosis factor (TNF)-α-induced chemokines (C-C motif chemokine ligand (CCL)2, CCL20, C-X-C motif chemokine ligand (CXCL)10, IL-8, and IL-6) production and intercellular adhesion molecule (ICAM)-1 expression in HPDLCs. In addition, berteroin inhibited phosphorylation of IκB kinase (IKK)- α/ ß, nuclear factor (NF)- κB p65, and IκB- α and degradation of IκB- α in the NF-κB pathway induced by IL-1 ß or TNF- α stimulation. Moreover, berteroin could inhibit signal transducer and activator of transcription (STAT)3 phosphorylation in TNF- α -stimulated HPDLC. Furthermore, berteroin increased the expression of the antioxidant enzymes, heme oxygenase (HO)-1 and NAD(P)H quinone dehydrogenase (NQO)1, in IL-1 ß or TNF- α -stimulated HPDLCs. These results suggest that berteroin may decrease the production of inflammatory mediators in HPDLCs by suppressing the NF-κB pathway, and may also decrease the local reactive oxygen species (ROS) production in periodontal lesions by increasing the production of antioxidant enzymes.

Eldest gastric cancer patient with high microsatellite instability responding to pembrolizumab.

Pembrolizumab has been associated with a high tumor response rate among high microsatellite instability (MSI-H) cancer patients. The efficacy and safety of pembrolizumab in the treatment of MSI-H gastric cancer (GC) patients aged ≥ 85 years have not been reported. This study reports the case of an 89-year-old woman diagnosed with stage IIA MSI-H GC based on her chief complaint of abdominal pain. We considered surgery, but it was contraindicated due to the patient's age and cardiovascular comorbidity. Therefore, we administered pembrolizumab after receiving approval from the ethics committee, and no significant adverse events were noted. The tumor was markedly responsive to pembrolizumab, and the computed tomography and endoscopic findings revealed a complete response. This is the first report on the efficacy and safety of pembrolizumab in the treatment of GC in an "oldest old" patient with MSI-H.

[A case of lymph node metastasis from esophageal small-cell-type endocrine cell carcinoma diagnosed by endoscopic ultrasound-guided fine-needle aspiration].

A 70-year-old man presented with a 0-IIc lesion in the lower esophagus. He underwent esophageal endoscopic submucosal dissection (ESD). The pathological diagnosis was squamous cell carcinoma with small cell carcinoma. The patient received chemoradiotherapy after ESD, but this was interrupted because of myelosuppression. Follow-up endoscopic ultrasonography (EUS) at 5 months after the ESD revealed paracardial lymph node swelling (17mm). He underwent EUS-guided fine-needle aspiration (FNA), and lymph node metastasis of small-cell-type endocrine cell carcinoma was diagnosed. He received VP-16 chemotherapy but died 22 months after the ESD.

Cerebral Venous Thrombosis as an Initial Presentation of Ulcerative Colitis.

Cerebral venous thrombosis (CVT) is a rare complication of ulcerative colitis (UC) that is potentially fatal once it occurs. This report describes a case of CVT that led to a diagnosis of UC. A 48-year-old woman was diagnosed with CVT due to paresthesia and weakness and was hospitalized for treatment. She developed bloody diarrhea on admission and was further diagnosed with UC based on endoscopic and pathologic findings. Treatment of UC with steroids and sulfasalazine was administered immediately. Her condition improved significantly within several days following treatment. After discharge, the patient experienced no recurrence of either CVT or UC flare-up over the last five years. This report describes CVT as an initial presentation of UC. This is also the first report of a long-term follow-up following successful treatment of CVT with concomitant UC.

Endoscopic submucosal dissection using a grasping-type scissors forceps for early gastric cancers and adenomas.

AIM: To reduce the risk of complications related to endoscopic submucosal dissection (ESD) using knives, we developed a new grasping-type scissors forceps (GSF) that can grasp and incise the target tissue using electrosurgical current. The aim of the present study was to evaluate the efficacy and safety of ESD using GSF for the removal of early gastric cancers and adenomas. METHODS: ESD using GSF was carried out on 35 consecutive patients with early gastric cancers or adenomas who had preoperative EUS diagnoses of mucosal tumor without lymph node involvement. Therapeutic efficacy and safety were assessed. RESULTS: All lesions were treated easily and safely without unexpected incision. The mean size of epithelial tumors and resected specimens was 15.6mm and 32.7mm, respectively. Curative en-bloc resection rates according to tumor size and location were 96% (26/27) in tumors ≤20mm, 100% (8/8) in tumors >20mm, 100% (18/18) of tumors in the lower portion, 100% (8/8) of tumors in the middle portion, 89% (8/9) of tumors in the upper portion, and 97% (34/35) overall. The mean operating time according to tumor size and location was 93.4min in tumors ≤20mm, 140min in tumors >20mm, 77.6min for tumors in the lower portion, 113.4min for tumors in the middle portion, 148.6min for tumors in the upper portion, and 104.1min overall. No intraoperative complication occurred, and postoperative bleeding was seen in 3% (1/35). CONCLUSIONS: ESD using GSF allows simple and safe en-bloc resection of early gastric cancer or adenoma irrespective of tumor size and location.

Perfluorinated compounds (PFCs) in Yodo River system, Japan.

There is increasing concern about occurrences of perfluorinated compounds (PFCs) in the environment due to their persistent, bioaccumulation, and potentially toxic effects. We investigated contamination of 11 PFCs (C4-C12) in the Yodo River system, which is a major source of drinking water for more than 11 million people of Kansai region in Japan. PFCs were detected in higher concentration even exceeding more than 1000 ng/L in some cases. Composition profile of total PFCs concentration showed that PFOS, PFHxA, PFOA, and PFNA were occurring dominantly in the river system. PFOS and PFOA concentration were nearly proportional to a previous study in the same river system, indicating continuous sink of the compounds into the river system. Although discharge of PFCs from wastewater effluents at upstream could be one of the major point sources, concentration did not exceed 29.4 ng/L at the downstream of main Yodo River, possibly, due to dilution effects. Total mass load of PFCs in mainstream of Yodo River was estimated 451.7 g/d based on observed river discharge and PFCs concentration. Results showed that Yodo River system could have a unique upstream-downstream linkage of PFCs contamination and the river could be a continuous pathway of PFCs exposure to the people of Kansai region.

Is double-balloon colonoscopy a better aid to unskilled colonoscopists than conventional colonoscopy?

BACKGROUND AND STUDY AIMS: To compare the clinical outcome of double-balloon colonoscopy (DBC) with conventional colonoscopy (CC) for colon evaluation performed by an unskilled colonoscopist. PATIENTS AND METHODS: Between June 2008 and November 2008, 1544 colonoscopies were performed in our hospital. Fifty-eight patients, (29 male and 29 female ; 19-86 years; mean age, 63 years) involving 60 intubations, were enrolled in this study and were assigned randomly to the DBC or CC group. One first-year GI fellow was enrolled and performed these 60 consecutive colonoscopies (30 DBCs, 30 CCs). Completion rate of colonoscopy, cecal intubation time, and rate of analgesic agent usage were analyzed. RESULTS: Completion of DBC was 100% (30/30), while completion of CC was 73% (22/30). There was a statistically significant difference (p < 0.05). The mean cecal intubation time was 36.2 +/- 14.4 minutes (DBC) and 36.5 +/- 15.2 minutes (CC). There was no statistically significant difference. Analgesic agent was used with 19 intubations (63%) (DBC) and with 27 intubations (90%) (CC) (p < 0.05). CONCLUSIONS: For inexpert endoscopists, using DBC has a higher rate of effectiveness than using CC and can decrease the discomfort of patients during colonoscopic procedures.

An incidentally identified 15 cm cavernous hemangioma of the small intestine: Case report and literature review.

INTRODUCTION: Hemangiomas of the small intestine are rare, usually present with symptoms such as anemia, gastrointestinal bleeding or abdominal pain and are resected. We report resection of an incidentally identified cavernous hemangioma of the small intestine that did not present symptoms referable to the hemangioma. Although it was a large lesion, it was resected using laparoscopy and a mini-laparotomy. PRESENTATION OF CASE: A 29-year-old otherwise healthy man was referred for evaluation of ileal wall thickening found on a contrast-enhanced computed tomography scan obtained for the workup of chronic diarrhea. Double balloon enteroscopy (DBE) showed a cavernous hemangioma of the small intestine. The lesion was 15 cm and resected using laparoscopy and a mini-laparotomy to prevent future bleeding. The histopathological diagnosis was a cavernous hemangioma of the ileum. DISCUSSION: Though there have been no reports of the asymptomatic patients of the disease, the recent spread of double balloon enteroscopy and capsule endoscopy will allow us to diagnose more asymptomatic hemangiomas like this patient. Also, this large lesion was able to be resected through a small incision due to its compressible nature. CONCLUSION: Future studies in asymptomatic patients of the disease may help to determine the optimal management for these patients. Even large hemangiomas are compressible, facilitating minimally invasive resection.

Randomized controlled trial: roxatidine vs omeprazole for non-erosive reflux disease.

BACKGROUND/AIMS: Proton pump inhibitor (PPI) therapy is considered as the first choice for treatment of non-erosive reflux disease (NERD). However, NERD is less sensitive to PPIs than erosive gastroesophageal reflux disease (GERD) and the differences between PPIs and H2 receptor antagonists are less evident in NERD than in erosive GERD. Since gastric acid secretion is lower in the Japanese population than in Western populations, we aimed to investigate whether PPI therapy is really necessary for NERD patients in Japan. METHODOLOGY: Thirty-three symptomatic endoscopically diagnosed NERD patients were randomly assigned to receive roxatidine acetate 75 mg twice daily (n = 16) or omeprazole 20 mg once daily (n = 17). Gastrointestinal symptoms were assessed using the Gastrointestinal Symptom Rating Scale at baseline and after 4 and 8 weeks of treatment. RESULTS: Both roxatidine and omeprazole significantly improved the heartburn score at 4 and 8 weeks. The clinical response rates did not differ between roxatidine and omeprazole. Both roxatidine and omeprazole significantly relieved not only reflux but also abdominal pain and indigestion. The degrees of improvement did not differ between the two groups. CONCLUSION: Roxatidine relieved the symptoms of NERD patients with similar effectiveness to omeprazole. Therefore, roxatidine may be a good choice for NERD treatment.

[A case of protein-losing gastroenteropathy accompanied by Sjögren syndrome and mixed connective tissue disease].

Case reports of protein-losing gastroenteropathy (PLGE) associated with not only mixed connective tissue disease (MCTD) but also Sjögren syndrome (SjS) are very rare. We report a first case of PLGE in a patient with both MCTD and SjS. A 58-year-old Japanese woman was referred and admitted to our hospital because of abdominal fullness and lower leg edema. Her past medical history revealed SjS at age 40. Physical examination demonstrated lower leg edema and Raynaud's phenomenon. Blood chemistry data showed severe hypoproteinemia. Anti RNP antibody was positive. MCTD was diagnosed. The alpha-1 antitrypsin clearance level was high. The (99m)Tc-DTPA human serum albumin scintigraphy demonstrated abnormal accumulation in the intestine. PLGE associated with both MCTD and SjS was diagnosed, but she was successfully treated by prednisolone.

Endoscopic submucosal dissection of a rectal carcinoid tumor using grasping type scissors forceps.

Endoscopic submucosal dissection (ESD) with a knife is a technically demanding procedure associated with a high complication rate. The shortcomings of this method are the inability to fix the knife to the target lesion, and compression of the lesion. These can lead to major complications such as perforation and bleeding. To reduce the risk of complications related to ESD, we developed a new grasping type scissors forceps (GSF), which can grasp and incise the targeted tissue using electrosurgical current. Colonoscopy on a 55-year-old woman revealed a 10-mm rectal submucosal nodule. The histological diagnosis of the specimen obtained by biopsy was carcinoid tumor. Endoscopic ultrasonography demonstrated a hypoechoic solid tumor limited to the submucosa without lymph node involvement. It was safely and accurately resected without unexpected incision by ESD using a GSF. No delayed hemorrhage or perforation occurred. Histological examination confirmed the carcinoid tumor was completely excised with negative resection margin.