A short-term intervention of ingesting iron along with methionine and threonine leads to a higher hemoglobin level than that with iron alone in young healthy women: a randomized, double-blind, parallel-group, comparative study.

PURPOSE: Enhancing iron absorption and utilization is important for amelioration iron status faster and thereby, for improving quality of life. Dietary protein and amino acids, including methionine and threonine, have been reported to facilitate the absorption and utilization of dietary iron. Here, we investigated the effect of combined ingestion of methionine, threonine, and iron on the improvement of iron status during a short-term intervention, by comparing that with iron ingestion alone in healthy young women. METHODS: This was a randomized, double-blind, parallel-group, comparative study with 45 participants (aged 20-39) randomly assigned to three groups (n = 15 each): one group was administered 200 mg methionine, 400 mg threonine, and 6 mg iron once daily (FEMT); another ingested 6 mg iron alone (FE); and the third group ingested a placebo (PCG). Blood samples and dietary nutrient data were collected before the intervention (week 0) and after 2, 4, and 6 weeks. Serum iron, hemoglobin, transferrin, and ferritin levels were measured. RESULTS: Blood hemoglobin levels were significantly higher in the FEMT than in the FE group (P < 0.05) at week 4. Serum iron, transferrin, and ferritin levels were not changed across groups. In addition, our analyses showed that the observed increase in hemoglobin levels was affected by the intervention rather than changes in dietary nutrient intake. CONCLUSIONS: Ingestion of methionine and threonine with low doses of iron leads to a higher hemoglobin levels than that with iron alone in a short period of 4 weeks. TRIAL REGISTRATION: University Hospital Medical Information Network Clinical Trial Registry (UMIN000046621).

[A Case of Rectal Cancer That Could Be Safely Resected by Laparoscopic Surgery, after Capecitabine plus L-OHP plus Cetuximab as Pre-Operative Chemotherapy].

A 56-year-old woman. She was underwent a lower gastrointestinal endoscopy for bloody stool, and type 2 advanced rectal cancer was found. In CT scan, although distant metastasis is not found, the tumor has been expanded to the dorsal side. So, infiltration into the sacrum was suspected. For the risk of bleeding and residual tumor in circumferential resection surface, it was decided to perform pre-operative adjuvant chemotherapy. Because RAS gene has no mutation, the regimen chose CAPOX plus cetuximab. Although skin damage and cytopenia were observed, there was no appearance of adverse events that were intolerant, and 4 courses were performed. Although scar stenosis was observed in the endoscope after 4 courses, tumor size decreased. Even in CT, the wall thickening was significantly reduced, and progress to the tumor dorsal side was also reduced, so laparoscopic lower anterior resection was performed. During surgery, the tumor dorsal side sacral infiltration was suspected, although observed a sclerotic change, it is relatively easily peelable, it was possible to safely complete the laparoscopic operation. Even after the operation, the course was good, and it was discharged from the hospital lightly on the 12th day after the operation. In pathological diagnosis, medium-differentiated adenocarcinoma, T3, N0, histological therapeutic effect of chemotherapy was grade 2. Cetuximab combination regimen was considered to be an effective option.

[Goblet Cell Carcinoid of the Appendix with Complicated Appendicitis-A Case Report].

A 50-year-old male was referred to our hospital for the further evaluation and treatment of abdominal pain. He was diagnosed with complicated appendicitis using computed tomography. After conservative treatment, he underwent an interval appendectomy. A histopathological examination revealed a goblet cell carcinoid(GCC)of the appendix with subserosal invasion. He underwent laparoscopic ileocecal resection with D3 lymph node dissection. Histopathological findings showed neither residual tumor nor lymph node metastasis. The patients is currently followed as an outpatient without recurrence. Here we report our experience with GCC, a rare disease.

Interaction between Angiotensin Receptor and ß-Adrenergic Receptor Regulates the Production of Amyloid ß-Protein.

Alzheimer's disease (AD) is characterized by the formation of extracellular amyloid plaques containing the amyloid ß-protein (Aß) within the parenchyma of the brain. Aß is considered to be the key pathogenic factor of AD. Recently, we showed that Angiotensin II type 1 receptor (AT1R), which regulates blood pressure, is involved in Aß production, and that telmisartan (Telm), which is an angiotensin II receptor blocker (ARB), increased Aß production via AT1R. However, the precise mechanism underlying how AT1R is involved in Aß production is unknown. Interestingly, AT1R, a G protein-coupled receptor, was strongly suggested to be involved in signal transduction by heterodimerization with ß2-adrenergic receptor (ß2-AR), which is also shown to be involved in Aß generation. Therefore, in this study, we aimed to clarify whether the interaction between AT1R and ß2-AR is involved in the regulation of Aß production. To address this, we analyzed whether the increase in Aß production by Telm treatment is affected by ß-AR antagonist using fibroblasts overexpressing amyloid precursor protein (APP). We found that the increase in Aß production by Telm treatment was decreased by the treatment of ß2-AR selective antagonist ICI-118551 more strongly than the treatment of ß1-AR selective antagonists. Furthermore, deficiency of AT1R abolished the effect of ß2-AR antagonist on the stimulation of Aß production caused by Telm. Taken together, the interaction between AT1R and ß2-AR is likely to be involved in Aß production.

[A Case Report with R0 Resection for Locally Advanced Appendiceal Cancer Invading to the Right External Iliac Vessels].

A 46-year-old man presented with right lower quadrant pain. Enhanced CT revealed a 30 mm sized irregular shaped mass originating from the appendix and invading the abdominal wall. We performed the laparoscopic appendectomy. Finally, the pathologic finding revealed a local advanced appendiceal carcinoma. From the intraoperative findings, the residual tumor was existed around the right external iliac vessels and abdominal wall. The patient was treated with chemotherapy for 13 months. Although the residual tumor encased the right external iliac vessels, the abdominal wall, and psoas major muscle, there were no signs of distant metastasis. We performed ileocecal resection with D3 lymph node dissection. The right external iliac vessels, abdominal wall, and psoas major muscle were resected simultaneously, and reconstructed by femoral- femoral bypass. As a result, R0 resection was achieved pathologically.

[A Case of Advanced Hepatocellular Carcinoma Successfully Treated with Liver Resection after Administration of Sorafenib].

CASE: A 76-year-old man was referred to our hospital for advanced hepatocellular carcinoma(HCC)with chronic hepatitis type B. Although he underwent right anterior sectionectomy and S3 segmentectomy, multiple recurrences were found in the hepatic remnant after 2 months. Transcatheter arterial chemoembolization(TACE)and transcatheter arterial infusion (TAI)were performed separately. One and a half month after the last TAI, AFP and PIVKA-Ⅱ levels markedly elevated, and multiple early enhancing nodules with portal vein tumor thrombosis were detected on CT. A half dose of sorafenib(400mg/ day)was administered to the patient who was refractory to TACE. Sorafenib was discontinued after 4 weeks because the patient developed general fatigue and anorexia(Grade 3). Furthermore, these adverse events became worse, and ascites appeared. He was hospitalized in the palliative care unit for best supportive care for 3 weeks and also received outpatient treatment for more than 14 months. Fifteen months after discontinuing sorafenib administration, his condition improved dramatically, and CT revealed that the multiple HCC had reduced in size. Moreover, the portal vein tumor thrombosis disappeared. As his performance status and liver function were well preserved, he underwent partial hepatectomy for residual HCC. The patient remains alive without recurrence at 18 months, despite no administration of sorafenib. CONCLUSION: This case demonstrates that sorafenib administration combined with surgical treatment could possibly cure advanced HCC refractory to TACE.

Impaired insulin signaling in the B10.D2-Hc0 H2d H2-T18c/oSnJ mouse model of complement factor 5 deficiency.

Given the chemoattractant potential of complement factor 5 (C5) and its increased expression in adipose tissue (AT) of obese mice, we determined whether this protein of the innate immune system impacts insulin action. C5 control (C5cont) and spontaneously C5-deficient (C5def, B10.D2-Hc0 H2d H2-T18c/oSnJ) mice were placed on low- and high-fat diets to investigate their inflammatory and metabolic phenotypes. Adenoviral delivery was used to evaluate the effects of exogenous C5 on systemic metabolism. C5def mice gained less weight than controls while fed a high-fat diet, accompanied by reduced AT inflammation, liver mass, and liver triglyceride content. Despite these beneficial metabolic effects, C5def mice demonstrated severe glucose intolerance and systemic insulin resistance, as well as impaired insulin signaling in liver and AT. C5def mice also exhibited decreased expression of insulin receptor (INSR) gene and protein, as well as improper processing of pro-INSR. These changes were not due to the C5 deficiency alone as other C5-deficient models did not recapitulate the INSR processing defect; rather, in addition to the mutation in the C5 gene, whole genome sequencing revealed an intronic 31-bp deletion in the Insr gene in the B10.D2-Hc0 H2d H2-T18c/oSnJ model. Irrespective of the genetic defect, adenoviral delivery of C5 improved insulin sensitivity in both C5cont and C5def mice, indicating an insulin-sensitizing function of C5.

Anesthetic management with sevoflurane combined with alfaxalone-medetomidine constant rate infusion in a Thoroughbred racehorse undergoing a long-time orthopedic surgery.

A three-year old Thoroughbred racehorse was anesthetized with sevoflurane and oxygen inhalation anesthesia combined with constant rate infusion (CRI) of alfaxalone-medetomidine for internal fixation of a third metacarpal bone fracture. After premedication with intravenous (IV) injections of medetomidine (6.0 µg/kg IV), butorphanol (25 µg/kg IV), and midazolam (20 µg/kg IV), anesthesia was induced with 5% guaifenesin (500 ml/head IV) followed immediately by alfaxalone (1.0 mg/kg IV). Anesthesia was maintained with sevoflurane and CRIs of alfaxalone (1.0 mg/kg/hr) and medetomidine (3.0 µg/kg/hr). The total surgical time was 180 min, and the total inhalation anesthesia time was 230 min. The average end-tidal sevoflurane concentration during surgery was 1.8%. The mean arterial blood pressure was maintained above 70 mmHg throughout anesthesia, and the recovery time was 65 min. In conclusion, this anesthetic technique may be clinically applicable for Thoroughbred racehorses undergoing a long-time orthopedic surgery.

Getah Virus Infection among Racehorses, Japan, 2014.

An outbreak of Getah virus infection occurred among racehorses in Japan during September and October 2014. Of 49 febrile horses tested by reverse transcription PCR, 25 were positive for Getah virus. Viruses detected in 2014 were phylogenetically different from the virus isolated in Japan in 1978.

Epizootiological Investigation of Getah Virus Infection among Racehorses in Japan in 2014.

To clarify the factors causing an outbreak in 2014 of Getah virus infection among racehorses at the Miho training center, Japan, we isolated virus strains and performed an epizootiological investigation of affected horses and related horse populations. Three Getah virus isolates were recovered from clinical samples, and one of them (14-I-605) was used in a virus-neutralizing test. Of the affected horses (n = 33), 20 (60.6%) were 2-year-olds. We investigated the histories of Getah virus vaccination of the affected horses and the whole population at the Miho training center. Among the 2-year-old population, the prevalence of the disease in horses that had been vaccinated once was 14.1%. This was significantly higher than that in horses that had been vaccinated twice or more (1.3%; P < 0.01). Among horses that had entered the training center from farms in Ibaraki Prefecture surrounding the training center and from neighboring Chiba Prefecture, the rate of seropositivity for Getah virus was 13.0% in September 2014 and 42.9% in October 2014; that in the corresponding periods in 2010 and 2013 was 0%. In conclusion, we identified two possible causes of the outbreak of Getah virus infection in the training center in 2014: (i) the existence of susceptible horses that had received only one dose of vaccination before the outbreak and (ii) increased risk of exposure to the virus because of epizootic Getah virus infection among horses on surrounding farms in Ibaraki and Chiba prefectures.

Cellular differentiation assessments by measuring the degree of cellular internalization and membrane adsorption using designed peptides.

We demonstrate examples of cellular differentiation assessments, including cellular neurite outgrowth and fat cell maturation, by measuring the degree of membrane adsorption or cellular internalization using designed peptides. Because changes in the cellular membrane and cytosol during differentiation were shown to influence membrane adsorption and cellular internalization, we could successfully evaluate the extent of differentiation simply like stain indicators.

Reactive Lymphoid Hyperplasia of the Liver Incidentally Found in a 55-Year-Old Woman with a History of Ulcerative Colitis.

Reactive lymphoid hyperplasia (RLH) is a benign disease, rarely occurring in the liver. Reactive immune phenomenon has been reported in association with its occurrence, but the true pathogenesis is unknown. No case was reported in association with inflammatory bowel disease. We report a case of RLH of the liver in a patient with ulcerative colitis (UC). A 55-year-old woman with UC went to the outpatient clinic with abdominal pain, and antibiotics were prescribed with diagnosis of acute appendicitis. Imaging study detected a mass in the liver but ruled out appendicitis. She was referred to our hospital for further examination after pain improving. A 12 mm hypoechoic mass was detected in the liver on ultrasonography. There were no typical malignant findings on computed tomography and magnetic resonance imaging. Regular image follow-up was recommended, but the patient strongly requested surgery because of family history of malignant disease. Laparoscopic partial hepatectomy was performed. Histopathological findings revealed a conglomerate hyperplasia of lymphoid follicles with germinal centers. Infiltrating lymphocytes were non-neoplastic. Final diagnosis was RLH of the liver. UC is chronic inflammatory bowel disease and may be related to RLH, but there is no clear explanation at this point. This is the first known reported case of RLH of the liver in a patient with UC. But the relationship between the RLH and UC remains uncertain. Further investigation and case accumulation are necessary.

Systematic screening of the cellular uptake of designed alpha-helix peptides.

The cellular penetration (CP) activity of functional molecules has attracted significant attention as one of the most promising new approaches for drug delivery. In particular, cell-penetrating peptides (CPPs) have been studied extensively in cellular engineering. Because there have been few large-scale systematic studies to identify peptide sequences with optimal CP activity or that are suitable for further applications in cell engineering, such as cell-specific penetration and cell-selective culture, we screened and compared the cellular uptake (CU) activity of 54 systematically designed α-helical peptides in HeLa cells. Furthermore, the CU activity of 24 designed peptides was examined in four cell lines using a cell fingerprinting technique and statistical approaches. The CU activities in various cells depended on amino acid residues of peptide sequences as well as charge, α-helical content and hydrophobicity of the peptides. Notably, the mutation of a single residue significantly altered the CU ability of a peptide, highlighting the variability of cell uptake mechanisms. Moreover, these results demonstrated the feasibility of cell-selective culture by conducting cell-selective permeation and death in cultures containing two cell types. These studies may lead to further peptide library design and screening for new classes of CPPs with useful functions.

Determination of human FaFg of polyphenols using allometric scaling.

Certain polyphenols exhibit low permeability; precise prediction of their intestinal absorption is important for understanding internal exposure in humans. Intestinal availability, which represents the fraction of administered compounds that reach the portal blood (FaFg), is calculated by dividing bioavailability (F) by hepatic availability (Fh), and F is obtained from pharmacokinetic data from both intravenous (i.v.) and oral (p.o.) administration. However, human FaFg of polyphenols is hardly reported, as human i.v. data are extremely scarce. In this study, we developed an estimation method for FaFg of polyphenols in humans based on the extrapolation of rat clearance using allometric scaling (allometric scaling-based FaFg calculation method, AS- FaFgCM). First, for quercetin, for which human i.v. data have been reported, we compared the FaFg obtained by AS-FaFgCM with the traditional approach using human i.v. and p.o. data. Less than two-fold difference in FaFg values was observed between the two approaches. Next, we obtained FaFg of structurally diverse polyphenols (genistein, baicalein, resveratrol, and epicatechin) using AS-FaFgCM, demonstrating that all of them were poorly absorbable. Furthermore, to utilize the pharmacokinetic data of the total concentration, including aglycones and metabolites, we modified the AS-FaFgCM to focus on their excretion. The FaFg value of naringenin was obtained using modified AS-FaFgCM and was nearly equal to that of baicalein, a structural isomer of naringenin. This study provides quantitative information on the intestinal absorption of polyphenols using comprehensive estimation methods.

Adult Intestinal Malrotation Treated with Laparoscopic Ladd Procedure.

Intestinal malrotation is a rare congenital disease caused by abnormal intestinal rotation and fixation of the intestinal tract in the early embryonic state. Adult cases are rare. A laparoscopic Ladd procedure for adult intestinal malrotation is increasingly reported, but owing to the rarity, some important aspects of the disease and its treatment may be overlooked. Three adult cases of intestinal malrotation that underwent surgery at our hospital between January 2019 and October 2020 were retrospectively examined about patient backgrounds, short-term results, and complications. All patients were male, median age was 54.6 years, and the complaints were abdominal pain and/or distention. No midgut volvulus was observed. The laparoscopic Ladd procedure was performed for all cases. One patient underwent reoperation (duodenoduodenostomy) because of impaired passage of the duodenal descending section due to postoperative pancreatic fistula. The postoperative courses of the other two patients were good. No recurrence of symptoms was observed in any of the cases. The reason for reoperation in one of the cases is considered to be pancreatic injury when the severe curve from the duodenum to the upper jejunum near the pancreatic head was straightened. Correction of the curve is important to improve passage disorder of the duodenum, but special care is required to avoid organ damage, especially during a laparoscopic procedure with forceps. The laparoscopic Ladd procedure for adult intestinal malrotation is recommended if there is no midgut volvulus; it is minimally invasive and a comparatively simple technique, but surgeons should take special care to avoid organ damage.

Multidrug resistance-associated protein 2 (MRP2) is an efflux transporter of EGCG and its metabolites in the human small intestine.

Green tea polyphenols have various beneficial effects on human health, such as antiobesity and anti-carcinogenesis. (-)-Epigallocatechin-gallate (EGCG) is one of the major potent green tea catechins; however, detailed mechanisms of EGCG transport and metabolism in the human small intestine remain unknown due to lack of a suitable model. We investigated metabolite profiles of EGCG in the fresh human duodenal biopsy, cryopreserved human duodenal mucosal enterocytes and Caco-2 cells, and found that EGCG was readily metabolized into methylated and sulphate conjugates, which are major metabolites in these models. Next, we examined possible efflux transporters of EGCG and its metabolites using specific inhibitors of MRP2, P-gp and BCRP in Caco-2 cell monolayers. MRP2 was thereby identified as an efflux transporter, and further analysis using MRP2-knockout Caco-2 cells and vesicular transport assays confirmed that MRP2 is a selective efflux transporter of EGCG and its metabolites. Assuming that functional inhibition of MRP2 would result in efficient uptake of EGCG, we screened for MRP2 functional blockade and identified quercetin, which led to increased intracellular accumulation and basal transport of EGCG in Caco-2 cells. This result suggested that co-administration of quercetin and EGCG would enable efficient transport of EGCG in the human intestine. Therefore, we performed co-oral administration of quercetin and EGCG in human subjects to examine whether this occurred in humans. These studies demonstrated that MRP2 is a selective transporter of EGCG and conjugates and Caco-2 is a model to examine transport mechanisms and metabolites of polyphenols in the human small intestine.

Total synthesis of the bicyclic depsipeptide HDAC inhibitors spiruchostatins A and B, 5''-epi-spiruchostatin B, FK228 (FR901228) and preliminary evaluation of their biological activity.

The bicyclic depsipeptide histone deacetylase (HDAC) inhibitors spiruchostatins A and B, 5''-epi-spiruchostatin B and FK228 were efficiently synthesized in a convergent and unified manner. The synthetic method involved the following crucial steps: i) a Julia-Kocienski olefination of a 1,3-propanediol-derived sulfone and a L- or D-malic acid-derived aldehyde to access the most synthetically challenging unit, (3S or 3R,4E)-3-hydroxy-7-mercaptohept-4-enoic acid, present in a D-alanine- or D-valine-containing segment; ii) a condensation of a D-valine-D-cysteine- or D-allo-isoleucine-D-cysteine-containing segment with a D-alanine- or D-valine-containing segment to directly assemble the corresponding seco-acids; and iii) a macrocyclization of a seco-acid using the Shiina method or the Mitsunobu method to construct the requisite 15- or 16-membered macrolactone. The present synthesis has established the C5'' stereochemistry of spiruchostatin B. In addition, HDAC inhibitory assay and the cell-growth inhibition analysis of the synthesized depsipeptides determined the order of their potency and revealed some novel aspects of structure-activity relationships. It was also found that unnatural 5''-epi-spiruchostatin B shows extremely high selectivity (ca. 1600-fold) for class I HDAC1 (IC(50)=2.4 nM) over class II HDAC6 (IC(50)=3900 nM) with potent cell-growth-inhibitory activity at nanomolar levels of IC(50) values.

Crystal structure of benzyl 2-naphthyl ether, a sensitiser for thermal paper.

The title compound [systematic name: 2-(benz-yloxy)naphthalene], C17H14O, which is used as a sensitiser for thermal paper, has a twisted conformation with a dihedral angle of 48.71 (12)° between the phenyl ring and the naphthyl ring system. In the crystal, one mol-ecule inter-acts with six neighbouring mol-ecules via inter-molecular C-H⋯π inter-actions to form a herringbone mol-ecular arrangement.

The enhancement of antibody concentration and achievement of high cell density CHO cell cultivation by adding nucleoside.

Recently, with the dramatic increase in demand for therapeutic antibodies, Chinese hamster ovary (CHO) cell culture systems have made significant progress in recombinant antibody production. Over the past two decades, recombinant antibody productivity has been improved by more than 100-fold. Medium optimization has been identified as an important key approach for increasing product concentrations. In this study, we evaluated the effects of deoxyuridine addition to fed-batch cultures of antibody-expressing CHO cell lines. Furthermore, we investigated the effects of combined addition of deoxyuridine, thymidine, and deoxycytidine. Our results suggest that addition of these pyrimidine nucleosides can increase CHO cell growth, with no significant change in the specific production rate. As a result of the increased cell growth, the antibody concentration was elevated and we were able to achieve more than 9 g/L during 16 days of culture. Similar effects of nucleoside addition were observed in fed-batch cultures of a Fab fragment-expressing CHO cell line, and the final Fab fragment concentration was more than 4 g/L. This nucleoside addition strategy could be a powerful platform for efficient antibody production.