Influence of Endogenous Substances on Site-II to Site-I Displacement of Diclofenac Bound to Albumin in the Aqueous Humor of Patients with Cataract.

Diclofenac instillation is useful in preventing intraoperative miosis and macular edema caused by postoperative inflammation in cataract surgery; however, optimum efficacy is not attained when the instilled diclofenac strongly binds to albumin in patients' aqueous humor. Therefore, a method that inhibits diclofenac binding and increases the concentration of its free fraction is needed. We conducted a basic study regarding the effects of inhibitors on the binding of instilled diclofenac to albumin and endogenous substances in aqueous humor. Aqueous humor samples from 16 patients were pooled together for analysis. The free fraction of diclofenac was measured using ultrafiltration methods in various experiments with pooled and mimic aqueous humor. Free fraction of diclofenac, a site II drug, in pooled aqueous humor was 0.363 ± 0.013. The binding of diclofenac in the presence of phenylbutazone (PB), a site I inhibitor, was significantly inhibited (free fraction = 0.496 ± 0.013); however, no significant inhibition by ibuprofen, a site II inhibitor, (free fraction = 0.379 ± 0.004), was observed. The unexpected result was due to free fatty acids (FFAs; palmitic acid (PA)) and L-tryptophan (Trp). The inhibition of diclofenac binding by PB in the mimic aqueous humor containing these endogenous substances revealed significant binding inhibition in the presence of PA and Trp. Diclofenac is strongly rebound from site II to site I in the presence of FFAs and Trp in the aqueous humor because FFAs and Trp induce a conformational change in albumin. Therefore, PB significantly inhibits the binding of diclofenac to albumin.

Severity of constipation related to palonosetron during first-line chemotherapy: a retrospective observational study.

PURPOSE: Palonosetron, a long-acting 5-HT3 receptor antagonist, is an effective antiemetic agent for chemotherapy-induced nausea and vomiting; however, it sometimes causes severe constipation. The aim of the present study was to evaluate the severity of palonosetron-related constipation. METHODS: We retrospectively analyzed the incidence and severity of constipation after intravenous administration of 0.75-mg palonosetron in 150 chemotherapy-naïve patients who received first-line chemotherapy at Saga University Hospital. Constipation was classified into grades 1-5 according to the Common Terminology Criteria for Adverse Events version 5.0. Multiple logistic regression analysis was performed to identify factors associated with palonosetron-related worsening of constipation to grade 2 or higher. RESULTS: Palonosetron significantly increased the incidence and severity of constipation (incidence: before vs. after palonosetron, 35.4% vs. 74.0%, p < 0.0001, and severity: before vs. after palonosetron, 26.7% and 8.7% in grades 1 and 2, respectively, vs. 46.7%, 23.3%, and 4.0% in grades 1, 2, and 3, respectively, p < 0.0001). Despite the use of laxatives, 4.0% of patients had grade 3 constipation requiring manual evacuation. Combination treatment with aprepitant (odds ratio (OR), 10.9; 95% confidence interval (CI), 1.3-90.0; p = 0.026) and older age (OR, 1.25; 95% CI, 1.01-1.57; p = 0.039) were factors associated with the severity of constipation. CONCLUSION: Constipation was more severe in patients receiving combination treatment with aprepitant than in those treated with palonosetron alone. Older age was also associated with increased risk of severe palonosetron-related constipation. Identification of risk factors can help target risk-based laxative therapy.

A case of a rectal adenocarcinoma mimicking a neuroendocrine tumor in the background mucosa of amoebic colitis.

A 54-year-old man with diarrhea and hematochezia for 2 months was referred to our department. A total colonoscopy revealed amoebic colitis caused by Entamoeba histolytica. Concurrently, a submucosal tumor-like yellowish hemispheric polypoid lesion was incidentally detected in the rectum. We speculated that the lesion was a NET, which could be cured by endoscopic treatment. However, histopathological assessment of a biopsy specimen unexpectedly revealed a well- or moderately differentiated adenocarcinoma. After treatment of the amoebic colitis, endoscopic ultrasound revealed a low, hetero-echoic, 6-mm-diameter lesion mainly in the submucosal layer. We performed surgical resection because the invasion was estimated to be to the deeper submucosal layer. Histopathological assessment of the surgically resected specimen revealed a focal lesion of a well-differentiated adenocarcinoma in the granulation tissue of the submucosal layer. In cases accompanied by amoebic colitis, a tumor's initial gross type might change. Diagnostic endoscopic resection could be acceptable in such cases.

[Selection of Operative Procedure for Breast Cancer in Carriers of BRCA VUS in Japan].

Risk classification and clinical management of the DNA variant of unknown significance(VUS)in BRCA 1/2 remains unestablished. The Japanese hereditary breast and ovarian cancer(HBOC)consortium and myriad genetics reported that the VUS rate of BRCA is 6.5% in Japanese patients, but is <2% in the USA. The types of mutation supposedly differ between Asian and European ethnicities. Breast-conserving therapy(BCT)is not recommended in HBOC breast cancer, according to the 2017 Japanese guidelines by the Ministry of Health, because of the risk of ipsilateral breast recurrence(IBR)and carcinogenesis by radiation. In our hospital, we recommend an initial mastectomy and breast reconstruction with an implant for patients with HBOC breast cancer, considering future surgery on the contralateral side and symmetry of the reconstructed breast. However, the risk of IBR after BCT is not significantly high in patients with HBOC breast cancer, and BCT is a reasonable option even for definite HBOC breast cancer under low risk conditions. Hence, BCT is feasible for treating breast cancer in carriers of VUS following decision-making and informed consent from the patients.

Plasma homocysteine concentrations in novel microminipigs.

A novel microminipig has been recently developed for use in biomedical research. In the present study, age- and sex-related differences, as well as 24-h fluctuations in plasma total homocysteine concentrations (tHcy), were investigated in these microminipigs. tHcy (mean±SD) was 10.2±3.4 µM and significantly correlated with age. By contrast, neither the differences in tHcy between sexes nor the 24-h fluctuations in tHcy after feeding were significant. The kinetics of plasma tHcy after intravenous injection of reduced Hcy showed that its levels peaked within 5 min post-injection, as did the levels of tHcy. These results suggested that reduced Hcy is rapidly oxidized or metabolized. The half-lives of reduced Hcy, tHcy, and reduced cysteine in the blood were 47, 71, and 141 min, respectively. In conclusion, there was a significantly positive correlation between age and plasma tHcy in microminipigs. After intravenous injection of reduced Hcy, plasma tHcy quickly returned to pre-injection levels.

Albumin-binding of diclofenac and the effect of a site II inhibitor in the aqueous humor of cataract patients with the instillation of diclofenac.

Diclofenac instillation has been used widely in cataract surgery to prevent postoperative inflammation. Since diclofenac binds strongly to albumin in the circulation, it does not have a sufficient effect on patients in whom diclofenac binds strongly to albumin in the aqueous humor. A decrease in diclofenac binding and an increase in free diclofenac levels are necessary in these patients. The binding of diclofenac to albumin was investigated in the aqueous humor. In a diclofenac binding assay with albumin in the aqueous humor of individual patients, diclofenac was extracted from aliquots of the aqueous humor, and its total levels were measured using ultra high performance liquid chromatography (UHPLC). Free diclofenac levels were measured using ultrafiltration and UHPLC. The albumin-binding fraction of diclofenac was 0.8 or higher in the aqueous humor of some patients. Ibuprofen significantly inhibited diclofenac binding to site II of albumin in mimic aqueous humor, but not in pooled aqueous humor. This difference may have been due to the weak binding of diclofenac to site II in the pooled aqueous humor. Flurbiprofen was used instead of diclofenac. Flurbiprofen has been shown to bind more strongly than diclofenac to the same site of albumin. Thus, the inhibitory effect of ibuprofen on the binding of flurbiprofen to albumin was investigated in pooled aqueous humor. The results indicated that ibuprofen significantly inhibited the flurbiprofen binding. An effective diclofenac administration method may be established for clinical application by the instillation of an appropriate inhibitor of binding to the albumin site II.

The citrus flavonoids hesperetin and naringenin block the lipolytic actions of TNF-alpha in mouse adipocytes.

Obese adipose tissue is characterized by an excessive production of inflammatory adipokines including tumor necrosis factor-alpha (TNF-alpha). TNF-alpha stimulates free fatty acid (FFA) secretion through adipocyte lipolysis, and increased plasma levels of FFA promote insulin resistance. In this report, we show that hesperetin and naringenin, two citrus flavonoids, inhibit TNF-alpha-stimulated FFA secretion from mouse adipocytes. These flavonoids block the TNF-alpha-induced activation of the NF-kappaB and ERK pathways. Moreover, hesperetin and naringenin prevent TNF-alpha from downregulating the transcription of two antilipolytic genes, perilipin and PDE3B. These effects are mediated through the inhibition of the ERK pathway. In contrast, the inhibition of the NF-kappaB pathway by hesperetin and naringenin suppresses the transcription of IL-6, which induces FFA secretion in an autocrine manner. Our results provide novel evidence that hesperetin and naringenin directly inhibit TNF-alpha-stimulated FFA secretion. These findings may be useful for ameliorating FFA-induced insulin resistance.

Anti-human very late antigen-alpha4 (CD49d) monoclonal antibody (BU49) cross-reacts with the canine B-cell leukemia cell line GL-1, resulting in the induction of homotypic cell aggregation.

We have found that the anti-human very late antigen-alpha4 (VLA-alpha4) (CD49d) monoclonal antibody (mAb) BU49 cross-reacts with the canine B-cell leukemia cell line GL-1. Interestingly, the BU49 mAb specifically induced the homotypic cell aggregation of GL-1 cells accompanied by morphological changes. Homotypic cell aggregates induced by BU49 mAb were blocked by the presence of a protein kinase C inhibitor, a protein kinase A inhibitor, an actin filament formation inhibitor, and an EDTA. On the other hand, a protein tyrosine kinase inhibitor, a DNA-synthesis inhibitor, and an anti-canine CD45 mAb did not affect the GL-1 homotypic cell aggregation induced by BU49 mAb. The BU49 mAb immunoprecipitated at a molecular weight of about 150kDa in the GL-1 cells, similar to the results in the human monocyte-like cell line U937. Taken together, our results provide the first evidence that human CD49d recognized by BU49 mAb has unique immunological functions against canine cells.

Diagnosis of small pancreatic cancer by endoscopic balloon-catheter spot pancreatography: an analysis of 29 patients.

OBJECTIVES: The diagnosis of small pancreatic cancer remains difficult. The present study describes the diagnostic value of endoscopic balloon-catheter spot pancreatography for small pancreatic cancer. METHODS: Since April 1984, balloon spot pancreatography has been used to detect small-sized pancreatic cancer in patients having possible symptoms or findings of obstructive pancreatitis. RESULTS: A resection was performed on 175 of 416 patients with conditions diagnosed as pancreatic cancer. Of the 175 patients, 23 (13%) had invasive carcinoma 2 cm or smaller based on histological measurements, 3 intraductal papillotubular adenocarcinoma, and 3 carcinoma in situ (CIS). Regarding invasive carcinoma, balloon pancreatography displayed duct abnormalities diagnosed as carcinoma in 20 of 22 patients, whereas carcinoma was suggested in 2. A definite diagnosis was obtained based on the findings of main duct stenosis or obstruction with marked stricture of the branch ducts (n = 18) and a filling defect in the main duct (n = 2). Moreover, this pancreatogram demonstrated an intraductal filling defect in 2 of 3 with intraductal carcinoma and dead twiglike findings in the branch ducts in 1 of 3 with CIS. CONCLUSIONS: Balloon spot pancreatography is an essential tool for the diagnosis of small ductal pancreatic cancer, and it also makes it possible to locate CIS lesions of the branch ducts.

Dosage plan of a flurbiprofen injection product using inhibition of protein binding by lipid emulsion in rats.

Flurbiprofen-axetil (FP-ax), a bolus injection product of a non-steroidal anti-inflammatory drug (NSAID), is a prodrug of flurbiprofen, an NSAID. As flurbiprofen strongly binds to site II of human serum albumin (HSA), the free (unbound) concentration of flurbiprofen after injection of FP-ax is low. We have examined the inhibitory effect of free fatty acid (FFA), a binding inhibitor for site II of HSA, on the binding of flurbiprofen in-vitro and in-vivo by ultrafiltration, to establish an effective dosage of FP-ax. In-vitro, fatty acid mixtures (FAs) inhibited the binding of flurbiprofen to rat serum albumin. The free fraction of flurbiprofen was remarkably increased by FAs in rat serum. In-vivo, FP-ax was injected into a control group (low FFA concentration in serum) and a lipid emulsion group (high FFA concentration in serum). The area under the curve of the free concentration of flurbiprofen during the alpha phase and the distribution volume of the central compartment of flurbiprofen were significantly higher in the lipid emulsion group than the control group (5.0- and 1.2-times, respectively). When FP-ax was administered at high FFA concentration, the free concentration of flurbiprofen and distribution of flurbiprofen to tissues increased transiently. This administration method may be useful for patients with cancer pain, having a potent analgesic effect.

Microscopic observation and characterization of the oil bridge between dehulled-roasted sesame seeds.

The formation of an oil bridge between adhesive seeds was observed microscopically. The geometry of the oil bridge was affected by the shape of the adhesive seeds. The capillary force of the oil bridge was estimated from the image captured by the microscope. The average capillary force was 127 microN, which was five times higher than the average gravity of the seeds. It was observed that several oil bridges formed between two seeds. These results indicated an adequate ability of the seeds to adhere. The capillary force of the oil bridge increased with surface oil content. The probability of formation of an oil bridge increased with surface oil content when the surface oil content was above 0.63%. The probability of formation of an oil bridge markedly increased when sucrose was added to the seeds.

[Evaluation of diagnostic imagings of liver metastases].

With the advent of various therapeutic modalities for the management of metastatic liver tumor, the task of pretreatment imaging has become more demanding. US and CT are non-invasive, and the most widely used techniques for pretreatment imaging, but they are far from optimal. Recently, the most sensitive pretreatment imaging modality for the depiction of focal liver lesions is CT during arterial portography (CTAP); however, it is an invasive procedure with an established risk of false-positive results. CT and MRI are the sensitivities of these techniques have recently been improved with the development of multidetector CT, contrast agents for MRI that specifically accumulate in the liver, and other advances. Superparamagnetic iron oxide (Feridex) has been developed as a liver-specific particulate MRI contrast agent that is taken up by the Kupffer cells of the liver. Rational selection of appropriate modalities for a given purpose requires familiarity with the characteristics of each modality. The aim of the present study was to evaluate the sensitivities of diagnostic modalities in the pre- and post-treatment periods of metastatic liver tumor. Feridex-enhanced MRI (Ferumoxide MRI) is more sensitive than enhanced CT and MRI in the depiction of metastatic liver tumor. In terms of the ability to visualize residual tumor after MCT, dynamic MRI was superior to enhanced CT and Ferumoxide MRI. In conclusion, combined Ferumoxide MRI and dynamic MRI is a noninvasive method and clinically useful since it can change the therapeutic approach.

Role of cyclic nucleotides in ischemia and reperfusion injury of canine livers.

BACKGROUND: In a series of canine liver ischemia experiments, we have shown that amelioration of hepatic injury is achievable by the inhibition of vasoconstriction, cytokine production, platelet aggregation, and neutrophil infiltration. Cyclic adenosine diphosphate (cAMP) was considered to be involved in most of these events. In our study, we tested our hypothesis that augmentation of endogenous cAMP by phosphodiesterase (PDE) 3 inhibitor, amrinone (AM), or adenylate cyclase stimulator, NKH477 (NKH), could attenuate ischemia and reperfusion injury of the liver. METHODS: Thirty-six beagle dogs were used. They were divided into group CT (untreated control), group AM, group NKH, and group CB (treated by both agents). AM or NKH were administered i.v. 1 hr before ischemia (group preAM and group preNKH) or 15 min before reperfusion (pos-AM and postNKH). Combination group animals were treated only before ischemia. Animal survival, hepatic tissue blood flow, liver enzymes, platelet counts, energy metabolism, hepatic cAMP and cyclic guanosine 3',5'-cyclic monophosphate levels, and histopathology were analyzed. RESULTS: Two-week animal survival was significantly improved by pre- or posttreatment with either agent. After reperfusion, hepatic tissue blood flow, liver enzyme release, platelet counts, energy metabolism, tissue cAMP levels, and histological architecture were also ameliorated markedly. Combination of both agents induced severe liver damage and lethal hypotension. AM treatment exhibited more protective effects than NKH, particularly when it was given before ischemia. Interestingly, not only cyclic guanosine 3',5'-cyclic monophosphate, were also restored at higher levels after reperfusion by preischemia treatment. CONCLUSIONS: Administration of amrinone or NKH477 maintained hepatic tissue concentrations of cyclic nucleotides, and attenuated ischemia and reperfusion injury of the liver. Thus, regulation of hepatic tissue cyclic nucleotides is an important alternative for prevention of hepatic damage in liver preservation and surgery.