Short-chain fatty acids induce tissue plasminogen activator in airway epithelial cells via GPR41&43.

BACKGROUND: Chronic rhinosinusitis (CRS) is a heterogeneous chronic inflammatory disease generally divided based on the presence or absence of nasal polyps (NPs). One of the features of NPs is excessive fibrin deposition, which is associated with down-regulation of tissue plasminogen activator (t-PA) in NPs. As t-PA is expressed in epithelial cells, and epithelium is readily accessible to topical therapies, identifying compounds that can mediate the induction of t-PA would be a potential new strategy for the treatment of NPs. OBJECTIVE: The objective of this study was to determine whether short-chain fatty acids (SCFAs) can induce t-PA in airway epithelial cells via their known receptors GPR41 and GPR43. METHODS: We performed immunohistochemistry (IHC) to determine whether receptors for SCFAs, known as G protein-coupled receptor 41/free fatty acid receptor 3 (GPR41/FFAR3) and GPR43/FFAR2, are expressed in nasal tissue. Primary normal human bronchial epithelial (NHBE) cells were stimulated with different concentrations of SCFAs to test induction of t-PA, which was analysed by expression of mRNA and protein. Mediation of responses by SCFA receptors was evaluated by specific receptor gene silencing with siRNA. RESULTS: Immunohistochemistry study revealed that airway epithelial cells expressed GPR41 and GPR43. Acetic acid, propionic acid, butyric acid and valeric acid significantly induced t-PA expression from two- to tenfolds. The strongest inducer of t-PA from NHBE cells was propionic acid; cells stimulated with propionic acid released t-PA into the supernatant in its active form. Gene silencing of GPR41 and GPR43 revealed that induction of t-PA by SCFAs was dependent upon both GPR41 and GPR43. CONCLUSIONS AND CLINICAL RELEVANCE: Short-chain fatty acids were shown to induce airway epithelial cell expression of t-PA via GPR41 and GPR43. Topical delivery of potent compounds that activate these receptors may have value by reducing fibrin deposition and shrinking nasal polyp growth.

Novel scoring system and algorithm for classifying chronic rhinosinusitis: the JESREC Study.

BACKGROUND: Chronic rhinosinusitis (CRS) can be classified into CRS with nasal polyps (CRSwNP) and CRS without nasal polyps (CRSsNP). CRSwNP displays more intense eosinophilic infiltration and the presence of Th2 cytokines. Mucosal eosinophilia is associated with more severe symptoms and often requires multiple surgeries because of recurrence; however, even in eosinophilic CRS (ECRS), clinical course is variable. In this study, we wanted to set objective clinical criteria for the diagnosis of refractory CRS. METHODS: This was a retrospective study conducted by 15 institutions participating in the Japanese Epidemiological Survey of Refractory Eosinophilic Chronic Rhinosinusitis (JESREC). We evaluated patients with CRS treated with endoscopic sinus surgery (ESS), and risk of recurrence was estimated using Cox proportional hazard models. Multiple logistic regression models and receiver operating characteristics curves were constructed to create the diagnostic criterion for ECRS. RESULTS: We analyzed 1716 patients treated with ESS. To diagnose ECRS, the JESREC scoring system assessed unilateral or bilateral disease, the presence of nasal polyps, blood eosinophilia, and dominant shadow of ethmoid sinuses in computed tomography (CT) scans. The cutoff value of the score was 11 points (sensitivity: 83%, specificity: 66%). Blood eosinophilia (>5%), ethmoid sinus disease detected by CT scan, bronchial asthma, aspirin, and nonsteroidal anti-inflammatory drugs intolerance were associated significantly with recurrence. CONCLUSION: We subdivided CRSwNP in non-ECRS, mild, moderate, and severe ECRS according to our algorithm. This classification was significantly correlated with prognosis. It is notable that this algorithm may give useful information to clinicians in the refractoriness of CRS before ESS or biopsy.

Conventional tumor markers are prognostic indicators in patients with head and neck squamous cell carcinoma.

We tested for squamous cell carcinoma-related antigen (SCC), carcinoembryonic antigen (CEA), ferritin, immunosuppressive acid protein (IAP) and sialic acid in the serum from 247 patients with head and neck squamous cell carcinoma prior to therapy. Significant correlations were found between IAP and tumor size, lymph node metastasis, and clinical stage (P<0.0001, P<0.001, and P<0.0001). Also, sialic acid and SCC were also correlated with tumor size, lymph node metastasis, and clinical stage. Moreover IAP, sialic acid and SCC were strongly associated with survival rate (P<0.0001, P = 0.0230 and P = 0.0159). A multivariate Cox proportional hazard model demonstrated that being positive for IAP was an independent predictor for patients with H&NSCC (P = 0.0115). The results indicate that IAP, sialic acid and SCC are useful as prognostic factors.

Obstructive jaundice caused by a huge liver cyst riding on the hilum: report of a case.

A 71-year-old man presented to our hospital with obstructive jaundice, found to be caused by a huge liver cyst which was centrally located and riding on the hilum. Percutaneous transhepatic cyst drainage was performed, following which obstruction of the bile duct was relieved and the jaundice subsided. As jaundice recurred after removal of the drainage tube, the patient underwent deroofing, since when he has remained well. Only 13 cases of liver cysts producing obstructive jaundice have been reported in the English literature, most of which were characteristically enormous, located centrally, and riding on the hilum. Liver cysts possessing such features are likely to cause obstructive jaundice by compressing the hepatic hilum. Cyst drainage is helpful for ameliorating the jaundice and making an accurate diagnosis; however, subsequent deroofing or injection therapy is necessary to prevent recurrence.

Both C3a and C3a(desArg) regulate interleukin-6 synthesis in human peripheral blood mononuclear cells.

Synthesis of complement components is part of the acute-phase response. Interleukin-6 (IL-6) is a critical mediator of the acute-phase response during infections and injuries. Plasma levels of C3a and IL-6 have been proposed as prognostic indicators in sepsis and trauma. The effects of C3a and C3a(des)Arg on IL-6 gene expression and protein production in human peripheral blood mononuclear cells (PBMC) were investigated. Neither C3a nor C3a(des)Arg alone induced detectable IL-6 protein or mRNA levels. However, C3a and C3a(des)Arg affected endotoxin-induced IL-6 synthesis in a dose-dependent manner. In nonadherent PBMC, C3a or C3a(des)Arg suppressed, while in adherent PBMC, C3a or C3a(des)Arg enhanced IL-6 protein and mRNA levels. These results suggest that C3a and C3a(des)Arg may provide a control mechanism of acute-phase responses by enhancing IL-6 synthesis in adherent monocytes at local inflammatory sites and by inhibiting IL-6 synthesis in circulating monocytes.

Anti-interleukin-8 monoclonal antibody reduces free radical production and improves hemodynamics and survival rate in endotoxic shock in rabbits.

BACKGROUND: Although high levels of interleukin-8 (IL-8) have been found in patients with sepsis and a monoclonal antibody (MoAb) against IL-8 has been successfully used in some animal models of inflammation, no specific therapeutic agent against IL-8 has been tested for the treatment of sepsis. We studied the effects of a MoAb against IL-8 in the treatment of endotoxic shock with a prospective randomized rabbit endotoxic shock model. METHODS: Twenty New Zealand white rabbits were anesthetized and divided into four groups: normal, anti-IL-8, control-Ab, and lipopolysaccharide (LPS). Anti-IL-8 and control-Ab groups received a MoAb (immunoglobulin G, 3 mg/kg) 5 minutes before the LPS injection. All groups, except the normal group, received a continuous 20-minute infusion of LPS (500 micrograms/kg). The normal group received NaCl (0.9%) rather than LPS. RESULTS: The 7-day survival rates were 100% for normal group, 80% for anti-IL-8 group, 40% for control-Ab group, and 0% for LPS group. Compared with the LPS group, anti-IL-8 rabbits had a smaller decrease in mean arterial blood pressure (p < 0.05) and increased urinary volume (p < 0.05). Anti-IL-8 rabbits had lower plasmatic levels of IL-1 beta, less free radical production (p < 0.05), and a higher survival rate (p < 0.01). CONCLUSIONS: IL-8 plays a significant role in endotoxic shock, and IL-8 blockage results in attenuation of the hypotensive and tachypneic effects of LPS, reduced free radical production, and an increased survival rate after lethal endotoxic shock.

A new biologic role for C3a and C3a desArg: regulation of TNF-alpha and IL-1 beta synthesis.

The complement activation products C3a and C3a desArg are generated in the course of trauma, infection, tissue injury, and ischemia. We have investigated the effects of C3a and C3a desArg on gene expression and protein synthesis of TNF-alpha and IL-1 beta in PBMC. Neither C3a nor C3a desArg alone induced detectable protein or mRNA levels for TNF-alpha and IL-1 beta. C3a modulated LPS-induced TNF-alpha and IL-1 beta synthesis. In nonadherent PBMC, C3a suppressed LPS-induced synthesis of TNF-alpha (20-71% decrease by 0.2-10 microgram/ml of C3a, p less than 0.01) and IL-1 beta (19-57% decrease by 0.5-10 microgram/ml of C3a, p less than 0.01), independently of endogenous production of PGE2. C3a also suppressed LPS-induced mRNA levels for TNF-alpha and IL-1 beta. In contrast, in adherent PBMC, C3a at 5 to 20 microgram/ml enhanced LPS-induced TNF-alpha (75-188% increase, p less than 0.001) and IL-1 beta (119-274% increase, p less than 0.001) synthesis. C3a enhanced TNF-alpha and IL-1 beta mRNA levels in LPS-stimulated adherent cells. Furthermore, C3a desArg shared with C3a the ability to modulate LPS-induced mRNA and protein synthesis for TNF-alpha and IL-1 beta. These results suggest that C3a, thought to be proinflammatory, and C3a desArg, thought to be biologically inactive, are modulators of inflammation. Both C3a and C3a desArg may enhance cytokine synthesis by adherent monocytes at local inflammatory sites, while inhibiting the systemic synthesis of proinflammatory cytokines by circulating cells.

Western blot analysis of glycoproteins bearing Lewis(a) and sialyl-Lewis(a) antigens in human colorectal mucosa.

Glycoproteins (GPs) bearing Lewis(a) and sialyl-Lewis(a) antigens (Le(a), sialyl-Le(a)) derived from human colorectal carcinomas and their surrounding non-neoplastic mucosa (normal mucosa) were analyzed using Western blotting. GPs bearing Le(a) were detected mainly as segmental bands of M(r) 310, 220, 160, and 80 kDa in 80% of the normal mucosa, but these GPs were detected predominantly as broad bands ranging from high to low molecular weight (MW) in 71% of the carcinoma tissues. GPs bearing sialyl-Le(a) were detected only in 23% of the normal mucosa and limited on huge MW bands, i.e., more than 400 kDa, whereas these GPs were detected predominantly as broad bands in 49% of the carcinoma tissues. In the cases with lymph node metastasis, the MW of GPs bearing sialyl-Le(a) varied over a wide range and were detected as broad bands, compared with the cases without metastasis. In conclusion, the MW of GPs bearing Le(a) and sialyl-Le(a) in normal colorectal mucosa was different from that in colorectal carcinomas. That is, the MWs of GPs bearing Le(a) varied more in carcinoma tissues, and the GPs bearing sialyl-Le(a) from carcinoma tissues had lower MWs than those from normal mucosa. It was, furthermore, suggested that the increased expression of lower MW GPs bearing sialyl-Le(a) are associated with an increased metastatic potential of the tumor cells.

Sister chromatid exchanges in mouse after exposure to pulse-wave ultrasound in utero.

The induction of sister chromatid exchanges (SCEs) was investigated in mice after a ten min exposure, in vivo, to 2 MHz focused, pulse-wave ultrasound with a pulse repetition rate of 1000 Hz, pulse duration of 10 microseconds. The bone marrow cells of the pregnant female mice and the fetal liver cells were analyzed. The cell cycle specific metaphase patterns were additionally evaluated. In the bone marrow cells, the mean frequencies of SCEs were 2.77 in control, 3.56 in the cells exposed to ultrasound at 586.2 mW/cm2 (spatial average temporal average, SATA); in the fetal liver cells, 2.64 in control, 3.84 in the cells exposed. The frequencies of SCEs significantly were increased by the treatment. Faster cell kinetics was observed in fetal liver cells than bone marrow cells of pregnant female. But there was no cell-growth inhibitory effect of ultrasound on both bone marrow and fetal liver cells. In fetal liver cells, the critical acoustic power was 160.0-278.9 mW/cm2 (SATA).

[Hepatic arterial infusion chemotherapy using totally implantable reservoir in liver metastases in colorectal cancer].

Hepatic arterial infusion chemotherapy using totally implantable reservoir was performed for the treatment of liver metastases of colo-rectal cancers, and the therapeutic effects, side effects and complications were evaluated. Reservoir catheters were implanted into hepatic artery via gastroduodenal artery during operation. Mitomycin C (MMC), adriamycin (ADM), and 5-fluorouracil (5-FU) were used as chemotherapeutic agents. Eleven cases of H1 (metastases in one lobe only), 7 cases of H2 (a few scattered metastases in both lobes) and 12 cases of H3 (multiple metastases in both lobes) were treated intermittently with one-shot administration of MMC or ADM (A-group). Ten cases (H1: 1, H2: 2, H3: 7) were treated with intermittent one-shot administration of MMC or ADM following two-week continuous infusion of 5-FU through infusion pump after operation (B-group). In 5 of 10 cases of B-group, serum CEA level fell below the preoperative level, and the tumor size regressed in 3 of those 5 cases which were evaluated on the basis of CT scan. But no remarkable change in CEA level or tumor size on CT scan was seen in A-group. No particular side effect such as leucopenia, liver dysfunction or gastroduodenal symptom was noted except one case developing multiple gastric ulcers and pancreatitis in B-group. Five cases (25%) showed obstruction of catheter and 3 cases (14%) evidence leakage of chemotherapeutic agents in A-group. Three cases (30%) in agents in A-group. Three cases (30%) in B-group displayed obstruction of gastroduodenal artery beyond the tip of catheter. Median survival time of both groups (A, B) was 6 months and 12 months, respectively. The treatment seemed effective for the improvement of serum CEA level and tumor size, and there was a tendency toward prolongation of survival time in B-group.

[Transcatheter arterial chemoembolization and selective hepatic arterial infusion using totally implantable reservoir].

Transcatheter arterial chemoembolization (TAE) and hepatic arterial infusion using totally implantable reservoir were performed for the treatment of liver metastasis of colo-rectal cancers, and their therapeutic effects, side effects and complications were evaluated. Eleven cases of H1 (metastasis in one lobe only), 7 cases of H2 (a few scattered metastases in both lobes), 12 cases of H3 (numerous metastases in both lobes) were entered into the study and underwent TAE 45 times. Gel foam, Ivaron and Lipiodol were used as embolic materials in combination with chemotherapeutic agents such as mitomycin C and adriamycin. Serum CEA level was decreased less than 50% of pre-TAE level 20 out of 32 (61%). The tumor size was regressed in 25% of TAE cases which were evaluated on the basis of CT scan. Abdominal symptoms including abdominal pain, nausea and vomiting and fever, leukocytosis, elevated GOT, LDH and bilirubin level were seen after TAE therapy. Median survival of H1, H2 and H3 cases were 21 months, 8 months and 4.5 months, respectively. Another 21 cases (H1, 5 cases: H2, 3 cases: H3, 13 cases) of liver metastasis of colo-rectal cancers were treated with selective hepatic arterial infusion therapy using totally implantable reservoir. Reservoir catheters were implanted into hepatic artery via gastroduodenal artery under direct vision at laparotomy. Mitomycin C, adriamycin and fluorouracil (5-FU) were used as chemotherapeutic agents. No particular side effect such as leukopenia or liver dysfunction was noted. Median survival of H1, H2 and H3 cases treated with arterial infusion were 4 months, 9 months and 9 months, respectively. Median survival of TAE cases and arterial infusion cases was 10 and 6 months, respectively. Thus, the survival rate of cases treated with TAE was better than that of cases treated with arterial infusion.

A case of cebocephaly-holoprosencephaly with an aberrant adenohypophysis.

An autopsy case of cebocephaly-holoprosencephaly at 27 weeks' gestation is reported. Chromosome analysis revealed a 46, XX,-7, +der(7), t(7;13) (q32;q34) pat karyotype. Pathological examination disclosed a hypoplastic aberrant adenohypophysis, in which immunohistochemical localization of four anterior pituitary hormones (ACTH, GH, PRL, TSH-beta) was demonstrated. Malformation of the pituitary gland in holoprosencephaly can include ectopic adenohypophysis as well as pituitary dysgenesis, so a careful search for adenohypophysis should be made in future cases for a better understanding of endocrine dysgenesis associated with holoprosencephaly.

Spontaneous and MMC induced SCE in lymphocytes from patients with cervical cancer.

The frequencies of spontaneous and mitomycin C (MMC) induced sister chromatid exchanges (SCEs) were investigated in lymphocytes of the peripheral blood of women in different stages (O-III) of cervical cancer. Average generation time (AGT) was also examined by the replicative index (R.I.) method. The following results were obtained: The spontaneous SCE frequency was found to be significantly higher in the cancer group than in the control group; 8.21 +/- 1.42 (mean +/- S.D.) against 5.62 +/- 0.55. The SCE frequency gradually increased with the progression of the cervical cancer. The SCE frequency in patients with carcinoma in situ (CIS), the lowest stage of cervical cancer, was significantly higher than that of the controls. The frequency of MMC induced SCE was higher than that of spontaneous SCE in all groups, and the difference in frequency between MMC induced and spontaneous SCEs of cervical cancer group of the stages I-III was significant by different from that of the control group. The AGT of the cervical cancer group was shorter than that of the control group, but it was not statistically significant.

Cytoplasmic estrogen receptors in carcinoma of the uterine cervix.

The concentration of cytoplasmic estrogen receptors (ER) in cancer of the uterine cervix was measured in 30 cases (28 squamous cell carcinomas and 2 adenocarcinomas). The mean ER concentration in squamous cell carcinoma was 19.3 +/- 26.0 fmol/mg cytosol protein; for pre- and postmenopausal women 6.82 +/- 9.86 and 28.6 +/- 30.1 fmol/mg protein, respectively, were found, the latter being significantly higher. When 'ER-positive' was defined as concentrations greater than 10 fmol/mg protein, 12 of the 28 cases (43%) were found to be ER-positive. There were no significant differences between the ER concentrations of clinical stage I and II squamous cell carcinomas (19.2 +/- 24.2 and 20.9 +/- 27.2 fmol/mg, respectively). ER were detectable in the cervical tissue from all of the control cases of myoma of the uterus. There were, however, no differences in the ER content between the proliferative and secretory phases, but the concentration in premenopausal women was significantly lower than that in postmenopausal women. In comparison with the controls, the mean ER level in both pre- and postmenopausal women with squamous cell carcinoma was significantly lower, due to the fact that ER were not detectable in 57% of these cases. Both cases of cervical adenocarcinoma were ER-positive.

Ultraviolet light and mitomycin C induced sister-chromatid exchanges in fibroblasts from patients with retinoblastoma.

Ultraviolet light and mitomycin C (MMC) induced sister-chromatid exchanges (SCEs) were investigated in 6 diploid fibroblast strains derived from 3 patients with deletion 13 and retinoblastoma, one patient with a hereditary form of retinoblastoma, one patient with trisomy 13, and one normal control. Two fibroblast strains with del(13)(q14q22) showed a significant increase in SCEs compared to the control after UV and MMC treatments. In contrast, cell strains with del(13)(q12q14) and trisomy 13 did not show increased SCEs. The frequency of SCEs in fibroblasts from a patient with autosomal dominant retinoblastomas (no deletions) was significantly increased by UV, but not by MMC. The results suggest that cell strains with different deletions of chromosome 13 have different SCE responses to UV and MMC inductions. The cells with del(13)(q14q22) may have a DNA-repair defect.

An in vitro and in vivo study of a BrdU-sensitive fragile site in the Chinese hamster.

The frequencies of chromosome aberrations and development of the bromodeoxyuridine (BrdU)-sensitive fragile site were studied in vitro in Chinese hamster kidney and bone marrow cells and in vivo in Chinese hamster bone marrow cells. Chromosome aberrations in these cell systems were measured in response to different concentrations of BrdU, fluorodeoxyuridine, or both. The fragile site was found in both homologues of chromosome 1 at 1q22. Treatment with BrdU in vitro but not in vivo produces significant chromosome aberrations. About 50% of chromosome aberrations found after treatment in vitro were at the BrdU-sensitive fragile site compared with 12.5% after treatment in vivo. These results show that BrdU is much more potent in vitro than in vivo in inducing both chromosome aberrations and the expression of the BrdU-sensitive site.

Effects of radiotherapy and chemotherapy on sister chromatid exchanges in peripheral lymphocytes of the patients with gynecologic cancer.

Studies were done on the influence of radiotherapy or chemotherapy on peripheral lymphocyte SCE (sister chromatid exchange) frequency in patients with gynecologic cancer. The average value of spontaneous SCE frequency in 10 patients who had received radiotherapy was 6.94 +/- 2.57 (mean +/- S.D.), which was significantly lower than the control value of 8.59 +/- 2.99 ( p less than 0.001). The rate of decline of these frequency values was negatively correlated with the irradiation dosage. In contrast, the average spontaneous SCE frequency in the patients who had received combination chemotherapy was 12.48 +/- 4.02, which was significantly higher than the control value ( p less than 0.001). The average value of MMC-induced SCE frequency in patients under chemotherapy was much the same as the control value of 26.16 +/- 5.77. That is patients under radiotherapy was 23.88 +/- 5.65, which was slightly lower than the control value. There was, however, no significant difference in the MMC-induced SCE frequency value among these 3 groups.

Sister chromatid exchanges and chromosome aberrations in fibroblasts from patients with retinoblastoma.

The frequencies of sister chromatid exchanges (SCEs) and chromosome breaks were investigated in five diploid fibroblast strains derived from three patients with deletion 13 [del(13)] retinoblastoma, one patient with a hereditary form of retinoblastoma, and one trisomy 13. The fibroblasts with del(13)(q14q22) showed slightly increased SCEs (at a P level of 5-10%), but the others, including del(13)(q12q14), the hereditary form of retinoblastoma, and trisomy 13, did not have increased SCEs as compared to normal controls. No increase in chromosome breaks was found in these fibroblasts. The results suggest that retinoblastoma is not associated with spontaneous increased chromosomal instability.