Tea crude extracts effectively inactivate severe acute respiratory syndrome coronavirus 2.

It is well known that black and green tea extracts, particularly polyphenols, have antimicrobial activity against various pathogenic microbes including viruses. However, there is limited data on the antiviral activity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which emerged rapidly in China in late 2019 and which has been responsible for coronavirus disease 2019 (COVID-19) pandemic globally. In this study, 20 compounds and three extracts were obtained from black and green tea and found that three tea extracts showed significant antiviral activity against SARS-CoV-2, whereby the viral titre decreased about 5 logs TCID50 per ml by 1·375 mg ml-1 black tea extract and two-fold diluted tea bag infusion obtained from black tea when incubated at 25°C for 10 s. However, when concentrations of black and green tea extracts were equally adjusted to 344 µg ml-1 , green tea extracts showed more antiviral activity against SARS-CoV-2. This simple and highly respected beverage may be a cheap and widely acceptable means to reduce SARS-CoV-2 viral burden in the mouth and upper gastrointestinal and respiratory tracts in developed as well as developing countries.

MEG measurement of event-related brain activity evoked by emotional prosody recognition.

Cortical areas involved in processing of emotional prosody (EP) in spoken language, such as joy or sadness, have been found in functional magnetic resonance imaging (fMRI) studies bilaterally or dominantly in the right frontal or temporal lobes. In this study, we investigated spatiotemporal patterns of cortical activity related to EP processing using magnetoencephalography (MEG). In this experiment, a joyful face (JF) or a sad face (SF) was displayed after voices which had emotional features of joy (joy prosody: JP) or sadness (sad prosody: SP) were presented. Subjects were requested to judge whether emotional features of the voice and the face were identical or not. MEG signals evoked by emotional voices were measured and significant differences of cortical activities associated with processing of emotional feature were observed between the right and left hemisphere during the latency of 100-150 ms that includes the N1m component. Our study suggests that MEG is a useful method, in addition to fMRI and event-related scalp potentials (ERP) for studying non-invasively EP processing in the human brain.

The relationship between consumption of antimicrobial agents and the prevalence of primary Helicobacter pylori resistance.

BACKGROUND: Primary and acquired resistance to the antimicrobial agents is a primary reason for the failure of Helicobacter pylori eradication therapies. We assessed the primary antibiotic resistance rates of H. pylori to three different antibiotics and its relationship due to the annual antibiotic consumption in Japan during the period prior to approval of anti-H. pylori therapy in Japan. MATERIALS AND METHODS: Antibiotic susceptibility was tested using the agar dilution method for clarithromycin, amoxicillin and metronidazole. Isolates were considered resistant when the MIC value was > 8 mg/l for metronidazole, > 1 mg/l for clarithromycin and < 0.5 mg/l for amoxicillin. RESULTS: Helicobacter pylori isolates were obtained from 593 Japanese patients from 1995 to 2000. Primary resistance of H. pylori to clarithromycin, metronidazole and amoxicillin was found in 11%, 9% and 0.3% strains, respectively. The proportion with clarithromycin resistance significantly increased from 7% in 1997-98 to 15.2% in 1999-2000 (p =.003). During the same period the metronidazole resistance rate also increased from 6.6% in 1997-98 to 12% in 1999-2000 (p =.02). The prevalence of clarithromycin and metronidazole was related to the annual consumption of these antimicrobial agents. CONCLUSION: Resistance rates for both clarithromycin and metronidazole appear to reflect the annual consumption of these agents. The high rate of clarithromycin resistance in Japan suggests that the effectiveness of clarithromycin-based therapies may be compromised in the near future.

Ubiquitin-proteasome inhibitor enhances tumour necrosis factor-alpha-induced apoptosis in rat gastric epithelial cells.

BACKGROUND: Tumour necrosis factor (TNF-alpha) is a candidate factor for involvement in inflammation-mediated gastric mucosal injury. However, the effect of this cytokine on gastric epithelial cells has been poorly investigated. In the present study, we examined whether gastric epithelial cells are resistant to TNF-alpha-induced apoptosis, and whether this resistance is related to ubiquitin-proteasome-associated nuclear factor-kappaB (NF-kappaB) activation. METHODS: The rat gastric mucosal cell line RGM-1 was grown in DMEM/F12 medium supplemented with 10% FCS. Confluent monolayers of cells were pretreated or not for 60 min with PSI, a peptide aldehyde known to specifically inhibit the chymotrypsin-like activity of 26S proteasome. Cells were subsequently stimulated with recombinant rat TNF-alpha and their viability was determined by WST-1 assay. Apoptosis was confirmed by fluorescence microscopy after staining with Hoechst 33342 and propidium iodide, and DNA fragmentation was determined by flow cytometry using an APO-BRDU kit. IkappaB-alpha and the p65 binding subunit of NF-kappaB were detected by Western blots. RESULTS: Twenty-four-hour incubation with TNF-alpha alone or PSI alone did not affect the cell viability of RGM-1 cells. Pretreatment with PSI significantly enhanced the level of apoptosis induced by TNF-alpha. In RGM-1 cells treated with TNF-alpha, cytoplasmic IkappaB-alpha decreased and p65 in nuclear extracts increased markedly 30 min after cytokine stimulation. Pretreatment with PSI at 12.5 micromol/L blocked these TNF-alpha-induced changes. CONCLUSION: PSI enhances TNF-alpha-induced apoptosis through inhibition of NF-kappaB activation in RGM-1 cells.

Role of P-selectin and intercellular adhesion molecule-1 in TNB-induced colitis in rats.

It has been proposed that neutrophil-endothelial cell interactions mediated by adhesion molecules are involved in the pathogenesis of inflammatory bowel disease. The objective of the present study was to determine the effects of monoclonal antibodies (MAbs) directed against endothelial adhesion molecules, P-selectin and intercellular adhesion molecule-1 (ICAM-1), in rats with colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNB). Colonic inflammation was induced by administering an enema of TNB dissolved in 50% ethanol (120 mg/ml) to male Wistar rats (at a total volume of 0.25 ml per rat) after a 48-hour fast. Anti-P-selectin MAb or anti-ICAM-1 MAb was injected via the tail vein at a dose of 1 mg/kg after the induction of colitis. Rats in the control group received nonbinding mouse immunoglobulin G1. The plasma level of soluble P-selectin showed an increase within 48 h after the TNB enema. Colonic inflammation was assessed at 1 week after TNB administration. The colonic damage score and the wet weight of the colon were significantly decreased by treatment with either MAb. The increase of myeloperoxidase (MPO) activity, an index of neutrophil accumulation, and the increase of thiobarbituric acid-reactive substances (TBA-RS), an index of lipid peroxidation, in the colonic mucosa were inhibited by both MAbs. These results suggest that neutrophil-endothelial cell interactions via P-selectin and ICAM-1 play an important role in the development of TNB-induced colitis in rats.

Morphologic fate of diencephalic prosomeres and their subdivisions revealed by mapping cadherin expression.

The expression of four cadherins (cadherin-6B, cadherin-7, R-cadherin, and N-cadherin) was mapped in the diencephalon of chicken embryos at 11 days and 15 days of incubation and was compared with Nissl stains and radial glial topology. Results showed that each cadherin is expressed in a restricted manner by a different set of embryonic divisions, brain nuclei, and their subregions. An analysis of the segmental organization based on the prosomeric model indicated that, in the mature diencephalon, each prosomere persists and forms a coherent domain of gray matter extending across the entire transverse dimension of the neural tube, from the ventricular surface to the pial surface. Moreover, the results suggest the presence of a novel set of secondary subdivisions for the dorsal thalamus (dorsal, intermediate, and ventral tiers and anteroventral subregion). They also confirm the presence of secondary subdivisions in the pretectum (commissural, juxtacommissural, and precommissural). At most of the borders between the prosomeres and their secondary subdivisions, changes in radial glial fiber density were observed. The diencephalic brain nuclei that derive from each of the subdivisions were determined. In addition, a number of previously less well-characterized gray matter regions of the diencephalon were defined in more detail based on the mapping of cadherin expression. The results demonstrate in detail how the divisions of the early embryonic diencephalon persist and transform into mature gray matter architecture during brain morphogenesis, and they support the hypothesis that cadherins play a role in this process by providing a framework of potentially adhesive specificities.

Molecular design of conjugated tumor necrosis factor-alpha: synthesis and characteristics of polyvinyl pyrrolidone modified tumor necrosis factor-alpha.

We conjugated tumor necrosis factor-alpha (TNF-alpha) with the synthetic polymeric modifier polyvinyl pyrrolidone (PVP) to facilitate its clinical use for anti-tumor therapy. TNF-alpha was chemically conjugated with the terminal carboxyl-bearing PVP at one end of its main chain, which was radically polymerized via the formation of an amide bond between the lysine amino groups of TNF-alpha and carboxyl group of PVP. In vitro specific bioactivity of PVP-conjugated TNF-alpha (PVP-TNF-alpha) relative to that of native TNF-alpha gradually decreased with increases in the degree of PVP attachment. In contrast, PVP-TNF-alpha in which 40% of TNF-alpha lysine residues were coupled with PVP (MPVP-TNF-alpha) exhibited the highest anti-tumor activity among the conjugated derivatives examined. MPVP-TNF-alpha had more than 200-fold higher anti-tumor efficacy than native TNF-alpha, and the anti-tumor activity of MPVP- TNF-alpha was more than 5-fold stronger than that MPEG- TNF-alpha which had the highest anti-tumor activity among PEG-conjugated TNF-alphas examined. Additionally, a high dose of native TNF-alpha induced toxic side-effects such as body weight reduction, piloerection and tissue inflammation, while no side effects were observed following i.v. administration of MPVP-TNF-alpha. The plasma half-life of MPVP-TNF-alpha (360 min) was about 80 and 3-fold longer than those of native TNF-alpha (4.6 min) and MPEG-TNF-alpha (122 min), respectively. These results suggested that PVP is a useful polymeric modifier for increasing the anti-tumor activity of PVP.

Patients with ovarian carcinoma upstaged to stage III after systematic lymphadenctomy have similar survival to Stage I/II patients and superior survival to other Stage III patients.

BACKGROUND: Patients with epithelial ovarian carcinoma upstaged from Stage I/II to Stage IIIC based on lymph node involvement are known to have poor prognoses. The authors investigated whether systematic aortic and pelvic lymphadenectomy would affect the prognoses of these patients. METHODS: During the period 1987-1996, 103 patients in Stage I-III underwent optimal cytoreductive surgery with systematic aortic and pelvic lymphadenectomy at initial surgery. All patients except for those in Stage IA received adjuvant cisplatin-based chemotherapy after surgery. Of 67 patients with intraperitoneal tumors limited to the pelvis, 14 were upstaged to Stage III based on lymph node positivity (Group A). The authors compared the survival of Group A patients with that of 53 patients who had intraperitoneal tumors limited to the pelvis and negative lymph nodes (Group B), and also with that of 36 patients who had intraperitoneal tumors beyond the pelvis irrespective of lymph node status (Group C). RESULTS: The 5-year survival of Group A patients in Stage III based only on lymph node positivity had fairly good survival, although it was not significantly different from that of Group B patients in Stage I/II (84% vs. 96%, P=0.107). Group A had much better 5-year survival than Group C patients who were considered to be Stage III because they had intraperitoneal tumors beyond the pelvis (84% vs. 26%, P=0.042). CONCLUSIONS: Relatively good survival was observed for patients with intraperitoneal tumors limited to the pelvis and lymph node involvement who underwent systematic aortic and pelvic lymphadenectomy.

Novel S100 proteins in human esophageal epithelial cells: CAAF1 expression is associated with cell growth arrest.

CAAF1 and CAAF2, newly identified calcium-binding proteins from bovine amniotic fluid, have been revealed to be members of the S100 protein family preferentially produced by fetal squamous epithelial cells, including epidermal keratinocytes. Having previously cloned the cDNA of human CAAF1 protein from the esophageal epithelium, we report here on the characteristic expression pattern of CAAF1 and related S100 proteins in human esophageal epithelial cells. Normal cells of the human esophageal epithelium expressed CAAF1, and also expressed the homologous novel S100 proteins including CAAF2, MRP8, and MRP14, but not S100alpha. An immunohistochemical study with specific monoclonal antibodies against CAAF1 proteins demonstrated that CAAF1 proteins were produced by the esophageal epithelial cells in the process of cell differentiation. The immature proliferating cells in the epithelium did not produce CAAF1 proteins, but the differentiated cells expressed CAAF1, which overlay the immature cells and were stratifying in the epithelium. These CAA 1-producing cells did not show any proliferating activities. Esophageal carcinoma cells did not express CAAF1, except for the keratinized cells with no proliferating activity. In addition, the forced expression of CAAF1 proteins in the carcinoma cells resulted in a marked decrease in DNA synthesis. These findings indicate that human esophageal epithelial cells express the multiple genes of S100 proteins including CAAF proteins, and that CAAF1 is closely associated with the terminal differentiation of these cells. CAAF1 expression also might play some role in cell growth.

Therapeutic efficacy of BO-3482, a novel dithiocarbamate carbapenem, in mice infected with methicillin-resistant Staphylococcus aureus.

The in vivo activity of BO-3482, which has a dithiocarbamate chain at the C-2 position of 1beta-methyl-carbapenem, was compared with those of vancomycin and imipenem in murine models of septicemia and thigh infection with methicillin-resistant Staphylococcus aureus (MRSA). Because BO-3482 was more susceptible than imipenem to renal dehydropeptidase I in a kinetic study of hydrolysis by this renal enzyme, the therapeutic efficacy of BO-3482 was determined during coadministration with cilastatin. In the septicemia models, which involved two homogeneous MRSA strains and one heterogeneous MRSA strain, the 50% effective doses were, respectively, 4.80, 6.06, and 0.46 mg/kg of body weight for BO-3482; 5.56, 2.15, and 1.79 mg/kg for vancomycin; and >200, >200, and 15.9 mg/kg for imipenem. BO-3482 was also as effective as vancomycin in an MRSA septicemia model with mice with cyclophosphamide-induced immunosuppression. In the thigh infection model with a homogeneous MRSA strain, the bacterial counts in tissues treated with BO-3482-cilastatin were significantly reduced in a dose-dependent manner compared with the counts in those treated with vancomycin and imipenem-cilastatin (P < 0.001). These results indicate that BO-3482-cilastatin is as effective as vancomycin in murine systemic infections and is more bactericidal than vancomycin in local-tissue infections. The potent in vivo activity of BO-3482-cilastatin against such MRSA infections can be ascribed to the good in vitro anti-MRSA activity and improved pharmacokinetics in mice when BO-3482 is combined with cilastatin and to the bactericidal nature of the carbapenem.

Bone density and bone metabolic markers in active collegiate athletes: findings in long-distance runners, judoists, and swimmers.

We investigated the bone metabolic system status of 103 male and female volunteer collegiate athletes, who were actively pursuing one of three different sports: Long-distance running (LR); judo (JU); and swimming (SW). The following parameters were evaluated: total body bone mineral density (TMBD); bone-forming metabolic markers; serum procollagen type I C-peptide (PICP) levels; bone alkaline phosphatase (B-ALP) content; bone resorption markers, urinary pyridinoline (Pyd) and deoxypyridinoline (Dpd) levels. We found that the TBMD and urinary Dpd values in JU athletes were significantly higher (p < 0.001) than in athletes of the same sex in the other two groups. The urinary Pyd level in male JU athletes was also higher (p < 0.001) than that in the other two groups, but that in females JU athletes was only higher (p < 0.01) than that in female LR athletes. The PICP levels were similar to the TBMD values in all groups. No differences in bone density or in bone metabolic markers were seen in LR and SW athletes of the same sex. We thus conclude that differences in bone mineral density are in part due to the demands of the specific sport, and that they are reflected in bone metabolic markers. In addition, the status of bone metabolic turnover in male JU athletes in training may be hypermetabolic and as well as that of female JU athletes with regular menses cycles.

Effects of estrogen on cell growth and fibroblast growth factor receptor induction in MtT/Se cells.

The effects of estradiol (E2) on cell growth and the expression of fibroblast growth factor receptor (FGFR) were investigated in a cultured rat anterior pituitary cell line (MtT/Se) established from an E2-induced mammotropic pituitary tumor. E2 stimulated cell growth 2-3 fold as compared to the control. Basic fibroblast growth factor (bFGF) stimulated cell growth in the presence of E2 (10(-9) M) but not in the absence of E2. Tamoxifen as an antiestrogen agent inhibited the proliferation of MtT/Se cells which had been treated with E2 or E2 and bFGF. Numbers of FGFR in E2-stimulated MtT/Se cells significantly exceeded those in cells not treated with E2. These observations indicate that E2 plays an important role in the regulation of FGFR induction in the anterior pituitary cells.

Indonesian medicinal plants. XVII. Characterization of quassinoids from the stems of Quassia indica.

Four new quassinoids named samaderines X (1), Y (2) and Z (3), and indaquassin X (5), and a new C19 quassinoid glycoside, 2-O-glucosylsamaderine C (10), together with five known quassinoids, samaderines B (7), C (8), and E (4), indaquassin C (6), and simarinolide (9), were isolated form the stems of Quassia indica (Simaroubaceae), an Indonesian medicinal plant. The chemical structures of these quassinoids have been elucidated on the bases of their chemical and physiochemical properties. Samaderines X (1), Z (3), E (4), and B (7) were shown to exhibit significant growth-inhibitory activity against the cultured malarial parasite Plasmodium falciparum (a chloroquine- resistant K1 strain), and 1--8 were shown to exhibit in vitro cytotoxicity (IC50: 0.04--100 micrograms/ml) against KB cells. Samaderines X (1), B (7) and C (8), as well s indaquassin X (5), exhibited inhibitory activity in the in vitro endothelial cell-neutrophil leukocyte adhesion assay, whereas samaderines X (1) and B (7) were found to exhibit significant anti-inflammatory activity.

Evaluation of angiogenic inhibitors with an in vivo quantitative angiogenesis method using agarose microencapsulation and mouse hemoglobin enzyme-linked immunosorbent assay.

In the present work, using a previously reported in vivo quantitative tumor-angiogenesis model, we attempted to ascertain whether this animal model is suitable for practical use in monitoring inhibitors of tumor angiogenesis. Mouse sarcoma-180 cells, human A431 cells or rat C6 cells microencapsulated in agarose beads were implanted s.c. into C57BL/6 mice. The level of blood vessel induction at the agarose pellet site was evaluated using mouse hemoglobin enzyme-linked immunosorbent assay on day 10 after implantation. Hydrocortisone, tetrahydro-S, medroxyprogesterone acetate, pentosan polysulfate and suramin inhibited blood vessel growth in our in vivo tumor-angiogenesis assay system, and heparin enhanced the antiangiogenic effects of hydrocortisone and tetrahydro-S. These results are almost entirely consistent with those observed in common assay systems, and suggest that this method may be useful for the identification and quantitative evaluation of inhibitors of tumor angiogenesis.

[Successful collection of peripheral blood stem cells mobilized by high-dose etoposide for patients with chemotherapy-resistant and/or poor prognostic testicular cancer].

Clinical effects of peripheral blood stem cell autotransplantation (PBSCT) after ultra high-dose chemotherapy were evaluated in patients with chemotherapy-resistant and/or poor prognostic testicular cancer. Four patients with testicular cancer, who had high-risk malignancy, were treated with high-dose etoposide (500 mg/m2 x 4 days) in order to collect peripheral blood stem cells. After the administration of high-dose etoposide, rG-CSF (250 micrograms/body) was administered from nadir state. Blood mononuclear cells were collected using a Fenwall CS-3000 blood cell separator. Fractions enriched for stem cells were obtained by discontinuous Percoll gradient centrifugation and were stored in liquid nitrogen using patient's sera and DMSO. The mean number of peripheral blood granulocyte-macrophage-colony-forming units (CFU-GM) collected by one apheresis was 22.3 x 10(5)/kg body weight. In addition, CFU-GM more than 2.0 x 10(5)/kg body weight could be collected in each apheresis, which was though to be sufficient dosis to perform PBSCT in safe, based upon our previous studies. All the patients were treated by a combination of cisplatin (20 mg/m2 x 5 days), etoposide (100 mg/m2 x 5 days) and bleomycin (15 mg x 3 days). Three patients responded to BEP therapy and obtained a CR, however, remaining 1 patient failed to achieve CR, who was later treated by ultrahigh-dose chemotherapy including carboplatin (200 mg/m2 x 4 days), etoposide (250 mg/m2 x 4 days) and cyclophosphamide (50 mg/kg x 2 days) followed by PBSCT. He responded to this therapy and obtained a CR for 10 months. The results suggested the method was promising for patients with chemotherapy-resistant and/or poor prognostic testicular cancer.

[Ultra high-dose chemotherapy with peripheral blood stem cell autotransplantation for refractory testicular cancer].

This is a report of 45-year-old man with advanced nonseminomatous germ cell tumor (stage IIIB2: embryonal carcinoma, yolk sac tumor, seminoma), who had relapse after PVB (cisplatin, vinblastine, bleomycin) chemotherapy. Peripheral blood stem cells (PBSCs) were taken by two consecutive apheresis using a CS-3000 blood separator after high-dose chemotherapy of cytarabine and mitoxantrone. In total, 6.4 x 10(5)/kg of granulocytic cells (CFU-GM) was collected. He was treated with ultra high-dose chemotherapy consisting of carboplatin (800 mg/m2), etoposide (1,000 mg/m2) and cyclophosphamide (100 mg/kg) from day 1, followed by peripheral blood stem cell autotransplantation (PBSCT) on day 9. We transfused 2.4 x 10(5)/kg CFU-GM, which was enough number of stem cells for safe PBSCT. No serious side effects or complications were encountered. The patient achieved partial remission for more than two months. However, he died of respiratory dysfunction caused by metastatic lung cancer 5 months later. It was thought that ultra high-dose chemotherapy with PBSCT might be a new therapy for refractory testicular cancer.

Gomisin A inhibits tumor promotion by 12-O-tetradecanoylphorbol-13-acetate in two-stage carcinogenesis in mouse skin.

Gomisin A, isolated from the fruits of Schisandra chinensis, is one of the dibenzocyclooctadiene lignans. Application of 12-O-tetradecanoylphorbol-13-acetate (TPA, 1 microgram/ear), a tumor-promoting agent, to the ears of mice induces inflammation. Among seven dibenzocyclooctadiene lignans assayed, gomisin A, gomisin J, and wuweizisu C inhibited the inflammatory activity induced by TPA in mice. The ED50 of these compounds for TPA-induced inflammation was 1.4-4.4 mumol. Gomisin A, with an ED50 of 1.4 mumol, showed the strongest inhibitory effect. Furthermore, at 5 mumol/mouse, it markedly suppressed the promotion effect of TPA (2.5 micrograms/mouse) on skin tumor formation in mice following initiation with 7,12-dimethylbenz[a]anthracene (50 micrograms/mouse). It is assumed that the inhibition of tumor promotion by gomisin A is due to its anti-inflammatory activity.

Topographical projections from the thalamus, subthalamic nucleus and pedunculopontine tegmental nucleus to the striatum in the Japanese monkey, Macaca fuscata.

Topographical projections from the thalamus, subthalamic nucleus (STN) and pedunculopontine tegmental nucleus (PPN) to the striatum were examined in the Japanese monkey (Macaca fuscata) by using the retrograde axonal transport technique of WGA-HRP (wheat germ agglutinin-conjugated horseradish peroxidase). After WGA-HRP injection in the head of the caudate nucleus (CN) or putamen (Put), labeled neuronal cell bodies in the thalamus were distributed mainly in the nucleus ventralis anterior (VA)-nucleus ventralis lateralis (VL) complex and the nucleus centrum medianum (CM)-nucleus parafascicularis (Pf) complex, and additionally in the paraventricular, parataenial, rhomboid, reuniens, centrodorsal, centrolateral, paracentral, and centromedial nuclei. The data indicated that the pars principalis of VA (VApc) projected mainly to CN and additionally to Put, and that the pars magnocellularis of VA (VAmc) or pars oralis of VL (VLo) projected selectively to CN or Put, respectively. It was also indicated that CM projected to the middle and caudal parts of Put, while Pf projected to CN and the rostral part of the Put. The data further indicated that the dorsomedial, ventromedial, or lateral part of CM projected respectively to the dorsolateral, ventromedial, or intermediate part of Put, and that the medial or lateral part of Pf projected respectively to the medial or lateral part of the head of CN. Direct projections from STN and PPN to the striatum were confirmed. The subthalamostriatal projections showed a mediolateral topography. The PPN was shown to project bilaterally to the striatum with an ipsilateral predominance.