[A Case Report of Uterine Metastasis Four Years after Breast Surgery for Breast Cancer].

We report a case of uterine metastasis of the breast cancer. The patient was diagnosed with invasive ductal carcinoma of the breast and underwent partial right mastectomy and sentinel lymph node biopsy. Tamoxifen was administered as adjuvant endocrine therapy. Four years after the surgery, she had irregular genital bleeding, and was referred to our hospital for cytological diagnosis of uterine cancer. Postoperative pathological diagnosis showed uterine metastasis of breast cancer, and it was decided to treat the recurrence of breast cancer with aromatase inhibitors and CDK4/6 inhibitors, a molecular targeted therapy. The patient has been progression-free for 5 months.

[A case of rectum metastasis of breast cancer].

A 53-year-old woman visited the hospital of this study complaining of constipation. Colonoscopy revealed a circumferential tumor with severe stenosis, and a computed tomography scan showed neoplastic lesions in the rectum and right breast area. Histology was poorly differentiated adenocarcinoma, requiring differentiation between type 4 and metastatic rectal cancer. Additional immunohistochemical tests were performed and a rectal metastasis of breast cancer diagnosis was made. Hormonal therapy was effective and the tumor volume was significantly reduced. Rectal metastasis of breast cancer is said to be rare. However, in the case of patients diagnosed with breast cancer or with a history of breast cancer, considering the possibility of gastrointestinal metastasis using histopathological examination is important.

Synthesis, anti-HIV and anti-oxidant activities of caffeoyl 5,6-anhydroquinic acid derivatives.

In our continued research on chlorogenic acid analogues and derivatives with improved bioactivity, we have synthesized some caffeoyl 5,6-anhydroquinic acid derivatives. The 1,7 acetonides of chlorogenic acid (15), and of the mono-caffeoyl 5,6-anhydroquinic acids (7-8) showed appreciable anti-HIV activity. The 3,4-dicaffeoyl 5,6-anhydroquinic acid (12) exhibited an anti-HIV activity twice as that of 3,5-dicaffeoylquinic acid (22). The caffeoyl 5,6-anhydroquinic acid derivatives displayed potent anti-oxidant activities. The mono-caffeoyl 5,6-anhydroquinic acids (10-11) were more than twice stronger than chlorogenic acid (21) on SOD-like activity.

Evaluation of the antiviral activity of chlorine dioxide and sodium hypochlorite against feline calicivirus, human influenza virus, measles virus, canine distemper virus, human herpesvirus, human adenovirus, canine adenovirus and canine parvovirus.

We evaluated the antiviral activity of a chlorine dioxide gas solution (CD) and sodium hypochlorite (SH) against feline calicivirus, human influenza virus, measles virus, canine distemper virus, human herpesvirus, human adenovirus, canine adenovirus and canine parvovirus. CD at concentrations ranging from 1 to 100 ppm produced potent antiviral activity, inactivating >or= 99.9% of the viruses with a 15 sec treatment for sensitization. The antiviral activity of CD was approximately 10 times higher than that of SH.

[Therapeutic effects of vinorelbine-based chemotherapy--retrospective study of 65 patients with metastatic or relapsed breast cancer].

PATIENTS: The subjects were 65 stage IV or relapsed breast cancer patients who received vinorelbine monotherapy or combination therapy at Fukushima Medical University between May 2002 and November 2006. METHOD: Chemotherapy was divided into the following 3 groups Vinorelbine monotherapy (group A), Trastuzumab combination therapy (group B), MMVC (mitomycin C, methotrexate, vinorelbine and cyclophosphamide) therapy(group C). RESULTS: There were 33 patients in group A, 15 patients in group B, and 17 patients in group C. The percentage of patients who had received more than 3 prior chemotherapy regimens were 69. 7% in group A, 66. 7% in group B, and 82. 4% in group C. No complete responses were observed. Partial responses was observed 6 patients in group A, 5 patients in group B, and 2 patients in group C. Overall response rate was 18. 2% in group A, 33. 3% in group B, and 11. 8% in group C. Clinical benefit rate was 24. 2% in group A, 46. 7% in group B, and 23. 5% in group C. The median overall survival time was 267, 522, and 275 days in group A, B, and C respectively. DISCUSSION: Vinorelbine-based chemotherapy was considered as a new treatment option for patients with metastatic or relapsed breast cancer. It is necessary to examine the use at the early stage or more to obtain a high response rate and duration of response.

[Two cases of HIV-1 acute infection with antibody negative by immunochromatography method].

We found two cases of HIV-1 acute infection, confirmed by nucleic amplification test (NAT) and/or RT-PCR, with HIV-1 antibody negative by immunochromatography (IC) method but weakly positive by particle agglutination (PA) test. These cases suggested that IC method was less sensitive than PA test in the detection of acute infections. It is necessary to execute the post counseling that considers the possibility of the acute infection in public health centers and testing places where IC method is used for the screening test. It is also important to recommend taking the following re-examination after a certain period to a person who seems to have had a chance of infection in a short time before testing.

Higher frequency of premature stop codon mutations at vpu gene of human immunodeficiency virus type 1 CRF01_AE compared with those of other subtypes.

Our previous study demonstrated the anti-apoptosis function of the human immunodeficiency virus type 1 (HIV-1) vpu gene product in normal CD4+ T lymphocytes. In this study, using sequences obtained from the HIV sequence database, we compared vpu sequences from 184 preparations of various subtypes of HIV-1 from diverse geographical regions. Our analysis revealed that CRF01_AE isolates had premature stop codon mutations at the vpu gene at a much higher rate (36%) than other subtypes (0-9%). The premature stop codon mutations in vpu existed mostly at two amino acid residues: the methionine initiation codon and the boundary between the transmembrane (TM) and cytoplasmic domains. The mutations at the latter site were more often detected in CRF01_AE. The higher mutation rates at vpu in CRF01_AE were confirmed by sequence comparison of polymerase chain reaction products newly obtained directly from the DNA extracted from peripheral blood mononuclear cells (PBMCs), but not from the RNA from the plasma, in CRF01_AE- and subtype B-infected individuals. This finding may indicate the possibility that the more abundant population of HIV-1 CRF01_AE is able to induce apoptosis in CD4+ T lymphocytes than the populations of other subtypes.

[A study on enterovirus infection in Osaka prefecture --comparison with virus isolation and RT-PCR amplifying viral protein 1 region].

Detection of viral genomes with RT-PCR, which amplifies viral protein 1 region, as well as virus isolation was performed on 860 patients (996 specimens) who had been suspected for enterovirus (EV) infection in Osaka Prefecture from April 2003 to Jury 2004. The viral positive rates of the clinical materials, combining above two procedures, were as follows: 48.2% from the feces, 38.3% from the throat swabs, and 18.0% from the cerebrospinal fluids. The positive rate by the clinical diagnosis indicated that herpangina was the highest at 44.7%, while encephalitis was the lowest at 13.4%. Out of the all specimens, the viral positive rate of RT-PCR varied from 22.3 to 24.7%, while that of virus isolation was 15.6%. The total viral positive rate of both procedures combined was 29.8%. Since the detection of EV by RT-PCR amplifing viral protein 1 region was more rapid than virus isolation and superior in detection of coxsackie group A virus, RT-PCR is an effective procedure for diagnosis of herpangina.

Ability to induce p53 and caspase-mediated apoptosis in primary CD4+ T cells is variable among primary isolates of human immunodeficiency virus type 1.

Infection with human immunodeficiency virus type 1 (HIV-1) is associated with dramatic depletion of CD4(+) T cells, the major HIV-1-induced pathogenesis. Apoptosis has been suggested to play an important role for the T cell depletion and a number of mechanisms have been proposed for the apoptosis in T cells. Here, we compared the levels for apoptosis induction in primary peripheral blood mononuclear cells (PBMCs) among several laboratory strains and primary isolates of the HIV-1 subtypes B and E. The results showed that apoptosis in infected PBMCs, preferentially in CD4+ T cell population, became detectable around the time for virus production by flow cytometric terminal transferase dUTP nick end labeling (TUNEL) technique and staining with the nuclear dye Hoechst 33342. The abilities to induce apoptosis in PBMCs were highly variable in individual isolates. The increase of p53 protein in infected PBMCs, which was initiated before virus production, was observed in infected PBMCs and the levels of p53 protein were almost proportional to the rates of the isolates to induced apoptosis. The cells infected and cultured in the presence of Z-VAD-FMK had significantly decreased cell mortalities, indicating that activated caspases also played a significant role in the apoptosis. Thus, HIV-1-induced apoptosis in primary T cells was accompanied by the p53 protein and caspase activation at varied levels in primary isolates.

Anti-human immunodeficiency virus-type 1 activity of constituents from Juglans mandshurica.

Three naphthalene glycosides (1-3), four flavonoids (4-7), and two galloyl glycosides (8-9) were isolated from the stem-bark of Juglans mandshurica (Juglandaceae). Their structures were determined by chemical and spectral means, including to 2D-NMR (COSY, HMQC, and HMBC) experiments. Amongst the isolated compounds, taxifolin (4) showed the most potent HIV-induced cytopethic activity against MT-4 cells with complete inhibitory concentration (IC100) value of 25 microg/ml and maximum cytotoxic concentration (CC100) value of above 100 microg/ml. However, naphthalene glycosides (1-3), flavonoids (5-7), and galloyl tannins (8-9) were inactive against anti-HIV-1 activity.

Down-regulation of p73 correlates with high histological grade in Japanese with breast carcinomas.

BACKGROUND: p73, a homologue of p53, has been located at chromosome 1p36-33, a region of frequently observed loss of heterozygosity in breast cancers. The objective of the present study was to investigate the function of p73 in Japanese with breast cancers. METHODS: Sixty Japanese patients with breast cancer were assessed by polymerase chain reaction single strand confirmation polymorphism analysis and direct sequencing to detect the p73 allele. p73 mRNA levels were also determined in 40 out of 60 patients by reverse-transcriptional polymerase chain reaction. RESULTS: We analyzed the entire open reading frame of the p73 gene by polymerase chain reaction single strand confirmation polymorphism and sequencing, and failed to identify any mutations of p73 in the encoding regions detected. Loss of heterozygosity of p73 was infrequent and only found in 9% of breast carcinomas. We revealed a few polymorphisms with a frequency of 13% - 29%, which had been reported previously. Down-regulation of p73 mRNA expression was observed in tumor tissues in comparison to the normal breast tissues. A significant inverse correlation was found between p73 transcripts and high histological grade, suggesting that down-regulated p73 expression could be related to poor prognosis in those patients. CONCLUSION: Our results suggest that p73 may serve as a tumor suppressor gene and its expression plays a role in tumorigenesis in Japanese patients with breast cancer.

Lignan, sesquilignans and dilignans, novel HIV-1 protease and cytopathic effect inhibitors purified from the rhizomes of Saururus chinensis.

Five lignans were isolated from the ethyl acetate extracts of Saururus chinensis rhizomes and evaluated for anti-HIV-1 activity. Their structures were elucidated as two dilignans, manassantin A (1), manassantin B (2), two sesquilignans, saucerneol B (3) and saucerneol C (4), and a new lignan, saururin B (5) by spectroscopic analysis. Of these components, manassantin A (1) and saururin B (5) showed dose-dependent inhibitory activities on HIV-1 protease with IC(50) values of 38.9 and 5.6 microM. In addition, manassantins A (1), B (2) and saucerneol B (3) inhibited HIV-1-induced cytopathic effects in a human T lymphoblastoid cell line with IC(100) values of 1.0, 1.0 and 0.2 microM, respectively. Of these active constituents, saucerneol B (3) showed the most potent and selective anti-HIV-1 activity (IC(100) of 0.2 microM, CC(0) of >125.0 microM, and SI of >520.8).

PDZK1 regulates organic anion transporting polypeptide Oatp1a in mouse small intestine.

Recent studies indicate that various members of the organic anion transporting polypeptide (OATP) family are expressed on apical membranes of the small intestine. In the present study, we investigated possible interaction of Oatp with the PDZ protein PDZK1 in mouse small intestine, using [³H]estrone-3-sulfate (E3S) as a typical substrate. After intraduodenal administration, the level of [³H]E3S appearing in the portal vein of pdzk1 gene knockout (pdzk1(-/-)) mice was much lower than that in wild-type mice. Lower intestinal absorption of [³H]E3S in pdzk1(-/-) mice was confirmed in Ussing-type chamber experiments, which showed smaller uptake of [³H]E3S from the apical side in intestinal tissues of pdzk1(-/-) mice compared with wild-type mice. The kinetics and inhibition profile of [³H]E3S uptake in the Ussing-type chamber were similar to those in HEK293 cells stably expressing Oatp1a5, suggesting involvement of Oatp1a5 in [³H]E3S uptake. Immunoreactivity to anti-Oatp1a antibody was colocalized with PDZK1 in the small intestine of wild-type mice, whereas apical localization of Oatp1a protein was reduced in pdzk1(-/-) mice. An immunoprecipitation study revealed physical interaction of PDZK1 with Oatp1a. Thus, PDZK1 appears to act as an adaptor for Oatp1a. This is the first demonstration of a regulatory protein directly interacting with small-intestinal OATP.

Recent diversity of human immunodeficiency virus type 1 in individuals who visited sexually transmitted infection-related clinics in Osaka, Japan.

By human immunodeficiency virus type 1 (HIV-1) antibody screening of people who visited sexually transmitted infection (STI)-related clinics (venereology, urology, and gynecology) and were considered to conduct high-risk sexual activities for HIV-1 infection in Osaka, Japan, during 1992 to 2004, a total of 54 HIV-1 infected individuals (51 Japanese males and 3 non-Japanese females) were identified. Based on the sequencing at env-C2V3 and pol regions, Japanese males were mostly of subtype B (50/51 cases), with the one remaining case being a recombinant circulating form, CRF01_AE, while 3/3 viruses in non-Japanese females were of CRF01_AE. Analysis of subtype B cases since 2001 showed that these viruses became wider in their genetic variation, including amino acid insertions and also deletions, than that of the cases before 2000. Thus, it was suggested that HIV-1 spreading in Osaka has been increasing in genetic variability. Although all these infected individuals were first recognized to be infected with HIV-1 by our screening, some of them were carriers of HIV-1 with drug-resistant pol sequences, indicating that they could be infected with drug-resistant HIV-1 mutants.

Synthesis and properties of G-quartet oligonucleotide-HIV-1 tat peptide conjugate.

Zintevir is a single strand DNA that forms an intramolecular quadruplex structure and shows potent anti-human immunodeficiency virus type 1 (HIV-1) activity. Zintevir was discovered as a potent inhibitor for HIV-1 integrase. Recently, the primary molecular target of Zintevir, however, was shown to be the HIV-1 gp120. In fact, in our previous study, Zintevir was shown to inhibit the only processes of the viral adsorption and the entry into the cell. This result suggests that Zintevir is not able to penetrate through the cell membranes. Therefore, we designed and synthesized the complex of D-17mer with HIV-1 tat peptide that has the cell membrane permeability.

Inhibitory effects of triterpene-azidothymidine conjugates on proliferation of human immunodeficiency virus type 1 and its protease.

The conjugates of some dicarboxylic acid hemiesters of triterpenes which show potent inhibition against human immunodeficiency virus type 1 protease (HIV-1 PR) with a reverse transcriptase inhibitor azidothymidine (AZT) or anti-HIV alkaloid FK 3000 were prepared, and their inhibitory activities were investigated against HIV-induced cytopathic effects (CPE) and HIV-1 PR. Most of the triterpene-AZT conjugates showed potent anti-HIV activity as well as moderate to potent PR inhibitory activity, though AZT itself showed no PR inhibitory activity at all. However, the triterpene-FK 3000 conjugates showed neither PR inhibitory activity nor anti-HIV activity.

Anti-HIV-1 activity and mode of action of mirror image oligodeoxynucleotide analogue of Zintevir.

Zintevir is an oligonucleotide analogue, which has the phosphorothioate modification at both termini, that forms a K(+)-induced quadruplex structure and shows potent anti-human immunodeficiency virus (HIV)-1 activity. We synthesized the non-modified analogue (D-17mer) of Zintevir and its enantiomer (L-17mer), and compared their anti-HIV-1 activity and molecular mechanism of action. Although L-17mer forms the exact mirror image quadruplex structure of D-17mer, which has a very similar structure with Zintevir, L-17mer showed comparable anti-HIV-1 activity with Zintevir. The results obtained by the time-of-addition experiments and the immunofluorescence binding assay strongly suggest that the primary molecular target of L-17mer is the viral gp120 envelope protein as well as Zintevir, regardless of their reciprocal chirality.