Search

The purpose of this retrospective study was to compare the toxicity and disease control rate of radiotherapy for prostate cancer in salvage settings after high-intensity focused ultrasound (HIFU) therapy (HIFU cohort) with those in radical settings (non-HIFU cohort). From 2012 to 2020, 215 patients were identified for this study and 17 were treated in the salvage settings after HIFU. The median follow-up time was 34.5 months (range: 7–102 months, inter-quartile range [IQR]: 16–64 months). Genitourinary (GU) and gastrointestinal (GI) adverse events were evaluated in acute and late periods with Common Terminology Criteria for Adverse Events version 5, and the rates of biochemical-clinical failure free survival (BCFS) and overall survival (OS) were estimated. The cumulative incidence of late GU Grade 2 or greater toxicity after five years was significantly different between the non-HIFU and HIFU cohorts with rates of 7.3% and 26.2%, respectively (P= 0.03). Regarding GI Grade 2 or greater toxicity, there was no significant difference between the two cohorts. The 5y-BCFS was 84.2% in the non-HIFU cohort and 69.5% in the HIFU cohort with no significant difference (P= 0.10) and the 5y-OS was 95.9% and 92.3%, respectively (P= 0.47). We concluded that the possibility of increased late GU Grade 2 or greater should be considered when applying salvage radiotherapy for local recurrence after HIFU.

The complete hydrolysis of konjac glucomannan (KGM) with an acid or enzyme generally takes a long time. To accelerate KGM hydrolysis without diminishing the conventional quality, a diluted acid hydrolysis of KGM with sulfuric acid was conducted using a microwave digestion system. The optimum conditions of microwave-assisted acid hydrolysis for KGM were: 10 mL of 0.25 M sulfuric acid, hydrolysis temperature of 135°C (microwave power of 600 W), and total microwave-irradiation time of 45 min. The yields of the component sugars, mannose and glucose, from two konjac powders were similar to those by conventional acid hydrolysis with 1 M sulfuric acid in a boiling water bath for 5 h. Furthermore, a pretest for microwave-assisted acid hydrolysis using mixtures of konjac powder and starch at different ratios proved that their konjac content can be calculated by determining the mannose generated by the new rapid hydrolysis method, if the raw materials are provided.

Three lipopeptides, ciliatamides A−C ( 1– 3), were isolated from the deep-sea sponge Aaptos ciliata, and their structures were elucidated on the basis of spectroscopic and chemical methods. Ciliatamides A ( 1) and B ( 2) were found to be antileishmanial, while 2also exhibited marginal cytotoxicity to HeLa cells.

Previous studies have shown that cellular prion protein (PrPC) plays anti‐apoptotic and anti‐oxidative role against cell death induced by serum‐deprivation (SDP) in an immortalized prion protein gene‐deficient neuronal cell line derived from Rikn prion protein (PrP) gene‐deficient (Prnp–/–) mice, which ectopically produce excess Doppel (Dpl) (PrP‐like glycoprotein). To investigate whether PrPCinhibits apoptotic neuronal cell death without Dpl, an immortalized cell line was established from the brain of ZrchI Prnp–/–mice, which do not show ectopic expression of Dpl. The results using a ZrchI neuronal Prnp–/–cell line (NpL2) showed that PrPCpotently inhibited SDP‐induced apoptotic cell death. Furthermore, PrPCexpression enhanced the superoxide dismutase (SOD) activity in NpL2 cells. These results indicate that Dpl production did not affect anti‐apoptotic and anti‐oxidative functions of PrP, suggesting that PrPCmay be directly correlated with protection against oxidative stress.

Our previous studies have shown an essential role played by the octapeptide repeat region (OR) and the N‐terminal half of hydrophobic region (HR) in the anti‐apoptotic activity of prion protein (PrP). As PrP‐like protein Doppel (Dpl), which structurally resembles an N‐terminally truncated PrP, did not show any anti‐apoptotic activity, we examined apoptosis of HpL3–4 cells expressing Dpl fused to various lengths of the N‐terminal region of PrP to investigate whether the PrP/Dpl fusion proteins retain anti‐apoptotic function. HpL3–4 cells expressing Dpl fused to PrP(1–124) with the OR and N‐terminal half of HR of PrP showed anti‐apoptotic function, whereas Dpl fused to PrP(1–95) with OR did not rescue cells from apoptotic cell death induced by serum deprivation. These results indicate that the OR and N‐terminal half of HR of PrP retains anti‐apoptotic activity similar to full‐length PrP.

The protective effect of synthetic serum thymic factor (FTS) nonapeptide on lipopolysaccharide (LPS)‐induced pancreatic cell damage in 10‐week‐old BALB/c male mice was investigated. Mice were divided into three groups. Group I was treated with LPS (10 μg/head; i.p.) (LPS‐treated mice). Group II was administered with FTS (50 μg/head; i.p.) 24 hr before treatment with LPS and complemented immediately before LPS injection with FTS (50 μg/head; i.p.) (FTS‐administered mice). Group III was only treated with the same volume of saline (control mice). Treatment of LPS in vivoresulted in the destruction of pancreatic acinar cells. In those cells, many apoptotic cells were detected by immunohistochemistry using an anti‐single stranded DNA antibody. Immunohistochemistry and reverse transcription‐polymerase chain reaction (RT‐PCR) revealed that LPS treatment also caused low or a lack of insulin expression in pancreatic islets. In contrast, morphological change was not seen and apoptotic cell death was suppressed in pancreatic cells of FTS‐administered mice. Moreover, insulin expression was normal in those mice. FTS administration enhanced expression of Bcl‐2 mRNA levels in pancreatic tissues and IL‐6 mRNA levels in splenocytes significantly compared with those of LPS treatment at 3 hr after LPS injection. These findings suggest that FTS prevents LPS‐induced cell damage via enhancing Bcl‐2 expression in the pancreas and systemic IL‐6 production.

1. Cellular prion protein, PrPC, is a ubiquitous glycoprotein strongly expressed in neurons with an as yet unknown biological function. In previous studies, we demonstrated that PrPCcould be regulated by heat shock stress, implying that it might be a stress-responsive protein. Hyperbaric oxygen (HBO) administration is a well-defined model for the study of oxidative stress. 2. This study investigated the effect of HBO on PrPCand Hsp 70 expression in mouse neuroblastoma cell lines (N18), assessing the expression of PrPCand Hsp 70 using RT-PCR and Western blotting. HBO administration resulted in a time- and dose-dependent increase in PrPCand Hsp70 expression in N18 cells at both mRNA and protein levels, with a concomitant upregulation of c-Jun N-terminal kinase (JNK). 3. Under HBO treatment, luciferase reporter constructs of the rat PrPCpromoter, containing the heat shock element (HSE) also present in Hsp70, expressed higher luciferase activity (3- to 10-fold) than those constructs without HSE. 4. In summary, these data suggest that PrPCand Hsp 70 may be regulated by HBO, through the activation of JNK. Thus, the activated heat shock transcriptional factor 1, phosphorylated by JNK interacted with HSE in the promoter of PrPCresulted in increased gene expression. These findings are vital for future therapeutic approaches in transmissible spongiform encephalopathies and the understanding of the function of the PrPC.

The effects of tacrolimus on insulin‐dependent diabetes mellitus (IDDM) induced by the D‐variant of encephalomyocarditis virus (D‐EMCV) have been investigated. Male BALB/c mice were treated with tacrolimus before viral inoculation, and then were inoculated with 10 plaque forming units (PFU) of D‐EMCV. The mice continued to be treated with tacrolimus until the animals were sacrificed. D‐EMCV‐infected mice, which were treated with saline as controls, showed abnormal glucose tolerance test (GTT) values, whereas all infected mice with tacrolimus pretreatment were normal on 7 days‐post inoculation (DPI). Histological observations revealed that non‐treated tacrolimus D‐EMCV‐infected mice and which developed diabetes showed severe insulitis in their islets of Langerhans. On the other hand, D‐EMCV‐infected mice treated with tacrolimus were normal. In D‐EMCV‐infected mice, viruses in the pancreata were detected at the same level regardless of treatment with tacrolimus or saline. Expressions of TNF‐α and IFN‐γ mRNA in spleens of tacrolimus‐treated D‐EMCV‐infected mice were lower than that of non‐treated tacrolimus D‐EMCV‐infected mice on 7 DPI. The results suggest that tacrolimus suppresses expressions of TNF‐α and IFN‐γ mRNAs to prevent the onset of D‐EMCV‐induced IDDM.

The prion protein (PrP) nucleotide sequences of two ruminants were determined in order to elucidate the differences in susceptibility to spongiform encephalopathy agents in each species. The nucleotide sequences of PrP coding regions of the moufflon and the golden takin encompassed 771 bp in length. The PrP gene sequences of the golden takin were closely related to those of sheep with one amino acid difference. The PrP gene sequence of the moufflon was identical to that of sheep. The similarities between the PrP genes of these two animals and sheep imply that the species barriers between these animals are small or non-existent. These PrP genes could be used to establish transgenic mice with higher susceptibility to prion-related diseases.

Islet antigen (IA)-2 is a novel autoantigen of insulin-dependent diabetes mellitus (IDDM), and belongs to a new class within the receptor-type protein tyrosine phosphatase (PTP) family characterized by lack of PTP enzymatic activity with conventional substrates. Its expression is restricted primarily to the pancreas, pituitary, and brain with the highest level in the brain. IA-2 mRNA expressions in the brain, pituitary and pancreas of 1-, 4-, and 8-week-old mice were examined. In situ hybridization of the brain revealed that IA-2 mRNA was expressed in the cerebral cortex, hippocampus, thalamus, choroid plexus, hypothalamus, Purkinje cells, and granular layer of the cerebellum. In the pituitary, IA-2 mRNA was located in the anterior and posterior pituitary by in situ hybridization. The pattern of IA-2 mRNA expression in normal male mouse brain at 1, 4, and 8 weeks of age by the Northern blot analysis was similar to that in the pituitary by RT-PCR analysis. The expression level was higher at 4 weeks and lower at 1 week of age. In the pancreas, IA-2 mRNA expressions detected by RT-PCR were highest at 8 weeks of age. These results indicated that the amount of mRNA expression increased in accordance to development in brain, pituitary, and pancreas.

In this study, the authors investigated normal cellular prion protein (PrP) expression on murine immune systems using prion protein gene-deficient mouse as negative control. Immunocytes expressing PrP in adult and fetal mice were detected by flow cytometry with the monoclonal antibody against PrP, 6H4. Cells from thymus and bone marrow reacted positively with 6H4, while spleen cells, peritoneal cells, peripheral blood leukocytes, and intestinal intraepithelial lymphocytes were nonreactive. In thymus, PrP was observed in CD4CD8 double-negative thymocytes. PrP cells of double-negative thymocytes belonged to the CD3 subset, but not to the CD3 subset. Triple-negative PrP thymocytes expressed CD44 or CD25 antigens. Furthermore, PrP was observed in c-kit bone marrow cells. In fetuses, PrP cells were observed in the liver and thymus at day 16.0 and 15.0 of gestation, respectively. These results demonstrated that PrP is expressed on immature immunocytes.

ABSTRACTOne of the most characteristic clinical features in cutaneous leishmaniasis is the development of nodules followed by ulcerations at the site of infection. Leishmania amazonensis-infected mice show similar ulcerative lesions. Leishmania-infected severe combined immunodeficiency (SCID) mice, however, have been shown to develop nonulcerative nodules. In the present study, the roles of T cells in ulceration were examined using SCID mice in cell reconstitution experiments. After development of nonulcerative nodules, SCID mice were inoculated with splenocytes from eitherLeishmania-infected or naive immunocompetent mice, resulting in ulceration in all mice. When naive splenocytes were depleted of CD4+, CD8+, or B220+cell populations and the remaining cells were injected intoLeishmania-infected SCID mice after the development of nodules, only SCID mice inoculated with splenocytes depleted of CD4+cells did not show ulceration. The evidence obtained in this study clearly shows that the CD4+cell population is indispensable for ulceration in leishmaniasis lesions of SCID mice.

In order to understand the fundamental and putative roles of PrPc in the central nervous system, neuronal cell lines were established. Cells were immortalized by recombinant retrovirus vector-mediated transduction of SV40 T-antigen gene. Among these, two cell lines were selected based on their RT-PCR expressions of neuron-specific neurofilament (NF-H, NF-M) and cell morphology. These cell lines showed the properties of neuronal progenitor cells in antigenicity, morphology and responses to differentiating agents. Expression of PrPc was detected by immunocytochemical analysis. These cell lines responded to differentiating agents such as dibutyl cyclic AMP (dcAMP) and phorbol 12-myristate 13-acetate (PMA) before developing into neuronal-like cells. Neurite extensions were observed 20 min after incubation with the differentiating agents. Treatment with nerve growth factor (NGF) and insulin induced cell differentiation and enhanced expression of PrP gene (Prnp) mRNA and protein. The latter phenomenon was not inhibited by wortmannin, which is a specific inhibitor of phosphatidylinositol 3-kinase. These results suggest that PrPc plays an important role in the differentiation-mediated classic signaling pathway of neuronal cell.

Equine infectious anemia virus (EIAV) core proteins were obtained from a baculovirus expression system. Recombinant baculoviruses (rBVs) highly expressed the Gag precursor and p26 antigens in an rBV‐infected Sf21 cell culture supernatant. Enzyme‐linked immunosorbent assay (ELISA) and agar gel immunodiffusion (AGID) were conducted using the expressed proteins to detect antibodies from experimentally infected horses. The expressed antigens showed low background levels, high specificity and sensitivity in ELISA and AGID. The results of the serological tests using the expressed antigens were identical to those using a manufactured trial antigen. rBVs containing gagand p26 genes were found to express high quality and large quantities of Gag and p26 antigens, respectively. The antigens were quite useful for detecting anti‐EIAV antibodies from virus‐infected horses.

We have demonstrated the presence of a rat prion protein (RaPrP) gene promoter upstream of multiple initiation sites. A 0.1-kb fragment upstream of the 5′-untranslated region contains specific DNA motifs characteristic of promoter elements including an AP-1 binding site, an inverted CCAAT motif and three inverted Sp-1 binding sites. This fragment directs transcription of a luciferase reporter gene in pheochromocytoma cells (PC12) and rat glioma cells (C6), suggesting that it contains the promoter for the RaPrP gene. To more precisely localize the transcription regulatory elements in this region, a series of 5′-deletion mutants were generated. Deletion analysis showed that an inverted CCAAT and adjoining Sp-1 binding sequences may play an important role in transcription of the RaPrP gene.

The prion protein (PrP), encoded by a chromosomal gene, is associated with development of the neurodegeneration of prion-induced diseases. Since determination of the complete structure of the gene encoding PrP is important for understanding gene expression in the central nervous system (CNS), the nucleotide (nt) sequence of the isolated whole gene encoding rat PrP (raPrP) was determined. The rat PrP gene (raPrP) spans 16 kilobases (kb) of the rat genome and contains three exons of 19–47 base pairs (bp), 98 bp, and 2 kb separated by two introns of 2.2 kb and 11 kb. The first and second exons are noncoding, while the third exon contains a short 5' untranslated region, the entire 762-bp open reading frame (ORF), and a 3' untranslated region. The putative raPrP promoter in the 5' flanking region contains putative Sp1, AP-1, and AP-2 binding sites without a consensus TATA box. This TATA box-deficient feature, coupled with the presence of a high G+C content and Sp1-binding sites in the raPrP promoter, characterizes it as a housekeeping gene. Analysis of the raPrP cDNA 5'-end showed that raPrP mRNA transcription was initiated at multiple sites. Northern blot analysis showed that the levels of raPrP mRNA varied among rat tissues, with the highest levels found in the brain and placenta. This determination of raPrP nt sequences, including the introns and the 5' and 3' flanking regions, may make it possible to elucidate cis-acting elements that regulate the expression of this gene in different tissues and cell lines.

ABSTRACTTo diagnose visceral leishmaniasis (kala-azar), we have developed a nested PCR method based on amplification of the mini-exon gene, which is unique and tandomly repeated in the Leishmaniagenome. Nested PCR was sufficiently sensitive for the detection of DNA in an amount equivalent to a single Leishmaniaparasite or less. We examined the usefulness of this PCR method using bone marrow aspirates and buffy coat cells collected from kala-azar patients who had or had not received chemotherapy in northwest China. We obtained PCR positivity for all of the parasitologically positive bone marrow samples from the patients. Some ambiguities with the primary PCR results were eliminated by the subsequent nested PCR. The buffy coat samples from 7 of 12 patients with splenomegaly were positive by the nested PCR, although only 2 of them were positive for parasites by culture. However, buffy coat samples from nine children, whose splenomegaly has been reduced and clinically cured by antimony treatment, were all negative. Thus, this nested PCR method represents a new tool for the diagnosis of kala-azar with patient blood samples instead of bone marrow or spleen aspirates obtained by more invasive procedures.

Schistosomes are blood-dwelling trematode parasites that infect 200 million people in developing countries. The critical role served by the tegument in immune evasion and parasite homeostasis suggests that a detailed knowledge of tegumental components would be helpful in the design of new drugs and the production of vaccines. We demonstrate here, by immunoelectron microscopy, that the cytoskeletal proteins actin and paramyosin are organized into major tegumental structures of Schistosoma mansoni. The surface spines are composed of paracrystalline arrays of actin filaments. Actin is also present in areas recovering from damage, implying an important role for this structural protein in tegumental repair. Paramyosin exists predominantly in the tegument in a non-filamentous form, the membrane-bounded elongate bodies. The localization of this protein to the tegument of the parasite is the likely basis for resistance to S. mansoni observed in mice immunized with paramyosin (refs 1, 2 and T. P. Flanigen et al., in preparation).

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 200 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

Malaria, caused by plasmodium species has great impact on human generations. Each year 300–500 million of people are diagnosed and 1 million die of the disease. Malaria probably become endemic about 10,000 years ago and it has affected human genome by strong selective pressure. Several human genetic diseases are thought to be the outcome of this pressure. Our current research is focused on understanding natural host defense mechanism of this disease, especially effect to human fetal hemoglobin expression to plasmodium species. Other investigator indicated growth of Plasmodium falciparum is retarded in the cell containing human fetal hemoglobin in vivo (Nature, 270:171), and transgenic mouse (transgenic of human fetal hemoglobin and knockout of mouse beta-major gene) infected with Plasmodium has longer survival compare to control.(Blood, 92:2520). Here we use transgenic mouse express human fetal hemoglobin (human Gamma globin) which simulate human hereditary persistence of fetal hemoglobin (PNAS86:7033) to investigate fetal hemoglobin itself has a protective effect on Plasmodium infection in vivo. We injected 1,000 Plasmodium.yoelii infected RBCs to transgenic mouse expressing human fetal hemoglobin (n=5) and C57BL6J mouse (n=8). Blood smears obtained daily and smears were stained with Giemsa. Parasitemia is calculated from about 1000 RBCs. 5 days after injection, infected RBCs was 26.4 ±4.7% for C57BL6J and 32.6± 17.3% for human fetal hemoglobin transgenic mouse. We couldn’t show significant difference between these groups. And 60% of C57BL6J mice (3 of 5) were recovered from illness. In contrast, 20% of transgenic mouse (1of 5) was recovered from illness. These results demonstrates only human fetal hemoglobin expression is not enough to contribute defense mechanism against plasmodium species.

Malaria, caused by plasmodium species has great impact on human generations. Each year 300–500 million of people are diagnosed and 1 million die of the disease. Malaria probably become endemic about 10,000 years ago and it has affected human genome by strong selective pressure. Several human genetic diseases are thought to be the outcome of this pressure. Our current research is focused on understanding natural host defense mechanism of this disease, especially effect to human fetal hemoglobin expression to plasmodium species.