Search

Spinach (Spinacia oleracea L.) is widely known to be dioecious. However, monoecious plants can also occur in this species. Sex expression in dioecious spinach plants is controlled by a single gene pair termed X and Y. Our previous study showed that a single, incompletely dominant gene, which controls the monoecious condition in spinach line 03-336, should be allelic or linked to X/Y. Here, we developed 19 AFLP markers closely linked to the monoecious gene. The AFLP markers were mapped to a 38.2-cM chromosomal region that included the monoecious gene, which is bracketed between flanking markers with a distance of 7.1 cM. The four AFLP markers developed in our studies were converted into sequence-characterized amplified region (SCAR) markers, which are linked to both the monoecious gene and Y and are common to both populations segregating for the genes. Linkage analysis using the SCAR markers suggested that the monoecious gene (M) and Y are located in different intervals, between different marker pairs. Analysis of populations segregating for both M and Y also directly demonstrates linkage of the genes at a distance of ∼12 cM. The data presented in this study may be useful for breeding dioecious and highly male monoecious lines utilized as the pollen parents for hybrid seed production, as well as for studies of the evolutionary history of sexual systems in this species, and can provide a molecular basis for positional cloning of the sex-determining genes. [ABSTRACT FROM AUTHOR]

Background: Ulcerative colitis (UC) is a chronic relapsing inflammatory bowel disease, known to be associated with a markedly increased risk of colorectal carcinoma development.Methods: Using proteomic analysis with two-dimensional gel electrophoresis and mass spectrometry, differentially expressed proteins were assessed between UC-associated cancer and sporadic colon cancer cell lines. Western blot and immunostaining were performed for confirming the expression.Results: Heat-shock protein of 47 kDa (HSP47) was identified as one of the proteins expressed more highly in UC-associated cancer cell lines, and an immunohistochemical examination confirmed significantly higher levels of HSP47 in UC-associated colon cancers than in sporadic counterparts, the expression increasing with a progression of neoplastic lesions. Heat-shock protein of 47 kDa was further found to be coexpressed with type I collagen in the cytoplasm, and both HSP47 and type I collagen were released from cultured cells into the culture medium.Conclusion: These results suggest that overexpression of HSP47 is a unique characteristic of UC-associated carcinoma related to type I collagen synthesis, with possible clinical applications. [ABSTRACT FROM AUTHOR]

Background: Recently, preoperative chemoradiation therapy (CRT) for rectal cancer has been increasingly used as a neoadjuvant treatment. In the present study, the relation between histological response to CRT and immunohistochemical markers in biopsy specimens was investigated.Methods: Biopsy specimens from a total of 60 patients were collected before preoperative CRT with S-1 and irinotecan, and liniac 45 Gy. Immunohistochemical staining for Ki67, Mcm3, Bax, Bcl-2, ssDNA, Grp78, thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), CD34, vascular endothelial growth factor, nestin, and L-type amino-acid transporter 1 was performed to allow comparison of the Ki67 labelling index (LI), Bax score, TS score, DPD score, microvessel density by CD34, and Grp78 score with cancer regression.Results: When the cases were divided into responders (Dworak grades 3 and 4) and non-responders (grades 1 and 2) groups, good correlations were evident with Ki67 LI, Bax, Grp78, and TS expression. On multiple logistic regression analysis, Ki67 LI, Bax, and TS scores were found to be independent factors. With their use in a logistic model, P-values could predict responder cases with a sensitivity of 82.8% and a specificity of 83.9%.Conclusion: Using this system, treatment strategy for locally advanced rectal cancers can be determined before chemoradiation. [ABSTRACT FROM AUTHOR]

In this study, we have analyzed the temporal oscillations of precipitation in meso-scale zones of Sri Lanka to examine potential existence of periodic oscillatory behavior in rainfall. Only a few statistically significant cycles were identified: a 3.5-year cycle in most of central Sri Lanka during the January–March rainfall regime and a cycle of the same length in southwestern Sri Lanka during the October regime. A 2.1-year cycle marks the northeastern parts of Sri Lanka during the December/April contrast rainfall regime. This cycle is shown to be strongly related to Quasi-Biennial Oscillation. October and November rainfall are found to be coupled with ENSO fluctuations, and on average, more than 900 mm more rainfall is observed per month over all stations during El Niño than during La Niña years. Analysis of relationships between the observed meso-scale rainfall regions and the Sea Surface Temperatures (SSTs) in the Indian Ocean north of the Equator showed that the northern Indian Ocean can be divided into three particular regions based on similarity in the SST fluctuations: (a) a region with cool upwelling water, (b) non upwelling water, and (c) the Indian Ocean Warm Pool. We found that there are no statistically significant relationships between the observed SST regions in the Indian Ocean and the meso-scale precipitation patterns in Sri Lanka. [ABSTRACT FROM AUTHOR]

Background: We have shown previously that stromal genetic alteration may make a greater contribution to early genesis of ulcerative colitis-associated tumors than sporadic colon cancers. We assessed whether similar differences in genetic alteration might exist between squamous cell carcinomas (SCCs) and Barrett's adenocarcinomas (BACs) of the esophagus.Methods: We investigated epithelial and stromal genetic instability with five National Cancer Institute standard (NCI), four chromosome 17 (Chr. 17), and six tumor suppressor gene (TSG) microsatellite markers in 26 SCC and 12 BAC cases and in 11 normal controls, using a novel combination of microdissection, polymerase chain reaction, and GeneScan.Results: Frequency of epithelial loss of heterozygosity (LOH) increased in the order background mucosa, to precursor lesions, to tumors with both types of carcinoma, especially for the Chr. 17 and TSG markers, while stromal LOH was relatively high but consistent from background mucosa to carcinoma. Epithelial LOH of D17S796 demonstrated a significantly higher frequency in SCCs than in BACs, without significant variation in p53 overexpression. The frequency of microsatellite instability (MSI) showed constant high levels in both epithelium and stroma of background, dysplasia, and carcinomas in the SCC series, and rather low frequencies in the BAC series. Although epithelial hMLH1 and hMSH2 expression decreased with tumor progression, no correlation was found with the individual MSI status.Conclusions: Although epithelial LOH exists similarly in both lesion types, whereas epithelial and stromal MSI may occur in a relatively early phase of SCC development, stromal MSI is rare in BACs, strongly suggesting differences in tumorigenesis between the two types. [ABSTRACT FROM AUTHOR]

We report here the molecular mapping of a fertility restorer gene (named Rf1) for Owen cytoplasmic male sterility in sugar beet. Eight AFLP and two RAPD markers, tightly linked to the Rf1 locus, were identified using bulked segregant analysis. Three AFLP markers, mAFEM972, mAFEM976 and mAFEM985, were found to co-segregate with the Rf1 allele in our mapping populations. With the help of RFLP markers, previously mapped on the sugar beet genome, we showed that Rf1 is positioned in the terminal region of linkage group Kiel III/Koeln IV. This map location agrees well with that found for the restorer gene X, which suggests that the Rf1 locus corresponds to the X locus. The availability of the molecular markers will facilitate the selection of maintainer–pollinator lines in breeding program and provide the foundation for map-based cloning of the Rf1 gene. [ABSTRACT FROM AUTHOR]

In ulcerative colitis (UC)-associated tumours, p53 gene mutations and p53 protein overexpression are frequently found in early stages, but the two types of alteration do not always coincide. To clarify this discrepancy, p53 mutations and expression of p53-associated molecules were analysed in UC-associated dysplasias by a combination of microdissection, polymerase chain reaction-direct sequencing and immunohistochemistry at the single crypt level. Mismatch of p53 protein overexpression (+)/mutation (-) or p53 overexpression (-)/gene mutation (+) was found in nine crypts in regenerative mucosa (19 crypts), in 27 in low-grade dysplasia (41), in one in high-grade dysplasia (5) and in 12 in invasive carcinomas (17). Regarding these mismatched crypts of the first type, significant increase in p16INK4A and Bax expression was found. The Ki-67 labelling index was depressed in such p53-diffusely positive lesions with the wild-type p53 gene, compared to their p53-diffusely positive and mutant type counterparts. p16INK4A was upregulated indirectly as part of the negative feedback, and increase in Bax, directly controlled by wild-type p53, indicates upregulation of apoptosis. No significant relation with p53-related gene products was detected with the p53 protein overexpression (-)/p53 mutation (+) mismatch. Therefore, a tumorigenesis pathway independent of p53 dysfunction appears to exist in association with ulcerative colitis.British Journal of Cancer (2004) 91, 1081-1088. doi:10.1038/sj.bjc.6602050 www.bjcancer.com Published online 3 August 2004 [ABSTRACT FROM AUTHOR]

A computational model of the fluid dynamics of intraventricular flow was used to investigate the importance of the effects of flow disturbances existing within the left ventricle (LV) at the onset of diastole on a diastolic flow field. The simulation started with a quiescent flow state; it continued for a number of cardiac cycles to obtain a cyclically repeatable flow. After the flow became periodic, the initial diastolic flow was not quiescent: flow disturbances, remnants of a systolic flow, were present within the LV. Nevertheless, they faded away during an acceleration phase of diastole and almost ceased by the end of this phase. Consequently, a flow field during a deceleration phase of diastole, characterised by the formation of a vortex ring, was hardly affected by the initial flow disturbances. The propagation velocity of a colour M-mode Doppler echocardiogram obtained by scanning velocity along the LV long axis was 0.58 m s(-1) in the case where diastolic flow was initially quiescent and 0.56 m s(-1) in the case where flow disturbances existed at the beginning of diastole. These results indicated that the colour M-mode Doppler echocardiographic technique captures flow dynamics produced purely by ventricular expansion, with little influence from initial diastolic flow disturbances. [ABSTRACT FROM AUTHOR]

Summary. We studied the effect of buthionine sulfoximine (BSO) on the replication of an isolate of human echovirus 9 (EV9) and the apoptosis induced by it in GMK cells. One hundred μM BSO markedly inhibited the cytopathic effect (CPE) induced by EV9. BSO also significantly inhibited apoptosis induced by EV9. BSO did not influence replication of EV9 genome, but inhibited virion formation. These results suggest that the inhibition by BSO of CPE and apoptosis induced by EV9 may be associated with the impairment of virion formation. Moreover, apoptosis induced by infections of human poliovirus 3, human coxsackievirus B5, A10 and A16, which, like EV9, belong to the genus Enterovirus, was markedly abolished by BSO. This finding suggests that enteroviral infections cause apoptosis through the activation of a common pathway that can be inhibited by BSO. [ABSTRACT FROM AUTHOR]

Summary. Phylogenetic relationships of novel Vietnamese strains of feline immunodeficiency virus (FIV) were analysed. One Vietnamese strain was found to cluster with subtype D, which was previously known only in Japan, while the other seven strains were placed with members of subtype C. Calculation of the relative numbers of mutations resulting in amino acid and silent changes in FIV env subtypes suggested that subtype C isolates may be less structurally constrained (potentially more pathogenic) than subtype B. [ABSTRACT FROM AUTHOR]

Effects of neuronal nitric oxide synthase (nNOS) inhibitor (7-nitroindazole), nonselective NOS inhibitor (NG-nitro-L-arginine methyl ester; L-NAME), and monoamine oxidase inhibitor (pargyline) were studied on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice. The mice received four intraperitoneal injections of MPTP at 1-h intervals. A significant depletion in dopamine and DOPAC concentration was observed in the striatum from 1 day after MPTP treatment. The pretreatment of 7-nitroindazole and pargyline, but not L-NAME, dose-dependently protected against MPTP-induced depletion in dopamine content 3 days after MPTP treatment. Our histochemical study also showed that 7-nitroindazole and pargyline can prevent a marked decrease in the nigral cells and a marked increase in astrocytes in striatum 7 days after MPTP treatment. The protective effect of 7-nitroindazole against MPTP-induced dopamine and DOPAC depletion in the striatum was not attenuated by intraperitoneal pretreatment with L-arginine. Furthermore, the posttreatment of 7-nitroindazole or pargyline protected against MPTP-induced depletion of dopamine content. These results demonstrate that the protective mechanism by which 7-nitroindazole counteracts MPTP neurotoxicity in mice may be due not only to inhibition of nNOS, but also to MAO-B inhibition. Furthermore, our study suggests that the posttreatment of 7-nitroindazole and pargyline can prevent a significant decrease in dopamine levels in the striatum of MPTP-treated mice. These findings have important implications for the therapeutic time window and choice of nNOS or MAO inhibitors in patients with Parkinson's disease. [ABSTRACT FROM AUTHOR]

As part of a study to identify the characteristics of cord blood cells, we examined their morphological features by electron microscopy. Additionally, we cultured CD34-positive cells derived from cord blood and from bone marrow to perform morphological observations, as well as cytochemical examinations following the peroxidase reaction. Compared with normal peripheral blood cells, cord blood cells frequently showed immature morphology and a unique ultrastructure, such as nuclear pockets in neutrophils, several crystalloids in a single eosinophilic granule, and deformed nuclei in lymphoctytes. In contrast to bone marrow cells, cord blood cells yielded a large number of cells of immature myelo-monocytic lineages in cell culture, and demonstrated a weaker peroxidase reaction. We identified that cord blood cells were different from normal peripheral blood cells and bone marrow cells, confirming the functional differences that were previously assumed. [ABSTRACT FROM AUTHOR]

The sugar beet mitochondrial gene for subunit IV of NADH dehydrogenase (nad4) has been characterized. Unlike the corresponding genes in wheat and turnip, sugar beet nad4 lacks the second intron (nad4-i2). Northern-blot analysis demonstrates transcription of the gene. A total of 19 RNA editing sites were identified in the sugar beet nad4 transcripts; interestingly, there is no editing in the region which flanks the lost intron. This observation is in favour of intron loss via homologous recombination of an edited RNA intermediate. We also found that the nad4-i2 intron is absent from the mitochondrial genomes of all examined members of the Caryophyllales, but present in the closely related orders, Polygonales and Plumbaginales, which suggests that the intron was lost in the common ancestor of the Caryophyllales. [ABSTRACT FROM AUTHOR]

Copyright of Materials & Structures is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background. Defensins are antimicrobial peptides that constitute 30% to 50% of total azurophil granule proteins. Although the activation of azurophils during hemodialysis has been reported, including the release of granulocyte lactoferrin, myeloperoxidase, and elastase, the release of defensins during hemodialysis has not been investigated. The purpose of this study was to determine the effects of hemodialysis on defensin release. Methods. Plasma defensin levels and excreted defensins in dialyzer eluates and dialysates during hemodialysis were determined by radioimmunoassay. Thirty hemodialysis patients were divided into three groups, each group using a different dialyzer membrane: cuprophan (CU), polymethylmethacrylate (PMMA), or polysulfone (PS). Results. The postdialysis plasma defensin level in the CU group was significantly increased compared with the predialysis level. In contrast, defensin levels in the PMMA and PS groups showed significant decreases postdialysis. The predialysis neutrophil count and defensin level exhibited a significant positive correlation in all groups. However, postdialysis, a significant positive correlation was found only in the CU group. Interestingly, defensin excretion in the dialyzer eluates of the PMMA group and in the dialysates of the PS group was higher than that in each of the other two groups respectively. Conclusions. No significant statistical correlation between postdialysis neutrophil count and defensin level was observed in the PMMA and PS groups, a finding that may be explained by the increased excretion of defensins during hemodialysis in these groups. These results suggested that plasma defensin level may be a marker for granulocyte activation in patients dialyzed with membranes showing lower levels of excreted defensins. This study also revealed the importance of determining both plasma changes and excreted volumes of granule proteins to define neutrophil degranulation during hemodialysis. [ABSTRACT FROM AUTHOR]

Summary. We have previously reported the construction of a recombinant feline herpesvirus type 1 (FHV-1), designated C7301ddlTK-gag, expressing the Gag precursor protein of feline immunodeficiency virus (FIV). In this study, we report the construction of a further recombinant FHV-1 (ddlTK(gBp)-gag) which carries an FHV-1 gB promoter sequence upstream of the FIV gag gene of C7301ddlTK-gag. Strong expression of the FIV Gag protein by ddlTK(gBp)-gag was confirmed by immunoblot analyses and enzyme-linked immunosorbent assays. Although C7301ddlTK-gag and ddlTK(gBp)-gag failed to induce anti-FIV Gag antibodies in cats, we confirmed the infectivity and stability of these recombinants in cats. [ABSTRACT FROM AUTHOR]

Abstract. R. Jordan, D. Kinderlehrer, and E Otto proposed the discrete-time approximation of the Fokker-Planck equation by the variational formulation. It is determined by the Wasserstein metric, an energy functional, and the Gibbs-Boltzmann entropy functional. In this paper we study the asymptotic behavior of the dynamical systems which describe their approximation of the Fokker-Planck equation and characterize the limit as a solution to a class of variational problems. [ABSTRACT FROM AUTHOR]

Summary. In order to identify the products of the equine herpesvirus type 4 (EHV-4) gI and gE genes, we have constructed recombinant vaccinia viruses containing the putative gI or gE genes. These recombinant viruses synthesized EHV-4 gI and gE with apparent molecular masses of 75 and 80 kDa, respectively. Antibodies raised against both recombinant viruses detected a 75 kDa gI and a 95 kDa gE in EHV-4-infected cells. The results also suggest that the EHV-4 gI and gE would form a complex like in other herpesviruses. [ABSTRACT FROM AUTHOR]

Pharmacologic stress testing is recommended to elderly patients as a valuable alternative to exercise testing. We examined whether exercise testing is as useful for evaluating myocardial ischemia in the elderly as in the young. The consecutive 1,508 patients who underwent exercise 201Tl single-photon emission computed tomography (SPECT) were divided into six age groups: 6-29 years (n = 56), 30-44 (n = 143), 45-54 (n = 311), 55-64 (n = 498), 65-74 (n = 402), and 75-88 (n = 98). Both heart rate and rate-pressure product at peak exercise were significantly lower in patients aged 75-88 than in the other five groups. The frequency of ischemic ST depression was higher in patients aged 75-88 than in those aged 6-74, although the difference was not significant. Moreover, the frequency of 201Tl transient defect was significantly higher in patients aged 75-88 than in those aged 6-74. On the other hand, the sensitivity of ischemic ST depression for 201Tl transient defect was similar among the six groups, but the specificity was significantly lower in patients aged 75-88 than in those aged 6-74. In conclusion, exercise 201Tl SPECT is useful for evaluating myocardial ischemia even in the elderly, but exercise electrocardiography has limitations such as lower specificity in the elderly than 201Tl SPECT. [ABSTRACT FROM AUTHOR]

We describe the expression of SAG-1 cDNA in B6C3F1 mice by microinjecting a 3.3 kbp DNA fragment, consisting of the cytomegalovirus enhancer-chicken β-actin hybrid promoter and SAG-1 into the pronucleus of a fertilized egg at the one-cell stage. Offspring derived from this microinjection were analyzed for the integration and functional expression of the SAG-1 transgene. Steady-state expressions of both the mRNA for SAG-1 and SAG-1 protein product were detected in the brain, thymus, spleen and liver. Approximately 50% of F1 and F2 progeny inherited the SAG-1 transgene from SAG-1 transgenic mice in Mendelian fashion. These results indicated that SAG-1 transgenic lines were established. Transgenic mice harboring the SAG-1 gene will contribute a critical tool of defining the molecular mechanisms of SAG-1 in pathogenesis and host immune response. [ABSTRACT FROM AUTHOR]

We have found that a gene coding for NADH dehydrogenase subunit 4L and a presumed gene, orf25, are linked and co-transcribed with each other in sugar beet mitochondria. Ten and twelve C-to-U editing events were observed in the mRNAs of nad4L and orf25, respectively; the amino-acid sequence specified after editing is better-conserved in comparison with the homologues of other organisms. It is interesting to note that the translation initiation codon of nad4L is created by editing. The conservation of the nad4L-orf25 linkage was examined by PCR-amplification of the intergenic region. We obtained successful PCR products from five dicots (spinach, apple, snapdragon, petunia and tobacco) and two monocots (tulip and pineapple), but not in two poaceous plants, rice and maize. The intergenic region, when present, was found to be well-conserved in its sequence, suggesting a monophyletic origin of this linkage. Our result, together with previous reports of Arabidopsis and four poaceous species, favour the argument that the nad4L-orf25 linkage is conserved throughout angiosperms except in the Poaceae. [ABSTRACT FROM AUTHOR]

To map the transcripts encoding the equine herpesvirus type 4 (EHV-4) glycoproteins I (gI) and E (gE), transcriptional analyses were performed at the right part of the unique short segment of EHV-4 genome. The results revealed that the gI gene is encoded by a 1.6-kb transcript which is 3′ coterminal with a 3.0-kb gD mRNA while the gE gene is encoded by two transcripts of 3.5- and 2.4-kb in size. The transcriptional patterns described in this study for the EHV-4 gI and gE are similar to those found in the equivalent region of herpes simplex virus type 1 and feline herpesvirus type 1. Characterization of EHV-4 gI and gE glycoprotein genes may facilitate future studies to define their roles in the EHV-4 infection. [ABSTRACT FROM AUTHOR]

We have constructed a physical map of the mitochondrial DNA of a cytoplasmic male sterile (CMS) sugar beet line, TK81-MS, and compared it with that published for normal fertile sugar beet (cv. TK81-O) to clarify the differences between the CMS and normal mitochondrial genomes. The TK81-MS genome is present as a single circular molecule of 481.8 kb, or as two molecules of 184.9 and 296.9 kb. The CMS genome was found to be highly rearranged relative to the normal mitochondrial genome, with at least fifteen rearrangement and/or inversion events being required to align the two DNAs. Analysis of transcription patterns of known mitochondrial genes and rearranged regions revealed six genes, coxI, coxII, atpA, atp6, rps3, and orf324, whose expression is altered in the CMS line relative to the normal line. Of these six, only the coxI transcript pattern differs between male-sterile and fertility-restored genotypes, making it likely that the coxI locus is involved in mediating CMS in sugar beet. [ABSTRACT FROM AUTHOR]

Background: Diabetic cardiac autonomic dysfunction often causes lethal arrhythmia and sudden cardiac death. 123I-Metaiodobenzylguanidine (MIBG) can evaluate cardiac sympathetic dysfunction, and analysis of heart rate variability (HRV) can reflect cardiac parasympathetic activity. We examined whether cardiac parasympathetic dysfunction assessed by HRV may correlate with sympathetic dysfunction assessed by MIBG in diabetic patients.Methods and Results: In 24-hour electrocardiography, we analyzed 4 HRV parameters: high-frequency power (HF), HF in the early morning (EMHF), rMSSD and pNN50. MIBG planar images and SPECT were obtained 15 minutes (early) and 150 minutes (late) after injection and the heart washout rate was calculated. The defect score in 9 left ventricular regions was scored on a 4 point scale (0 = normal approximately 3 = severe defect). In 20 selected diabetic patients without congestive heart failure, coronary artery disease and renal failure, parasympathetic HRV parameters had a negative correlation with the sum of defect scores (DS) in the late images (R = -0.47 approximately -0.59, p < 0.05) and some parameters had a negative correlation with the washout rate (R = -0.50 approximately -0.55, p < 0.05). In a total of 64 diabetic patients also, these parameters had a negative correlation with late DS (R = -0.28 approximately -0.35, p < 0.05) and early DS (R = -0.27 approximately -0.32, p < 0.05).Conclusions: The progress of diabetic cardiac parasympathetic dysfunction may parallel the sympathetic one. [ABSTRACT FROM AUTHOR]

We induced apoptosis in cells of the human leukemia cell line HL-60 using an antitumor agent, docetaxel (Taxotere), and investigated apoptosis in various aspects using in situ end-labeling (ISEL) of DNA, DNA fragmentation assay, flow cytometry, and electron microscopy. Because it inhibits depolymerization of tubulin, docetaxel is thought to arrest the cell cycle at the mitotic stage and to exert an antitumor effect. In this study, accumulation of docetaxel-treated cells at the G2/M phase was detected using flow cytometry. On ISEL of DNA, DNA fragmentation was observed at the mitotic stage. On electron microscopy, the nuclei of apoptotic cells lost their nuclear membranes, as do cells at mitosis, demonstrating that the cells were arrested mainly at the M phase in the cell cycle. [ABSTRACT FROM AUTHOR]

The paper describes steady-state and transient responses of transcutaneous pO and pCO by mass spectrometry. The effects of factors affecting the responses were estimated from a mathematical model comprising the skin, a membrane of a sampler and a tube connected to a mass spectrometer. The effect of heading temperature, membrane thickness, gas permeabilities of the membrane and the dimensions of the connecting tube were verified experimentally. Transcutaneous tension (tcp) in O in a steady state was much more dependent on temperature and membrane thickness than in CO. The transient response of the tcp was found to be improved, without decreasing the magnitude of the steady-state tcp, by a thinner membrane with a lower permeability and by a shorter connecting tube of a larger inner diameter. The method of calibration was also examined because the magnitude of tcp by mass spectrometry was influenced in particular by water vapour. [ABSTRACT FROM AUTHOR]

A microcomputer-based system for the breath-by-breath assessment of respiratory and metabolic function during exercise was developed. To minimise the load for subjects ventilation was measured by use of an impedance plethysmograph instead of a pneumotachograph. The computer calculated respiratory frequency, tidal volume, minute ventilation, end-tidal pO and pCO, O input and CO output, and gas exchange ratio of each breath from the impedance pneumogram and the gas composition of the expired air. To avoid the error due to the cardiogenic oscillations appearing to overlap the impedance pneumogram, discrimination of the inspiratory and expiratory phases was achieved by detecting the temperature difference between ambient and expired air. The impedance pneumogram could follow tidal volume variations during a moderate bicycle exercise (120 W) with satisfactory accuracy, but its sensitivity is apt to fall at the severe exercise (180 W). Possible factors causing errors such as the distribution of regional ventilation and perfusion during exercise are discussed. [ABSTRACT FROM AUTHOR]

Copyright of Medical & Biological Engineering & Computing is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Five hundred and thirteen cases of gastric carcinoma were investigated for the presence of viral RNA, and the clinico-pathological data, geno-type, BamHIF restriction fragment polymorphism (RFLP) and specific LMP-1 30 bp gene deletion were also examined. EBVs detected in lymphocytes in 20 normal gastric mucosa, 7 lymphoma cell lines (LCLs) maintained in severe combined immunodeficiency (SCID) mice and 18 non-Hodgkin's lymphomas were compared with those in the gastric carcinoma cases. Thirty-three cases (6.4%) were demonstrated to be positive for EBV by means of EBER-1 RNA in situ hybridization. Clinico-pathological data showed no statistically significant difference in histological grading, location of cancer and status of vessel and lymphatic invasion between the EBV-positive and -negative groups, although the former significantly predominated in the submucosal invasion group (submucosal vs mucosal P=0.021; submucosal vs advanced cancer P=0.033). Some of these data were different from corresponding data in earlier reports. In cases that were evaluated by molecular biology, type A, wild-type F and LMP-1 gene deletion predominated except one in 21 informative cases, one in 24 and two in 16, respectively. EBVs detected in lymphocytes in normal gastric mucosa, LCLs in SCID mice and non-Hodgkin's lymphoma were also predominantly affected by type A, wild-type F and LMP-1 gene deletion with few exceptions. The results indicate a lack of genetic differences among EBVs in gastric carcinoma, normal population, LCLs of SCID mice and non-Hodgkin lymphomas. Some EBV infections in gastric carcinomas may be transient, especially in the submucosal invasion group. [ABSTRACT FROM AUTHOR]

Copyright of International Journal of Legal Medicine is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The appearance of two kinds of membrane antigen (MA) in Marek's disease herpesvirus (MDHV) or herpesvirus of turkey (HVT) infected cells was examined using antiserum to the common antigen (common-Ag). No early-appearing MA (EMA) was detected in these cultures until 16 hours post-inoculation, after which a number of cells in these cultures had late-appearing membrane antigen (LMA) as detected by immunofluorescence (IF). Fluids from cultures infected with these viruses were examined for the presence of the common-Ag by agar-gel precipitation (AGP) test. No common-Ag was detected until 16 hours, but after 24 hours, it was detected by AGP test. These results suggest that the common-Ag is related to LMA but not to EMA. [ABSTRACT FROM AUTHOR]

An agar-gel precipitation antigen prepared from the skin (feather-Ag) of chicken infected with JM strain of Marek's disease herpesvirus (MDHV) and cell extracts of cultures infected with either Type 2 plaque producing agent (PPA) of MDHV or turkey herpesvirus (HVT) (Type 2-Ag, HVT-Ag) had 1 precipitation line in common. Hyperimmune sera to the common antigen (common-Ag) neutralized both cell-free virus of Type 2 PPA and HVT. However, these viruses were neutralized to a greater extent by homologous antiserum. Absorption of HVT antiserum or hyperimmune serum to Type 2 PPA with common-Ag reduced their neutralizing activity. This reduction was almost complete with homologous antigen but less complete with heterologous antigen. The location of antigen in Type 2 PPA and HVT infected cells was determined using hyperimmune serum to the common-Ag in fluorescent antibody tests. Antigen mainly occurred in the cytoplasm of cells corresponding to the rounded refractile cells in the plaques, whereas unfixed cells showed antigen on the cytoplasmic membrane. The common-Ag associated with MDHV or HVT infections might be a virus structural component which is associated with the envelope. [ABSTRACT FROM AUTHOR]

Following establishment, via the vaginal route, of infection with an AP-1 binding-site deleted mutant (ΔAP-1) of feline immunodeficiency virus (FIV), cats were challenged with a homologous intact strain (TM2) of FIV. The cats were observed for 23 weeks to evaluate the efficacy of the ΔAP-1 against the homologous TM2 strain challenge. These two viruses were differentiated by Southern blotting after amplification of proviral DNA by semi-nested polymerase chain reaction in DNAs of peripheral blood mononuclear cells and tissues. A TM2-specific band was detected in one cat exposed to but not infected with ΔAP-1, but not in two ΔAP-1-infected. These results indicate that ΔAP-1 could protect against subsequent challenge with homologous FIV TM2 strain. [ABSTRACT FROM AUTHOR]

To examine the in vivo roles of auxiliary genes and regulatory elements of feline immunodeficiency virus (FIV), the provirus load in various tissues of cats infected with each of the mutant viruses (Δ vif, ΔORF-A and ΔAP-1) was studied. Although all mutant viruses could infect various tissues, provirus loads in various tissues especially those in cats infected with Δ vif virus were lower than those with the wild-type virus. Our results indicate the significance of vif and ORF-A genes and AP-1 binding site of FIV for efficient viral replication and full pathogenicity in cats. [ABSTRACT FROM AUTHOR]

We constructed a deletion mutant of feline herpesvirus type 1 (FHV-1) and a recombinant FHV-1. The deletion mutant is the virus with a region (367 bp) deleted from the start codon of thymidine kinase (TK) gene to the SmaI site within the TK gene, and the other is a recombinant FHV-1 expressing Gag protein of feline immunodeficiency virus (FIV), in which a cDNA encoding the Gag protein of FIV was inserted at the TK deletion site of the former deletion mutant. These viruses were designated as C7301 ddlTK and C7301 ddlTK-gag, respectively. Growth kinetics of these viruses in Crandell feline kidney cells was similar to that of the parent C7301 strain. By immunoblot analysis, C7301 ddlTK- gag was confirmed to express the FIV Gag precursor protein in the cells. [ABSTRACT FROM AUTHOR]

The role of the integrase region of feline immunodeficiency virus (FIV) in viral replication was examined using an integrase mutant clone of FIV which carries a frameshift mutation in the region. Upon transfection, although the integrase mutant was able to release virus-like particles into the supernatant from the transfected cells, the virions produced by the mutant contained unprocessed gag precursor protein and undetectable levels of reverse transcriptase activity. Furthermore, the mutant virions were unable to direct the synthesis of viral DNA after infection in target cells. To understand this phenotype of the integrase mutant in more detail, we constructed a gag-pol expression plasmid from an FIV molecular clone and assayed roles of the integrase region on virus particle formation following transfection. When an inframe deletion was introduced into the protease region of the expression plasmid, the mutant was able to efficiently release gag- and gag-pol precursor proteins into the supernatant from the transfected cells. An expression plasmid with mutations in both the protease and integrase regions, however, failed to release the gag-pol precursor protein from the cells. These results suggested an essential role for the integrase region for efficient incorporation of the gag-pol precursor into the virions. [ABSTRACT FROM AUTHOR]

Feline immunodeficiency virus (FIV) is classified into five genotypes, A, B, C, D and E, based on the phylogenetic analysis of the env V3-V5 region. However, whether there is correlation between phylogenetic and antigenic diversities remains unknown. In this study, we examined the cross virus neutralization of FIV genotypes A through E by sera from cats infected with a single genotype. The results indicated some relationships between phylogenetic genotype and neutralization serotype, and that cross-clade virus neutralization is possible. For example, serum from a cat infected with genotype E virus neutralized all five FIV genotypes. Our results suggest that the FIV subtyping according to the sequence diversity is partially reflected by antigenic diversity and serum neutralization. [ABSTRACT FROM AUTHOR]

We investigated the possibility of in vitro recombination of three different trains of feline herpesvirus type 1 (FHV-1), a virulent strain (C7805), a vaccine strain (F2), and a candidate vaccine strain with a deletion in the thymidine kinase gene (C7301 dlTK). The results showed that recombination within different genotypes of FHV-1 strains could occur at a relatively high frequency ranging from 10.1% to 21.5% in vitro. These results indicate the generation of multiple mutant viruses by in vitro recombination experiments. The possibility for generating a virulent virus or novel type of FHV-1 by recombination in vivo is discussed. [ABSTRACT FROM AUTHOR]

Feline immunodeficiency virus was isolated from four cats from Taiwan. The isolates were designated TI-1, TI-2, TI-3 and TI-4. Each was isolated from PBMCs following co-cultivation of PBMCs with a feline T-lymphoblastoid cell line (MYA-1 cells). However, the Taiwanese isolates did not grow in a feline kidney cell line (CRFK cells). The nucleotide sequences of the V3-V5 region of the envelope gene of the Taiwanese isolates were determined and compared with those of previously described isolates. Phylogenetic analysis of this region indicates that Taiwanese isolates belong to subtype C. [ABSTRACT FROM AUTHOR]

Summary: The genes encoding the canine herpesvirus (CHV) glycoprotein B (gB), gC and gD homologues have been reported already. However, products of these genes have not been identified yet. Previously, we have identified three CHV glycoproteins, gp145/112, gp80 and gp47 using a panel of monoclonal antibodies (MAbs). To determine which CHV glycoprotein corresponds to gB, gC or gD, the putative genes of gB, gC, and gD of CHV were inserted into the thymidine kinase gene of vaccinia virus LC16mO strain under the control of the early-late promoter for the vaccinia virus 7.5-kilodalton polypeptide. We demonstrated here that gp145/112, gp80 and gp47 were the translation products of the CHV gB, gC and gD genes, respectively. The antigenic authenticity of recombinant gB, gC and gD were confirmed by a panel of MAbs specific for each glycoprotein produced in CHV-infected cells. Immunization of mice with these recombinants produced high titers of neutralizing antibodies against CHV. These results suggest that recombinant vaccinia viruses expressing CHV gB, gC and gD may be useful to develop a vaccine to control CHV infection. [ABSTRACT FROM AUTHOR]

Feline herpesvirus type 1 (FHV-1) produces a single 5.4 kb immediate-early (IE) transcript encoding FHV-1 ICP4 which acts as a trans-acting factor [Kawaguchi et al. (1994) Virology 204: 430–435]. Our earlier study has shown that the FHV-1 IE transcript is spliced in the leader region and the FHV-1 IE gene product (ICP4) down-regulates its own promoter through the region which includes its transcription initiation site [Kawaguchi et al. (1996) J Vet Med Sci 58: 715–721]. Here we investigated FHV-1 ICP4 gene exression throughout FHV-1 productive infection and demonstrated that (i) a novel promoter is located downstream of the IE promoter and an early transcript is transcribed from this promoter region, (ii) a negative regulatory element, which is composed of a 20 bp direct repeat unit repeated 20 times, is located between the two promoters and the repeat unit shows high homology to a motif called direct repeat 2 in “a” sequence of herpes simplex virus type 1, (iii) the IE promoter and synthesis of the IE mRNA appear to be turned off after IE phase and the second promoter becomes active during early and late stages, and (iv) a gene product expressed only by the second promoter also possesses regulatory function. These findings indicate that expression of the FHV-1 ICP4 gene is alternatively regulated by the two promoters. [ABSTRACT FROM AUTHOR]

We previously constructed a recombinant feline herpesvirus type 1 (FHV1), C7301 dlTK-Cap, which contains an entire open reading frame encoding the capsid protein of feline calicivirus (FCV) F4 strain in the deleted locus of the thymidine kinase (TK) deficient mutant (C7301 dlTK) of FHV 1. In this report, we carried out in vivo experiments to assess the vaccine efficacy of the recombinant C7301 dlTK-Cap against FCV and FHV 1 infections in cats. As a result, two vaccinations with the C7301 dlTK-Cap by intraocular, intranasal and oral routes protected cats to a significant degree against subsequent virulent challenges with both parent FCV F4 and FHV 1 C7301 strains. The results are applicable for the further development of a new genetically engineered polyvalent vaccine for cats. [ABSTRACT FROM AUTHOR]

The gene of Marek's disease virus (MDV) serotype 2 (MDV2) homologous to glycoprotein D (gD) of herpes simplex virus (HSV) was identified and characterized by its nucleotide and predicted amino acid sequences. The MDV2 gD homologous gene contains an open reading frame capable of specifying a polypeptide of 385 amino acids, which include N- and C-terminal hydrophobic domains consistent with signal and anchor regions, respectively, and two potential N-linked glycosylation sites, one of which was located in a highly conserved region when compared to MDV serotype 1 (MDV1) and herpesvirus of turkeys (HVT). By northern blot analysis using a MDV2 gD-specific DNA probe, two highly abundant polycistronic 6.0 and 4.2 kb transcripts were detected in MDV2-infected cells. The genes encoding MDV2 protein kinase (PK), gD, and glycoprotein I (gI) homologues are transcribed to form 3′ coterminal mRNAs of 6.0 kb (encoding PK, gD and gI) and 4.2 kb (encoding gD and gI), respectively. By using rapid amplification cDNA end (RACE) method, several RNA start sites, to be thought those of the 4.2 kb mRNA, were detected in the upstream of MDV2 gD homologue. [ABSTRACT FROM AUTHOR]

We constructed a recombinant feline herpesvirus type 1 (FHV-1) which was deleted in a defined region (450 bp) within the thymidine kinase (TK) gene (C7301 dlTK) [Yokoyama et al. (1995) J Vet Med Sci 57: 709-714]. In this report, we carried out two experiments to assess the pathogenicity and vaccine efficacy of the recombinant C7301 dlTK in cats. The first experiment showed that, following multiple inoculation of the recombinant C7301 dlTK by intraocular, intranasal and oral routes, the virus was sufficiently attenuated in cats, although a high titer of the virus was recovered from target organs (eye, nose, and mouth). In the second experiment, two intramuscular vaccinations with the recombinant C7301 dlTK protected cats to a significant degree against subsequent challenge with the parent FHV-1 strain C7301 at 4 weeks after the last vaccination. These results demonstrate that the recombinant C7301 dlTK is effective as a live attenuated vaccine with a clear genetic marker. [ABSTRACT FROM AUTHOR]

The nucleotide sequences of the long terminal repeat of five Japanese, five Argentine and three Australian isolates of feline immunodeficiency virus (FIV) were determined and compared with those of isolates previously described. The results revealed that the Japanese isolates were found to cluster with nucleotide sequence similarity of 95.6%-99.4%. The Australian isolates also clustered with nucleotide sequence similarity of 97.2%-99.4%. The Argentine isolates formed two groups; the LP9 isolate is closely related to the Japanese isolates, whereas the LP1, LP3, LP20 and LP24 isolates are distant from both the Japanese and Australian isolates. From these results, FIV can be divided into three groups, namely: (I) the Californian, Australian and British isolates; (II) the Japanese isolates and one Argentine LP9 isolate; (III) the other Argentine isolates. [ABSTRACT FROM AUTHOR]

An oligonucleotide containing multiple AP-1 binding sites was introduced into the regulatory sequence in the long terminal repeat (LTR) of feline immunodeficiency virus (FIV). Chloramphenicol acetyltransferase assay revealed that basal promoter activity of the mutated LTR was higher than that of the wild-type LTR in Crandell feline kidney (CRFK) cells. The mutated LTR was introduced into an infectious molecular clone of FIV and the clone was transfected into CRFK cells. The virus production of the mutant in the cells was as high as that of the wild-type when determined by the reverse transcriptase activity assay. The growth of the mutant virus obtained from the transfected CRFK cells was examined in feline T lymphoblastoid cell lines (MYA-1 and FeL-039 cells) and primary feline peripheral blood mononuclear cells (fPBMCs). The growth was delayed when compared with that of the wild-type virus in all the cells used. Upon examination by polymerase chain reaction, the length of the LTR of the mutant virus was shortened in both MYA-1 cells and fPBMCs. Sequence analysis revealed that the insertion was completely deleted 39 days after infection in the MYA-1 cells. [ABSTRACT FROM AUTHOR]

The mechanism of cell death induced by feline immunodeficiency virus (FIV) infection was investigated in an interleukin 2(IL-2)-dependent T-lymphoblastoid cell line (MYA-1). DNA extracted from FIV-infected MYA-1 cells showed a ladder of nucleosomal DNA, indicating that the cytopathic effect (CPE) observed in these cells was due to apoptosis. Infection of MYA-1 cells with FIV was associated with suppression of the proliferative response of the cells to exogenous IL-2 prior to DNA fragmentation. These findings suggest that FIV-induced CPE in these T-lymhoblastoid cells is associated with apoptosis possibly due to a defect in the IL-2 signal transduction pathway. [ABSTRACT FROM AUTHOR]

Feline immunodeficiency virus (FIV) is a member of the genus Lentivirus of the family Retroviridae. FIV can infect T lymphocytes and monocytes/macrophages in vitro and in vivo, and causes an acquired immunodeficiency syndrome-like disease in cats. Several isolates of FIV from geographically distant countries have been molecularly cloned. There is considerable heterogeneity especially in Env gene among the FIV isolates and they can be divided into two or more subgroups. Like other lentiviruses, FIV has a complex genome structure. Gag gene encodes matrix, capsid and nucleocapsid proteins, and Pol gene encodes protease, reverse transcriptase, dUTPase and integrase. The dUTPase is not present in the primate lentiviruses but present in the nonprimate lentiviruses. Env gene encodes surface and transmembrane envelope glycoproteins. In addition to the structural and enzymatic proteins, at least three more genes (Vif, ORF A, Rev) are present in FIV. Vif is related to the infectivity of the cell-free viruses. Rev functions in the stability and transport of incompletely spliced viral RNAs from the nucleus to cytoplasm and is indispensable for virus replication. Although the Tat protein of the primate lentiviruses is essential for virus replication, ORF A (putative Tat gene) of FIV is not essential for virus replication in established feline T lymphoblastoid cell lines. However, the ORF A gene product is related to the efficient replication of the virus in primary peripheral blood lymphocytes. In the long terminal repeat (LTR) of FIV, there are many putative binding sites for enhancer/promoter proteins. Among these binding sites, the putative AP-1 site is important for basal promoter activity of the LTR and responsible for the T cell activation signal through protein kinase C, however the site is not required for the virus replication in established feline T lymphoblastoid cell lines. Comparative study of the molecular biology of lentiviruses revealed that the genome structure, splicing pattern and functional enhancer protein-binding sites of FIV are more similar to those of the ruminant lentiviruses than those of the primate lentiviruses. [ABSTRACT FROM AUTHOR]

A gene encoding the glycoprotein H (gH) homologue of feline herpesvirus type 1 was identified and sequenced. It was located immediately downstream of the thymidine kinase gene within an Eco RI 6.6 kbp fragment. In addition, a partial UL 21 homologous gene was located downstream of the gH homologous gene. The primary translation product of the gH homologous gene is predicted to consist of 821 amino acids with a molecular weight of 92.5 kDa. It possesses several characteristics typical of transmembrane glycoproteins, including a N-terminal hydrophobic signal sequence, C-terminal transmembrane domain, and putative N-linked glycosylation sites. Analysis of this protein revealed amino acid sequence homologies of 33.1% with equine herpesvirus type 1 (EHV-1) gH, 32.6% with EHV-4 gH, 29.1% with varicella-zoster virus gIII, 28.5% with pseudorabies virus gH, and 25.1% with herpes simplex virus type 1 gH. By Northern blot analysis, one of the transcripts specific for the gH homologous gene might be a mRNA of approximately 3.0 kb. [ABSTRACT FROM AUTHOR]