We describe the embryonic expression of musculin, a new murine member of the bHLH family of transcription factors. Musculin protein is closely related to human ABF-1, which is expressed in activated B cells, and to epicardin/capsulin/Pod-1, which is expressed in branchial myoblasts, visceral and urogenital mesoderm and epicardium. In situ hybridisation revealed musculin expression in embryos was largely restricted to the embryonic skeletal muscle lineage. While all skeletal muscles expressed the gene, only a subset of myocytes within each muscle were positive, indicating molecular heterogeneity within fetal muscle.

Amino Acid Sequence↗

PubMed1998

DAN is a secreted glycoprotein related to Xenopus cerberus.

We report that DAN, a potential cell cycle regulator and tumour suppressor, is a secreted glycoprotein related to Xenopus cerberus. DAN, cerberus, its mouse relative Cer-1/cer-l/Cerberus-like/Cerr1, and the recently described factor DRM/Gremlin, appear to be members of the cystine knot superfamily, which includes TGFbetas and BMPs. Like cerberus and mCer-1, DAN-induced cement glands as well as markers of anterior neural tissue and endoderm in Xenopus animal cap assays, features of BMP signalling blockade. During mouse embryogenesis, Dan was expressed from E8.5 in cranial mesenchyme and somites, then later in limb and facial mesenchyme. The pattern in somites was highly dynamic, with transcripts initially localized to the caudal half of the nascent epithelial somite, then, after maturation, to sclerotomal cells adjacent to the neural tube. Dan was also expressed in the developing myotome. The expression domains include sites in which BMP inhibition is known to be important for development. Thus, DAN appears to be a secreted factor belonging to the cystine knot superfamily, and one of a growing number of antagonists acting to modulate BMP signalling during development.

Amino Acid Sequence↗

PubMed1998

Uncoupling of bradykinin-induced phosphoinositide hydrolysis and Ca2+ mobilization by phorbol ester in canine cultured tracheal epithelial cells.

1. Regulation of the increase in inositol phosphates (IPs) production and intracellular Ca2+ concentration ([Ca2+]i by protein kinase C (PKC) was investigated in canine cultured tracheal epithelial cells (TECs). Stimulation of TECs by bradykinin (BK) led to IPs formation and caused an initial transient [Ca2+]i peak in a concentration-dependent manner. 2. Pretreatment of TECs with phorbol 12-myristate 13-acetate (PMA, 1 microM) for 30 min attenuated the BK-induced IPs formation and Ca2+ mobilization. The maximal inhibition occurred after incubating the cells with PMA for 2 h. 3. The concentrations of PMA that gave half-maximal (pEC50) inhibition of BK-induced IPs accumulation and an increase in [Ca2+]i were 7.07 M and 7.11 M, respectively. Inactive phorbol ester, 4alpha-phorbol 12,13-didecanoate at 1 microM, did not inhibit these responses. Prior treatment of TECs with staurosporine (1 microM), a PKC inhibitor, inhibited the ability of PMA to attenuate BK-induced responses, suggesting that the inhibitory effect of PMA is mediated through the activation of PKC. 4. In parallel with the effect of PMA on the BK-induced IPs formation and Ca2+ mobilization, the translocation and down-regulation of PKC isozymes were determined. Analysis of cell extracts by Western blotting with antibodies against different PKC isozymes revealed that TECs expressed PKC-alpha, betaI, betaII, gamma, delta, epsilon, theta and zeta. With PMA treatment of the cells for various times, translocation of PKC-alpha, betaI, betaII, gamma, delta, epsilon and theta from cytosol to the membrane was seen after 5 min, 30 min, 2 h, and 4 h treatment. However, 6 h treatment caused a partial down-regulation of these PKC isozymes. PKC-zeta was not significantly translocated and down-regulated at any of the times tested. 5. Treatment of TECs with 1 microM PMA for either 30 min or 6 h did not significantly change the KD, and Bmax receptor for BK binding (control: KD=1.7+/-0.3 nM; Bmax=50.5+/-4.9 fmol/mg protein), indicating that BK receptors are not a site for the inhibitory effect of PMA on BK-induced responses. 6. In conclusion, these results suggest that activation of PKC may inhibit the phosphoinositide hydrolysis and consequently attenuate the [Ca2+]i increase or inhibit independently both responses to BK. The translocation of pKC-alpha, betaI, betaII, delta, epsilon, gamma, and theta induced by PMA caused an attenuation of BK-induced IPs accumulation and Ca2+ mobilization in TECs.

Animals↗

PubMed1998

Inhibition of an established allergic response to ovalbumin in BALB/c mice by killed Mycobacterium vaccae.

Allergic disorders are mediated by T lymphocytes secreting T helper 2 (Th2) cytokines, interleukin-4 (IL-4) and interleukin-5 (IL-5), resulting in high levels of serum immunoglobulin E (IgE) and recruitment of eosinophils. One of the treatment strategies is to downregulate the Th2 component by inducing a T helper 1 (Th1) response to the relevant allergen, because Th1 and Th2 cytokines are thought to be mutually antagonistic. In this study, we examined the effects of Mycobacterium vaccae, a potent inducer of Th1 immunity, on allergic responses in a murine model. A single injection of M. vaccae into ovalbumin (OVA)-preimmunized BALB/c mice suppressed serum IgE over a wide dose range (10(7), 10(8) or 10(9) M. vaccae). Further experiments, using 10(7) M. vaccae injected twice, showed that this treatment inhibited not only serum IgE, but also the potential for ovalbumin-induced IL-5 production by spleen cells. This non-specific ability of a mycobacterium to decrease Th2 activity, even when not presented together with the allergen, is in agreement with recent epidemiological studies on the impact of bacillus Calmette-Guérin (BCG) vaccination, and of other potent Th1 stimuli, on the incidence of atopy. The suppression of serum IgE and allergen-specific IL-5 synthesis by M. vaccae suggest that this organism is likely to have clinical application in the immunotherapy of allergy.

Animals↗

PubMed1998

Methods to improve a periodontally involved terminal abutment of a cantilever fixed partial denture--a finite element stress analysis.

The two-dimensional plane stress finite element stress analysis was used to evaluate the stress of mandibular posterior cantilevers for compromised periodontal involved distal terminal abutments. Some commonly practiced methods to alleviate the stress from mastication of the cantilever FPDs were also evaluated. From the analysis the following was concluded: (1) The most distal terminal abutment of a cantilever FPD generally experienced a high stress concentration. (2) The cantilever FPDs require at least two abutments. Optimum stress reduction occurred with a splinting of three abutments. To increase the number of abutments to more than three, would not result in a proportional and meaningful reduction of stress in the periodontium. (3) To alter the bridge material with more rigid material or to have a longer marginal preparation of the terminal abutment are ineffective for better stress distribution.

Alveolar Process↗

PubMed1998

Lipid peroxidation in cord blood at birth: the effect of labour.

OBJECTIVE: To determine the effect of labour on free oxygen radical activity in the fetus, as reflected by lipid peroxide levels in umbilical cord arterial blood. DESIGN: Prospective, observational study. SETTING: Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong. METHODS: Umbilical cord arterial and venous blood samples were collected from singleton term infants delivered by elective caesarean section. Base excess, PO2, pCO2 and pH were measured in both samples and compared to identify double venous samples. Cord arterial acid-base balance and concentrations of organic hydroperoxides and malondialdehyde were compared with those obtained from normal vaginal deliveries. RESULTS: Cord arterial blood samples, obtained from cases of uncomplicated labour followed by spontaneous vaginal delivery, had significantly higher lipid peroxide concentrations than those delivered following elective caesarean section. This was most marked for malondialdehyde with a median value increased by 105%, whilst organic hydroperoxide was increased by only 27%. Of the acid-base parameters, base excess was increased by 78%, with only minimal changes in pH, pCO2 and PO2. These differences remained highly significant after including other pregnancy characteristics in multivariate analysis. CONCLUSION: The findings indicate that high levels of free oxygen radical activity in the fetus are a function of the labour process, as are changes in acid-base balance.

Carbon Dioxide↗

PubMed1998

Substance P receptor expression in intestinal epithelium in clostridium difficile toxin A enteritis in rats.

We previously reported that the inflammatory effects of Clostridium difficile toxin A on rat intestine can be significantly inhibited with a specific neurokinin-1 receptor (NK-1R) antagonist. In this study we investigated the localization and expression of NK-1R mRNA and protein in rat intestine by in situ hybridization, Northern blot analysis, and immunohistochemistry, respectively, after exposure to toxin A. Northern blot analysis showed increased mucosal levels of NK-1R mRNA starting 30 min after toxin A administration. In situ hybridization showed that toxin A increased NK-1R mRNA expression in intestinal epithelial cells after 30, 120, and 180 min. In rats pretreated with the NK-1R antagonist CP-96345 the increase in NK-1R mRNA levels after exposure to toxin A was inhibited, indicating that NK-1R upregulation is substance P (SP) dependent. One hour after exposure to toxin A many of the intestinal epithelial cells showed staining for NK-1R compared with controls. Specific 125I-SP binding to purified epithelial cell membranes obtained from ileum exposed to toxin A for 15 min was increased twofold over control and persisted for 4 h. This report provides evidence that NK-1R expression is increased in the intestinal epithelium shortly after exposure to toxin A and may be important in toxin A-induced inflammation.

Animals↗

PubMed1998

Differential expression of fibroblast growth factor receptors 1 to 4 and ligand genes in late fetal and early postnatal rat lung.

To characterize fibroblast growth factor (FGF) gene expression in the late fetal (days E18 to E22) and early postnatal lung (days P0 to P28), when the alveolar region undergoes extensive growth and reorganization, we analyzed the expression of four FGF receptors and six ligands. FGF receptor 1 (FGFR1) RNA levels were first low (E18) before rising late in the postnatal period (P28). FGFR2 RNA levels were detected early (at E18) and then increased (E20-P0) before falling (P2) to below later postnatal levels (P6 to P28). FGFR3 RNA levels were low at first (E18) and then increased, with peak levels in the days after birth (P2 to P10). FGFR4 RNA levels, barely detected in fetal lung (E18 to E22), increased at birth (P0) and remained high postnatally (P2 to P28). In fetal lung, FGF2 (basic FGF) RNA expression levels were low and FGF1 (acidic FGF) RNA levels were not detected: low RNA levels of each ligand were detected postnatally (P7 to P28). FGF3 to 5 and FGF7 RNA were not detected in fetal or postnatal lung. With in situ hybridization, predominantly the smooth muscle cells of large vessels expressed FGFR1 and 4 mRNA; the epithelial cells of large airways expressed FGFR1, 2, and 4; and alveolar cells expressed FGFR2, 3, and 4. Analysis of protein expression first identified FGF2 localized to the basement membrane of large airways and branching epithelial buds, to mesenchymal cells associated with buds, to the putative smooth muscle cells of large airways and vessels, and to pleural- and mesenchymal-associated cells (E18). Immediately before birth, this pattern of expression persisted (E20 to E22), with FGF2 also being expressed by putative smooth muscle cells of smaller airways and vessels (E22). After birth (P0 to P28), FGF2 expression remained relatively high in the smooth muscle cells of large and small vessels and in pleural cells; in airway smooth muscle cells and in most cells in the alveolar region, however, although FGF2 expression persisted in some cells, its intensity decreased with time.

3T3 Cells↗

PubMed1998

Serum thyroglobulin autoantibodies: prevalence, influence on serum thyroglobulin measurement, and prognostic significance in patients with differentiated thyroid carcinoma.

The prevalence of circulating thyroid autoantibodies (TgAb or antithyroid peroxidase) was increased nearly 3-fold in patients with differentiated thyroid cancers (DTC) compared with the general population (40% vs. 14%, respectively). Serum TgAb (with or without antithyroid peroxidase) was present in 25% of DTC patients and 10% of the general population. Serial postsurgical serum TgAb and serum Tg patterns correlated with the presence or absence of disease. Measurements of serum Tg were made in 87 TgAb-positive sera by a RIA and two immunometric assay (IMA) methods to study TgAb interference. TgAb interference, defined as a significant intermethod discordance (>41.7% coefficient of variation) between the Tg RIA and Tg IMA values relative to TgAb-negative sera, was found in 69% of the TgAb-positive sera. TgAb interference was characterized by higher Tg RIA vs. IMA values and was, in general, more frequent and severe in sera containing high TgAb concentrations. However, some sera displayed marked interference when serum TgAb was low (1-2 IU/mL), whereas other sera with very high TgAb values (>1000 IU/mL) displayed no interference. An agglutination method was found to be too insensitive to detect low TgAb concentrations (1-10 IU/mL) causing interference. Exogenous Tg recovery tests were an unreliable means for detecting TgAb interference. Specifically, the exogenous Tg recovered varied with the type and amount of Tg added and the duration of incubation employed. Further, recoveries of more than 80% were found for some sera displaying gross serum RIA/IMA discordances. The measurement of serum Tg in DTC patients with circulating TgAb is currently problematic. It is important to use a Tg method that provides measurements that are concordant with tumor status. IMA methods are prone to underestimate serum when TgAb is present, increasing the risk that persistent or metastatic DTC will be missed. The RIA method used in this study provided more clinically appropriate serum Tg values in the group of TgAb-positive patients with metastatic DTC. Furthermore, as serial serum TgAb measurements paralleled serial serum Tg RIA measurements, TgAb concentrations may be an additional clinically useful tumor marker parameter for following TgAb-positive patients. Disparities between serial serum Tg and TgAb measurements might alert the physician to the possibility of TgAb interference with the serum Tg measurement and prompt a more cautious use of such data for clinical decision-making.

Adolescent↗

PubMed1998

Reducing pregnancy and induced abortion rates in China: family planning with husband participation.

OBJECTIVES: This study assessed the effectiveness of a family planning intervention with and without husband's participation in reducing pregnancy and abortion rates in Shanghai, China. METHODS: In this 3-arm randomized trial among 1800 nonsterilized married women, educational interventions targeting both spouses and targeting the wife only were compared with usual family planning care. RESULTS: Among women not using intrauterine devices (IUDs), the intervention with husband's participation had an effect in reducing pregnancy rates (adjusted odds ratio [OR] = 0.36, 95% confidence interval [CI] = 0.12, 1.1) and abortion rates (adjusted OR = 0.29, CI = 0.09, 0.94) compared with control subjects, and a significant effect in reducing pregnancy rates (adjusted OR = 0.29, CI = 0.10, 0.85) and abortion rates (adjusted OR = 0.24, CI = 0.07, 0.77) compared with wife-only subjects. CONCLUSIONS: Family planning interventions involving husbands may reduce pregnancy and abortion rates among non-IUD users.

Abortion, Induced↗

PubMed1998

Ability of azithromycin in combination with quinine for the elimination of babesial infection in humans.

We report the ability of azithromycin in combination with quinine to eliminate the Babesia infection in a native Taiwanese woman. Failure of elimination of the babesial infection was observed two weeks after treating with standard regimen of oral quinine plus intravenous clindamycin for a 10-day course of therapy. Azithromycin in place of clindamycin was administered for another 10-day course of therapy two months following initial treatment. Clearance of Babesia parasites was observed and verified by hamster inoculation. These results suggest that azithromycin plus quinine should be considered as an alternative therapy for human babesiosis, especially in the failure of treatment with standard regimens.

Animals↗

PubMed1998

Effects of dexamethasone on antigen expressions and proliferation of amoeboid microglial cells in fetal rat brain.

The present study examined the effect of maternal administration of dexamethasone (DEX) on amoeboid microglial cells (AMC) in fetal rats extending from 16 to 20 days postconception (E16 to E20). After an intraperitoneal injection of DEX into pregnant rats at E10, the external morphology and distribution of immunolabelled AMC as detected with OX-42 and ED1 monoclonal antibodies remained unaltered when compared with those of the controls. The major effect of dexamethasone was on microglial cell population. Thus, with OX-42 and ED1, the numbers of immunolabelled AMC in the intermediate zone lateral to the striatum (IZS) of DEX-treated fetuses which remained relatively unchanged at E16 were significantly reduced at E18. However, OX-42 labelled cells showed an unexpected increase in number at E20 following DEX treatment. Microglial response to DEX was also analyzed in sections stained with the isolectin, GSA I-B4, which specifically binds alpha-D-galactosyl glycoproteins on microglia. The number of GSA I-B4 labelled AMC was significantly increased at E16, declined at E18 and remained constant thereafter in DEX-treated rats when compared with that of the controls. A major finding after DEX treatment was the wider occurrence of AMC double labelled with anti-BrdU antibody and GSA I-B4 or OX-42 at E16 compared with those in the controls suggesting that the initial increase of GSA I-B4 labelled AMC may be attributed to their proliferation. The drastic reduction of OX-42 and ED1 positive microglial cells notably at E18 may be due to the downregulation of surface antigens as a result of possible suppressive action of dexamethasone. On the basis of present findings, it is concluded that the antigenic expressions of fetal AMC may be modulated by DEX administrated maternally. Such however appeared to be extremely selective as reflected by the varied expression for certain immune molecules at different stages of brain development. This information would be useful in potential use of glucocorticoids in prenatal therapy of brain pathology via maternal circulation.

Animals↗

PubMed1998

Recurrent acute suppurative thyroiditis in a child: case report.

A 23-month-old girl presented with a history of persistent fever and growing left anterior neck mass following an upper respiratory tract infection. Laboratory studies revealed leukocytosis, elevated levels of the erythrocyte sedimentation rate and C-reactive protein, and a mild impairment of thyroid function. Thyroid scan showed a decreased radioactive iodine uptake of the left thyroid gland. Culture of the thyroid aspirate grew the mixed flora, viridans streptococci, Prevotella spp, and Peptostreptococcus magnus. She was discharged after a surgical drainage and a 14-day course of penicillin-G therapy. Unfortunately, she was readmitted for the resembling problems, the fever and progressing left anterior neck mass 3 months later. Culture of thyroid aspirate also grew the viridans streptococci. She recovered after a 14-day course of penicillin-G therapy. A left pyriform sinus fistula was found by barium esophagogram. A selective operation was performed 8 weeks later.

Acute Disease↗

PubMed1998

Isomerase and chaperone activities of protein disulfide isomerase are both required for its function as a foldase.

Protein disulfide isomerase (PDI) is not only an isomerase catalyzing the formation of native disulfide bond(s) of nascent peptide, but also a molecular chaperone assisting chain folding. The intrinsic chaperone activity of PDI is independent of its isomerase activity as shown by its ability of promoting in vitro reactivation and suppressing aggregation during refolding of denatured proteins containing no disulfide. The -CGHC- active sites of PDI are not required for its chaperone activity and a mutant PDI with no isomerase activity does function in vitro and in vivo. The peptide binding site of PDI is responsible for its chaperone activity. Both isomerase and chaperone activities are required for PDI to function as a foldase in assisting protein folding, in other words, the foldase activity of PDI consists of both isomerase and chaperone activities.

Animals↗