Indocyanine green fluorescence lymphography during gastrectomy after initial endoscopic submucosal dissection for early gastric cancer.

BACKGROUND: Indocyanine green (ICG) fluorescence lymphography can be used to visualize the lymphatic drainage of gastric cancer. Few studies have been performed to identify lymphatic drainage patterns after endoscopic submucosal dissection (ESD). ESD results in changes to lymphatics owing to fibrosis of the submucosal layer. This study aimed to evaluate the efficacy of ICG fluorescence lymphography for visualization of lymphatic drainage after ESD, and to assess its clinical application in additional gastrectomy after ESD for early gastric cancer. METHODS: All patients who underwent gastrectomy after ESD between 2014 and 2017 in a single centre were reviewed. ICG was injected endoscopically into the submucosal layer around the ESD scar the day before surgery. At the time of surgery, lymph nodes (LNs) were visualized and lymphadenectomy was performed with near-infrared imaging. Ex vivo, all LNs were examined for the presence of fluorescence. Number of LNs resected and number of tumour-positive LNs were compared between patients who underwent near-infrared imaging and those who had conventional lymphadenectomy without intraoperative imaging. RESULTS: Some 290 patients underwent gastrectomy after ESD, 98 with fluorescence lymphography-guided lymphadenectomy and 192 with conventional lymphadenectomy. Fluorescence lymphography visualized lymphatic drainage in all patients, without complications related to ICG injection or near-infrared imaging. Fluorescence lymphography visualized all stations containing metastatic LNs. The sensitivity for detecting LN metastasis in fluorescent stations was 100 per cent (9 of 9 stations), and the negative predictive value was 100 per cent (209 of 209). One patient with LN metastasis had one non-fluorescent metastatic LN within a fluorescent station. CONCLUSION: Fluorescence lymphography successfully visualized all draining LNs after ESD, with high sensitivity and negative predictive value for detecting LN metastasis. Fluorescence lymphography-guided lymphadenectomy could be an alternative to systematic lymphadenectomy during additional surgery after ESD.

ANTECEDENTES: La linfografía de fluorescencia con verde de indocianina (indocyanine green, ICG) visualiza el drenaje linfático del cáncer gástrico. Se han realizado pocos estudios para identificar los patrones de drenaje linfático tras una disección submucosa endoscópica (endoscopic submucosal dissection, ESD). La ESD introduce cambios de los linfáticos debido a la fibrosis de la capa submucosa. El objetivo de este estudio era valorar la eficacia de la linfografía con ICG para visualizar el drenaje linfático tras ESD y evaluar su aplicación clínica en la gastrectomía adicional después de ESD por carcinoma precoz gástrico (early gastric cancer, EGC). MÉTODOS: Se revisaron todos los pacientes sometidos a gastrectomía tras ESD entre 2014 y 2017 en un único centro. El ICG se inyectó por vía endoscópica en la capa submucosa alrededor de la cicatriz tras ESD el día antes de la cirugía. En el momento de la cirugía, se visualizaron los ganglios linfáticos (lymph nodes, LNs) y se realizó la linfadenectomía siguiendo las imágenes de infrarrojo. Ex vivo, todos los LNs se examinaron para detectar la presencia de fluorescencia. Se compararon el número de LNs resecados y el número de LNs afectados por el tumor entre pacientes sometidos a imágenes de infrarrojo y pacientes a los que se les realizó una linfadenectomía convencional sin imágenes intraoperatorias. RESULTADOS: Un total de 290 pacientes fueron sometidos a gastrectomía tras ESD (98 con linfadenectomía por linfografía con ICG y 192 con linfadenectomía convencional). La linfografía con ICG visualizó el drenaje linfático en todos los pacientes, sin complicaciones relacionadas con la inyección de ICG o con las imágenes de infrarrojo. La linfografía con ICG permitió visualizar todas las estaciones ganglionares en las que había LNs metastásicos. La sensibilidad para detectar los LN con metástasis en las estaciones con fluorescencia fue del 100% (9 de 9 estaciones), y el valor predictivo negativo (negative predictive value, NPV) del 100% (209 de 209 estaciones). Un paciente con metástasis en LN tenía un ganglio metastásico sin fluorescencia en el seno de una estación con fluorescencia. CONCLUSIÓN: La linfografía con fluorescencia visualiza satisfactoriamente todos los LNs que drenan después de ESD, con una elevada sensibilidad y NPV para detectar metástasis en LN. La linfadenectomía guiada por fluorescencia podría ser una alternativa a la linfadenectomía convencional durante la cirugía adicional después de ESD.

Nanoscale heat transport through the hetero-interface of SrRuO3 thin films.

A SrRuO3 thin film has been widely used as a metal electrode in electronic devices based on transition metal oxides, and hence it is important to understand its thermal transport properties to minimize a thermal degradation problem during the device operation. Using the time-domain thermoreflectance measurement technique, we investigate the cross-plane thermal conductivity of the SrRuO3 thin films with a thickness variation from 1 µm to 8 nm. We find that the thermal conductivity is reduced from about 6 W m-1 K-1 for the 1 µm thick film to about 1.2 W m-1 K-1 for the 8 nm thick film, and attribute this behavior to the boundary scattering of thermal carriers which originally have the mean free path of about 20 nm in a bulk state. Also, we observe a clear dip behavior of the thermal conductivity in the intermediate thickness around 30 nm which suggests an existence of a strong scattering source other than the film boundary. We explain this result by considering an additional interfacial scattering at the tetragonal-orthorhombic phase boundary which is formed during the strain relaxation with an increase of the film thickness.

Covalent-bonding-induced strong phonon scattering in the atomically thin WSe2 layer.

In nano-device applications using two-dimensional (2D) van der Waals materials, a heat dissipation through nano-scale interfaces can be a critical issue for optimizing device performances. By using a time-domain thermoreflectance measurement technique, we examine a cross-plane thermal transport through mono-layered (n = 1) and bi-layered (n = 2) WSe2 flakes which are sandwiched by top metal layers of Al, Au, and Ti and the bottom Al2O3 substrate. In these nanoscale structures with hetero- and homo-junctions, we observe that the thermal boundary resistance (TBR) is significantly enhanced as the number of WSe2 layers increases. In particular, as the metal is changed from Al, to Au, and to Ti, we find an interesting trend of TBR depending on the WSe2 thickness; when referenced to TBR for a system without WSe2, TBR for n = 1 decreases, but that for n = 2 increases. This result clearly demonstrates that the stronger bonding for Ti leads to a better thermal conduction between the metal and the WSe2 layer, but in return gives rise to a large mismatch in the phonon density of states between the first and second WSe2 layers so that the WSe2-WSe2 interface becomes a major thermal resistance for n = 2. By using photoemission spectroscopy and optical second harmonic generation technique, we confirm that the metallization induces a change in the valence state of W-ions, and also recovers a non-centrosymmetry for the bi-layered WSe2.

Hydroxychloroquine treatment during pregnancy in lupus patients is associated with lower risk of preeclampsia.

BACKGROUND: Hydroxychloroquine (HCQ) is regarded as a mainstay in the treatment of systemic lupus erythematosus (SLE) because of its efficacy in preventing flares, achieving remission, and reducing overall mortality. However, the impact of HCQ on pregnancy outcomes remains controversial. OBJECTIVE: We aimed to investigate the effect of HCQ on pregnancy outcomes in patients with SLE. METHODS: We performed a retrospective cohort study of 151 pregnancies in 122 patients with SLE (80 pregnancies in the HCQ treatment group and 71 pregnancies in the HCQ nontreatment group). We reviewed baseline characteristics including maternal comorbidities such as antiphospholipid syndrome, lupus nephritis, and autoimmune hepatitis. Pregnancy outcomes (preeclampsia, preterm delivery, and fetal growth restriction) and neonatal outcomes (gestational age at delivery and birth weight) were compared between HCQ treatment and nontreatment groups. RESULTS: Preeclampsia was significantly less complicated (7.5% vs 19.7%, p = 0.032) and neonatal birth weight was significantly greater (2757.0 ± 583.5 g vs 2542.3 ± 908.3 g, p = 0.001) in the HCQ treatment group than in the HCQ nontreatment group. Multiple logistic analysis adjusting for body mass index (BMI), lupus nephritis, serum uric acid, and estimated glomerular filtration rate revealed HCQ treatment was associated with exceedingly lower risk of preeclampsia in SLE pregnancy (odds ratio (OR) 0.106 (confidence interval (CI) 0.017-0.671)). Other independent risk factors for preeclampsia were a high prepregnancy BMI (OR 1.575 (CI 1.114-2.227)) and low eGFR level (OR 0.931 (CI 0.886-0.979)) before pregnancy. CONCLUSION: Our data showed pregnancy outcomes in SLE patients can be improved in the HCQ treatment group with about 90% reduction of preeclampsia.

Revisiting the diagnostic criteria of clinical chorioamnionitis in preterm birth.

OBJECTIVE: To re-evaluate the utility of the conventional criteria for clinical chorioamnionitis in the prediction of early-onset neonatal sepsis (EONS) in preterm birth. DESIGN: Retrospective cohort study. SETTING: Seoul, Republic of Korea. SAMPLE: A total of 1468 singleton births between 24 and 34 weeks due to preterm labour (n = 713) or preterm prelabour rupture of membranes (n = 755). METHOD: We evaluated three diagnostic categories of clinical chorioamnionitis: Criteria 1, conventional criteria; Criteria 2, combination of any three conventional parameters without prerequisite fever; Criteria 3, Criteria 1 plus positive maternal C-reactive protein and neutrophil left-shift into minor criteria. EONS included proven or suspected sepsis within 7 days following birth. Neonatal morbidity and mortality of EONS were also reviewed. MAIN OUTCOME MEASURES: Diagnostic performance of three combinations. RESULTS: The prevalence of EONS was 13.8%. Among 203 cases of EONS, maternal manifestation of clinical chorioamnionitis by criteria 1 was evident in only one out of seven, indicating 15.3% sensitivity for EONS prediction. However, with application of criteria 2, sensitivity significantly increased to 34.0%, while compromising specificity from 92.3% to 78.7%. Criteria 3 showed similar diagnostic performance compared with criteria 1 (sensitivity 16.7%, specificity 91.6%). Overall, neonatal mortality and neonatal composite morbidity in EONS were 14.9% and 67.8%, respectively, and there was no difference in neonatal morbidity and mortality between neonates whose mothers showed fever as a sign of clinical chorioamnionitis and those whose mothers did not. CONCLUSION: The renouncement of fever as a prerequisite for the criteria of clinical chorioamnionitis could increase sensitivity for the identification of EONS, a serious outcome of preterm birth. TWEETABLE ABSTRACT: The renouncement of fever as an essential can increase sensitivity for prediction of neonatal sepsis.

The efficacy of cefazolin plus macrolide (erythromycin or clarithromycin) versus cefazolin alone in neonatal morbidity and placental inflammation for women with preterm premature rupture of membranes.

INTRODUCTION: Although the use of broad-spectrum antibiotics in women with preterm premature rupture of membranes (PPROM) is recommended to prolong pregnancy and decrease short-term neonatal complications, the optimal regimen remains undetermined. The objective of this study was to compare the efficacy of cefazolin plus macrolide (erythromycin or clarithromycin) versus cefazolin alone in reducing neonatal morbidity and placental inflammation for women with PPROM. METHODS: This prospective study included singleton pregnancies with PPROM (23-33 weeks gestation). The primary outcome was neonatal composite morbidity and the secondary outcomes were the incidence of abnormal brain sonography and infant neurological outcome at one year of age. The presence and the stage of acute histological chorioamnionitis and funisitis were also reviewed blinded to all clinical information. RESULTS: 102 women were randomly assigned to cefazolin (n = 35), cefazolin plus erythromycin (n = 31), or cefazolin plus clarithromycin (n = 36). The neonatal composite morbidity, the incidence of abnormal brain sonography, and infant neurological outcome at one year of age were similar between the comparison treatments (combination of cefazolin plus erythromycin or clarithromycin) and cefazolin. However, the presence and stage of histological funisitis showed significant difference between cefazolin plus clarithromycin versus cefazolin alone (p = 0.023). DISCUSSION: This study is the first clinical trial of the use of cefazolin with either clarithromycin or erythromycin compared to cefazolin alone in the management of PPROM in which the primary and secondary analyses showed no difference among the three antibiotic regimens. The only noted difference was from a lesser degree of histological funisitis associated with clarithromycin exposure. CONCLUSION: Our data suggests that clarithromycin may be an alternative worth considering with potentially beneficial effects compared to erythromycin in PPROM.

The effect of human fetal liver-derived mesenchymal stem cells on CD34+ hematopoietic stem cell repopulation in NOD/Shi-scid/IL-2Rã(null) mice.

Mesenchymal stem cells (MSCs) are progenitors that are capable of differentiating into mesenchymal tissues. They are known to support allogeneic hematopoietic stem cell transplantation by facilitating engraftment without increasing the risk of graft-versus-host disease. We optimized culture conditions for human fetal liver-derived MSCs (hFL-MSCs) to investigate the role of hFL-MSCs on repopulation of hematopoietic stem cells in NOD/Shi-scid/IL-2Rγ(null) (NOG) mice using CD34(+) hematopoietic stem cells (HSCs) derived from umbilical cord blood (UCB). FL-MSCs and CD34(+) HSCs were prepared from fetal liver and UCB, respectively. Twenty-four hours after irradiation, CD34(+) HSCs and hFL-MSCs were injected intravenously and intratibially into NOG mice. During 24 weeks posttransplantation, engraftment levels of human cells were analyzed in bone marrow, peripheral blood, and spleen of transplanted mice by flow cytometry. hFL-MSCs showed a fibroblast-like morphology and immunophenotypic characteristics appropriate for MSCs. hFL-MSCs prolonged the survival of NOG mice that had been cotransplanted with UCB CD34(+) cells. Fluorescence-activated cell-sorting analysis showed that engraftment of human cells was increased by cotransplantation of hFL-MSCs. However, significant enhancement of human cell engraftment was not detected in NOG mice regardless of the number of cotransplanted MSCs. Although survival of repopulating NOG mice and engraftment of human cells were prolonged by cotransplantation of hFL-MSCs, 8.0 × 10(6) MSCs were not sufficient to increase HSC engraftment in irradiated NOG mice in vivo.

The expression of connective tissue growth factor in pregnancies complicated by severe preeclampsia or fetal growth restriction.

We tested the hypothesis that the expression of placental connective tissue growth factor (CTGF) is enhanced in pregnancies complicated by severe preeclampsia (PE) or fetal growth restriction (FGR). CTGF expression was analyzed using immunostaining, western blot and real-time quantitative PCR in placental samples obtained after third trimester cesarean deliveries without labor from women with severe PE (n=11), idiopathic FGR (n=14), or healthy controls (n=14). Serum CTGF concentrations were analyzed using ELISA. We found that CTGF was stably expressed in villous trophoblasts throughout pregnancy. The expression of CTGF mRNA in placentas from severe PE or FGR was higher than placentas from controls. Whereas the levels of placental CTGF protein were similar between normal and severe PE, maternal and fetal serum CTGF levels were elevated in severe PE. Maternal CTGF levels were also distinctively elevated in women with PE or FGR with histological evidence of placental injury. The enhancement of CTGF expression as well as serum CTGF levels in clinical conditions attributed to placental dysfunction suggests a role for this secretary protein in the pathophysiology of placental injury or its sequelae.

Isolation of a low-temperature adapted lipolytic enzyme from uncultivated micro-organism.

AIMS: The aim of the study was to isolate a novel lipolytic enzyme from the activated sludge of uncultured micro-organisms. METHODS AND RESULTS: The metagenomic DNA was directly extracted from the activated sludge, and a metagenomic library was constructed by using the pUC vector. The library was screened for lipolytic enzyme activity on 1% tributyrin agar plate. A clone among c. 100 000 recombinant libraries showed the lipolytic activity. The putative lipolytic gene encoding lipo1 from the metagenomic library was subcloned and expressed in Escherichia coli BL21 using the pET expression system. The expressed recombinant enzyme was purified by Ni-nitrilotriacetic acid affinity chromatography and characterized using general substrates of lipolytic property. The gene consisted of 972 bp encoding a polypeptide of 324 amino acids with a molecular mass of 35.6 kDa. Typical residues essential for lipolytic activity such as penta-peptide (GXSXG) and catalytic triad sequences (Ser166, Asp221 and His258) were detected. The deduced amino acid sequence of lipo1 showed low identity with amino acid sequences of esterase/lipase (32%, ZP_01528487) from Pseudomonas mendocina ymp and esterase (31%, AAY45707) from uncultured bacterium. This lipolytic enzyme exhibited the highest activity at pH 7.5 and 10 degrees C. At thermal stability analysis, lipo1 was more unstable at 40 degrees C than 10 degrees C. CONCLUSIONS: An activity based strategy has been an effective method for fishing out a low-temperature adapted lipolytic enzyme from the metagenomic library. This lipo1 enzyme can be considered to belong to the hormone-sensitive lipase family due to the enzyme's oxyanion hole by the sequence HGGG. SIGNIFICANCE AND IMPACT OF THE STUDY: Lipo1 is a novel psychrophilic esterase obtained directly from the metagenomic library. Owing its support of significant activity at low temperature, this enzyme is expected to be useful for potential application as a biocatalyst in organic chemistry.

Screening and purification for novel cytochrome b5 from uncultured environmental micro-organisms.

AIMS: We describe a sequence-based PCR method suitable for the isolation of a novel soluble heme-binding domain of cytochrome b(5) (cyt b(5)) gene directly from metagenomic DNA is described. METHODS AND RESULTS: Using the degenerate primer set, a cyt b(5) gene was isolated directly from metagenomic DNA. Based on the sequence-based PCR method, the similar conserved motif of cyt b(5) from Rhodopseudomonas palustris strain makes the novel target gene. The gene encoding cyt b(5) was cloned and expressed in Escherichia coli BL21 (DE3) using pET expression system. The expressed recombinant enzyme was purified by Ni-nitrilotriacetic acid affinity chromatography and characterized. CONCLUSIONS: Sequence-based strategy is an effective method for application of the novel gene from metagenomic DNA. SIGNIFICANCE AND IMPACT OF THE STUDY: Investigation of novel genes from metagenome, most of the micro-organism species are largely untapped, could represent an interesting and useful reservoir for biological processes.

Hypoxia regulates the expression of PHLDA2 in primary term human trophoblasts.

Hypoxia influences gene expression in placental trophoblasts. We sought to examine the effect of hypoxia on trophoblast expression of human PHLDA2 (also termed IPL, TSSC3 or BWR-1C), a product of an imprinted gene on human chromosome 11p15.5 whose murine ortholog plays a pivotal role in placental development. We initially confirmed that PHLDA2 was expressed in term placental villi, primarily in the trophoblast layer. Using quantitative PCR we found that the expression of PHLDA2 gradually declined during differentiation of primary term human trophoblasts. A similar expression pattern was seen for p57(Kip2) and IGF-II, both products of imprinted genes on chromosome 11p15.5. Exposure of trophoblasts to hypoxia in vitro (O(2)

The correlation between sampling site and gene expression in the term human placenta.

Using oligonucleotide microarrays we recently identified a set of transcripts that were up-regulated in hypoxic human trophoblasts. To test the hypothesis that expression of hypoxia-related placental transcripts depends on sampling site we analyzed nine different sites from term human placentas (n=6), obtained after uncomplicated pregnancies. These sites spanned the placental center to the lateral border and the basal to the chorionic plate. Relative gene expression at each site, determined using quantitative PCR, was correlated with villous histology. The expression of vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF), the cytoskeleton proteins lamininA3 and alpha-tubulin, and the signal transduction protein Rad was enhanced in the subchorionic lateral border compared to medial basal site (1.6-2.9 fold, p<0.05). In contrast, the expression of NDRG1, adipophilin and human placental lactogen was unchanged. Enhanced villous maturation, syncytial knots and fibrin deposits were more frequent in the subchorionic placental lateral border, and correlated with up-regulation of hypoxia-related transcripts (p<0.05). The association between sample site and expression level was not observed in placentas with marginal cord insertion. The expression of hypoxia-related genes in the term human placenta is dependent on sampling site within the placental disk, likely reflecting local differences in villous perfusion.

Microarray-based identification of differentially expressed genes in hypoxic term human trophoblasts and in placental villi of pregnancies with growth restricted fetuses.

Hypoxia adversely influences the function of the human placenta. We sought to identify a set of hypoxia-regulated transcripts in both term human trophoblasts in vitro and in villous trophoblasts in vivo. Using high-density oligonucleotide microarrays we initially examined differences in gene expression between trophoblast cultured in standard conditions (20% oxygen) vs. hypoxic conditions (< or =1% oxygen), as well as in placental tissues from pregnancies complicated by intrauterine growth restriction vs. matched controls. We used a novel computation method to compile data from the two approaches and identify transcripts that exhibited a marked expression change. Using quantitative PCR we confirmed an up-regulation of transcripts for vascular endothelial growth factor, connective tissue growth factor, follistatin-related protein, N-Myc down-regulated gene 1 and adipophilin in hypoxic term trophoblasts. In contrast, the expression of human placental lactogen and Beckwith-Wiedemann region 1 C was reduced in hypoxic trophoblast. Using in situ hybridization we validated the expression of each transcript in cultured term villous trophoblasts, and determined transcript expression in placental samples derived from four sets of dichorionic twins complicated by growth restriction of one twin. The identification of hypoxic trophoblast signature transcripts may implicate new mediators in pathways underlying trophoblast hypoxic injury and adaptation.

Lipoprotein lipase and leptin are accumulated in different secretory compartments in rat adipocytes.

Adipose cells produce and secrete several physiologically important proteins, such as lipoprotein lipase (LPL), leptin, adipsin, Acrp30, etc. However, secretory pathways in adipocytes have not been characterized, and vesicular carriers responsible for the accumulation and transport of secreted proteins have not been identified. We have compared the intracellular localization of two proteins secreted from adipose cells: leptin and LPL. Adipocytes accumulate large amounts of both proteins, suggesting that neither of them is targeted to the constitutive secretory pathway. By means of velocity centrifugation in sucrose gradients, equilibrium density centrifugation in iodixanol gradients, and immunofluorescence confocal microscopy, we determined that LPL and leptin were localized in different membrane structures. LPL was found mainly in the endoplasmic reticulum with a small pool being present in low density membrane vesicles that may represent a secretory compartment in adipose cells. Virtually all intracellular leptin was localized in these low density secretory vesicles. Insulin-sensitive Glut4 vesicles did not contain either LPL or leptin. Thus, secretion from adipose cells is controlled both at the exit from the endoplasmic reticulum as well as at the level of "downstream" secretory vesicles.

17Beta-estradiol inhibits proliferation of cultured vascular smooth muscle cells induced by lysophosphatidylcholine via a nongenomic antioxidant mechanism.

OBJECTIVE: Lysophosphatidylcholine (lysoPC), an active component of oxidized low-density lipoprotein, stimulates proliferation of vascular smooth muscle cells (VSMC). We investigated the direct impact of 17beta-estradiol (E2) on the proliferation of VSMC from rat aorta. RESULTS: VSMC derived from both female and male rats expressed estrogen receptors alpha and beta. Treatments with 1% fetal bovine serum or 5 microM lysoPC increased the incorporation of [3H]-thymidine in VSMC obtained from female rats. 17Beta-E2 did not alter the response to fetal bovine serum, but significantly suppressed the enhanced deoxyribonucleic acid synthesis which had been induced by lysoPC in a dose-dependent manner (10(-4)-10(-6) M). Estrogen also inhibited the proliferation of VSMC from male animals. ICI 182,780, a specific estrogen receptor antagonist, and 17alpha-E2, an inactive form of estradiol, also decreased the mitogenic response to lysoPC in VSMC. In addition, N-acetyl-L-cysteine, a potent antioxidant, inhibited the lysoPC effect. Flow cytometric analysis using the oxidation-sensitive probe 2',7'-dichlorofluorescin diacetate revealed that elevated intracellular formation of reactive oxygen species elicited with lysoPC was depressed significantly by 17beta-E2, ICI 182,780, or 17alpha-E2 as well as by N-acetyl-L-cysteine. CONCLUSION: 17Beta-E2 inhibits in vitro VSMC proliferation induced by lysoPC via a nongenomic antioxidant mechanism.

Identification and characterization of leptin-containing intracellular compartment in rat adipose cells.

The major leptin-containing membrane compartment was identified and characterized in rat adipose cells by means of equilibrium density and velocity sucrose gradient centrifugation. This compartment appears to be different from peptide-containing secretory granules present in neuronal, endocrine, and exocrine cells, as well as from insulin-sensitive GLUT-4-containing vesicles abundant in adipocytes. Exocytosis of both leptin- and GLUT-4-containing vesicles can be induced by insulin; however, only leptin secretion is responsive to serum stimulation. This latter effect is resistant to cycloheximide, suggesting that serum triggers the release of a stored pool of presynthesized leptin molecules. We conclude that regulated secretion of leptin and insulin-dependent translocation of GLUT-4 represent different pathways of membrane trafficking in rat adipose cells. NIH 3T3 cells ectopically expressing CAAT box enhancer binding protein-alpha and Swiss 3T3 cells expressing peroxisome proliferator-activated receptor-gamma undergo differentiation in vitro and acquire adipocyte morphology and insulin-responsive glucose uptake. Only the former cell line, however, is capable of leptin secretion. Thus different transcriptional mechanisms control the developmental onset of these two major and independent physiological functions in adipose cells.

Changes in biochemical bone markers during pregnancy and puerperium.

To elucidate the changes in bone turnover during pregnancy and puerperium, we measured serially the levels of serum osteocalcin and urine deoxypyridinoline (Dpy) as markers of bone formation and bone resorption, respectively, in 22 healthy women with normal pregnancy. Nineteen non-pregnant women served as control. The Dpy levels increased significantly at 16 weeks of pregnancy and remained elevated thereafter. The levels of osteocalcin, however, were significantly decreased at 16 weeks of pregnancy and elevated later at 6 weeks postpartum. Bone turnover ratio (Dpy/osteocalcin) continued to rise during pregnancy, but returned to control levels 6 weeks after delivery. Dpy levels and bone turnover ratio during puerperium tended to be higher in 17 breast-feeding women than those of 5 exclusive bottle-feeders. In conclusion, bone resorption begins to increase from the second trimester of pregnancy and calcium release from bone tissue might play a major role in calcium homeostasis during the whole period of pregnancy as well as during lactation.

Serologic responses of Korean soldiers serving in malaria-endemic areas during a recent outbreak of Plasmodium vivax.

Anti-Pv200 antibody levels were assessed in samples from endemic areas of Plasmodium vivax malaria in the Republic of Korea (ROK), using an indirect enzyme-linked immunosorbent assay (ELISA) method. Asymptomatic carriers of P. vivax were detected using nested polymerase chain reaction (PCR) of blood samples. Anti-Pv200 antibody levels in 20 vivax malaria patients (optical density +/- standard deviation [OD +/- SD] values 1.85 +/- 0.29 of IgG isotype and 1.33 +/- 1.33 of IgM isotype) were markedly higher than those of uninfected, malaria-naive controls (0.08 +/- 0.16 of IgG isotype and 0.04 +/- 0.04 of IgM isotype). Antibody levels for 7 out of 8 soldiers with a recent malaria infection were sustained above the cut-off values for 4 months after successful treatment. Analysis of serum collected from 40 healthy, asymptomatic soldiers who had a P. vivax malaria attack within 3 months after our sampling, revealed 11 antibody-positive samples (27.5%), compared to 5 positive samples (12.5%) collected from a random selection of 40 soldiers. Among a larger pool of 1,713 soldiers who had served in high-risk areas for P. vivax transmission, 15% were antibody positive. Among 1,000 blood samples from asymptomatic soldiers who had served in the high-risk areas, 4 samples (0.4%) were parasite positive, as determined by nested PCR. Our results show that anti-Pv200 antibody levels can provide useful information in the late diagnosis of P. vivax malaria infection in a previously naive population and also in large seroepidemiologic studies. Furthermore, our results suggest that asymptomatic P. vivax carriers could be important in the current outbreak of malaria in Korea.