Tissue plasminogen activator levels change with plasma fibrinogen concentrations during pregnancy.

To investigate the relationship between coagulation activities and the fibrinolytic system during normal pregnancy, we measured the plasma concentrations of coagulation factors, antithrombin III (AT III), D-dimer, tissue plasminogen activator (tPA), total protein S (TPS), and plasminogen activator inhibitor type 1 (PAI-1) in 436 apparently healthy pregnant, postpartum, and nonpregnant women. There were no significant changes in AT III, TPS, and factor XI concentrations during pregnancy and puerperium. However, factor VII, VIII, IX, and XII activities increased gradually as pregnancy progressed, reached maximum values in the third trimester, and returned to nonpregnant levels by 5-8 weeks postpartum. Plasma D-dimer levels in the third trimester of pregnancy were 1.23+/-0.42 micro g/ml, significantly higher than for the first trimester (0.34+/-0.16 micro g/ml, P<0.01). The tPA antigen levels averaged 1.8-fold higher in the late third trimester than in the first trimester; the plasma fibrinogen concentrations averaged 1.6-fold higher in the late third trimester than in the first trimester. Compared to the peak values during pregnancy, tPA levels averaged 39.8% lower and plasma fibrinogen concentrations averaged 40.0% lower at 5-8 weeks postpartum. The tPA levels correlated strongly with the plasma fibrinogen concentrations ( r=0.52, P<0.01). In short, this study shows that tPA levels change in parallel with plasma fibrinogen concentrations during and after normal pregnancy.

Iron deficiency anemia increases nitric oxide production in healthy adolescents.

To investigate the influence of iron deficiency and iron supplementation on nitric oxide (NO) production, we measured serum iron markers, serum nitrate and nitrite (NOx) concentrations, reticulocyte maturity index (RMI), and serum transferrin receptor (sTfR) levels in 369 females aged 14-19 years. RMI was analyzed by flow cytometry, sTfR concentrations were measured by enzyme immunoassay, and serum NOx levels were tested by the Griess reaction. NOx concentrations of the subjects in iron depletion phase were significantly higher than those of healthy controls. NOx concentrations increased gradually as iron deficiency progressed and were threefold higher than for the healthy controls, when the subjects attained a frank iron deficiency anemia. In particular, the NOx concentrations were 7.5-fold higher in the patients with severe iron deficiency anemia (Hb<80 g/l) than for the subjects with high hemoglobin value (Hb>or=140 g/l). The increased NOx concentrations (132.6+/-42.1 microM) observed in the group with severe anemia decreased significantly (46.3+/-15.8 microM) after hemoglobin levels were normalized by iron supplementation ( P<0.01). NOx concentrations correlated inversely with hemoglobin levels ( r(2)=0.202, P<0.01), but correlated positively with the sTfR concentrations ( r(2)=0.322, P<0.01) and the RMI ( r(2)=0.369, P<0.01). In conclusion, iron deficiency anemia increases NO production, and elevated NOx concentrations in iron deficiency anemia return to normal with iron supplementation.

Changes in serum lipid concentrations during iron depletion and after iron supplementation.

To investigate the effects of body iron depletion and iron supplementation on serum lipid concentrations, hematologic indices, iron markers, and serum lipid profiles were measured in 427 girls, age 14-19 yr. There were no significant differences in serum lipid concentrations between subjects with moderate iron deficiency anemia (blood Hb < 12.0 g/dL) and healthy controls. However, serum total cholesterol concentration (mean +/- SD, 148 +/- 16 mg/dL) in severely anemic subjects with blood Hb < 8.0 g/dL was significantly lower than in subjects with blood Hb > or = 14.0 g/dL (170 +/- 17 mg/dL) (p < 0.01). Moreover, serum triglyceride concentration in subjects with blood Hb > 14.0 g/dL was 2-fold higher than in the severely anemic subjects. Mean values of serum total cholesterol and triglyceride (149 +/- 17 mg/dL and 58 +/- 22 mg/dL) in girls with severe anemia were significantly elevated after iron supplementation (164 +/- 17 mg/dL and 98 +/- 26 mg/dL) (p < 0.01, respectively). In the severely anemic subjects, blood Hb concentration was correlated with serum total cholesterol (r = 0.49, p < 0.01) and triglyceride concentrations (r = 0.51, p < 0.01). These findings indicate that severe iron deficiency anemia in girls is attended by decreased concentrations of serum total cholesterol and triglyceride, and that these reduced serum lipid levels return to normal following iron supplementation.

Reticulocyte subpopulations and reticulocyte maturity index (RMI) rise as body iron status falls.

To investigate the influence of body iron status on reticulocyte subpopulations and reticulocyte maturity index (RMI), we measured serum iron markers, fluorescent intensity of reticulocytes, and serum transferrin receptor (sTfR) concentrations in 374 females aged 14-19 years. Reticulocyte subpopulations were analyzed by flow cytometry, and sTfR concentration was measured by enzyme immunoassay. There were no significant differences in the values of reticulocyte subpopulations and RMI between ferritin alone-depleted group and healthy controls. However, middle- and high-fluorescence reticulocytes and RMI were significantly higher in both the serum iron- and serum ferritin-depleted groups than in the ferritin alone-depleted group. Middle-fluorescence reticulocytes and RMI increased gradually as the body iron store was depleted and were 3.4- and 3.6-fold higher, respectively, than normal controls, when the subjects attained a frank iron-deficiency anemia. There were no significant changes in the values of red blood cells or total reticulocyte counts during iron-depleted states. The mean value of sTfR (3.98 mg/l) in the subjects with RMI > or = 1.5% was significantly higher than that (2.26 mg/l) in the subjects with RMI < 1.5% (P < 0.01). The sTfR concentration correlated significantly with RMI (r = 0.61, P < 0.01) and middle-fluorescence reticulocytes (r = 0.59, P < 0.01). In short, body iron depletion induces elevation of immature reticulocyte fractions and RMI.

Change in erythropoiesis with gestational age during pregnancy.

To investigate the changes in erythropoiesis during pregnancy, 342 pregnant and postpartum women were examined for reticulocyte subpopulations, reticulocyte maturity index (RMI), and serum transferrin receptor (sTfR) levels. The reticulocyte subpopulations were analyzed by flow cytometry, and the sTfR concentration was measured by an immunoenzymometric method. There were no significant differences in the values of reticulocyte subpopulations, RMI, and sTfR between first trimester and non-pregnant women. However, the RMI and sTfR concentration increased gradually from the second trimester of pregnancy, during which time values were twofold higher than those in the first trimester, and peaked in the third trimester. Then the RMI and sTfR values decreased abruptly 1-4 weeks postpartum and finally dropped to the levels of non-pregnant women 5 weeks after delivery. The maternal sTfR concentration correlated significantly with the corrected reticulocyte count (r = 0.52, P < 0.01), RMI (r = 0.67, P < 0.01), and high fluorescence reticulocytes (HFR) (r = 0.62, P < 0.01); however, the serum ferritin level correlated poorly with the reticulocyte subpopulations during pregnancy. In conclusion, erythropoiesis increases with gestational age during pregnancy and returns to normal 5 weeks after delivery. Measurement of sTfR in combination with RMI may improve the assessment of erythropoietic activity during pregnancy.

Serum lipid concentrations change with serum alkaline phosphatase activity during pregnancy.

To investigate the relationship between serum lipids and alkaline phosphatase during normal pregnancy, we measured triglyceride, total cholesterol, HDL-cholesterol, and LDL-cholesterol concentrations and alkaline phosphatase activity in serum samples from 546 apparently healthy pregnant, postpartum, and nonpregnant women. Serum HDL-cholesterol levels did not change significantly during pregnancy, but serum triglyceride, total cholesterol, LDL-cholesterol, and alkaline phosphatase levels increased gradually as pregnancy proceeded, reached maximum values in the third trimester, and returned to nonpregnant levels by 20-24 wk postpartum. The serum alkaline phosphatase activity averaged 2.1-fold higher in the late third trimester than in the first trimester; the serum triglyceride concentration averaged 2.3-fold higher in the late third trimester than in the first trimester. Compared to the peak values during pregnancy, serum alkaline phosphatase activity averaged 45% lower and serum triglyceride level averaged 47% lower at 12-16 wk postpartum. The serum alkaline phosphatase activity was correlated with the serum concentrations of total cholesterol (r = 0.68, p < 0.01) and triglyceride (r = 0.71, p < 0.01). In short, this study shows that serum triglyceride and total cholesterol levels change in parallel with serum alkaline phosphatase activity during and after normal pregnancy.

Erythropoietic activity and soluble transferrin receptor level in neonates and maternal blood.

UNLABELLED: Serum transferrin receptor (sTfR) concentration reflects functional iron status and erythropoietic activity. The aims of this study were to examine gender differences of erythropoiesis in newborns and to evaluate the influences of maternal anaemia or iron deficiency on foetal cord blood parameters for iron status and sTfR. In total, 527 newborns and their mothers were examined. Reticulocytes were analysed by flow cytometry and sTfR was measured by an immunoenzymometric method. There were no sex differences in haematological or iron parameters. However, the reticulocyte maturity index (RMI) of male neonates was 37.45%, significantly higher than the 26.81% in female neonates (p <0.01). The high fluorescence reticulocytes (HFR) and middle fluorescence reticulocytes (MFR) of male neonates were 4.91% and 22.36%, respectively, while those of female neonates were 3.31% and 17.83%, respectively (p < 0.01 for each gender). The sTfR concentrations of male and female neonates were 6.27 mg/l and 5.09 mg/l, respectively (p < 0.01). Values for serum iron, ferritin and reticulocyte subpopulations were significantly lower in the newborns of anaemic mothers. However, newborns of iron-deficient mothers showed no differences in iron parameters from those of non-iron-deficient mothers. CONCLUSIONS: The higher values of RMI and sTfR observed in male neonates indicate that erythropoietic activity is higher in this group. Iron transport to the foetus appears to be independent of maternal iron deficiency. However, iron transport and erythropoiesis in newborns seems to decline from the time that the mothers acquire frank iron deficiency anaemia.

Randomized placebo-controlled trial of Helicobacter pylori eradication for iron-deficiency anemia in preadolescent children and adolescents.

BACKGROUND: A few cases relating H. pylori infection to iron-deficiency anemia have been described recently. We investigated the role of H. pylori infection in iron-deficiency anemia in preadolescent children and adolescents. PATIENTS AND METHODS: We conducted a double-blind, placebo-controlled therapeutic trial in 43 subjects (mean age, 15.4 years) with iron-deficiency anemia. Endoscopy was performed, and biopsy specimens were examined by urease test and histological analysis. Twenty-two of 25 H. pylori-positive patients were assigned randomly to three groups. Group A patients were given oral ferrous sulfate and a 2-week course of bismuth subcitrate, amoxicillin, and metronidazole. Group B patients were given placebo for iron and a 2-week course of triple therapy. Group C patients were given oral ferrous sulfate and a 2-week course of placebo. Iron status was reassessed 4 weeks and 8 weeks after the 2-week regimen ended. RESULTS: Of the 43 subjects with iron-deficiency anemia, 25 (58.1%) had H. pylori in the antrum. Group A and B subjects, who received eradication therapy, showed a significant increase in hemoglobin level as compared with group C subjects at 8 weeks after therapy (p = .0086). CONCLUSIONS: Treatment of H. pylori infection was associated with more rapid response to oral iron therapy as compared with the use of iron therapy alone. Such treatment also led to enhanced iron absorption even in those subjects who did not receive oral iron therapy.

Two cases of Chromobacterium violaceum infection after injury in a subtropical region.

Chromobacterium violaceum is a gram-negative rod and is isolated from soil and water in tropical and subtropical regions. The species have pigmented and nonpigmented colony types. Infections caused by nonpigmented strains are rare. We report on two cases of infection caused by both pigmented and nonpigmented strains of C. violaceum. Two 24-year-old Korea Airline stewardesses were admitted to Inha University Hospital, Inchon, South Korea, on 9 August 1997, 3 days after an airplane accident in Guam. Both had multiple lacerations on exposed parts of their bodies. There was swelling, tenderness, and pus discharge. The wounds contained many small fragments of stones and weeds. A pigmented strain was isolated from the left hand and a nonpigmented strain was isolated from the left knee of one patient. For the other patient only a nonpigmented strain was isolated from a foot wound. The nonpigmented colonies from the left-knee and the left-foot wounds did not produce any pigment even after an extended period of incubation. The biochemical characteristics were the same for each strain except for oxidase and indole reactions. The pigmented strain was oxidase negative and indole positive, whereas the nonpigmented strains were oxidase positive and indole negative. The patients were successfully treated by debridement and with appropriate antibiotics.

Isolation and characterization of a newly identified type II restriction endonuclease from a local Streptomyces sp. in Taiwan.

Streptomyces chusanensis ZS-2, isolated from a soil sample in Chusan in Taiwan, was found to produce a new Type II restriction endonuclease. This restriction enzyme was designated as SchI. The purified enzyme was characterized as having a subunit mol wt of 28 kDa, and was apparently free from exonuclease activities. It cleaves the phosphodiester bond between the fourth C and the fifth G on the 5'-CCGCGG-3' sequence of DNAs, leaving a 2-nucleotide protruding end at its 3' site. This data suggests that SchI is an isoschizomer of SacII. In addition, based on the comparison between SchI and SacII regarding reaction parameters, it seems that SchI is a better choice of restriction enzyme for genetic analysis and mapping.

A relapsed case of imported tertian malaria after a standard course of hydroxychloroquine and primaquine therapy.

Resistance of Plasmodium species to antimalarial agents has become increasingly challenging to the management and prevention of malaria. We experienced an imported case of tertian malaria due to Plasmodium vivax relapsed after a seemingly successful treatment with conventional course of hydroxychloroquine and primaquine. A 35-year-old man developed fever three days after return from India and mainland China. After his illness was diagnosed as tertian malaria, he was managed with hydroxychloroquine and then primaquine (primaquine base 15 mg/day for 14 days). Thereafter peripheral blood smears showed no malarial parasites, and there was no relapse of symptom until the 55th post-treatment day, however, six months after the above treatment tertian malaria relapsed. He was managed with the same medications again and malaria did not relapse for 10 months.

[Five human cases of Diphyllobothrium latum infection through eating raw flesh of redlip mullet, Liza haematocheila].

A total of 32 human diphyllobothriasis cases have been reported so far in Korea, excluding 21 egg-positive cases from stool examinations. Authors experienced five more human cases of Diphyllobothrium latum infection, especially infected due to eating raw flesh of redlip mullet, Liza haematocheila. Five cases were neighbors residing in the Puchon area, Kyonggi-do, who ate raw mullets (L. haematocheila) in a party in February 1996. The mullets were purchased at the Noryangjin fisheries market in Seoul. All of cases (2 males and 3 females) were 35 to 43 years old and healthy with the body weight range of 56-62 kg. They complained about gastrointestinal trouble and abdominal discomfort, but were in normal ranges of their hematology and urinalysis data. None revealed any sign of anemia. The patients experienced natural discharge of a chain of segments before, and showed diphyllobothriid eggs in their stool specimens when they visited our laboratory. They were administered with praziquantel (15 mg/kg of body weight) and 30 g of magnesium sulfate as a purgative. Two whole worms with the scolices (310-340 cm in length; 8-13 mm in width) were expelled each from two out of five cases after anthelmintic treatment, and the others expelled the parts of strobilae without scolex. The worms were identified as D. latum, based on the following biological characters: external morphologies, coiling of uterus, the number of uterine loops, position of genital opening, morphologies of cirrus, cirrus sac and seminal vesicle on the histological sections, position of vagina and uterine pore, and microscopical and SEM morphologies of the eggs.

Isolation and molecular cloning of human sorcin a calcium-binding protein in vincristine-resistant HOB1 lymphoma cells.

A vincristine-resistant lymphoma cell line (HOB1/VCR1.0) that is resistant to 1.0 microM of vincristine has been established from a human immunoblastic B lymphoma cell line, HOB1. HOB1/VCR1.0 cells demonstrated the typical multidrug resistant phenotypes. Using two-dimensional gel electrophoresis, we discovered one protein with a molecular mass of 22 kDa and pI 5.7 that was overexpressed in HOB1/VCR1.0 cells. This protein was purified to the degree of apparent homogeneity by preparative isoelectric focusing and sodium dodecylsulfate-polyacrylamide gel electrophoresis. The identification of this protein with sorcin was revealed by comparing the internal amino acid sequence of three Lys-C digested peptides from the purified protein with the sequence previously determined for hamster sorcin. The complete primary structure of the human sorcin was deduced from nucleotide sequence analysis of its cDNA clones. It is composed of 198 amino acid residues with a calculated molecular weight of 21,676, and its sequence is highly similar to that of hamster sorcin (95%). Direct-binding assay with calcium showed that human sorcin is a calcium-binding protein with four 'E-F hand' structures typical of calcium-binding sites. Like the sorcin of hamster, two of the calcium-binding sites of human sorcin contain putative recognition sites for cAMP-dependent protein kinase. Southern and Northern blot analyses showed that the human sorcin gene was greatly amplified and overexpressed in resistant HOB1/VCR1.0 cells but not detected in the parental HOB1 cells. The overproduction of this protein in resistant cells implies that sorcin plays a role in expression of the resistant phenotype.

Comparison of 5' region sequences of human and rabbit phosphofructokinase genes.

A phosphofructokinase gene was screened and cloned from a human genomic library prepared in the lambda EMBL4 phage vector. DNA sequencing shows that the first exon of this human phosphofructokinase gene is identical in length and highly homologous in sequence to that of a rabbit phosphofructokinase gene. Two amino acid replacements are indicated, an Arg----Lys and a Val----Ile at positions 9 and 13, respectively. Eleven base substitutions, 8 of them silent, are identified. Surprisingly, at ten of these sites, complete bias for A's and T's in the human gene and C's and G's in the rabbit gene are seen. Strong conservation is also observed in the 5' untranslated region and for the first 15 base pairs in the intron. All the nine variant nucleotides in these regions are, again, A's and T's in the human gene and G's and C's in the rabbit gene. The unit evolutionary period of change between the first exon of rabbit and human phosphofructokinase genes is estimated as 2.3 million years at silent sites and 15.6 million years at replacement sites.

Technetium 99m-DTPA microcapsules: a new preparation for gastric emptying studies.

Technetium 99m-DTPA microcapsules have been developed to measure gastric emptying. Such capsules not only provide high labeling efficiency in vitro, but demonstrate limited dissociation in vivo, resulting in decreased error during measurement. In normal control subjects, the half-life ranged from 40 to 80 minutes under the aforementioned conditions.

Purification of Eco R1 endonuclease on heparin Sepharose-4B column chromatography.

Restriction endonucleases play a very important role in genetic engineering and DNA mapping. Among hundreds of restriction endonucleases, the Eco R1 enzyme is the most useful and widely investigated enzyme. After sonication and ultracentrifugation, crude extracts of E. coli RY 13 were purified by employing the polyethyleneimine precipitate, ammonium sulfate precipitate and heparin Sepharose-4B affinity column chromatography. The Eco R1 enzyme were purified at about 42 folds and the specific activity was about 100,000 U/mg of protein. The whole purification procedure was finished within two days. The recovery was about 42%. The enzyme was sufficiently concentrated for direct specific DNA hydrolysis.