Pre-analytical parameters associated with unsuccessful karyotyping in myeloid neoplasm: a study of 421 samples.

Cytogenetics is essential in myeloid neoplasms (MN) and pre-analytical variables are important for karyotyping. We assessed the relationship between pre-analytical variables (time from collection to sample processing, material type, sample cellularity, and diagnosis) and failures of karyotyping. Bone marrow (BM, n=352) and peripheral blood (PB, n=69) samples were analyzed from acute myeloid leukemia (n=113), myelodysplastic syndromes (n=73), myelodysplastic syndromes/myeloproliferative neoplasms (n=17), myeloproliferative neoplasms (n=137), and other with conclusive diagnosis (n=6), and reactive disorders/no conclusive diagnosis (n=75). The rate of unsuccessful karyotyping was 18.5% and was associated with the use of PB and a low number of nucleated cells (≤7×103/µL) in the sample. High and low cellularity in BM and high and low cellularity in PB samples showed no metaphases in 3.9, 39.7, 41.9, and 84.6% of cases, respectively. Collecting a good BM sample is the key for the success of karyotyping in MN and avoids the use of expensive molecular techniques.

A novel nuclear protein in rat ventral prostate. Androgen-dependent and age-related change.

A novel protein was found in the nuclei of rat ventral prostate. This protein has a molecular weight of about 21 kDa as measured by SDS-polyacrylamide gel electrophoresis. It showed a characteristic change between 3 and 84 weeks after birth in close association with the level of testosterone in the blood. After castration, the level of the 21-kDa protein decreased to 1/60 of normal in 7 days, but on daily injection of testosterone the level was restored to normal in 8 days and to twice the normal level in 14 days. Unlike H1 and H1(0) histone and high mobility group proteins, the 21-kDa protein was not extracted with 5% HClO4, but was partially extracted with 0.35 M NaCl. The 21-kDa protein was not found in kidney, liver, or bain, suggesting that it is specific to the ventral prostate.

Androgen receptor in human placental villi.

In studies with a synthetic androgen, R 1881, an androgen-binding component was found in the cytosol of human placental villi. Kinetic analysis indicated that the Kd value of this component was 1.4 nM at 0-4 degrees C and that binding of R 1881 amounted to 277 +/- 73 fmol/mg protein. glycerol density gradient ultracentrifugation showed a peak of binding activity in the 8S region in a medium of low ionic strength, but in the 4.5S region in a medium containing 9.5 M KCl. The R 1881-binding component was inactivated by mild heat- or trypsin-treatment, but not by treatment with DNase or RNase. Most of the R 1881-binding activity was sedimented at 20 to 40% saturation of ammonium sulfate. These findings indicate that the R 1881-binding component in human placental cytosol is quite similar in its characteristics to androgen receptors, which are present in various androgen-responsive organs. Testosterone was a more potent competitor of R 1881-binding than DHT or cyproterone acetate. Scatchard plots indicated that the binding site of testosterone was identical with that of R 1881. These findings suggest that the androgen receptor in placental cytosol is specific for testosterone. The Kd value for testosterone was calculated to be 3.2 nM.

Demonstration of the distal end of the oesophagus by transabdominal ultrasound.

This study aimed to demonstrate the distal end of the oesophagus using a transabdominal ultrasound technique (TUS) and to measure the normal oesophageal wall thickness in adults. 65 patients without oesophageal disease and 38 normal volunteers were examined by TUS. A left subcostal approach was used to demonstrate the oesophagus. The wall thickness and length were measured in both the supine and 45 degrees right side up oblique (RUO) positions. The abdominal oesophagus was visualized in 80% of patients in the supine position and 92% in the RUO position. Satisfactory demonstration was obtained in 67% of patients in the supine and 85% in the RUO position. The oesophageal wall thickness averaged 2.8 mm (range 2.0-4.0 mm, SD 0.7 mm). The visualized length in these subjects averaged 2.3 cm in the supine position and 3.0 cm in the RUO position, which included approximately 1.5 cm of the lowest portion of the thoracic oesophagus in addition to the abdominal oesophagus. TUS can demonstrate the abdominal oesophagus in the majority of patients and has the potential to provide information on disorders of structure and motility.

Radical scavenging properties of tryptophan metabolites. Estimation of their radical reactivity.

Radical scavenging properties of tryptophan metabolites were estimated using their radical reactivity. Metabolites of the kynurenine and the melatonin biosynthesis pathway were mainly examined by use of a kinetical model. Their radical reactivity was determined as the reaction rate constant with a stable free radical, such as galvinoxyl; that is a phenoxy radical. The rate constants of the metabolites have a widely ranged spectrum, which can be divided into three groups. The first group (3-hydroxykynurenine, 3-hydroxyanthranilic acid, and indole-3-pyruvic acid) is more reactive than alpha-tocopherol; the reactivity of the second group (xanthurenic acid, serotonin, N-acetylserotonin) is similar to that of butylated hydroxytoluene (BHT); the third group (kynurenic acid, melatonin, and other ones) is less reactive than BHT.

Changes in hepatic enzyme activities related to ethanol metabolism in mice following chronic ethanol administration.

Male mice of three strains, C57BL, DBA and C3H/He, were fed on commercial food with 10% (v/v) ethanol solution as drinking liquid ad libitum for eighty days, and the changes in the activities of enzymes in the metabolic pathway of ethanol in the liver were examined. C57BL and C3H/He mice showed a preference for drinking the 10% (v/v) ethanol solution, while DBA mice did not. The ethanol intake g/g of body weight of C3H/He mice showed the highest value among all three strains and that of C57BL mice tended to show higher value than that of DBA mice. The liver weights of C57BL and C3H/He mice increased significantly following chronic ethanol administration, but that of DBA did not. The cytosolic enzyme alcohol dehydrogenase (ADH) showed no changes in any of the strains following chronic ethanol administration. The microsomal ethanol-oxidizing system (MEOS) of C57BL mice exhibited approximately 2-fold higher activity compared to that of DBA and C3H/He mice but did not increase in any strain following chronic ethanol administration. However, the microsomal aniline hydroxylase activity in the liver increased significantly in C57BL and C3H/He mice following chronic administration of ethanol. The microsomal cytochrome P-450 content also tended to slightly increase in the same strains of mice. It seemed that cytochrome P-450IIE1 was induced in the liver microsomes of these strains. Total aldehyde dehydrogenase (ALDH) activities together with high-Km ALDH activity increased markedly in the microsomes of C57BL mice and tended to increase in C3H/He mice, while it did not change in DBA mice following chronic ethanol administration. In the mitochondria of C57BL, total ALDH activities increased slightly and high-Km ALDH activities tended to increase. These mitochondrial ALDH activities of C3H/He and DBA mice tended to increase following chronic ethanol administration. The cytosolic ALDH activity showed no changes in any strain of mice following chronic ethanol administration. It seemed that in the microsomes, the activities of enzymes related to oxidation of ethanol increased in C57BL and C3H/He mice, which tended to consume a large amount of ethanol, and did not in DBA mice which tended to consume a small amount of it. It seemed that the increases in activities of enzymes related to oxidation of acetaldehyde in the microsomes and in the mitochondria were responsible for the strain difference.

Combined effects of ethanol and garlic on hepatic ethanol metabolism in mice.

The combined effects of ethanol and components in fresh garlic on ethanol metabolism were investigated in the livers of mice. Male, 11-wk-old C3H/HeNCrj mice were intragastrically administered 2 g ethanol/kg body weight after being administered fresh garlic juice for 8 d (garlic group), and changes in the concentrations of ethanol, acetaldehyde and acetate in the serum, and changes in the activity of hepatic enzymes related to ethanol metabolism in mice were examined. The increases in the concentrations of acetaldehyde and acetate in the serum after ethanol administration tended to be diminished following garlic administration. The microsomal ethanol-oxidizing system (MEOS) in the livers of the garlic groups was significantly lower than that of the control microsomes at 2 h after ethanol administration. It therefore seems that the decrease of MEOS in hepatic microsomes caused a smaller increase in the acetaldehyde concentration in the serum of the garlic groups because cytosolic alcohol dehydrogenase showed no significant difference between the control and garlic groups. After ethanol administration, the content of cytochrome P-450 in the hepatic microsomes of the control groups increased, while that of the garlic groups did not change although cytochrome P-450 (CYP) 2E1 and 1A2 in the hepatic microsomes of the garlic groups increased. These results indicate that the induction of isozymes of cytochrome P-450 other than CYP 2E1 and 1A2 was inhibited following garlic administration. Cytosolic high Km and total aldehyde dehydrogenase (AIDH) in the liver of the garlic groups tended to be lower than those activities of the control groups at 1 and 2 h after ethanol administration. It therefore seems that the decreases of AIDH in the hepatic cytosols diminished the increase of acetate in the serum of the garlic groups after ethanol administration. These results suggest that the ethanol metabolism in the mouse liver is controlled by components in fresh garlic juice.

Effect of chronic administration of alcoholic beverages and seasoning containing alcohol on hepatic ethanol metabolism in mice.

Five-week-old male mice, C3H/HeNCrj (C3H/He), were given a 5% (v/v) ethanol solution, commercial alcoholic beverages (Japanese sake (sake) or red wine) or a Japanese seasoning (mirin [containing ethanol and a large amount of glucose]) ad libitum for 45 d, and were then examined for changes in the hepatic enzymes related to ethanol metabolism 2 h after oral administration of 5 g of ethanol/kg body weight. The specific activity of aniline hydroxylase (ANH) in the hepatic microsome increased significantly in all groups chronically administered ethanol solution, sake, red wine or mirin, and the greatest increase was in the hepatic microsome of mirin-administered mice. The cytochrome P-450 (CYP) 2E1 increased in the hepatic microsome of the mice administered ethanol solution, red wine or mirin where accompanied by high ANH activity. The immunoreactive band for CYP1A1 showed high specificity in the microsome of mice given sake, red wine or mirin. It was assumed that CYP1A1 was induced by unknown component(s) other than ethanol in these solutions. In the cytosolic fraction, following the chronic administration of sake and mirin, the total aldehyde dehydrogenase (A1DH) activity with high-Km decreased significantly. In the mitochondrial fraction, the activity of high-Km A1DH increased significantly in the mirin-administered mice which drank a large amount of ethanol, whereas that in the red wine-administered group tended to decrease. These results indicate that the enzyme activities related to the oxidation of both ethanol and acetaldehyde in the cytosolic, mitochondrial and microsomal fractions of the liver were affected by either the action of ethanol or its interaction with other constituents of sake, red wine and mirin.

Recovery of alveolar macrophages from rhesus and cynomolgus macaques by lung lavage.

A lung lavage technique was developed to recover alveolar macrophages from rhesus and cynomolgus macaques. Sterile saline solution was injected through an endotracheal tube in anesthetized macaques; lung wash fluids containing leukocytes were withdrawn. The lung wash fluids from each animal routinely contained more than 16 x 10(6) leukocytes. The predominant cell type was the alveolar macrophage; lung wash fluids contained more than 53% and 80% alveolar macrophages from rhesus and cynomolgus macaques, respectively. Lung lavage was performed each week for 6 weeks in both species with no ill effects. This technique has many applications in the study of infection and of pulmonary defense mechanisms.

[Sexually transmitted disease infection in HIV carriers].

Heterosexual transmission of the human immunodeficiency virus (HIV) appears to occur readily all over the world. Recent reports have suggested an association between HIV and sexually transmitted disease (STD). We conducted a case study among foreign female prostitutes who were seropositive to HIV to determine the prevalence of STD. In this study, we checked the prevalence of STD infection in 19 HIV seropositive female prostitutes. Overall, 84 percent were seropositive for Chlamydia trachomatis, 37% for HBs, 32% for Treponema pallidum (Tp) and 6% for hepatitis C virus (HCV). The high frequency of a history of STD may be associated with an increased risk of HIV infection acquired through heterosexual contact. Prevention of heterosexually transmitted HIV infection will require an extensive educational program aimed specifically at the risk associated with the number and selection of sexual partners and at promoting safer sexual practices.

Pre-analytical parameters associated with unsuccessful karyotyping in myeloid neoplasm: a study of 421 samples

Cytogenetics is essential in myeloid neoplasms (MN) and pre-analytical variables are important for karyotyping. We assessed the relationship between pre-analytical variables (time from collection to sample processing, material type, sample cellularity, and diagnosis) and failures of karyotyping. Bone marrow (BM, n=352) and peripheral blood (PB, n=69) samples were analyzed from acute myeloid leukemia (n=113), myelodysplastic syndromes (n=73), myelodysplastic syndromes/myeloproliferative neoplasms (n=17), myeloproliferative neoplasms (n=137), and other with conclusive diagnosis (n=6), and reactive disorders/no conclusive diagnosis (n=75). The rate of unsuccessful karyotyping was 18.5% and was associated with the use of PB and a low number of nucleated cells (≤7×103/µL) in the sample. High and low cellularity in BM and high and low cellularity in PB samples showed no metaphases in 3.9, 39.7, 41.9, and 84.6% of cases, respectively. Collecting a good BM sample is the key for the success of karyotyping in MN and avoids the use of expensive molecular techniques.

[Quantification of blood flow using MR imaging with pre-saturation band].

A velocity profile was obtained by magnetic resonance (MR) imaging with a pre-saturation band (PSB) and automatically analyzed using a personal computer with the data transferred through the picture archiving communication system (PACS). A phantom study showed a good correlation of the measured flow rate to the actual volumetric flow rate (R¿2 = .984; MR rate = 0.11 + 0.90 x actual rate ml/min). Images were also obtained in 11 healthy fasting male volunteers during a 16-sec breath-hold. The average portal blood flow of these subjects was 16.2 + 5.0 mil/min/kg body weight. Quantitative MR imaging with PSB is a practical and noninvasive technique for measuring volumetric flow rate in the portal venous system.

Prevalence of rickettsial antibody and cell-mediated reaction in cynomolgus monkeys (Macaca fascicularis).

Serologic studies on feral, colony-held cynomolgus monkeys indicated that 61% reacted to Coxiella burnetii antigens, and 36% reacted to Rickettsia conorii antigens. The results suggest that a high percentage of cynomolgus monkeys have been exposed to these organisms.