Okayama University Medical School

Affinity electrophoresis of human alpha-fetoprotein (AFP) in amniotic fluid from pregnant women between 6 to 42 weeks of gestation and in the serum of a yolk sac tumor was performed with concanavalin A (Con A), lentil lectin (LCA), erythroagglutinating phytohemagglutinin (E-PHA) and Allomyrina dichotoma lectin (allo A). Separated AFP bands were detected by sensitive antibody-affinity blotting. In the first trimester, amniotic fluid AFP showed elevated percentages of Con A-nonreacting AFP (AFP-C1) and LCA weakly-reacting AFP (AFP-L2) as previously reported. Additionally, high percentages of E-PHA strongly-reacting AFP (AFP-P5) and E-PHA-reacting AFP (AFP-P4) were observed. E-PHA-nonreacting AFP (AFP-P1), E-PHA weakly-reacting AFP (AFP-P3f), allo A-nonreacting AFP (AFP-A1) and asialo-AFP, AFP-A1 s, were present only in amniotic fluids from 6 to 17 weeks of gestation. With advancing gestation, percentages of AFP-C1, AFP-P4 and AFP-P5 decreased and AFP-L2, AFP-P3f, AFP-A1, and AFP-A1 s disappeared by the end of 18 weeks. The glycoforms of serum AFP of the yolk sac tumor resembled those of amniotic fluid AFP in the early gestational stages.

There are few reports on skeletal reconstruction using the bone transport technique to repair bone defects caused by resections of tumors associated with osteosarcoma. We attempted to reconstruct a 23 cm bone defect after resection of an osteosarcoma of the left femur, and succeeded in gaining 17 cm by bone transport. Five years after surgery, this patient remains alive without metastasis or local recurrence.

Mast cells in osteoarthritic and rheumatoid arthritic synovial tissues of the human knee.

The fibrous components of the metacarpophalangeal (MP) joint including the palmar plate, the collateral ligament and the dorsal plate were studied with particular attention paid to the fibrous structure of the fibrous tendon sheath and the deep transverse metacarpal ligament. The tough fibrillar structure around the MP joint, especially the force nucleus, consisted of three types of mixed fibers: the fibrous tendon sheath of the A1 pulley, the deep transverse metacarpal ligament, and the palmar plate. The tendon sheath was located on the ulnar side in the index and middle fingers, on the central position in the ring finger, and on the radial side in the little finger. These fibrous connections among the fingers formed a transverse arch in the hand. The palmar plate of the MP joint was relatively rigid and appears to function as a cushion when flexed. A fold-like protrusion of the synovial layer of the palmar plate of the MP joint had a meniscoid function, which was larger than that of the proximal interphalangeal joint. The capsule of the MP joint was thicker at the dorsal area, forming a dorsal plate, which is a sliding floor of the extensor mechanism and has a meniscoid function for joint congruity. The main lateral stabilizer consisted of collateral ligaments and accessory collateral ligaments anchored to the palmar plate. These structures act together as a "phalangeal cuff", connecting the proximal phalanx to the metacarpal head and sta

Helicobacter pylori (H. pylori) infection in the stomach is etiologically closely associated with chronic active gastritis, peptic ulcer, gastric cancer and gastric mucosa-associated lymphoid tissue lymphoma. In this study, we examined the antibody responses and cytokine profiles of three strains of mice (BALB/c, C3H/He, and C57BL/6) infected with H. pylori. Following this, correlations between host-immune reactions and intensity of inflammation were analyzed. H. pylori (ATCC43504) was intragastrically administered once a week to the mice from 4 weeks of age, and they were sacrificed at the ages of 4 and 7 months. In these mice, we examined the histology of the stomach, antibody titers against H. pylori, and serum levels of cytokines (IL-4, IL-10, TNF-alpha, IL-2 and Interferon-gamma). In BALB/c mice, inflammation of the stomach was minimal. Inflammation was observed in 63.6% of C57BL/6 mice and 33.3% of C3h/He mice. In C57BL/6 and C3H/He mice, all the cytokines tended to increase. In contrast, BALB/c mice were inactive in cytokine production except for IL-2. Two C3H/He mice developed severe inflammation with lymph follicles; one showed a response largely typical of Th-1, and the other showed a response largely typical of Th-2. Although a definite correlation was not shown between Th-1/Th-2 response evaluated by cytokine production and intensity of inflammation, it appears that in H. pylori-induced inflammation both cell-mediated (Th-1) and humoral (Th-2) immunity play a role in pathogenesis.

To establish a method for estimating the time between the last consumption of alcohol and death, we examined the ethanol levels in body fluids and tissues of rats that had been orally administered 1 g/kg ethanol. We observed the following relationships between ethanol levels in the cardiac blood (blood in the heart itself), vitreous humor, and urine: cardiac blood > vitreous humor > urine at 10 min (early absorption stage); vitreous humor > cardiac blood > urine from 20 to 50 min (late absorption stage); vitreous humor > urine > cardiac blood from 60 to 120 min (distribution stage); and urine > vitreous humor > cardiac blood at 180 min (excretion stage). It was also observed that, in cases of death immediately following drinking, ethanol levels in the stomach contents are very high, and the following ratios of ethanol levels were observed: skeletal muscle to cardiac blood--less than 1; liver to cardiac blood--around 1. buccal mucosa to cardiac blood-greater than 1. These ratios at equilibrium after drinking were around 1, lower than 1 and around 1, respectively. We also measured alcohol levels in the cardiac blood, urine, vitreous humor and stomach contents of nine cadavers who had consumed alcohol prior to death. The relationships between the time since last consumption of alcohol and relative ethanol levels in these specimens were in good accordance with the results of the animal experiments.

There are few reports on skeletal reconstruction using the bone transport technique to repair bone defects caused by resections of tumors associated with osteosarcoma. We attempted to reconstruct a 23 cm bone defect after resection of an osteosarcoma of the left femur, and succeeded in gaining 17 cm by bone transport. Five years after surgery, this patient remains alive without metastasis or local recurrence.

The main purpose of this study was to evaluate the inhibitory effects of 5-fluorouracil antineoplastics, cephem antibiotics containing the methyltetrazolylthiol (MTT) group and antidiabetics on aldehyde dehydrogenase (ALDH) activity in vivo and in vitro. In in vivo experiments, rats were given a 100 mg/kg dose of drugs (10 mg/kg for glibenclamide) orally or intraperitoneally. When each drug was administered singly immediately after an oral administration of 1.5 g/kg ethanol, only carmofur, an antineoplastic, produced marked increases in blood acetaldehyde concentrations. This action was also noted when ethanol was ingested 15 h after administration. The remaining drugs did not increase blood acetaldehyde concentrations. When rats were treated with carmofur at 12 h intervals for 3 consecutive days and were given 1.5 g/kg ethanol after the final treatment, blood acetaldehyde concentrations were elevated more significantly than with a single administration of carmofur. Furthermore, daily administration of cephem antibiotics containing the MTT group, latamoxef, cefamandole, cefoperazone and cefbuperazone, significantly increased blood acetaldehyde concentrations. Daily administration of sulfonylurea antidiabetics, chlorpropamide and acetohexamide, slightly increased blood acetaldehyde concentrations. Drugs causing increases in blood acetaldehyde concentrations when administration was combined with ethanol ingestion also inhibited ALDH activity in vitro. The results of the in vitro experiments roughly correlated with those of the in vivo experiments. The inhibitory effects of drugs on ALDH activity were in the following order: carmofur >> cephem antibiotics containing the MTT group > sulfonylurea antidiabetics.

The hematological findings of 233 patients with brucellosis are presented and the possible pathologies discussed. Anemia was present in 128 patients (55%), leukopenia in 49 (21%) and thrombocytopenia in 59 (26%). Bone marrow aspirates of 18 patients (8%) with pancytopenia were examined. The bone marrow was hypercellular in 15 and normocellular in 3 patients. Granulomatous lesions were detected in 12 cases (67%), and slight to moderate cytophagocytosis of erythrocytes, granulocytes and platelets existed in all patients. Blood cell counts reverted to normal within 2-3 weeks of initiating chemotherapy with recovery from the disease.

With advances in lectin affinity electrophoresis of alpha-fetoprotein (AFP), the detection of significant changes in serum AFP at low levels in cirrhotics has become important for early detection of hepatocellular carcinoma. Serum AFP levels of 616 healthy individuals without abnormal liver function tests or virus markers of hepatitis B and C were determined by enzyme immunoassay with IMx-AFP Dainapack using automated IMx apparatus set at twice the ordinary sensitivity and compared with those of 241 individuals with abnormal liver function tests and/or positive hepatitis virus markers. The coefficient of variation in this assay was less than 10% at AFP levels as low as 0.2 ng/ml with a lower detection limit of 0.1 ng/ml. The AFP level of healthy population showed a Gaussian distribution curve after logarithmic transformation with a median and 2.5-97.5 percentile reference range of 2.2 (0.6-5.6) ng/ml. There was no significant difference in the AFP level between males and females. Individuals with abnormal liver function tests alone showed no significant increase in serum AFP unless they were associated with positive hepatitis virus markers.

The cross-sensitization to stereotyped behavior between mazindol (MZD) and methamphetamine (MAP) was investigated in rats. MZD (5 and 10 mg/kg/day, p.o.), MAP (5 and 10 mg/kg/day, p.o.) and saline (1 ml/kg, p.o.) were administered once daily for a week. Challenge with MZD (10 mg/kg, p.o.) on the 8th day caused markedly stereotyped behavior in MAP-pretreated group compared with the saline-pretreated control group. MAP (10 mg/kg, p.o.)-induced stereotyped behavior on the 8th day was also greater in MZD-pretreated group rather than the saline-pretreated control group. These results suggest that repeated MZD and MAP administration cross-sensitizes to their stereotype-producing effects.

To test transdifferentiation of retinal pigment epithelial (RPE) cells in suspension culture, chick embryonic RPE sheets and dissociated RPE cells were cultured for two months in a non-adherent dish for suspension culture. RPE cells, isolated as a sheet, aggregated immediately and remained the same size with their differentiated characteristics for two months. The presence of basic fibroblast growth factor (bFGF) at concentrations of 10 ng/ml or higher induced the formation of a spherical lentoid structure which was positive for crystallin and bFGF receptor. In contrast, dissociated RPE cells did reaggregate but did not develop the lentoid structure even in the presence of bFGF. The transdifferentiation of RPE cells to the lentoid structure in this study was in contrast to their transdifferentiation to the retina, as reported in a previous study.

Steady-state serum concentrations of carbamazepine (CBZ) and valproic acid (VPA) were investigated in normal weight (body mass index; BMI 20 to 25), lean (smaller than 20 BMI) and moderately obese subjects (greater than 25 BMI) who received either 400 mg/day of CBZ or 800 mg/day of VPA. The CBZ serum concentration in lean subjects was significantly higher than that in normal weight subjects. However, no significant differences in VPA serum concentration were found between the three groups. The CBZ serum concentration decreased with increases in total body weight, and the VPA serum concentration decreased with increases in ideal body weight. However, both serum concentrations were not correlated with BMI. These results suggest that VPA doses should be calculated using ideal body weight and that degree of obesity may affect CBZ serum concentration rather than VPA serum concentration.

To study the effect of partial liquid ventilation (PLV) with perfluorocarbon on acute respiratory failure, 3 groups of 17 rabbits were examined to compare. After acute respiratory failure was induced by lung lavage with sea water in 12 of the 17 rabbits, 7 of the 12 rabbits were treated with conventional mechanical ventilation (AC group) and 5 of the 12 rabbits were treated with PLV using perfluorocarbon (AP group). The remaining 5 normal rabbits without acute respiratory failure were treated with PLV with perfluorocarbon as a control group (PL group). In the PL group, PaO2, PaCO2, blood pH, pulmonary compliance or pathological findings were not so changed after PLV. In the AC and AP groups, PaCO2 significantly increased, and in contrast, PaO2 and pulmonary compliance significantly decreased after lung lavage. However, these findings improved to almost the same levels as those of a control group within 2 h after the PLV treatment in the AP group, but in the AC group, these gradually deteriorated over time. As for the pathological findings, pulmonary vascular congestion, alveolar hemorrhage and inflammatory infiltration were observed in the AC group. However, these findings were not observed in the specimens of the AP group. From these results, PLV with perfluorocarbon was shown to be useful to improve gas exchange and pulmonary functions without major side effects.

An exposure to GB virus C/hepatitis G virus (GBV-C/HGV) was studied among populations at risk for blood and sexual exposure to analyze risk factor of the transmission of the virus. Blood samples were drawn from 98 intravenous drug users (IVDU), 100 female high-class commercial sex workers (CSW) and 50 male outpatients (MOP) at a sexually transmitted diseases (STD) clinic in Chiang Mai, Thailand. These blood samples were analyzed for GBV-C/HGV RNA; antibodies against second envelope protein of GBV-C/HGV (anti-E2); anti-hepatitis C virus antibody (HCV-Ab); hepatitis B core antibody (HBcAb); and antibodies against human immunodeficiency virus (HIV-Ab). Prevalences of GBV-C/HGV RNA, anti-E2, HCV-Ab, HBcAb and HIV-Ab were 27.6%, 16.3%, 84.7%, 76.5% and 45.0% in IVDU; 0%, 21.5%, 2.0%, 72.0% and 11.0% in CSW; 6.0%, 13.6%, 0%, 64.0% and 14.0% in MOP. While the prevalence of GBV-C/HGV RNA was higher in IVDU than in CSW and MOP, comparable prevalences of anti-E2 among the three populations were found. Intravenous drug injection showed association with GBV-C/HGV RNA, while history of STD associated with anti-E2. In conclusion, intravenous drug injection and STD were found to be risk factors for the previous exposure to GBV-C/HGV, but STD did not increase the risk of the GBV-C/HGV viraemia.

We investigated the degree of local heat and swelling of the thigh muscles produced by exercise. Eleven university athletes aged from 19 to 23 years old performed isokinetic exercise of the right knee on a Cybex II. Then serial determination of thigh circumference and thigh temperature (up to 120 min after exercise) as well as serial magnetic resonance (MR) imaging (up to 60 min after exercise) was performed on both thighs. The circumference of the right thigh peaked at 5.6 +/- 2.1 min after exercise and returned to normal at 38.6 +/- 9.2 min. The temperature of the right thigh peaked at 14.2 +/- 5.7 min after exercise and was not normalized after 120 min except in two subjects. T2-weighted MR images showed a marked increase in the signal intensity of the right knee flexor and extensor muscles. The signal intensity peaked immediately after exercise and subsequently decreased gradually but did not return to normal after 60 min in some muscles. Changes in the thigh circumference were closely correlated with changes in the MR findings. The changes in the thigh muscles after knee exercise could be demonstrated using MR imaging, thigh circumference, and thigh temperature data. These parameters may provide indicators for managing muscle fatigue and recovery.

A new method for the determination of glutathione peroxidase activity in erythrocytes was developed. The present method was applied to the measurement of hydrogen peroxide removal rates by glutathione peroxidase in erythrocytes at 70 microM hydrogen peroxide under simulated in vivo conditions. The removal rates by glutathione peroxidase in mouse erythrocytes were twenty-times faster than those in human ones and were 5.2 mumol/sec/g of Hb. The removal rates in acatalasemic mouse erythrocytes indicate that glutathione peroxidase is the main means of hydrogen peroxide removal in acatalasemic mouse erythrocytes. Based on these results, we concluded that glutathione peroxidase in mouse erythrocytes had sufficient ability to remove hydrogen peroxide at even relatively high concentrations. This may be one of the reasons why acatalasemic mice suffer no health problems while Japanese acatalasemic patients suffer from Takahara disease when infected with hydrogen peroxide-generating bacteria.