Screening of the Men1 gene and discovery of germ-line and somatic mutations in apparently sporadic parathyroid tumors.

Hyperparathyroidism is the first manifestation in a majority of multiple endocrine neoplasia (MEN1) patients. To discriminate between sporadic and hereditary parathyroid tumors and characterize MEN1 somatic mutations, we examined MEN1 gene mutations in patients who had undergone surgery for sporadic parathyroid tumors. DNA was extracted from fresh frozen parathyroid tumor specimens from 112 patients as well as from peripheral blood leukocytes from 64 of the 112 patients. Sequence analysis was performed to examine exons 2-10 of the MEN1 gene for mutations. Loss of heterozygosity (LOH) was also examined by an analysis of codon 418 and 541, which lie within a polymorphic region of MEN1. Somatic MEN1 mutations were found in 25 of the 112 patients (22%). Two patients had two point mutations (508del33 and Y341X and 363insT and 1767delT, respectively). A total of 27 mutations were characterized, 20 of which have not been reported previously. There were 7 nonsense mutations, 10 frameshift mutations, 2 splice site deletions, 5 missense mutations, and 3 in-frame mutations. Nineteen mutations (70%) predicted truncation of the menin protein. Germ-line MEN1 mutations were found in 3 of 64 patients (5%) who had no family history of endocrine tumors associated with MEN1, and these patients were identified as MEN1 gene probands. LOH at the MEN1 locus was detected in three parathyroid tumors showing germ-line mutation. LOH was significantly frequent in parathyroid tumors with somatic MEN1 mutations (15 of 22 tumors, 68%) but not in those without germ-line or somatic MEN1 mutations (14 of 51 tumors, 28%; P = 0.0011). Our findings suggest that alterations of both alleles of the MEN1 gene may be associated not only with endocrine tumors of affected MEN1 patients but also with sporadic parathyroid tumors. Germ-line MEN1 gene analysis can distinguish heritable from nonheritable parathyroid tumors, and MEN1 gene evaluation of patients with apparently sporadic parathyroid tumor is recommended before parathyroid surgery.

Absence of somatic RET gene mutation in sporadic parathyroid tumors and hyperplasia secondary to uremia, and absence of somatic Men1 gene mutation in MEN2A-associated hyperplasia.

Germline mutations of the MEN1 gene are found in more than 85% of multiple endocrine neoplasia type 1 (MEN 1) patients, and germline mutations of the RET gene are found in more than 95% of multiple endocrine neoplasia type 2 (MEN2) patients. Parathyroid hyperplasia is seen in more than 90% of MEN 1 and about 15% of MEN2A patients. To date, somatic MEN1 mutations are reported in about 20% of sporadic parathyroid tumors. To elucidate the genetic basis of parathyroid tumor development, we examined somatic RET gene mutations in sporadic parathyroid tumors and hyperplasia secondary to uremia, and somatic MEN1 gene mutations in parathyroid hyperplasia from MEN2A patients. A total of 145 parathyroid tumors comprising 129 sporadic parathyroid tumors, 14 hyperplastic lesions secondary to uremia, and two hyperplastic lesions from MEN2A patients were examined. DNA was extracted from fresh frozen parathyroid tissue. Exons 2-10 of the MEN1 gene and exons 10 and 11 of the RET gene were sequenced. No somatic RET gene mutations were found in the 129 sporadic parathyroid tumors or 14 parathyroid hyperplastic lesions secondary to uremia. No somatic MEN1 gene mutations were found in the two parathyroid hyperplasia from MEN2A patients. These data suggest that RET gene mutation may not be involved in the development of sporadic parathyroid tumors and hyperplasia secondary to uremia and that MEN1 gene mutation may not be or is rarely associated with development of parathyroid hyperplasia in MEN2A patients.

A portable multi-channel recording system for analysis of acceleration and angular velocity in six dimension.

A portable recording system was developed for analysis of more than three analog signals collected in field works. Stereo audio recorder, available as consumer products, was made use for a core cornponent of the system. For the two tracks of recording, a multiplexed analog signal is stored on one track, and reference code on the other track. The reference code indicates the start of one cycle for multiplexing and swiching point of each channel. Multiplexed signal is playbacked and decoded with a reference of the code to reconstruct original profiles of the signal. Since commercial stereo recorders have cut DC component off, a fixed reference voltage is inserted in the sequence of multiplexing. Change of voltage at switching from the reference to the data channel is measured from playbacked signal to get the original data with its DC component. Movement of vehicles and human head were analyzed by the system. It was verified to be capable to record and analyze multi-channel signal at a sampling rate more than 10Hz.

Increased macrophage-colony stimulating factor levels in neonates with perinatal complications.

OBJECTIVE: To examine whether perinatal complications induce the production of macrophage-colony stimulating factor (M-CSF), we have compared M-CSF levels in the cord blood between normal neonates and neonates with complications. METHODS: The M-CSF levels were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: In 54 normal neonates, the M-CSF level was 1859 +/- 287 U/ml (mean +/- S.D.), being significantly higher than the serum M-CSF level in normal adults (697 +/- 132 U/ml). Compared with M-CSF levels in normal neonates, significantly higher levels were evidenced in neonates with perinatal complications including premature rupture of the membranes, neonatal asphyxia, meconium staining of the amniotic fluid and maternal anemia. However, no difference in M-CSF concentrations was observed irrespective of complication types; furthermore, the M-CSF level was highly correlated with the leukocyte counts in the neonates with complications, but not in normal neonates. Incidentally, CRP levels were within normal limits in most of these neonates, CONCLUSION: M-CSF levels in the cord blood from neonates with premature rupture of the membranes, neonatal asphyxia, meconium staining of the amniotic fluid and maternal anemia were significantly higher than those in the cord blood sampled from normal neonates. The stress given to neonates may account for the higher M-CSF levels rather than infections.

Detection of HBV-DNA and HCV-RNA viral sequences by polymerase chain reaction in selected Kenyan samples and the relationship to HBV seromarkers.

We undertook a study on selected samples from patients who had presented with viral hepatitis and conditions of the liver (liver cirrhosis, chronic hepatitis and hepatocellular carcinoma). Diagnosis, screening and confirmation for viral hepatitis was done using a battery of techniques: ultrasound, conventional serological methods (Hepatitis B surface Antigen [HBsAg] - Reverse Passive Haemagglutination [RPHA], Hepatitis B core Antibody [HBcAb] - Passive Haemagglutination [PHA], Alpha-feto Protein - RPHA), Hepatitis B e Antigen/Antibody [HBeAg/Ab] - Radioimmunoassay [RIA], Hepatitis C antibody [HCV-Ab] - Enzyme Immunosorbent Assay [EIA]. Due to the high specificity and sensitivity of the Polymerase Chain Reaction technique [PCR] in detecting the viral genomes, it was used to establish the presence of the HBV-DNA and HCV-RNA to correlate the serological diagnosis of their respective seromarkers. A total of 39 serum samples were tested comprising 11 blood donors, 8 chronic liver disease patients and 20 hepatocellular carcinoma cases. 4/19 (21%) HCV-antibody (C-l) reactive samples were found to be positive for HCV-RNA by PCR. 14 of the 19 (73.7%) including the 4 HCV-RNA positive cases tested positive for HBcAb. 6 of 11 (55%) HBsAg positive cases also tested positive for HBV-DNA by PCR, In 8 of 20 (40%) hepatocellular carcinoma cases, no aetiological role could be assigned to hepatitis B or C as only HBcAb was demonstrated in those cases.

Evidence of sensory conflict and recovery in carp exposed to prolonged weightlessness.

BACKGROUND: Evidence in support of the sensory conflict hypothesis for space motion sickness (SMS) is still needed. HYPOTHESIS: We hypothesized that sensory conflict and recovery processes should be demonstrated in intact fish during initial days of microgravity exposure, as a disturbance and restoration of the dorsal light response (DLR; a functional model of visual-graviceptor interaction), respectively. We also expected that this would be true in an otolith-removed fish if it had been fully compensated for dysfunction before the exposure. METHODS: The DLR of carp (Cyprinus carpio) was examined intermittently during the 8-d mission of Spacelab-J. Two carp, normal and labyrinthectomized (LB), made the flight. RESULTS: In the normal carp, the DLR was unstable for the first 3 d inflight but gradually recovered thereafter. The recovery was characterized by gradual restoration of the DLR tilt speed. The LB fish whose otoliths had been removed 2 mo before the flight maintained DLR at the first inflight test session (22 h after launch), but DLR was disrupted at 2 d as in normal fish. The recovery process could not be evaluated in this fish, because the EEG cable which was attached to the fish for supplementary study became tightly twisted and thus immobilized the fish for the remainder of the experiment. CONCLUSION: These findings provided additional evidence in fish for sensory-motor disorder and readjustment during the early phase of microgravity, thus supporting the sensory conflict hypothesis for SMS.

Serum level of macrophage colony-stimulating factor as a marker for gynecologic malignancies.

We determined the serum levels of macrophage colony-stimulating factor (M-CSF) in 441 women with gynecologic diseases to evaluate its role as a marker for gynecologic malignancy. Serum M-CSF levels were above the normal baseline level of 1,056 U/ml in 64% (56/88) of patients with ovarian cancer, 27% (16/60) of those with cervical cancer, and 25% (15/61) of those with endometrial cancer. M-CSF was significantly elevated in the serum of patients with advanced as compared with early stage cancer (stage I) of the ovary (p < 0.01), cervix (p < 0.05), and endometrium (p < 0.05). Only 5.6% of the patients with benign ovarian tumors and 7.0% of those with endometrial cysts had serum levels of M-CSF that exceeded 1,056 U/ml. M-CSF was localized in the glandular epithelial cells as well as in the stromal macrophages and the endothelial cells of the ovarian cancers. M-CSF thus appeared to be a marker with high specificity for ovarian cancer.

Branch retinal vein occlusion decreases the potential of the inner retinal layer.

To identify the affected retinal layer in cases of branch retinal vein occlusion (BRVO), we analyzed the scotopic electroretinogram (ERG) and electrooculogram (EOG) obtained on the same day in 30 patients with unilateral BRVO. Patients with eye diseases other than unilateral BRVO were excluded. A dome-shaped Ganzfeld illuminator was used for scotopic ERG stimulation and EOG illumination under conventional recording conditions. There was no significant difference in the mean values of scotopic ERG variables between the eyes with BRVO and the normal eyes, but the EOG light peak and L/D ratio were significantly lower in the eyes with BRVO (p < 0.05). However, there was no significant difference between eyes with BRVO and normal eyes regarding the dark through value. Since the light rise potential is induced by background illumination, the low values for light peak and L/D ratio in the eyes with BRVO represent a decrease in the light rise potential compared with that of normal eyes. In addition, since the b wave amplitude of the ERG was normal in the eyes with BRVO, the scotopic ERG and EOG tests suggest that generalized mild hypoxia affects the inner retinal layer of the eyes with BRVO.

Space experiment using large-sized fish: in case of carp in Spacelab-J mission.

Two carp of 26 cm in size, intact and otolith-removed (LB), were flown on the Spacelab-J for 8 days in September 12-20, 1992. Light-dependent reaction to alternating direction of illumination was recorded for 10 min twice a day together with the cerebellar EEG activity, on 2 days before the launch, during the flight and for 4 days after the landing, in same fish chamber. Reproducing the video image, it was revealed that both carp were healthy during the mission, but the LB fish was almost immobilized from the 3rd test session (48 hours in flight) by tight twisting of the EEG cable. Both fish after landing tended to stay still on the bottom of the fish chamber. Findings that the body weight reduced remarkably in both fish and that nitrite and nitrate levels of the fish water were usually high, suggested that the fish metabolism might have been enhanced during the flight.

Mortality among inhabitants of an HTLV-I endemic area in Japan.

A community-based cohort study was conducted to clarify the risk of human T-cell leukemia virus type I (HTLV-I) infection for cause-specific deaths. A total of 1,997 individuals (751 men and 1,246 women) aged 30 or older in A-Island, Nagasaki Prefecture, Japan who had voluntarily attended annual mass health examinations, including serum HTLV-I antibody test, were followed up for a mean period of 5.3 years. In a Cox proportional hazards analysis adjusted for age at baseline, the HTLV-I seropositivity was found to be associated with mortality from all causes in men (hazard ratio (HR) 1.89; 95% confidence interval (CI) 1.01-3.54) and women (HR 1.94; 95% CI 1.16-3.22). When the effects of 2 deaths (1 man and 1 woman) from adult T-cell leukemia/lymphoma (ATL) were excluded, the mortality risk decreased slightly but was still significantly or marginally significantly greater than 1 in both men (HR 1.77; 95% CI 0.93-3.37) and women (HR 1.87; 95% CI 1.12-3.12). Further analysis of cause-specific deaths revealed a significant increase in the risk for non-neoplastic diseases but not for neoplasms excluding ATL. These findings suggest that long-term HTLV-I infection represents a health hazard greater than just that for the development of ATL. It was difficult, however, to draw a conclusion regarding the association between HTLV-I infection and cancer risk, because the number of cancer deaths was small and the incidence of cancer was not investigated.

Macrophage colony-stimulating factor as a tumor marker for epithelial ovarian cancer.

OBJECTIVE: To determine the serum level of macrophage colony-stimulating factor in ovarian cancer patients in order to evaluate its role as a marker for ovarian cancer. METHODS: Serum macrophage colony-stimulating factor levels were assayed in 69 patients with epithelial ovarian cancer, 55 with benign ovarian tumors, and 634 healthy individuals, including 398 women, using an enzyme-linked immunosorbent assay. RESULTS: The average serum macrophage colony-stimulating factor level was 754.4 +/- 153.9 U/mL in healthy females; 1056 U/mL (mean plus 1.96 standard deviations) was considered to be the upper limit of normal. Serum macrophage colony-stimulating factor levels were significantly elevated in patients with ovarian cancer (average 1460.5 +/- 1006.2 U/mL; P < .001) and exceeded 1056 U/mL in 42 of the 69 patients with ovarian cancer (61%). No differences in levels were observed among the histologic types. No definite relationship was found between serum levels of macrophage colony-stimulating factor and those of CA 125. We found that 96% of the patients with ovarian cancer had high serum levels of macrophage colony-stimulating factor and/or CA 125 values. There was no significant difference in the levels of macrophage colony-stimulating factor between patients with benign ovarian tumors and healthy controls. Only 7.3% of the group with benign tumors had levels exceeding 1056 U/mL. CONCLUSION: Macrophage colony-stimulating factor is a marker for ovarian cancer. Determination of serum levels can be useful in detecting ovarian cancer, particularly in combination with CA 125.

Absence of HTLV-I infection among seronegative subjects in an endemic area of Japan.

To examine the prevalence of infection by human T-cell leukemia virus type I (HTLV-I) among seronegative subjects, healthy subjects on Tsushima Island, Japan, where the infection is endemic, were evaluated. A total of 209 healthy adults were examined for HTLV-I provirus in peripheral blood mononuclear cells by the polymerase chain reaction (PCR), as well as for anti-HTLV-I antibodies by the particle agglutination (PA) method, the enzyme-linked immunosorbent assay (ELISA) and by immunofluorescence analysis (IF). A total of 76 subjects were positive and 133 were negative for the provirus, showing a close correlation with the results of 3 assays for anti-HTLV-I serum antibodies. None of the seronegative subjects reacted positively on PCR analysis. These observations indicate that seronegative HTLV-I carriers are rare in an area of Japan in which this viral infection is endemic.

Magnetic resonance imaging of retained surgical sponges. Case report.

The magnetic resonance imaging (MRI) features of sponges retained postsurgically in three patients are described. MRI depicted these as well-defined round masses, which were of low signal intensity on T1-weighted images and very high signal intensity on T2-weighted images. We scrutinized these masses on T2-weighted images. MRI revealed the low-signal-intensity structures to have wavy, striped, and/or spotted appearances, which raised the suspicion that they might be retained surgical sponges within mass lesions.

Cloning and nucleotide sequences of the AccI restriction-modification genes in Acinetobacter calcoaceticus.

The genes of the AccI restriction-modification system specific for GT(A/C) (G/T)AC were cloned from the chromosomal DNA of Acinetobacter calcoaceticus, and their nucleotides sequenced. The restriction and modification genes coded for polypeptides with calculated molecular weights of 42,494 and 63,078, respectively. Both the enzymes were coded by the same DNA strand and the restriction gene was upstream of the methylase gene, separated by 2 bp. The restriction gene was significantly expressed in E. coli cells, so that the AccI restriction endonuclease could be purified to homogeneity. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration indicated that the catalytically active form of the endonuclease was tetrameric. Sequence comparison with related enzymes indicated that AccI methylase contained a segment of tetra-amino acids, NPPY, characteristic of N6-adenine methylases. In addition, some homologous regions were found in the sequence of HincII methylase specific for GT(C/T) (A/G)AC.

Epidemic patterns of hepatitis type B virus (HBV) and human T lymphotropic virus type I (HTLV-I) in two ATL-endemic islands in Kyushu, Japan.

To evaluate the relationships between human T lymphotropic virus type I (HTLV-I) and hepatitis virus type B (HBV) infections, we compared both individual and geographical distributions of carriers of HTLV-I and HBV, and antibody positives to HBV in two ATL-endemic islands in Kyushu, Japan. The positive rates of antibodies to HTLV-I (anti-HTLV-I-Ab) in sera among healthy inhabitants older than 30 years of age were 27.5% (617/2,232) in Nakadohri, Goto Island and 24.0% (500/2,048) in Shimo-agata, Tsushima Island. The positive rates for surface antigen of hepatitis B virus (HBs-Ag) in sera among the same subjects were 6.4% and 2.5%, respectively. In Nakadohri, the age-adjusted positive rates of HBs-Ag and antibody to HBs-Ag (anti-HBs-Ab) in anti-HTLV-I-Ab positives (so-called HTLV-I carriers) differed little from those in negatives. In Shimo-agata, the geographical distribution of HBs-Ag positives (so-called HBV carriers) showed no positive association with that of HTLV-I carriers. These results did not support the positive correlations between HTLV-I and HBV infections among the general population in ATL-endemic areas in Japan.

Epidemiological features of HTLV-I carriers and incidence of ATL in an ATL-endemic island: a report of the community-based co-operative study in Tsushima, Japan.

To extend the epidemiological study on adult T-cell leukemia-lymphoma (ATL) in Japan, the geographical and demographic characteristics of carriers of human T-lymphotropic virus type I (HTLV-I) and patients with ATL were analyzed in Tsushima Island, which is one of the typical endemic areas of ATL in Kyushu, Japan. Even on the small island of Tsushima (710 km2; pop. 48,875; 123 villages), the positive rates of anti-HTLV-I antibody among the 58 villages studied varied from 2% to 50%, a fact that is probably associated with the historical events affecting the movement of the indigenous population of Tsushima Island. The positive rate of anti-HTLV-I antibody in males increased little with age; however, the female rate increased distinctly with age in moderate and high HTLV-I-endemic villages where more than 15% of the inhabitants had positive anti-HTLV-I antibody. Analysis of anti-HTLV-I antibody positivity between spouses confirmed that HTLV-I was more contagious from husband to wife than from wife to husband, which corresponded to the fact that the positive rate of anti-HTLV-I antibody in females older than 30 years was higher than that in males. Recently the rate of carrier children from HTLV-I carrier mothers was estimated at around 20%. The recent annual incidence rates of ATL among 1,000 HTLV-I carriers older than 40 years living in Tsushima Island was estimated at 2.2 in males, 0.8 in females.