Molecular cloning and characterization of two sets of alpha-theta genes in the rat alpha-like globin gene cluster.

The highly heterogeneous rat hemoglobin system was investigated at the gene level. Two regions of the alpha-like globin gene cluster from a Wistar rat were isolated. Four lambda Dash recombinant clones carrying rat alpha-like globin genes were localized on two distinct gene regions. A region of approximately 16kb was found to contain the 5'-IIalpha1-psi theta 1-3' loci, and another of approximately 24kb the 5'-IIalpha2-psi theta2-psiI alpha3-3' loci. Both IIalpha1 and IIalpha2 are considered to be active, coding the IIalpha-globin chain. The nt sequences of IIalpha1 and IIalpha2 are identical except for six nt in the non-coding region. The psiI alpha3 locus is a truncated pseudogene. The putative promoter region of an alpha-like globin gene is joined directly to the third exon, homologous to that of Ialpha-globin cDNA. psi theta1 and psi theta2 are also pseudogenes, as evidenced by several deletions located in the protein-coding regions of these loci. The psi theta1 and psi theta2 loci exhibit extensive homology, but the restriction maps of these genes and their flanking regions differ considerably. Genomic Southern blot analyses of the total liver DNA from six rats showed the existence of three theta-globin-related genes, including psi theta1 and psi theta2. These results indicate that the two gene regions investigated are not allelic variants, but may be generated by block duplication. This is the first report of the existence of rodent theta-globin genes.

Cardiodynamic and neurohormonal importance of atrial contribution in rate-responsive pacing.

To elucidate the physiologic importance of atrial contribution in recently developed rate-responsive pacing, changes in cardiodynamics and neurohormonal factors were analyzed during exercise in patients with respiratory rate-dependent, rate-responsive atrial (AAIR; n = 6) and ventricular (VVIR; n = 9) demand mode pacemakers implanted for sick sinus syndrome. With increasing pacing rate during bicycle ergometer exercise, the AAIR group had significant increases in cardiac index (p < 0.05), left ventricular ejection fraction (p < 0.05), and ejection (p < 0.05) and peak filling (p < 0.05) rates; however, the VVIR group had a significant decrease in ejection fraction (p < 0.05), and an increase in cardiac index (p < 0.05) that was significantly less than in the AAIR group. At rest, the mean plasma concentrations of atrial natriuretic peptide (p < 0.005) and cyclic guanosine monophosphate (p < 0.05) were significantly greater in the VVIR group than in the AAIR group and normal subjects (n = 8). Atrial natriuretic peptide, norepinephrine, and cyclic adenosine and guanosine monophosphates were significantly greater (p < 0.05) during exercise, and atrial natriuretic peptide was significantly greater in the VVIR group (207.5 +/- 8.3 pg/ml) than in the AAIR group (116.4 +/- 51.5) and normal subjects (30.8 +/- 19.2; p < 0.05); this suggested a further increase in the nonphysiologic atrial overload with VVIR pacing. The data show both the neurohormonal and cardiodynamic importance of atrioventricular synchrony in rate-responsive pacing.

Memory of chirality in diastereoselective alpha-alkylation of isoleucine and allo-isoleucine derivatives.

[reaction: see text] alpha-Methylation of 3 gave 5 as a major product whereas 4 gave 6 predominantly, although both 3 and 4 have an (S)-chiral center at C(3). This indicates that chirality at C(2) in 3 and 4 was memorized in the corresponding intermediate enolates and the induced chirality made a major contribution in the stereochemical course of the reaction, while chirality at the adjacent chiral center C(3) had little effect.

Micronucleus test and erythropoiesis: effect of cobalt on the induction of micronuclei by mutagens.

The micronucleus test is used widely as an in vivo short-term assay for potential carcinogens. In the present study, results of the micronucleus test were affected by cobalt dichloride pretreatment. Cobalt dichloride was used to induce erythropoietin, a growth factor for erythropoiesis. The increase in mutagen-induced micronucleus response following cobalt pretreatment, therefore, may have been due to a change in the rate of erythropoiesis. The greatest interaction between cobalt pretreatment and mutagen treatment for the induction of micronucleated polychromatic erythrocytes (MPCE) occurred when mice were injected with 1,1-dimethylhydrazine (DMH) 12-24 hr after pretreatment with cobalt dichloride and killed 30 hr later. Increased sensitivity of the micronucleus test was attributable to the administration of mutagen during the differentiation and multiplication of erythroblast, which is presumed to have been accelerated by pretreatment with cobalt dichloride. An increased induction of MPCE in the bone marrow by two chemicals--benzo(a)pyrene, 2-naphthylamine--was also observed following pretreatment with cobalt dichloride.

Effect of erythropoietin on the micronucleus test.

The micronucleus test is used widely as an in vivo short-term assay for potential carcinogens. In the present study, results of the micronucleus test were affected by the rate of erythropoiesis in the bone marrow erythropoietin, a growth factor for the erythroblast, which was used to induce erythropoiesis. The highest frequency of micronucleated polychromatic erythrocytes (MPCE) and a dose-response relationship between erythropoietin doses and MPCE frequency were seen 30 hr after injection of 1,1-dimethylhydrazine (DMH) to mice administered 24 hr previously with erythropoietin. The effect of erythropoietin was maximal when erythropoietin was given 24 hr before DMH, indicating that accelerating the multiplication of erythroblasts will increase the frequency of micronuclei induced by mutagens. Induction of MPCE in the bone marrow by four other compounds--benzo(a)pyrene, 2-naphthylamine, mitomycin C, and vincristine--was also increased by pretreatment with erythropoietin.

Progressive outer retinal necrosis after bone marrow transplantation.

PURPOSE: To describe a case of progressive outer retinal necrosis in a human immunodeficiency virus-negative immunosuppressed patient undergoing treatment for graft-versus-host disease after bone marrow transplantation. METHOD: Onset, course, and outcome of the patient's eye disease were analyzed. RESULTS: Findings and course were typical for the recently described progressive outer retinal necrosis syndrome. CONCLUSION: Progressive outer retinal necrosis is not limited to the acquired immunodeficiency syndrome but may occur in patients immunocompromised because of other conditions.

Optical coherence tomography patterns of choroidal osteoma.

PURPOSE: To determine the optical coherence tomographic images that are commonly observed in eyes with choroidal osteoma (choroidal ossification). METHODS: Three patients with choroidal osteoma were examined by optical coherence tomography. RESULTS: We found two optical coherence tomographic patterns in the eyes with choroidal osteoma. First, multiple tracks of high refractivity were present posterior to the tumor lesion. Second, thick and irregular plate-like, high-signal intensity areas were present in the choroid in the region of the tumor. CONCLUSIONS: Optical coherence tomography can be useful in the diagnosis of choroidal osteoma.

Effect of dietary alpha-linolenate/linoleate balance on mean survival time, incidence of stroke and blood pressure of spontaneously hypertensive rats.

Following the suckling period, stroke-prone spontaneously hypertensive rats (SHR-SP) were fed semi-purified diets supplemented either with safflower seed oil (rich in linoleic acid) or with perilla seed oil (rich in alpha-linolenic acid). The mean survival time of male SHR-SP fed the perilla diet was longer than that fed the safflower diet by 17% (p less than 0.001) while the difference was 15% in female SHR-SP (p less than 0.05). The mean survival times of female SHR-SP were more than 40% longer than those of male SHR-SP in both dietary groups. Post-mortem examinations of brains revealed apoplexy-related symptoms as the major cause of the death in both dietary groups. The systolic blood pressure was lower by ca. 10% (21 mmHg) in the perilla group than in both the safflower group and conventional diet group. The eicosapentaenoate (20:5 n-3)/arachidonate (20:4 n-6) ratio of platelet phospholipids in spontaneously hypertensive rat (SHR), a measure of platelet aggregability, was much higher in the perilla group than in the safflower group. Thus, increasing the dietary alpha-linolenate/linoleate ratio resulted in an increased mean survival time of SHR-SP rats, possibly by lowering blood pressure and platelet aggregability.

Effect of estrogen on induction of micronuclei by mutagens in male mice.

The effect of estrogen on the induction of micronucleated polychromatic erythrocytes (MPCE) by mutagens was examined in male mice. In the dose-response study, a dose-related inhibition of the mitomycin C (MMC)-induced MPCE frequency by estradiol (E2) treatment was observed. In the time study, inhibitory effects of E2 on MPCE frequency by MMC were observed when MMC was administered at 0 or 1 day after injection of E2. The most effective protocol for inhibition was when E2 and MMC were used on the same day. Of mutagens other than MMC, only vincristine (VCR) showed a significant decrease in MPCE frequency when used together with E2. Benzo[a]pyrene (BaP) and 5-fluorouracil (5-FU) showed no significant decrease in MPCE frequency. The data suggest that the induction of micronuclei by mutagens is inhibited by treatment with estrogen, and this could result in a sex difference in the sensitivity of mice employed in the micronucleus test. Mechanisms of the inhibitory effects of estrogen are discussed; these might include a suppression of erythropoiesis and a possible effect on the cell membrane permeability of erythroblasts.

Effect of estrogen on liver plasma membrane in rats.

The effect of estrogen on plasma membrane was investigated using the primary cultured rat hepatocytes treated with carbon tetrachloride (CCl4) and the isolated plasma membrane of rat liver. 17 beta-Estradiol (E2), at concentrations of 10(-10) M to 10(-4) M, 10(-8) M to 10(-6) M and 10(-12) M to 10(-4) M, had an inhibitory effect on the CCl4-induced leakage of glutamic oxaloacetic transaminase, glutamic pyruvic transaminase and lactate dehydrogenase, respectively from primary cultured rat hepatocytes. Diethylstilbestrol, which caused inhibition at a dose of 10(-4) M, did not inhibit any enzyme leakage at any further concentrations of 10(-12) M to 10(-6) M. In the isolated plasma membrane of rat liver, Mg(2+)- and Na+,K(+)-adenosine triphosphatase activity was increased by E2 treatment at concentrations of 10(-6) M and 10(-4) M.

Effect of estrogen replacement on liver function in ovariectomized rats.

The effect of estrogen (17 beta-estradiol; E2) replacement on the liver function was examined in ovariectomized rats. Ovariectomy decreased serum transaminase activity and physiological dosage levels of E2 replacement returned the activity up to or above non-ovariectomized levels. Under the conditions of this experiment the elevated serum transaminase levels in the E2 replacement were not coincident with histological changes suggestive of liver dysfunction. The elevations of both serum glutamic oxaloacetic transaminase (GOT) and serum mitochondrial GOT were accompanied by increased hepatic tissue activity, suggesting that the elevated serum levels can be attributed to the accelerated synthesis of the enzymes. On the other hand, the elevation of serum glutamic pyruvic transaminase was not accompanied by increased tissue activity in the original organs, namely liver, kidney, heart and skeletal muscle. Ovariectomy decreased serum albumin and inhibited development of rough endoplasmic reticulum in the liver, and E2 replacement returned them up to or above the non-ovariectomized levels.

Collaborative work to evaluate toxicity on male reproductive organs by repeated dose studies in rats 4). Fadrozole hydrochloride: an oral 2/4-week male reproductive organ toxicity study.

Doses of 0, 30 and 60 mg/kg/day of fadrozole hydrochloride (Afema: non-steroidal aromatase inhibitor, antitumor agent) were given perorally by gavage to HanIbm WIST male rats from 6 or 8 weeks of age for 2 weeks, and from 6 weeks of age for 4 weeks. In all treatment groups, reduced weights of seminal vesicle, prostate and epididymis, and degeneration/necrosis of the pachytene spermatocytes in stages VII or VIII seminiferous tubules, were dose-relatedly observed. Effects could also be assessed quantitatively by staging analysis with the result of a reduction in the numbers of stage VII pachytene spermatocytes at 30 and 60 mg/kg/day. Epididymal sperm examination revealed no treatment-related changes in any groups. The effects of 4-week treatment on male reproductive organs were similar to those of 2-week treatment at the same dose levels, except for the weights of seminal vesicle and prostate, which were more reduced by 4-week treatment than by 2-week treatment. There was no notable difference in detectability of toxicity in male reproductive organs between 2-week treatment from 6 weeks of age and 2-week treatment from 8 weeks of age. It was concluded that the changes observed in the rat male reproductive organs following 4 weeks of treatment with fadrozole hydrochloride could be detected also with 2 weeks of treatment.

Ligneous conjunctivitis: a case report.

BACKGROUND: Ligneous conjunctivitis is a rare condition characterized by chronic, recurrent conjunctivitis associated with pseudomembrane, and it may involve other mucous membranes in the mouth, nasopharynx, trachea, and vagina. We examined and treated a case of presumed ligneous conjunctivitis. CASE: The patient was a 10-year-old boy. His chief complaints were visual impairment, discomfort, and discharge, but no itching in his eyes. His upper eyelids appeared thick without swelling. He had a past history of surgery for lid entropion. His two siblings had similar follicular conjunctivitis. OBSERVATIONS: This case exhibited several characteristics of ligneous conjunctivitis, such as large follicles, recurrent pseudomembrane and normal level IgE in the serum. Indispensable characteristics of vernal keratoconjunctivitis, strong itching, and extensive papillary formation, were not found. In spite of the lack of woody hardness of the conjunctiva, other clinical findings led to the diagnosis of ligneous conjunctivitis. Definite histological diagnosis was not obtained, because of the lack of common histological characteristics among previously reported cases with ligneous conjunctivitis. The boy had developed corticosteroid glaucoma after instillation of dexamethasone 0.1% for 7 months at a previous time. We successfully treated this case with combined instillation of fluorometholon and cyclosporin after trabeculotomy. CONCLUSIONS: Ligneous conjunctivitis must be considered as one type of differential diagnosis of vernal keratoconjunctivitis. Cyclosporin is an effective alternative for the treatment of ligneous conjunctivitis, especially in a case with a possible history of corticosteroid glaucoma.

[The diagnostic significance of polymerase chain reaction for ocular samples in viral retinitis].

During the past two years we studied the incidence of infection by eight members of the herpesvirus family in ocular samples (tear fluid, the aqueous, and the vitreous) using the polymerase chain reaction (PCR). A total of 31 ocular samples were collected from 22 eyes of 13 patients. The series comprised five cases of acute retinal necrosis, four of cytomegalovirus retinitis, three of proliferative diabetic retinopathy, and one of ocular malignant lymphoma. In 12 eyes of 9 patients with viral retinitis, causative viral DNA was detected either from the tear fluid (1/12, 8%), the aqueous (6/7, 86%), or the vitreous (1/1, 100%). In the fellow eyes free of viral retinitis, no viral DNA was detected in the samples (6 of tear fluid and one of the aqueous). Out of the other 4 patients without viral retinitis, viral DNA was detected in one patient with ocular malignant lymphoma. The findings show that PCR is a useful and sensitive method in the diagnosis of viral retinitis, but that it may give false positive results. The aqueous and the vitreous samples gave more positive results than tear fluid.

[The effect of recombinant epidermal growth factor in corneal angiogenesis].

Recent advances in genetic engineering techniques have enabled large-scale manufacture of human epidermal growth factor (hEGF), making possible the clinical use of this particular agent in treating a variety of corneal epithelial disorders. In view of future application to humans, it has to be determined whether hEGF could induce neovascularization in the cornea upon topical instillation, since the angiogenic effect of mouse EGF on the cornea in vivo has been reported. For this, a sheet of slow-release form polymer (EVA) containing hEGF was surgically implanted into the rabbit corneal stroma in search for subsequent corneal neovascularization. EVA sheets contained one of the following agents: (1) 250ng hEGF, (2) 500ng hEGF, (3) 250ng bFGF (positive control), (4) vehicle alone (negative control). On 5 and 14 days after implantation, the corneas were excised, sectioned, and stained with hematoxylin and eosin for histological evaluation. Slit lamp examination revealed that marked neovascularization developed in the corneas when EVA sheets containing bFGF were implanted. A number of polymorphonuclear leukocytes were accumulated around the implants. However, neovascularization did not occur in the corneas when EVA sheets containing either concentrations of hEGF or vehicle alone were implanted. Only a few polymorphonuclear leukocytes infiltrated. This result indicates that as much as 500ng hEGF does not induce corneal neovascularization.

Highly time-resolved evaluation technique of instantaneous amplitude and phase difference using analytic signals for multi-channel diagnostics.

A fluctuation analysis technique using analytic signals is proposed. Analytic signals are suitable to characterize a single mode with time-dependent amplitude and frequency, such as an MHD mode observed in fusion plasmas since the technique can evaluate amplitude and frequency at a specific moment without limitations of temporal and frequency resolutions, which is problematic in Fourier-based analyses. Moreover, a concept of instantaneous phase difference is newly introduced, and error of the evaluated phase difference and its error reduction techniques using conditional/ensemble averaging are discussed. These techniques are applied to experimental data of the beam emission spectroscopic measurement in the Heliotron J device, which demonstrates that the technique can describe nonlinear evolution of MHD instabilities. This technique is widely applicable to other diagnostics having necessity to evaluate phase difference.

Density fluctuation measurements using beam emission spectroscopy on Heliotron J.

This paper describes the measurement of the density fluctuation using beam emission spectroscopy in Heliotron J, having the non-symmetrical helical-magnetic-axis configuration. In order to optimize the sightlines, the numerical calculations are carried out to estimate the spatial resolution and the observation location. When a tangential neutral beam is used as diagnostic one, suitable sightlines from the newly installed diagnostic port are selected whose spatial resolution Δρ is less than ± 0.07 over the entire plasma region. Modification of the interference filter and the detection systems enables us to measure the radial profile of the density fluctuation. Each of the three coherent modes due to the fast-ion-driven magnetohydrodynamic instabilities has different radial structure of the density fluctuation.