Circulating levels of Interferon-Gamma in patients with neovascular age-related macular degeneration in Yogyakarta.

INTRODUCTION: Neovascular age-related macular degeneration (nAMD) is a major factor contributing to blindness and impaired visual acuity in elderly people. The pathophysiology of nAMD involves excessive inflammation events in the macula. Thus, it is crucial to study the dynamics of an important pro-inflammatory cytokine, interferon-gamma (IFN-γ). MATERIALS AND METHODS: This research is aimed to investigate plasma IFN-γ profiles of patients with nAMD. In this crosssectional study, blood plasma samples of 16 patients with AMD and 23 age-matched controls were collected. Samples were examined for two inflammatory cytokines (IFN-γ) using a commercially available enzyme-linked immunosorbent assay.Acquired data were log transformed to normalize any outliers before conducting student'st-test using the SPSS software. RESULTS: IFN-γ levels were higher in the control group, without statistically significant difference between the two groups. CONCLUSION: IFN-γ levels were not significantly different between patients with AMD and controls.

Loss of immunostaining of the RBP-J kappa transcription factor upon F9 cell differentiation induced by retinoic acid.

RBP-Jkappa is a novel type of transcriptional regulatory protein that does not contain any known DNA-binding motif. We raised anti-RBP-Jkappa monoclonal antibodies (K0043 and T6709) to investigate the roles of RBP-Jkappa in cell differentiation. These antibodies stained nuclei of undifferentiated embryonic stem cells and F9 cells but not those of the other differentiated cell lines tested so far although the RBP-Jkappa protein is expressed at similar levels. Interestingly, differentiated F9 cells lost the immunostaining reaction with the anti-RBP-Jkappa monoclonal antibodies. Biochemical subcellular fractionation study showed that the majority of RBP-Jkappa was localized in nuclei of F9 cells and that there are at least two forms of the RBP-Jkapppa protein in the nuclei of undifferentiated F9 cells, a free form and a chromatin-bound form. Upon induction of F9 cell differentiation, free nuclear RBP-Jkappa disappeared concomitantly with the loss of immunostaining, suggesting that the anti-RBP-Jkapppa antibodies cannot recognize chromatin-bound RBP-Jkappa. Since there is no evidence to indicate covalent modification of RBP-Jkappa, we assume that chromatin-bound RBP-Jkappa interacts with a large number of proteins which block the exposure of RBP-Jkappa epitopes to the monoclonal antibodies.

Neovascularization of the iris in rhegmatogenous retinal detachment.

To identify conditions associated with neovascularization of the iris in rhegmatogenous retinal detachment, we examined 36 eyes with this disorder seen at our hospital between 1979 and 1990. Clinical courses of disease were divided into the following three groups: (1) neovascularization of the iris without a history of a vitreoretinal operation (four eyes), (2) neovascularization of the iris after an unsuccessful vitreoretinal operation (26 eyes), and (3) neovascularization of the iris after surgical complications (six eyes). In all eyes of Groups 1 and 2, retinal detachment persisted at the onset of iris neovascularization; however, in six eyes, iris neovascularization subsided after retinal reattachment. Characteristic features of Groups 2 and 3 were patient age of 50 years or more, severe myopia, a history of increased intraocular pressure, a history of choroidal detachment, and a large scleral buckle.

Rapid serum vancomycin assay by high-performance liquid chromatography using a semipermeable surface packing material column.

A new isocratic high-performance liquid chromatographic (HPLC) assay has been developed for vancomycin that uses direct injection of microquantities of serum into a separation column filled with octyl-C(8) silica support that has a semipermeable surface. A mixture of disodium hydrogen phosphate buffer (pH 7.0) and acetonitrile is used as the mobile phase, and vancomycin is directly detected at 240 nm. The minimum limit of detection was 0.5 microg/ml at a signal-to-noise ratio of 3:1. Linearity was established from 0 to 100 microg/ml. The coefficient of variation for within-run reproducibility was 1.1-2.7% for a concentration range of 2.9-52.5 microg/ml; for day-to-day reproducibility it was 4.0% and 3.1% for a concentration range of 5.8-26.4 microg/ml, and the recovery rate was 94-105%. There was no interference from 41 antibiotics or other drugs currently in use. The correlation coefficient between the fluorescence polarization immunoassay (x) and this method (y) was 0.995 with a linear equation, y = 1.06x - 0.924. This method is simple, rapid, and provides an economical quantification of serum vancomycin.

Identification of trypanosomes isolated by KIVI from wild mammals in Côte d'Ivoire: diagnostic, taxonomic and epidemiological considerations.

In Côte d'Ivoire, a comparative study was carried out on 122 wild mammals by parasitological and serological examination and by in vitro isolation of trypanosomes from fresh blood (KIVI). Thirteen isolated stocks were studied by isoenzymes and compared with Trypanosoma congolense and T. brucei bouaflé group reference stocks. Of the 122 animals, only 22 were positive on blood smears while 88 were KIVI positive and 92 were CATT/T. b. gambiense positive. For six stocks identified by isoenzymes as T. congolense, the agreement between ELISA and CATT was good (75%). As compared with CATT, antigen detection ELISA was not satisfactory for T. brucei (20%). Out of 18, 16 stocks represented a separate zymodeme (seven T. congolense and nine T. brucei) and a high genetic heterogeneity was observed. For T. congolense, savanna, kilifi and forest groups were represented by one zymodeme each. The four remaining zymodemes while put into this T. congolense group, were strongly independent of each other. Morphology indicated that those new zymodemes correspond to T. congolense. In the other hand, five new zymodemes fit into T. brucei classification.

Survey of 1120 Japanese women with a history of recurrent spontaneous abortions.

Our recent 7-year clinical survey showed that among the 1120 women with repeated spontaneous abortions registered in this clinic, 2898 out of a total of 3216 pregnancies (90.1%) had terminated in spontaneous abortion. Among these wastages, 84.2% occurred before 12 weeks of gestation, and 11.1 percent occurred between 12 and 15 weeks. Through routine examination of reproductive wastage, 82 (9.9%) of the 825 Japanese couples examined were shown to have either a chromosomal abnormality or normal variants in the wife and/or husband, thus demonstrating no racial difference in the incidence of chromosomal abnormalities in infertile patients in comparison with studies performed in other countries. One hundred and forty-seven congenital uterine anomalies (14.7%) were found in 1000 hysterosalpingographies, and 12 of 148 examined females were positive for anti-cardiolipin antibody. 393 other females with no major abnormalities likely to induce spontaneous abortions were indicated for immunotherapy. Ample time spent on genetic counseling prevented further reproductive wastage, and ideal metroplasty resulted in a successful post-operative pregnancy course in more than 85% of cases. Immunosuppressant and anticoagulant therapy decreased the serum titer of anti-cardiolipin antibody, enabling pregnancies to be maintained to term. Immunotherapy utilizing the husband's lymphocytes also brought more than 80% of pregnancies to successful completion, with 200 deliveries achieved with this therapy. In contrast, 64.1% of pregnancies again terminated spontaneously in patients who were indicated for immunotherapy but did not receive treatment. The findings of the present study suggest that the causes of reproductive wastage, especially the etiology of early recurrent spontaneous abortion, are complex.(ABSTRACT TRUNCATED AT 250 WORDS)

Seasonal epidemiology of ticks and aspects of cowdriosis in N'Dama village cattle in the Central Guinea savannah of Côte d'Ivoire.

In the Central Guinea savannah of Côte d'Ivoire, cattle breeding started only approximately 30 years ago. The impact of parasitism on the overall health status and productivity of the trypanotolerant N'Dama cattle in this area is unknown. In close collaboration with national veterinary institutions and local farmers, we studied spectrum, burden and seasonal dynamics of ticks (including aspects of cowdriosis) on N'Dama village cattle. In a longitudinal study, three randomly selected cattle herds (traditional farming type) of one village were examined repeatedly for ticks. Spectrum, burden, seasonal epidemiology of ticks were assessed. In these traditional herds (which lack (ecto)parasite management), all animals were infested by ticks at monthly counts. Five different tick species were identified; the four genera in order of frequency were: Amblyomma (overall prevalence 96%), Boophilus (47%), Hyalomma (<1%) and Rhipicephalus (<1%). Amblyomma variegatum was the most-abundant tick on cattle in all seasons. Seroprevalence of Cowdria ruminantium was 31% (95% CI: 26, 36%). Most of the animals typically carried low tick burdens. N'Dama cattle seem well adapted to their environment and can resist the tick burdens under this traditional farming system.

Risk of retinal detachment in patients with lattice degeneration.

To determine the risk of retinal detachment in patients with lattice degeneration of the retina, we statistically analyzed the incidence of retinal detachment in these patients. The data of hospital patients with retinal detachment associated with lattice degeneration in Kumamoto Prefecture, Japan, in 1990 were collected. The prevalence of lattice degeneration in Kumamoto was reported to be 9.5% in 1980. Based on population data from the 1990 census, the cumulative incidence of retinal detachment associated with lattice degeneration was calculated in this study. Among 1,840,000 residents in Kumamoto, there were 110 patients with retinal detachment associated with lattice degeneration; 72 with detachment resulting from tractional tears (tears), and 38 with detachment from atrophic holes. The cumulative incidence of retinal detachment from atrophic holes was 1.5% at the age of 40 years; from tears it was 3.6% at the age of 80 years. The cumulative incidence of detachment from both atrophic holes and tears was 5.3% at the age of 80 years. The results of this study are useful for clarifying the natural course of lattice degeneration.

Artificial shrinkage of blastocoeles using either a micro-needle or a laser pulse prior to the cooling steps of vitrification improves survival rate and pregnancy outcome of vitrified human blastocysts.

BACKGROUND: Since we reported the first successful birth from a blastocyst vitrified using a cryoloop technique, our results showed that the survival rate of vitrified blastocysts was negatively correlated with the expansion of the blastocoele. We speculated that a large blastocoele may disturb the efficacy of vitrification. Therefore, we evaluated the effectiveness of artificial shrinkage (AS) of blastocoeles before vitrification, on increasing the survival rate of vitrified blastocysts. METHODS: Supernumerary expanded blastocysts on day 5 were vitrified after AS, which was performed by puncturing the blastocoele with a micro-needle, or by making a hole in the blastocoele with a laser pulse. After warming, viable blastocysts (confirmed by re-expansion of the blastocoele) were transferred to patients with hormone replacement cycle. We compared these data with those of our previous report where AS was not carried out. RESULTS: The survival rate was significantly higher (97.2%, 488/502) in this study than that of the previous report (86%). After 266 transferable cycles, 160 patients became pregnant (60.2%), which was significantly higher than our previous results (34.1%, 29/85). The implantation rate was 46.7% (209/448). CONCLUSIONS: Our results revealed that the survival rate and the pregnancy rate of vitrified expanded and hatching blastocysts can be improved by using AS to collapse the blastocele before vitrification.

V(D)J recombination of immunoglobulin genes.

We have constructed transgenic mice carrying an artificial substrate of V(D)J recombination. In this substrate, the only DNA fragments derived from Ig genes were short stretches of recombination signal sequences. This artificial substrate was rearranged at high frequency in lymphocytes, although in non-lymphoid cells no rearrangement was detected even by a sensitive PCR assay. This result indicates that the V(D)J recombination requires only the signal sequences and that a recombination similar to the V(D)J recombination does not occur in non-lymphoid tissues including the central nervous tissue. A protein binding to the V(D)J recombination signals was purified and its cDNA was cloned. This protein, termed RBP-J kappa, was initially considered to be involved in V(D)J recombination because of its DNA binding specificity and structural similarity to site-specific recombinases known as the integrase family. However, further study on the Drosophila homolog of RBP-J kappa indicated that RBP-J kappa probably functions as a transcription factor in the differentiation of the peripheral nervous tissues. The exact function of RBP-J kappa is still unknown. Analogous to the Drosophila gene, it is suggested that mouse RBP-J kappa participates in the regulation of differentiation of various tissues.

Isolation of citrate-positive variants of Escherichia coli from domestic pigeons, pigs, cattle, and horses.

Twenty-seven isolates of citrate-positive variants of Escherichia coli were obtained from domestic pigeons, pigs, cattle, and horses. With the exception of citrate utilization, all isolates closely resembled typical E. coli in their biochemical reactions. These isolates were multiply resistant to antibiotics in in vitro susceptibility tests. Transfer experiments of multiple-drug resistance to the E. coli K-12 strain showed that all citrate-positive isolates from domestic pigeons, pigs, and cattle, resistant to three or more drugs, carried R plasmids showing temperature-sensitive transfer.

Detection of conjugative R plasmids conferring chloramphenicol resistance in Escherichia coli isolated from domestic and feral pigeons and crows.

A total of 87 domestic pigeons of 2 lots and 184 feral pigeons of 15 lots were examined from 1975 to 1977 for the presence of drug-resistant (especially chloramphenicol resistant) Escherichia coli. 20 (23.0%) of the domestic pigeons of the 2 lots, and 39 (21.2%) of the feral pigeons of 5 lots (33,3%) showed resistant E. coli. Usage of selective media containing chloramphenicol, streptomycin or tetracycline resulted in the increase in isolation frequency of resistant E. coli excepting one lot of domestic pigeons in which isolation of chloramphenicol resistant E. coli was very frequent without selection by the drug. Among a total of 106 resistant E. coli isolates from pigeons, 64 (60.4%) were multiply resistant and 58 of the 64 isolates were resistant to chloramphenicol. 58 (90.6%) of the multiply resistant E. coli carried conjugative R plasmids, including 13 thermosensitive R plasmids. 8 (19.0%) of 42 singly resistant E. coli isolates had conjugative R plasmids. 10 crows of 2 lots were examined similarly. Half of them had resistant E. coli. 15 (78.9%) of a total of 19 resistant E. coli isolates were multiply resistant. Of the 15 multiply resistant E. coli isolates, 5, from 2 Japanese jungle crows, were resistant to chloramphenicol. 11 (73.3%) of the 15 multiply resistant isolates carried conjugative R plasmids, including one thermosensitive R plasmid. Difference of drug resistance status between Salmonella and E. coli isolated from pigeons was discussed.

Plasmids in Escherichia coli controlling citrate-utilizing ability.

The citrate-utilizing ability of 19 out of 22 citrate-positive Escherichia coli strains isolated from pig sewage was transferred via conjugation to E. coli K-12. The conjugal transfer of citrate-utilizing (Cit) abilities was thermosensitive and concurrent with transfer of drug resistance. Weakly citrate-positive colonies were readily obtained in conjugation experiments. Their Cit characters could be transmitted to the other E. coli strains at a similar frequency in the retransfer experiments, and the transconjugants obtained still showed same characteristic growth on Simmons citrate agar plates. The 19 thermosensitive plasmids conferring citrate utilization and drug resistance were Fi-, and 16 of these plasmids belonged to incompatibility group H1. However, occasionally two conjugative plasmids (pOH3122-1 and pOH3124-1) carrying only the citrate utilization were also obtained in the conjugation experiments, and they were Fi+ and compatible with 19 reference R plasmids. In the two citrate-positive E. coli strains, it was suggested that the conjugative Cit plasmid showing Fi+ character and the more thermosensitive H1 plasmid conferring both the Cit character and drug resistance coexisted in the strain. The characterization of citrate utilization plasmids derived from pig farm sewage is discussed.

Retinal detachment with atopic dermatitis similar to traumatic retinal detachment.

PURPOSE: The cause of retinal detachment (RD) with atopic dermatitis remains uncertain. The purpose of this study is to show that the probable cause of RD with atopic dermatitis is ocular contusion. METHODS: The authors retrospectively compared the fundus findings of 24 eyes (16 patients) that had RD and atopic dermatitis with 36 eyes (36 patients) that had traumatic RD. RESULTS: The authors found similar characteristics. Retinal breaks at vitreous base borders characterized by ocular contusion occurred in 79.2% of eyes with RD and atopic dermatitis and in 75.0% of eyes with traumatic RD. Irregular retinal traumatic breaks in the equatorial zone occurred in 20.8% of eyes with RD and atopic dermatitis and in 47.2% of eyes with traumatic RD. Objective signs of ocular contusion outside the retina appeared in 54.2% of eyes with RD and atopic dermatitis. CONCLUSIONS: The authors' data support the conclusion that self-inflected ocular contusion by tapping the eyes can cause RD with atopic dermatitis.

A yeast gene necessary for bud-site selection encodes a protein similar to insulin-degrading enzymes.

Cells of the yeast Saccharomyces cerevisiae choose bud sites in a non-random spatial pattern that depends on mating type: axial for haploid cells and bipolar for a/alpha diploid cells. We identified a mutant yeast, axl 1, in which the budding pattern is altered from axial to bipolar. Expression of the AXL1 gene is repressed in a/alpha diploid cells. With the ectopic expression of AXL1, a/alpha cells exhibited an axial budding pattern, thus AXL1 is a key morphological determinant that distinguishes the budding pattern of haploid cells from that of a/alpha diploid cells. AXL1 encodes a protein similar in sequence of the human and Drosophila insulin-degrading enzymes and to the Escherichia coli ptr gene product. The axial budding pattern might result from degradation of a target protein by the putative Axl1 protease.

Vitrification of human blastocysts using cryoloops: clinical outcome of 223 cycles.

BACKGROUND: The need to cryopreserve human blastocysts is increasing. The successful birth has been reported of a baby from a blastocyst vitrified using the cryoloop technique. The present study expands on this earlier report to confirm the effectiveness of this vitrification procedure. METHODS: In patients undergoing IVF at one of three clinics, supernumerary blastocysts on day 5 or 6 at various stages of development were vitrified using cryoloops. RESULTS: Of 725 vitrified blastocysts, 583 (80.4%) survived. After the transfer of 493 blastocysts in 207 cycles, 76 women (37%) became clinically pregnant. Among these women, 21 pregnancies ended in miscarriage, 23 healthy babies were born in 18 deliveries, and 37 pregnancies are ongoing. The survival rate of day 5 blastocysts (87%) was higher than that of day 6 blastocysts (55%), but implantation rates and pregnancy rates were not statistically significantly different. CONCLUSIONS: Clinical outcomes with 725 blastocysts and 207 transfers showed that vitrification using cryoloops is effective and practical for the cryopreservation of human blastocysts. Early blastocysts on day 5 seem to be the most suitable in terms of stage and age for cryopreservation, but developed and day 6 blastocysts can also be cryopreserved.

Recombination of exogenous interleukin 2 receptor gene flanked by immunoglobulin recombination signal sequences in a pre-B cell line and transgenic mice.

We have constructed a plasmid, pLTR100, which contains human interleukin 2 receptor light (IL-2R L) chain cDNA in the inverted orientation relative to the upstream SV40 promoter. The cDNA segment is flanked by the immunoglobulin gene recombination signal sequences so that the cDNA segment can invert and the human IL-2R L chain is subsequently expressed under the control of the SV40 promoter. A murine pre-B cell line, 38B9, transfected with pLTR100 began to express the human IL-2R L chain on the cell surface. The frequency of human IL-2R L chain positive cells increased almost linearly up to 50% for 60 days of culture after transfection. Southern blot analysis and sequencing of the DNA fragments at the recombination junction confirmed that the cDNA segment was inverted in a signal sequence-dependent manner by the variable-diversity-joining recombination process. Transgenic mice bearing the recombination substrate DNA similar to pLTR100 expressed the human IL-2 L chain in the spleen, thymus, and bone marrow, but not in the other tissues examined at the detectable level. Both IgM- and CD3-positive cells expressed the human IL-2R L chain, indicating that this artificial DNA can serve as a substrate for recombination both in B- and T-cells and that another DNA segment may be necessary to confer the cell-type specificity on the substrate DNA.

Epidemiologic characteristics of rhegmatogenous retinal detachment in Kumamoto, Japan.

BACKGROUND: The epidemiology of rhegmatogenous retinal detachment in Asians is not well known. We studied the epidemiologic characteristics of rhegmatogenous retinal detachment in Kumamoto, Japan. METHODS: The study was based on a retrospective chart review of hospital patients who were treated for primary rhegmatogenous retinal detachment in 1990. The data were collected from seven hospitals in the Kumamoto area. RESULTS: From a population of 1 840 000, 192 residents developed retinal detachment. The annual incidence was therefore 10.4 per 100 000 population (9.6 for males, 11.2 for females). The incidences of three types of detachment-nontraumatic phakic, aphakic, and blunt trauma--were 9.8, 0.5 and 0.2 per 100 000 population, respectively. In 109 of 180 patients (60.6%) with nontraumatic phakic detachment, retinal breaks were associated with lattice degeneration. In females, 14 of 106 nontraumatic phakic cases (13.2%) were secondary to macular holes. CONCLUSION: Compared with previously published studies from other countries, the incidence of detachments associated with lattice degeneration and macular hole was higher, while the incidences of aphakic detachment and detachment due to blunt trauma were lower in Japan. Racial factors and living habits may affect the development of retinal detachment.

Spontaneous emergence of an Hfr strain with a cit plasmid from natural isolates of citrate-positive Escherichia coli bovine origin.

From citrate-utilizing (Cit+) Escherichia coli strain C53 of bovine origin, strains C53A and C53B were obtained. Upon mating with recA+ but not with recA mutant recipients of K-12, C53A produced chromosomal recombinants at quite high frequencies, leading to the following conclusions: (i) C53A is an Hfr strain; (ii) the site of integration of the Cit plasmid (IncH1) is between metA (89 min) and ara (1 min); (iii) the direction of chromosome transfer is clockwise; and (iv) the plasmid-associated determinants are transferred as the terminal markers. A transductant of a dnaA(Ts) strain, CRT46, which acquired Cit determinants from a recombinant, SG13, was also an Hfr strain similar to SG13, and thermoresistant due to suppressive integration. On the other hand, unstable C53B did not produce recombinants, but the frequency of RecA-independent transfer of the Cit plasmid was high, indicating that the Cit plasmid (IncH1) exists autonomously in C53B. Attempts to isolate an Hfr strain from C53B failed.