Mutations of the KLF1 gene detected in Japanese with the In(Lu) phenotype.

BACKGROUND: In(Lu) is characterized by a reduced expression of antigens in the Lutheran blood group system as well as other blood group antigens. Mutations of the erythroid transcription factor, KLF1, have been reported to cause the In(Lu) phenotype, and we investigated Japanese In(Lu) to estimate the prevalence of the phenotype and KLF1 polymorphism. STUDY DESIGN AND METHODS: Blood samples were screened by monoclonal anti-CD44 and the In(Lu) phenotype was confirmed by tube tests including adsorption and elution tests using anti-Lua and anti-Lub . KLF1, LU, and A4GALT genes were analyzed by polymerase chain reaction and sequencing. RESULTS: We identified 100 of 481,322 blood donors (0.02%), and the previously characterized 20 donors, who had the In(Lu) phenotype with the LUB/LUB genotype. A total of 100 of the 120 In(Lu) individuals had mutant KLF1 alleles, and we identified 13 known and 21 novel alleles. The mutant KLF1 alleles with c.947G>A (p.Cys316Tyr), c.862A>G (p.Lys288Glu), or c.968C>G (p.Ser323Trp) were major in the In(Lu) individuals. The P1 antigen of 29 In(Lu) (two P1 /P1 , 27 P1 /P2 ) showed significantly weakened expression by hemagglutination. CONCLUSIONS: The prevalence of the In(Lu) phenotype in the Japanese population was 0.02%, and we identified 13 known and 21 novel KLF1 alleles. The KLF1 mutations cause the reduced expression of the P1 antigen.

A new ABCG2 null allele with a 27-kb deletion including the promoter region causing the Jr(a-) phenotype.

BACKGROUND: The high-prevalence antigen Jr(a) is carried on the ATP-binding cassette transporter ABCG2. The ABCG2 gene consists of 16 exons and its translation start codon is located on the second exon. Although the occurrence of the Jr(a-) phenotype is rare, several ABCG2 null alleles have been reported. We report a new ABCG2 null allele having a large deletion in this study. STUDY DESIGN AND METHODS: The Jr(a) status was determined by standard serologic tests and genomic DNA was isolated from whole blood. Exons 1 to 16 and the 5'-untranslated region of the ABCG2 gene were analyzed by polymerase chain reaction and sequencing. Expression of the ABCG2 protein on red blood cells was examined by immunoblotting. RESULTS: A Jr(a-) blood donor had a novel allele having a 27-kb deletion including noncoding Exon 1 and the promoter region of ABCG2, and the donor was apparently homozygous for the allele. In addition, we found three more individuals having heterozygosity for the same allele, with ABCG2*01N.01 having c.376C>T (p.Q126X), but did not find the allele having the 27-kb deletion in 3000 Jr(a+) individuals. Immunoblotting revealed that the ABCG2 protein was not found to be expressed in the individual with homozygosity for the ABCG2 27-kb deleted and in two individuals with an ABCG2 27-kb deleted/ABCG2*01N.01 genotype, which indirectly allows to conclude that the 27-kb deletion is responsible for a null ABCG2 allele. CONCLUSION: We first identified an ABCG2 null allele (provisional ISBT allele number ABCG2*01N.23) having a large deletion including the promoter region.

Effect of experimentally induced astigmatism on functional, conventional, and low-contrast visual acuity.

PURPOSE: To report the changes in functional visual acuity (FVA) in eyes with experimentally induced astigmatism. METHODS: This study included 26 right eyes from 26 healthy participants (mean age: 27.0±4.4 years). After confirming best correction under cycloplegia, the cylinder power ×90° and ×180° (against-the-rule [ATR] and with-the-rule [WTR] astigmatism, respectively) were added with 0.50-diopter (D) steps, from 0.50 to 2.50 D. Conventional visual acuity (VA), FVA as measured by the FVA Measurement System (NIDEK Co Ltd), and 10% low-contrast visual acuity (LCVA) were measured with 3-mm artificial pupil under each condition. RESULTS: The mean corrected logMAR VA, FVA, LCVA were -0.18±0.00 (20/13), -0.10±0.08 (20/16), and 0.06±0.06 (20/23), respectively. The VA, FVA, and LCVA had a significant linear negative correlation with the addition of cylinder irrespective of the axes. A significant decrease in FVA was noted on addition of ⩾0.50 D WTR astigmatism and ⩾1.00 D ATR astigmatism. With the addition of 1.00 D of cylindrical lens, 96% of eyes with WTR and ATR astigmatism could maintain 20/20 in VA testing, whereas only 50% and 62% of eyes with WTR and ATR astigmatism, respectively, could maintain 20/20 in FVA testing. CONCLUSIONS: Astigmatism may be associated with deterioration of visual function even when conventional VA of 20/20 is attained. Functional VA testing seems to be useful in evaluating the masked astigmatic visual impairment that cannot be detected by conventional VA testing.

Menstrual disturbances in various thyroid diseases.

The prevalence of menstrual disturbances, including secondary amenorrhea, hypomenorrhea, oligomenorrhea, hypermenorrhea, polymenorrhea and irregular menstrual cycle were prospectively examined in 586 patients with hyperthyroidism due to Graves' disease, 111 with hypothyroidism, 558 with euthyroid chronic thyroiditis, 202 with painless thyroiditis and 595 with thyroid tumor. In the overall patient group, the prevalence did not different from that in 105 healthy controls. However, patients with severe hyperthyroidism showed a higher prevalence of secondary amenorrhea (2.5%) and hypomenorrhea (3.7%) than those (0.2% and 0.9%, respectively) with mild or moderate hyperthyroidism. Moreover, patients with severe hypothyroidism had a higher prevalence (34.8%) of menstrual disturbances than mild-moderate cases (10.2%). Menstrual disturbances in thyroid dysfunction were less frequent than previously thought.

Light-sensitive vertical migration of the Japanese eel Anguilla japonica revealed by real-time tracking and its utilization for geolocation.

Short-time tracking (one to eight days) of the Japanese eel (Anguilla japonica) using ultrasonic transmitter was performed in the tropical-subtropical area adjacent to the spawning area and temperate area off the Japanese Archipelago. Of 16 eels (11 wild and five farmed) used, 10 wild eels displayed clear diel vertical migration (DVM) from the beginning, while the other five farmed eels tracked for 19 to 66 hours did not. During daytime, a significantly positive correlation between migration depth and light intensity recorded on the vessel was observed in the 10 wild eels, indicating that the eels were sensitive to sunlight even at the middle to lower mesopelagic zone (500 to 800 m). During nighttime, the eel migration depth was observed to be associated with the phase, rising and setting of the moon, indicating that the eels were sensitive to moonlight at the upper mesopelagic zone (<300 m). Two of 10 wild eels were in the yellow stage but shared similar DVM with the silver stage eels. Swimbladders of three silver stage eels were punctured before releasing, but very little effect on DVM was observed. The eels very punctually initiated descent upon nautical dawn and ascent upon sunset, enabling us to determine local times for sunrise and sunset, and hence this behavior may be used for geolocating eels. In fact, estimated positions of eels based on the depth trajectory data were comparable or even better than those obtained by light-based archival tag in other fish species.

[The acquisition of schemata about location of urban facilities in public space].

The purpose of this study is to investigate acquisition of schemata about location of urban facilities. Materials were photos of public space, in which one of three urban facilities (a mailbox, a public-telephone, and a coin--operated locker) were removed by photo-retouching software. The task was to choose the most suitable urban facility for each corrected-photo and to locate it at the most suitable position on the photo. Among three participant groups (primary school 3rd grade: n = 153, primary school 5th grade: n = 118, undergraduate: n = 250), undergraduates chose the most suitable urban facility, and their located position was more concentrated. The results suggest that participants acquire the schemata and use them for inference. Although even 3rd grade children have acquired the schema to some extent, the adults have more detailed schemata of urban facilities in public space.

Carbon dioxide­pneumoperitoneum in rats reduces ischemia/reperfusion­induced hepatic apoptosis and inflammatory responses by stimulating sensory neurons.

Laparoscopic surgery induces a milder inflammatory response than open surgery, however, the precise mechanisms underlying this phenomenon remain to be elucidated. Our previous study demonstrated that stimulation of sensory neurons inhibited hepatic apoptosis and inflammatory responses in rats subjected to hepatic ischemia/reperfusion (I/R). Since carbon dioxide (CO2) has been demonstrated to stimulate sensory neurons, it was hypothesized that CO2­pneumoperitoneum, as used in laparoscopic surgery, may attenuate inflammatory responses by stimulating sensory neurons. This hypothesis was examined using rats subjected to hepatic I/R. The rats were subjected to partial hepatic ischemia for 60 min followed by reperfusion. Abdominal insufflation with CO2 or air was performed for 30 min prior to hepatic I/R. Hepatic I/R­induced hepatocellular apoptosis and expression of the neutrophil chemoattractant endothelial monocyte­activated polypeptide­II, were inhibited by CO2­pneumoperitoneum, however, not by air­pneumoperitoneum. Pretreatment with the transient receptor potential vanilloid 1 antagonist SB366791 reversed the protective effects of CO2­pneumoperitoneum. The results from the present study demonstrated that CO2­pneumoperitoneum attenuates hepatic apoptosis and inflammatory responses in rats subjected to hepatic I/R, possibly by stimulating sensory neurons. These findings suggested that CO2­pneumoperitoneum contributed to the attenuated inflammatory response observed following laparoscopic surgery.

Endometrial glandular and stromal breakdown, Part 4: Cytomorphology of "condensed cluster of stromal cells including a light green body".

This study was undertaken to clarify the origin of the chromophilic substance that stained in light green [light green body (LGB)] was observed in the condensed cluster of stromal cells and in the background of endometrial glandular and stromal breakdown cases. The material consists of cytologic smears of endometrial glandular and stromal breakdown (EGBD) from 58 samples in which a histopathological diagnosis was subsequently obtained by endometrial curettage. The following parameters were examined. (1) The occurrence and number of a LGB. As for the "condensed cluster of stromal cells," "condensed cluster of stromal cells including LGB" and "LBG in background," it was observed with all fields on one slide per case. When it was observed more than one in a preparation, the occurrence was determined and the number was also calculated. (2) Immunostaining of paraffin-embedded tissue sections and cytologic preparations. 1 The occurrence of "condensed cluster of stromal cells," "condensed cluster of stromal cells including LGB" and "LGB in background" was 100%, 44.8% and 91.4%, respectively. The number of "condensed cluster of stromal cells," "condensed cluster of stromal cells including LGB" and "LGB in background" was 21.2, 2.0 and 4.0, respectively. 2 When histological sections from paraffin-embedded tissue were tested by immunocytochemistry, "LGB" stained weakly or negative for fibrinogen, whereas it stained positively for CD31 and factor VIII. In addition, it stained positively for CD42b. 3 When cytologic preparation was tested by immunocytochemistry, "LGB" stained positively for CD31, factor VIII and CD42b. However, as for the fibrinogen, quantitative and qualitative evaluation of the immunocytochemical stain was not feasible due to extensive nonspecific reaction in the whole preparation. The "LGB" in EGBD cases proved to be a thrombus mostly made up by platelets, and it seemed that the recognition of the LGB showed to be a useful cytomorphological criterion for an always more accurate diagnosis of EGBD cases.

Acute Visual Field Defect following Vitrectomy Determined to Originate from Optic Nerve by Electrophysiological Tests.

PURPOSE: To present our findings on the cause of an acute visual field defect (VFD) that developed in a patient on the day after vitrectomy for proliferative diabetic retinopathy. CASE: A 50-year-old man complained of a blind area in the superior visual field that developed one day after vitrectomy. The patient had undergone uncomplicated vitrectomy for a long-duration vitreous hemorrhage associated with proliferative diabetic retinopathy. Residual vitreous hemorrhage hampered a clear view of the fundus. Goldmann perimetry showed a horizontal VFD in the superior field. The area corresponding to the VFD was examined by multifocal electroretinograms (mfERGs) and multifocal visual evoked potentials (mfVEPs). The amplitudes of the mfVEPs were reduced with prolonged implicit times especially when the superior hemifield was stimulated, while the amplitudes and implicit times were within the normal range when other parts of the visual field were stimulated. In addition, the full-field photopic ERGs and photopic negative responses were attenuated in the right eye. These findings suggested that the VFD did not originate from alterations in the retinal inner and middle layer but in the ganglion cells. The visual acuity improved to 1.2 but his optic disc became pale and the VFD remained unchanged more than 12 years after the surgery. CONCLUSION: We suggest that vitrectomy can cause ischemic optic neuropathy by interfering with the circulation associated with diabetes mellitus. Evaluations by mfERGs, mfVEPs, and full-field photopic ERGs were helpful in making the diagnosis.

Neutrophil elastase contributes to the development of ischemia/reperfusion-induced liver injury by decreasing the production of insulin-like growth factor-I in rats.

Neutrophil elastase (NE) decreases the endothelial production of prostacyclin (PGI(2)) through the inhibition of endothelial nitric oxide synthase (NOS) activation and thereby contributes to the development of ischemia/reperfusion (I/R)-induced liver injury. We previously demonstrated that calcitonin gene-related peptide (CGRP) released from sensory neurons increases the insulin-like growth factor- I (IGF-I) production and thereby reduces I/R-induced liver injury. Because PGI(2) is capable of stimulating sensory neurons, we hypothesized that NE contributes to the development of I/R-induced liver injury by decreasing IGF-I production. In the present study, we examined this hypothesis in rats subjected to hepatic I/R. Ischemia/reperfusion-induced decreases of hepatic tissue levels of CGRP and IGF-I were prevented significantly by NE inhibitors, sivelestat, and L-658, 758, and these effects of NE inhibitors were reversed completely by the nonselective cyclooxygenase inhibitor indomethacin (IM) and the nonselective NOS inhibitor L-NAME but not by the selective inducible NOS inhibitor 1400W. I/R-induced increases of hepatic tissue levels of caspase-3, myeloperoxidase and the number of apoptotic cells were inhibited by NE inhibitors, and these effects of NE inhibitors were reversed by IM and L-NAME but not by 1400W. Administration of iloprost, a stable PGI(2) analog, produced effects similar to those induced by NE inhibitors. Taken together, these observations strongly suggest that NE may play a critical role in the development of I/R-induced liver injury by decreasing the IGF-I production through the inhibition of sensory neuron stimulation, which may lead to an increase of neutrophil accumulation and hepatic apoptosis through activation of caspase-3 in rats.

Identification and characterization of ectosymbionts of distinct lineages in Bacteroidales attached to flagellated protists in the gut of termites and a wood-feeding cockroach.

Bacterial attachments to nearly the entire surface of flagellated protists in the guts of termites and the wood-feeding cockroach Cryptocercus are often observed. Based on the polymerase chain reaction-amplified 16S rRNA gene sequences, we investigated the phylogenetic relationships of the rod-shaped, attached bacteria (ectosymbionts) of several protist species from five host taxa and confirmed their identity by fluorescence in situ hybridizations. These ectosymbionts are affiliated with the order Bacteroidales but formed three distinct lineages, each of which may represent novel bacterial genera. One lineage consisted of the closely related ectosymbionts of two species of the protist genus Devescovina (Cristamonadida). The second lineage comprised three phylotypes identified from the protist Streblomastix sp. (Oxymonadida). The third lineage included ectosymbionts of the three protist genera Hoplonympha, Barbulanympha and Urinympha in the family Hoplonymphidae (Trichonymphida). The ultrastructural observations indicated that these rod-shaped ectosymbionts share morphological similarities of their cell walls and their point of attachment with the protist but differ in shape. Elongated forms of the ectosymbionts appeared in all the three lineages. The protist cells Streblomastix sp. and Hoplonympha sp. display deep furrows and vane-like structures, but these impressive structures are probably evolutionarily convergent because both the host protists and their ectosymbionts are distantly related.