Measuring macular pigment optical density using reflective images of confocal scanning laser system.

PURPOSE: To develop a method to measure the macular pigment optical density (MPOD) using scanning laser ophthalmoscopic images in young adults and children. STUDY DESIGN: Cross-sectional study. METHODS: Blue light reflectance fundus images of 32 healthy subjects were used. A profile of the linear reflectance changes across the center of the fovea on a grayscale fundus image was generated. The ratio of the macula-to-periphery reflection was designated as the peak value of the MPOD (MPOD[FR]) based on established fundamentals. In the MPOD profile, the basal width of the pixels at MPOD < 0 (wMP) and width at one-half value of the MPOD[FR] (wMP0.5) were determined. The MOPD at eccentricity of 0.5° was measured by heterochromatic flicker photometry (MPOD[HFP]), and the correlation between the MPOD[FR] and MPOD[HFP] was evaluated. RESULTS: The MPOD[FR] ranged from 0.17 to 0.73 with a mean of 0.40 ± 0.13. The wMP ranged from 88 to 173 pixels with a mean of 121.7 ± 24.2 pixels, and the wMP0.5 ranged from 38 to 83 pixels with a mean of 54.1 ± 10.3 pixels. A significant correlation was found between the MPOD[FR] and MPOD[HFP] (r = 0.41, P = 0.02). CONCLUSIONS: This simplified method can provide accurate and reliable values of the MPOD comparable to heterochromatic flicker photometry. Obtaining the fundus images in this fast and easy way should be suitable for children thus enabling clinicians to determine the MPODs for children.

Autosomal dominant foveal hypoplasia without visible macular abnormalities and PAX6 mutations.

PURPOSE: Autosomal dominant foveal hypoplasia (FVH1) is a rare disorder associated with mutations in the PAX6 gene. As an isolated disease entity, FVH1 does not include ocular disorders such as aniridia, microphthalmia, albinism, and achromatopsia. However, it only includes isolated foveal hypoplasia and foveal hypoplasia with presenile cataract. The purpose of this report is to present our findings in four patients from two families with FVH1 without visible ophthalmic macular abnormalities. STUDY DESIGN: A review of the medical records of two families with FVH1 and genetic confirmation of mutations in the PAX6 gene. METHODS: Fundus photographs, optical coherence tomographic (OCT) and OCT angiographic (OCTA) images, and slit-lamp anterior segment findings were determined. The type of mutation of the PAX6 gene was determined. RESULTS: A 3-year-old girl (Patient 1) had signs and symptoms of an impairment in the development of vision without other retinal abnormalities OU. OCT images showed a shallow foveal pit, and OCTA showed the absence of the foveal avascular zone. The second patient (Patient 2) was a 6-year-old girl with unilateral mild cataract and shallow foveal pits OU. Similar shallow foveal pits were found in her asymptomatic mother (Patient 3) and maternal grandfather (Patient 4). Although the iris and posterior fundus were normal, all patients with FVH1 had goniodysgenesis. Genetic testing of the PAX6 gene revealed that Patient 1 had a novel heterozygous mutation (p.Asn365Lys) as a de novo mutation, and Patients 2, 3 and 4 had a novel heterozygous mutation (p.Pro20Ser). CONCLUSIONS: Heterozygous mutations in the PAX6 gene can cause FVH1 with nearly normal appearing macula. FVH1 is difficult to diagnose, but detailed observations of the foveal structure and vasculature, and detecting the presence of goniodysgenesis can be helpful in identifying patients with FVH1.

Blastomere movement post first cell division correlates with embryonic compaction and subsequent blastocyst formation.

BACKGROUND: Blastomere movement (BMov) occurs after the first cell division in human embryos. This movement has been suggested as a prognostic parameter for pregnancy outcome prediction following cleavage-stage embryo transfer. However, the effect of BMov on preimplantation development and pregnancy outcome after blastocyst transfer remains unclear. Therefore, this study aimed to evaluate whether BMov after the first cell division is correlated with blastocyst formation rate and live birth rate after single vitrified-warmed blastocyst transfer (SVBT). METHODS: Nine hundred and sixty-six embryos cultured in the EmbryoScope+® time-lapse system were retrospectively analyzed. The BMov type was categorized into three groups; namely, bouncing, wobbling, and twist-and-crumble. The BMov duration (dBMov) between the first (t2) and second cell division (t3) was monitored, and the ratio of dBMov to the duration of the 2-cell stage was calculated [dBMov/(t3-t2)]. Developmental rates to the 4-cell, 8-cell, morula, blastocyst, and expanded blastocyst stages were assessed, as well as blastocyst morphological grade. The correlations between dBMov and clinical pregnancy, ongoing pregnancy, and live birth rates were evaluated. RESULTS: Increased dBMov/(t3-t2) was significantly correlated with decreased developmental rates to the 8-cell, morula, blastocyst, and expanded blastocyst stages, especially from the 4-cell stage to the morula stage. Analysis of different types of BMov revealed that embryos with bouncing movement exhibited significantly higher developmental rates to the 8-cell, morula, blastocyst, and expanded blastocyst stages compared with embryos with twist-and-crumble movement. The morphological quality of blastocyst-stage embryos with twist-and-crumble movement was significantly lower than that of embryos with bouncing and wobbling movements. The rates of clinical pregnancy, ongoing pregnancy, and live birth after SVBT were not correlated with BMov type or duration. CONCLUSIONS: Embryonic compaction and subsequent blastocyst formation are adversely affected by twist-and-crumble movement and prolonged movement after the first cell division. Our results indicate that the preimplantation developmental competence of human embryos could be predicted by assessing BMov after the first cell division on day 1.

Prolonged blastomere movement induced by the delay of pronuclear fading and first cell division adversely affects pregnancy outcomes after fresh embryo transfer on Day 2: a time-lapse study.

RESEARCH QUESTION: What is the incidence, origin and clinical significance of blastomere movement after the first cell division in the human embryo? DESIGN: A total of 1096 embryos, cultured in the EmbryoScope+ ® time-lapse system and subjected to a single fresh cleaved embryo transfer, were retrospectively analysed. Type and duration of blastomere movement (dBMov) between the first (t2) and second cell division (t3) was monitored, and the ratio of dBMov during the 2-cell stage [dBMov/(t3-t2)] was calculated. Morphological evaluation of embryos was performed by referring to the size of the blastomere and fragmentation after first division in addition to Veeck's criteria on Day 2. The correlation between dBMov and ongoing pregnancy was evaluated and the association of dBMov with patient and embryonic characteristics was determined. RESULTS: Both movement type and the value of dBMov/(t3-t2) were significantly associated with asymmetrical first division, fragment formation and morphological grade on Day 2. Multivariate logistic regression analysis revealed that a higher value of dBMov/(t3-t2) significantly correlated with a decreased ongoing pregnancy rate, even after adjustment for co-founders (odds ratio 0.399, P = 0.0419). The time intervals of pronuclear (PN) alignment and PN fading were significantly correlated with the dBMov/(t3-t2) value. CONCLUSIONS: Embryos with extended blastomere movement after the first cell division, which is associated with the delay of PN fading and first cell division, have a lower competence to initiate an ongoing pregnancy after fresh embryo transfer on Day 2. Thus, blastomere movement could be a useful predictive parameter for selecting embryos at the early cleavage stage.

Development of a transgenic tobacco plant for phytoremediation of methylmercury pollution.

To develop the potential of plant for phytoremediation of methylmercury pollution, a genetically engineered tobacco plant that coexpresses organomercurial lyase (MerB) with the ppk-specified polyphosphate (polyP) and merT-encoding mercury transporter was constructed by integrating a bacterial merB gene into ppk/merT-transgenic tobacco. A large number of independent transgenic tobaccos was obtained, in some of which the merB gene was stably integrated in the plant genome and substantially translated to the expected MerB enzyme in the transgenic tobacco. The ppk/merT/merB-transgenic tobacco callus showed more resistance to methylmercury (CH3Hg+) and accumulated more mercury from CH3Hg+-containing medium than the ppk/merT-transgenic and wild-type progenitors. These results suggest that the MerB enzyme encoded by merB degraded the incorporated CH3Hg+ to Hg2+, which then accumulated as a less toxic Hg-polyP complex in the tobacco cells. Phytoremediation of CH3Hg+ and Hg2+ in the environment with this engineered ppk/merT/merB-transgenic plant, which prevents the release mercury vapor (Hg0) into the atmosphere in addition to generating potentially recyclable mercury-rich plant residues, is believed to be more acceptable to the public than other competing technologies, including phytovolatilization.

Histology of fibrovascular membranes of proliferative diabetic retinopathy after intravitreal injection of bevacizumab.

PURPOSE: The purpose was to study the histology of the fibrovascular membranes in proliferative diabetic retinopathy (PDR) with an intravitreal injection of bevacizumab. METHODS: Light and electron microscopic studies were performed on surgical specimens obtained during a pars plana vitrectomy from 6 PDR eyes after intravitreal injection of bevacizumab. The patients had preoperatively received no or scant retinal photocoagulations. The presence and distribution of CD34 was assessed as a marker of vascular endothelium using immunostaining. The presence of vascular endothelial growth factor was stained with a method of immunostaining. As controls, we examined 7 surgical specimens from 7 PDR eyes obtained during pars plana vitrectomy without bevacizumab therapy. All control patients had preoperatively received full or nearly full pan retinal photocoagulations. RESULTS: Light microscopy showed that the CD34-positive vascular endothelial cells formed capillarylike structures in the fibrovascular membranes of all 13 PDR eyes. Vascular endothelial growth factor was positively stained in the vascular endothelium of both groups; however, the number of vascular endothelial growth factor-positive vascular endothelial cells significantly decreased in the fibrovascular membranes with intravitreal injection of bevacizumab. Electron microscopy showed the newly formed vascular endothelial cells with junctional complex in both groups. CONCLUSION: The vascular endothelial cells with decreased expression of vascular endothelial growth factor are still present in the fibrovascular membranes of patients with PDR after intravitreal injection of bevacizumab.

Direct inhibition of N-methyl-D-aspartate (NMDA)-receptor function by antiglaucomatous beta-antagonists.

In the present study, we investigated the direct effects of antiglaucoma drugs (timolol, betaxolol, pilocarpine, and latanoprost) on N-methyl-D-aspartate (NMDA)-receptor function using a Xenopus oocytes expression system and electrophysiological techniques. In oocytes expressing wild-type NMDA (NR1a/NR2A) receptors, timolol and betaxolol significantly inhibited glutamate-evoked currents, whereas less inhibition was obtained with pilocarpine, and latanoprost had few effects. Moreover, the effect of timolol and betaxolol was noncompetitive with respect to glutamate. Mutations that changed Asn616 of the NR1a subunit, a critical residue for Mg(2+) blocking of NMDA receptors, to Arg (N616R) or Gln (N616Q) almost eliminated the inhibitory effects of timolol and betaxolol, as well as the blocking effect of Mg(2+). Experiments were also carried out to examine the protective effects of timolol and betaxolol against death of oocytes expressing NMDA receptors. During incubation of oocytes, especially in Mg(2+)-free medium, cell death was induced by addition of glutamate because of the continuous activation of the NMDA receptors expressed. Timolol and betaxolol significantly improved oocyte viability when they were added during the incubation period. These results suggest that timolol and betaxolol may have an additional role that they directly inhibit NMDA-receptor function, possibly via N616 of the NR1a subunit.

[Informed consent for colorectal cancer surgery].

There are two issues in informed consent for colorectal cancer surgery. One is the actual surgical technique. Recently intersphincteric resection (ISR) has been performed to avoid permanent colostomy, although it is not standard procedure. Regarding QOL score, Patients with colostomies do not necessarily have lower quality of life scores than patients who undergo sphincter-preserving surgery because of frequent bowel movements. Lateral lymphadenectomy for lower rectal cancer was standard procedure; however, its indications became limited due to urinary and sexual dysfunction. Preoperative radiotherapy is considered instead of lateral lymphadenectomy, as in the Western concept of the local recurrence of rectal cancer. Now laparoscopic surgery is accepted for stage I colon cancer according to the guidelines of the Japan Society for Cancer of the Colon and Rectum. The other issue is postoperative complications like anastomotic leakage, intestinal obstruction, etc. Frequent bowel movements and urinary and sexual dysfunction should also be explained before rectal cancer surgery.

Degeneration of the stria vascularis during development in melanocyte-deficient mutant rats (Ws/Ws rats).

Developmental changes in the stria vascularis of white spotting (Ws) rats were examined by scanning and transmission electron microscopes and by diaminobenzidine-staining techniques. The stria of Ws/Ws homozygote rats was found to have both pigmented and non-pigmented portions. While the pigmented portions possessed intermediate cells in the same manner as the stria of wild +/+ rats, the non-pigmented portions lacked the cells. Examination at 1, 2, 3, 4, 6, 8 and 14 weeks after birth revealed a progressive degeneration in the marginal cells and strial capillaries in the non-pigmented portions. At 1 week, no significant differences were seen in the marginal cells of any of the rats examined. At 2 weeks, the basolateral infoldings of the marginal cells were seen to be well developed and adult-like in the pigmented portions of Ws/Ws rats and in +/+ rats. In the non-pigmented portions, the basolateral infoldings of the marginal cells appeared well developed; however, vacant spaces were seen around the basolateral infoldings. At 3 weeks, the basolateral infoldings of the marginal cells in the non-pigmented portions had become more atrophic, and the empty spaces around the basolateral infoldings had enlarged. Also, the marginal cells themselves had become flatter or thinner. These findings became more prominent at 4 weeks and 6 weeks. At 8 weeks and 14 weeks after birth, the marginal cells appeared markedly flat, and no basolateral infoldings were seen in the non-pigmented portions. Pigmented portions of the stria in Ws/Ws rats, on the other hand, showed normal development throughout this period. A DAB-staining examination of the stria capillary net in Ws/Ws rats showed it to be well developed at 3 weeks in both pigmented and non-pigmented portions. At 8 weeks, a thickening of the capillary basement membrane was apparent. The above findings lead the authors to believe that intermediate cells play an important role in the development and maintenance of marginal cells and the strial capillary system.