Vitrified before and after genome editing via electroporation.

BACKGROUND: Cryopreservation of bovine zygotes allows for a flexible schedule of genome editing via electroporation. However, vitrification-induced cell membrane damage may not only affect embryonic development but also genome mutation. OBJECTIVE: To investigate the effects of vitrification of zygotes before and after electroporation treatments on the development and genome mutation of bovine presumptive zygotes. MATERIALS AND METHODS: In vitro-derived bovine zygotes were electroporated with the CRISPR/Cas9 system immediately (Vitrified-EP) or 2 h after incubation (Vitrified-2h-EP) following vitrification and warming, or electroporated before vitrification (EP-vitrified). RESULTS: The development rates of vitrified-warmed zygotes were significantly lower (p < 0.05) than those of control zygotes that were not vitrified. Moreover, no differences were observed in the mutation rates and mutation efficiency of the blastocysts resulting from electroporated zygotes, irrespective of the timing of electroporation treatment. CONCLUSION: Our results suggest that vitrification before and after electroporation treatments does not affect the genome editing of zygotes.

Topical phenylephrine decreases blood velocity in the optic nerve head and increases resistive index in the retinal arteries.

PURPOSE: To investigate the effects of topical phenylephrine on circulation in the optic nerve head (ONH), posterior choroids, or retina in rabbits and healthy humans. METHODS: Tissue blood velocity in the ONH and posterior choroid was measured using the laser speckle method as normalized blur (NB(ONH) and NB(CH)) in 28 anaesthetized albino rabbits. NB and intraocular pressure (IOP) in both eyes were measured for 180 min after unilateral single instillation of 5% phenylephrine and contralateral physiological saline as a control. In 11 normal volunteers aged 26.0+/-2.7 years, NB(ONH) was measured for 180 min after unilateral three drops of 5% phenylephrine and contralateral physiological saline in a double-masked manner. In the other 17 normal volunteers aged 25.5+/-2.4 years, blood velocity and blood flow in a major branch of the central retinal artery were measured using the laser Doppler blood flow metre and IOP and systemic circulatory parameters were monitored in similar fashion. Analysis of variance was applied for intergroup comparisons. RESULTS: NB(ONH) and NB(CH) decreased significantly only in the phenylephrine-treated eyes in rabbits by maximum of 9 and 20%, respectively (P=0.0046 and 0.0009), despite IOP decrease. In humans, NB(ONH) decreased significant by maximum of 13% (P=0.0047) and resistive index in the retinal arteries increased by 10% (P=0.0067) unilaterally; whereas IOP, diameter, blood velocity, or blood flow of the arteries was not significantly changed. CONCLUSION: Topical phenylephrine could exert a significant unfavourable effect on circulation in the ONH, choroidal tissue, and retinal arteries in rabbits and normal young humans.

Relation between retrobulbar circulation and progression of diabetic retinopathy.

AIMS: To investigate retrobulbar circulatory parameters in type 2 diabetic patients with and without diabetic retinopathy (DR) progression. METHODS: This was a prospective cohort study. One eye of 35 diabetic patients with background DR (BDR) were included in the study. Eyes without DR, with proliferative DR, photocoagulation, past surgical procedures, or other ophthalmic disease except BDR and cataract were excluded. The study was masked. Colour Doppler imaging (CDI) was used to measure the retrobulbar circulation at the beginning of the study and after a mean follow up interval of 21 months. Peak systolic velocity (PSV), end diastolic velocity (EDV), and resistivity index (RI) in the central retinal artery and vein and the posterior ciliary artery were measured. RESULTS: 18 patients who developed DR progression showed significantly increased central retinal vein PSV ( 5.6 (3.5-9.1) p = 0.003), EDV ( 3.4 (2.3-4.4) p = 0.04), and RI ( 0.43 (0.20-0.56) p = 0.02) at the final measurement compared to the initial measurement (PSV = 4.6 (3.2-7.0); EDV = 3.0 (2.3-3.7); RI = 0.40 (0.17-0.52)). Circulatory parameters in the central retinal artery and the posterior ciliary artery did not alter significantly after progression of DR. 17 patients were without DR progression and they did not show any significant differences in the measured circulatory parameters on entry compared to the final measurement. CONCLUSION: The authors suggest that the initial changes in the retrobulbar circulation during DR progression occur in the central retinal vein.

Retrobulbar circulation in myopic patients with or without myopic choroidal neovascularisation.

AIM: To investigate the effect of myopia and myopic choroidal neovascularisation (CNV) on retrobulbar circulation in central retinal artery (CRA) and vein (CRV) and posterior ciliary artery (PCA). METHODS: 52 subjects with and without myopia were included in the study. Retrobulbar circulation was measured using colour Doppler imaging. Analysis of correlation of degree of myopia with blood flow velocity parameters was done. Circulatory differences between eyes of patients with unilateral neovascular degenerative myopia were estimated. RESULTS: The analysis of correlation between dioptry and blood flow velocity in the CRA, CRV, and PCA showed a significant positive correlation. Axial length was also significantly correlated with CRA and CRV blood velocity and had a tendency to be correlated with PCA blood velocity. When compared with the fellow eye, the eye with myopic CNV had significantly higher resistivity index (RI) (p=0.048) in the PCA and no significant difference in the circulatory parameters of the CRA and CRV. CONCLUSION: Central retinal and posterior ciliary blood velocity decreases with the increase of the degree of myopia. PCA RI is higher in myopic CNV.

Choroidal circulation in diabetic patients.

PURPOSE: To evaluate choroidal circulatory changes in diabetic patients with and without background diabetic retinopathy (BDR) by measuring the retrobulbar circulation with colour Doppler imaging (CDI). METHODS: CDI was used to measure circulatory parameters of the PCA (posterior ciliary artery), CRA (central retinal artery), OA (ophthalmic artery) and the respective veins in 73 diabetic patients and 22 controls in a sitting posture. Among the diabetic patients, 38 patients were without diabetic retinopathy (NDR) and 35 had BDR. A non-parametric Kruskal-Wallis test with a Dunn correction was used for data analysis. RESULTS: End-diastolic velocity (EDV) in the PCA was decreased (2.55+/-0.80 cm/s) and resistivity index (RI) in the PCA was increased (0.70+/-0.08) in BDR patients compared with the control patients' EDV (3.23+/-1.08 cm/s, p = 0.01) and RI (0.62+/-0.06, p = 0.0003). RI in the CRA was significantly higher in the BDR group (0.74+/-0.09) than in the control group (0.68+/-0.08, p = 0.006). RI in the OA was significantly higher in the BDR group (0.87+/-0.06) compared both with the NDR group (0.83+/-0.07) and with the control group (0.81+/-0.06; p = 0.007, p = 0.004). NDR patients had a significantly higher RI in the PCA (0.67+/-0.08) than control patients (0.62+/-0.06, p = 0.01, while the CRA RI (0.71+/-0.09) did not show significant differences from the control group (0.69+/-0.08, p = 0.32). Decreased EDV in the CRA was detected in NDR patients (2.16+/-0.76 cm/s) compared with the controls (2.72+/-0.92 cm/s, p = 0.007). CONCLUSION: The results from this study suggest that not only the retinal but also the choroidal circulation is affected in NDR and BDR patients.

Effects of topical latanoprost on optic nerve head circulation in rabbits, monkeys, and humans.

PURPOSE: To evaluate the effect of topically administrated latanoprost on optic nerve head (ONH) circulation in Dutch rabbits, cynomolgus monkeys, and normal humans. METHODS: The ONH tissue blood velocity (NB(ONH)) was determined using the laser speckle method. Latanoprost (0.005%, 30 microl) was instilled into one eye, and vehicle into the other eye as a control. In rabbits, NB(ONH) was measured for 90 minutes after a single instillation and before and after a 7-day once-daily instillation regimen. In monkeys, NB(ONH) was measured before and after 1, 4, and 7 days of a once-daily instillation regimen. The effect of intravenous indomethacin on the latanoprost-induced NB(ONH) change was also studied in rabbits and monkeys. In humans, the time-course changes in NB(ONH) were measured for 4.5 hours before and after a 7-day once-daily instillation regimen. Intraocular pressure (IOP) and systemic parameters were simultaneously studied in each experiment. All measurements were performed by investigators masked to the experimental condition. RESULTS: Latanoprost significantly increased NB(ONH) 10% to 19% in treated eyes after a single instillation (P = 0.035) or 7-day instillation regimen (P = 0.035) in rabbits, after a 4-day (P = 0.035) or 7-day (P = 0.035) instillation regimen in monkeys, and after a 7-day (P = 0.013) instillation regimen in humans, whereas there were no significant changes in the vehicle-treated eyes in any of the experiments (P > 0.5). Pretreatment with indomethacin (5 mg/kg) abolished the NB(ONH) increase but not the IOP reduction in latanoprost-treated eyes in rabbits and monkeys. IOP remained unchanged in both eyes in rabbits (P > 0.4), whereas it significantly decreased only in latanoprost-treated eyes in monkeys (P < 0.05) and humans (P < 0.05). CONCLUSIONS: Topical latanoprost significantly increased ONH blood velocity only in treated eyes in rabbits, monkeys, and humans. This effect was independent of the IOP-reducing effect of latanoprost and probably was associated with local penetration of the drug and the production of endogenous prostaglandins.

Optic nerve head circulation after intraocular pressure reduction achieved by trabeculectomy.

OBJECTIVE: To study the effects of trabeculectomy and needling revision of poorly functioning blebs on the optic nerve head (ONH) circulation in patients with primary open-angle glaucoma (POAG). DESIGN: Prospective, nonrandomized, self-controlled trial. PARTICIPANTS: Nineteen POAG patients (age range, 52 +/- 12 years; mean +/- standard deviation) undergoing trabeculectomy and six POAG patients (age range, 62 +/- 14 years) undergoing needling revision of the bleb. METHODS: Using the laser speckle method, the normalized blur (NB) value, a quantitative index of blood velocity, was determined every 0.125 seconds and averaged more than three cardiac pulses in the optic nerve head (NB(ONH)). MAIN OUTCOME MEASURES: The NB(ONH) and intraocular pressure (IOP) in both eyes, and blood pressure (BP) and pulse rate (PR) were measured before and 2 days and 1, 4, and 8 weeks after trabeculectomy, and also before and 10 and 40 min after needling procedures. RESULTS: Intraocular pressure in the operated eye was significantly decreased after trabeculectomy or needling procedures, and the ocular perfusion pressure was significantly increased by a maximum of 38%. The IOP in the unoperated eye, BP, and PR did not significantly change. The NB(ONH) did not significantly change in either the operated or unoperated eye. CONCLUSIONS: Trabeculectomy and needling procedures induced little change in the ONH circulation.

The acute effects of stellate ganglion block on circulation in human ocular fundus.

PURPOSE: To study the acute effects of local-anesthetic stellate ganglion block (SGB) on tissue circulation in the human fundus. METHODS: Eleven patients with Bell's palsy (age 56+/-6 y, mean+/-SD) who underwent SGB for its treatment participated in the study. Using the laser speckle method, normalized blur (NB) value, a quantitative index of tissue blood velocity, was measured every 0.125 s over an area located halfway between the macula and the optic nerve head (ONH) with no discrete visible vessels and averaged over 3 pulses when fixation was satisfactory (NB(ch-ret)). NB(ch-ret) and intraocular pressure (IOP) in both eyes, blood pressure (BP), and pulse rate (PR) were measured before, and 10, 20, 30, and 60 min after SGB. SGB was induced by injecting 1% mepivacaine hydrochloride (5 ml) into the vicinity of the seventh cervical vertebra on the paralyzed side. RESULTS: The IOP in the blocked side significantly decreased between 20 and 60 min following SGB, compared to the baseline, while IOP in the unblocked side remained unchanged. The NB(ch-ret) was significantly increased after 10 min by about 8% in the blocked side, but its effect almost disappeared at 60 min. There was no significant change in NB(ch-ret) in the unblocked side, BP or PR throughout the experimental period. CONCLUSION: SGB increased tissue circulation in the fundus in the blocked side, but its effect was thought to be small and transient.

Topical latanoprost and optic nerve head and retinal circulation in humans.

The purpose of the present study was to study the effect of a single instillation of latanoprost on the human optic nerve head (ONH) and retinal circulation. Using laser-speckle tissue blood flow analysis, normalized blur (NB; a quantitative index of tissue blood velocity) was measured every 0.125 sec at a temporal ONH site free of visible surface vessels. Measurements were averaged for 3 cardiac cycles (NB(ONH)). Color Doppler Imaging (CDI) was also used to evaluate peak systolic blood velocity (PSV), endo-diastolic velocity (EDV), and resistive index (RI) in the central retinal artery (CRA) and mean blood velocity (MV) in the central retinal vein (CRV). One drop of 0.005% latanoprost was instilled into one eye and its vehicle into the other in eleven healthy volunteers in a double-blinded manner. Measurements of bilateral NB(ONH), CDI parameters, intraocular pressure (IOP), blood pressure (BP), and pulse rate (PR) were performed before, and 45, 90, 180, and 270 min after instillation. After a single instillation of latanoprost or the vehicle, there was no significant bilateral difference throughout the experimental period. The difference in NB(ONH) between that before and at each time point of measurement (delta NB(ONH)) in the latanoprost-treated eyes was significantly higher between 45 and 270 min after instillation than that in vehicle-treated eyes (P = 0.0003 to 0.0156); ANOVA for repeated measurements also revealed significant difference between both eyes (P < 0.00001). BP, PR, and NB(ONH) in the eye that received only the vehicle, PSV, EDV, and RI in the CRA in both eyes, and MV in the CRV in both eyes changed little. Tissue blood velocity in the ONH increased at least temporarily following a single instillation of topical latanoprost. Although the mechanism of the increase is unclear, the effects of latanoprost on ONH tissue circulation in humans may have clinical implications.

Two cases of frosted branch angiitis with central retinal vein occlusion.

BACKGROUND: Frosted branch angiitis usually occurs in children, and has a good prognosis. We report two cases of unilateral frosted branch angiitis in adults. Both had poor visual outcomes because of associated central retinal vein occlusion and neovascular glaucoma. CASES: Case 1 was a 36-year-old woman. Almost all retinal veins and some retinal arteries showed vasculitis in her right eye, and veins were slightly dilated and sheathed. Case 2 was a 23-year-old woman. Angle hypopyon was observed in her left eye. Retinal veins were dilated, meandering, and sheathed. Retinal hemorrhages were also observed. In both cases, after systemic steroid therapy the retinal vasculitis gradually decreased, but central retinal vein occlusions gradually developed. Despite systemic administration of urokinase and panretinal photocoagulation, neovascular glaucoma developed, and visual acuity diminished in both cases. CONCLUSIONS: Two cases of frosted branch angiitis complicated by retinal vein occlusion are reported. Careful observation of retinal blood flow is necessary in frosted branch angiitis in adults.

Effect of topical unoprostone on circulation of human optic nerve head and retina.

The purpose of the present study was to study the effect of topical unoprostone on the circulation of human optic nerve head (ONH) and retina in normal subjects. Using laser-speckle tissue blood flow analysis, normalized blur (NB), a quantitative index of tissue blood velocity, was measured every 0.125 sec at a temporal ONH site, free of visible surface vessels. Measurements were averaged for 3 cardiac cycles (NB(ONH)). Color Doppler imaging (CDI) was also used to evaluate peak systolic blood velocity (PSV), end-diastolic velocity (EDV), and resistive index (RI) in the central retinal artery (CRA) and mean blood velocity (MV) in the central retinal vein (CRV). For baseline comparison (Day 0), recordings of bilateral NB(ONH) and intraocular pressure (IOP), blood pressure (BP), and pulse rate (PR) were recorded in healthy volunteers before, and 45, 90, 180, and 270 min after instillation of one drop of unoprostone vehicle. On Day 1 (the day after baseline measurements), and twice daily for 7 days, one drop of 0.12% unoprostone was instilled into one eye and its vehicle into the other in a double-blinded manner. Measurements as on Day 0 were recorded on Days 1 and 7. CDI measurements were performed before and at 45 and 180 min after morning instillation on Days 1 and 7. During baseline recordings, there were no significant changes in any parameters. After administration of topical unoprostone, IOP was significantly lower bilaterally with more reduction in the unoprostone-treated eyes on Day 7. On Day 7, the NB(ONH) of the unoprostone-treated eyes was significantly higher 45 min after instillation than baseline (P = 0.035 with Bonferroni's correction). Analysis of variance for repeated measurements also revealed significant difference between Day 0 and Day 7 (P = 0.0017). BP, PR, NB(ONH) in the eye that received only the vehicle, PSV, EDV, and RI in the CRA in both eyes, and MV in the CRV in both eyes changed little. Tissue blood velocity in the ONH increased, at least temporarily, following instillation of unoprostone twice daily for 7 days. Although the clinical implication of the increase is unclear, the effects of topical unoprostone on human ONH circulation deserve further consideration.

The acute effects of cigarette smoking on human optic nerve head and posterior fundus circulation in light smokers.

PURPOSE: To study the acute effects of cigarette smoking on tissue circulation in the optic nerve head (ONH) and posterior fundus in smokers with a short smoking history. METHODS: Ten healthy smokers whose length of smoking history was within 2 years (age 25 +/- 1 years; smoking index (number of cigarettes smoked per day x length of smoking history in years) 16 +/- 4, mean +/- SE) were included in the study. Using the laser speckle method, normalised blur (NB) value, a quantitative index of tissue blood velocity, was measured every 0.125 s and averaged over three pulses across an area located in the temporal site of the ONH free of visible surface vessels (NBONH) and across an area located halfway between the macula and the ONH with no discrete vessels visible (NBch-ret). NBONH, NBch-ret and intraocular pressure (IOP) in one randomly chosen eye, and blood pressure (BP) and pulse rate (PR) were measured before, and 1, 5, 10, 15, 20, 25, 30, 45, 60 and 90 min after sham smoking. One week later, NBONH, NBch-ret and IOP in the same eye, and BP and PR were measured after cigarette smoking according to the same time schedule as in the control experiment. RESULTS: After sham smoking, no parameter showed a significant change during the experiment. Differences in NBONH from the baseline were not significant between the smoking experiment and sham smoking experiment, while NBch-ret showed a significant difference at 30 min. Inter-group difference in the time course of the difference from baseline was significant (ANOVA, p = 0.0246, 0.0021). BP and PR were significantly increased between 1 and 20 min after smoking while IOP showed no significant change at any time of measurement. CONCLUSIONS: In light smokers, cigarette smoking showed little effect on tissue blood velocity in the ONH and slightly decreased that in the posterior fundus, suggesting a significant increase in vascular resistance in these tissues.

Effects of scleral buckling and encircling procedures on human optic nerve head and retinochoroidal circulation.

AIMS: To study the effects of segmental scleral buckling and encircling procedures on tissue circulation in the human optic nerve head (ONH) and choroid and retina. METHODS: Using the laser speckle method, the normalised blur (NB) value, a quantitative index of tissue blood velocity, was measured every 0.125 seconds and averaged over three pulses in the optic nerve head (NB(ONH)) and choroid and retina (NB(ch-ret)) in 10 patients with unilateral rhegmatogenous retinal detachment (mean age 52 (SD 17)). NB(ONH), NB(ch-ret), and intraocular pressure (IOP) in both eyes, and blood pressure (BP) were measured before, and 1, 4, and 12 weeks after the scleral buckling and encircling procedure. RESULTS: NB(ch-ret) on the buckled side was significantly reduced after surgery and smaller than that in the unoperated contralateral eye throughout the study period (ANOVA, p<0.0001). NB(ch-ret) on the unbuckled side, in the foveal area, NB(ONH), IOP, and BP showed no significant change. CONCLUSIONS: It was indicated that the segmental scleral buckling procedure with encircling elements decreased tissue blood velocity in the choroid and retina on the buckled side but caused no significant change on tissue circulation in other areas of the fundus or ONH.

Effect of topical betaxolol on tissue circulation in the human optic nerve head.

There have been no reports to date on long-term betaxolol instillation effects on the human optic nerve head (ONH) tissue circulation. The purpose of this study was to study the effect of topical 0.5% betaxolol on tissue blood velocity in the human ONH. Using a laser-speckle tissue blood flow analyzer, normalized blur (NB; a quantitative index of tissue blood velocity) was measured every 0.125 seconds at a temporal ONH site free of visible surface vessels. Measurements were averaged for 3 cardiac cycles (NB(ONH)). For baseline comparison (day 0), recordings of bilateral NB(ONH) and intraocular pressure (IOP), blood pressure (BP) and pulse rate (PR) were recorded in healthy volunteers before, and 2, 4.5, and 7 hr after, instillation of 30 microL of betaxolol vehicle, and again on day 21; IOP was also recorded on days 7 and 14. On day 1 (the day after baseline measurements), and twice daily for 3 weeks, 30 microL of 0.5% betaxolol into one eye and 30 microL vehicle was instilled into the other in a double-blind study. Measurements as on day 0 were again recorded on day 21; IOP was also recorded on days 7 and 14. During baseline recordings, no significant changes were noted in any parameters. After administration of topical betaxolol, IOP was significantly reduced, bilaterally, with greater reduction in the betaxolol-treated eyes on day 21. Also on day 21, the NB(ONH) of the betaxolol-treated eyes was significantly higher 4.5 hr after instillation than that of the comparable baseline recording (p = 0.035 with Bonferroni's correction); BP, PR, and NB(ONH) in the eye which received only the vehicle showed little change. Tissue blood velocity in the human ONH was increased at least temporarily by instillation of topical betaxolol twice daily for 3 weeks. Although the obtained increase is small and may be clinically insignificant, the potential of betaxolol that can affect the ONH tissue circulation in humans after 21 days of instillation is thought to deserve further investigation.

[Two cases of frosted retinal angiitis with central retinal vein occlusion].

BACKGROUND: Frosted retinal angiitis usually occurs in children, and has a good prognosis. We report two cases of unilateral frosted retinal angiitis in adults. They resulted in visual degradation because of associated central retinal vein occlusion and neovascular glaucoma. CASES: Case 1 was a 36-year-old female. Almost all retinal veins and some retinal arteries had vasculitis in her right eye, and the veins were slightly dilated and sheathed. Case 2 was a 23-year-old female. Angle hypopyon was observed in her left eye. Retinal veins were dilated, meandered, and sheathed. Retinal hemorrhages were also observed. In both cases, systemic steroid therapy gradually improved the retinal vasculitis, but central retinal vein occlusions gradually developed, and in spite of systemic administration of urokinase and panretinal photocoagulation, neovascular glaucoma developed, and visual acuity became degraded in both cases. CONCLUSION: Two cases of frosted retinal angiitis complicated by retinal vein occlusion were reported. Careful observation of retinal blood flow is necessary in frosted retinal angiitis in adults.

Real-time blood velocity measurements in human retinal vein using the laser speckle phenomenon.

PURPOSE: To measure the in vivo blood velocity in human retinal veins using a laser speckle system. METHODS: The system consists of a fundus camera, a diode laser, an image sensor, and a personal computer system. The fundus area, including a target retinal vein, is illuminated with a diode laser through a fundus camera and the laser speckle pattern is imaged onto the area sensor. From the time change of the contrast of the speckle pattern, the normalized blur (NB) value, a quantitative index of blood velocity, was calculated using a logic board. RESULTS: In an in vitro experiment, the NB obtained from blood flow in 50-300 microm internal diameter glass capillary tubes, used as an analogue of a retinal vein, correlated with the diameter of the tube, the actual blood flow rate, and the background NB value, which was used as an analogue of choroidal circulation. In the in vivo experiment, the blood velocity in human retinal veins of approximately 50 microm in diameter was estimated in 16 normal human eyes using nomograms based on the result of the in vitro experiment. Velocity averaged 11.1+/-0.6 mm/s (mean +/- SD, n = 16) in retinal veins 53+/-6 microm in diameter. The coefficient of reproducibility of 5-minute interval measurements was 2.5+/-0.9%, and it took 63+/-15 seconds for one measurement. CONCLUSIONS: The present methodology is clinically valid for measuring blood velocity in retinal veins.

Effects of nilvadipine, a calcium antagonist, on rabbit ocular circulation and optic nerve head circulation in NTG subjects.

PURPOSE: To study the effects of nilvadipine, a Ca2+ antagonist, on tissue circulation in the optic nerve head (ONH), choroid, and retina in rabbits and on the ONH circulation in normal tension glaucoma (NTG) patients. METHODS: Nilvadipine (3.2 microg/kg) or vehicle solution was injected intravenously into urethane-anesthetized rabbits, and the normalized blur value (NB), a quantitative index of in vivo tissue blood velocity, was measured in the choroid and in an area of the ONH and retina free of visible surface vessels before and for 90 minutes after injection, using the laser speckle method. The effects of nilvadipine on the ONH circulation was also studied using the H2 gas clearance method in separate groups of rabbits. Oral nilvadipine (4 mg/d) or placebo was administered to NTG patients in a double-masked manner, and NB in an area of the ONH rim free of visible surface vessels was measured by the same method before and 2, 4, 8, and 12 weeks after administration. RESULTS: The NB obtained from the ONH, choroid, or retina during the experimental period was increased by approximately 10% to 25% in the nilvadipine group compared with the NB in the control group (P < 0.0001, ANOVA), although systemic condition parameters and intraocular pressure (IOP) showed no significant intergroup difference except for a transient decrease in blood pressure in the nilvadipine groups. Blood flow rate in the ONH determined by the H2 gas clearance method also showed an approximately 25% increase in the nilvadipine group. The NB in the ONH of the oral nilvadipine-treated patients was significantly increased, by approximately 20% compared with the placebo-treated patients throughout the follow-up period. No significant intergroup difference was seen in blood pressure, pulse rate, or IOP. CONCLUSIONS: Nilvadipine increased blood velocity and, probably, blood flow in the ONH, choroid, and retina of rabbits. It also increased blood velocity in the ONH of NTG patients.

Acute effects of cigarette smoking on tissue circulation in human optic nerve head and choroid-retina.

OBJECTIVE: To study the acute effects of cigarette smoking on tissue circulation in the human optic nerve head (ONH) and choroid-retina. DESIGN: Nonrandomized, comparative trial (sequential self-controlled). PARTICIPANTS: Nine healthy habitual smokers (age, 28 +/- 4 years; number of cigarettes smoked per day, 27 +/- 10; length of smoking history, 10 +/- 4 years; mean +/- standard deviation). INTERVENTION: Using the laser speckle method, normalized blur (NB) value, a quantitative index of tissue blood velocity, was measured every 0.125 second and averaged over three pulses across an area located in the temporal site of the ONH free of visible surface vessels (NB(ONH)) and across an area located halfway between the macula and the ONH with no discrete vessels visible (NB(ch-ret)). NB(ONH), NB(ch-ret), and intraocular pressure (IOP) in one randomly chosen eye, and blood pressure (BP) and pulse rate (PR) were measured before and 1, 5, 10, 15, 20, 25, 30, 45, 60, and 90 minutes after sham smoking using a short drinking straw as a cigarette substitute (control). One week later, NB(ONH), NB(ch-ret), and IOP in the same eye, and BP and PR were measured after cigarette smoking according to the same time schedule as in the control experiment. MAIN OUTCOME MEASURES: NB(ONH) and NB(ch-ret). RESULTS: In the control experiment, all parameters examined showed no significant change during the experimental period. Differences in NB(ONH) and NB(ch-ret) before and after actual smoking were significantly greater than those in the control experiment (analysis of variance, P = 0.0000, 0.0000). BP and PR were significantly increased between 1 and 30 minutes after actual smoking as compared with control data, while IOP showed no significant change at any time of measurement. CONCLUSIONS: These results indicated that cigarette smoking increased tissue blood velocity in the ONH and possibly in the choroid in habitual smokers.

[A case of neuroblastoma with abnormal LD isoenzyme].

We encountered a patient with neuroblastoma showing remarkably high total LD activity (12,585 IU/l). His liver and heart function was normal. In the serum LD isoenzyme pattern, LD1 was increased, and LD2 extra band (LD2 ex) was observed on the negative pole side of LD2. However, LD2 ex was absent in erythrocytes of the patient, which demonstrates the exclusion of genetic factors. Neither the enzyme counter current method or immunofixation showed immune complexes, excluding anomalies. The LD isoenzyme in a metastatic lymphnode lesion before treatment showed increased LD1. The LD isoenzymes in the tissue, removed after 5 courses of chemotherapy, showed predominance of LD3 and absence of LD2 ex. Rapid decreases in the serum total LD activity and LD1 fraction and the disappearance of LD2 ex after chemotherapy suggest that these changes in the two LD fractions are associated with production of abnormal LD (LD2 ex) by the tumor.