Requirement of Fas for the development of autoimmune diabetes in nonobese diabetic mice.

Insulin-dependent diabetes mellitus (IDDM) is assumed to be a T cell-mediated autoimmune disease. To investigate the role of Fas-mediated cytotoxicity in pancreatic beta cell destruction, we established nonobese diabetic (NOD)-lymphoproliferation (lpr)/lpr mice lacking Fas. Out of three genotypes, female NOD-+/+ and NOD-+/lpr developed spontaneous diabetes by the age of 10 mo with the incidence of 68 and 62%, respectively. In contrast, NOD-lpr/lpr did not develop diabetes or insulitis. To further explore the role of Fas, adoptive transfer experiments were performed. When splenocytes were transferred from diabetic NOD, male NOD-+/+ and NOD-+/lpr developed diabetes with the incidence of 89 and 83%, respectively, whereas NOD-lpr/lpr did not show glycosuria by 12 wk after transfer. Severe mononuclear cell infiltration was revealed in islets of NOD-+/+ and NOD-+/lpr, whereas islet morphology remained intact in NOD-lpr/lpr. These results suggest that Fas-mediated cytotoxicity is required to initiate beta cell autoimmunity in NOD mice. Fas-Fas ligand system might be critical for autoimmune beta cell destruction leading to IDDM.

The Effect of Branched Chain Amino Acids-Enriched Nutritional Supplements on Activities of Daily Living and Muscle Mass in Inpatients with Gait Impairments: A Randomized Controlled Trial.

OBJECTIVE: To investigate the effects of continuous intervention with branched chain amino acids-enriched nutritional supplements from the acute phase to convalescent rehabilitation wards in inpatients with gait impairments. DESIGN: Open-label, randomized, parallel-group comparison study (UMIN Clinical Trials Registry ID: UMIN000018640). SETTING: Acute care and convalescent rehabilitation wards. PARTICIPANTS: We studied 80 patients undergoing stand/gait training. INTERVENTIONS: Participants in the intervention group (RJ group) received nutritional supplements (jelly foods comprising 2500 mg BCAA and 20 IU vitamin D) twice a day until hospital discharge. MEASUREMENTS: The primary outcome was the motor components of the Functional Independence Measure (FIM-m), and the secondary outcome was skeletal muscle mass index. RESULTS: Analyses were conducted on 55 patients who were able to perform stand/gait training continuously from the acute until the recovery phases. FIM-m was significantly elevated in the RJ group and the control group , but no difference was noted between the two groups. Only the RJ group showed a significant increase in skeletal muscle mass index, and the amount of variation was significantly different between the two groups (the control group decreased an average of 2.2% and the RJ group increased an average of 4.3%; P = 0.014). A significant decrease in body weight was found only in the control group (P = 0.084). CONCLUSIONS: Nutritional interventions using branched chain amino acids (BCAA)-enriched nutritional supplements demonstrated no significant difference in activities of daily living; however, an increase in skeletal muscle mass was noted. Skeletal muscle mass and body weight differed significantly between the two groups, and BCAA-enriched nutritional supplements intake in acute and convalescent rehabilitation wards may be effective for the prevention of malnutrition and sarcopenia.

Weekly paclitaxel/5-fluorouracil followed by platinum retreatment for patients with recurrent ovarian cancer: a single institution experience.

PURPOSE: Since the prognosis of recurrent ovarian cancer patients is still poor, we need to establish a useful treatment strategy to achieve their long-term survival. We treated recurrent ovarian cancer patients with weekly paclitaxel (PTX)/5-fluorouracil (5-FU) followed by platinum retreatment to investigate its clinical efficacy in a preliminary manner. METHODS: Sixteen patients with recurrent ovarian cancer, pretreated with taxane and platinum, were treated with weekly paclitaxel (PTX)/5-fluorouracil (FU). PTX (80 mg/m2) on day 1, 8, and 15 was combined with a bolus injection of 5-FU (500 mg/m2) on day 2, 9, and 16. Chemotherapy was given every four weeks. Patients with stable disease or progressive disease were subsequently retreated with a platinum-containing regimen. Response was evaluated by RECIST criteria or CA125 criteria. Toxicities were evaluated according to the National Cancer Institute-common toxicity criteria (NCI-CTC) version 3. RESULTS: Among five patients with sensitive disease, one of four patients with measurable tumor and one without measurable tumor responded to weekly PTX/5-FU. Among 11 patients with resistant disease, none of five patients with measurable tumor and three of six patients without measurable tumor responded to weekly PTX/5-FU. Overall objective response rate by weekly PTX/5-FU was 31.3% (5/16). Among 16 patients, 13 patients who showed no response or progressive disease (three with sensitive disease, ten with resistant disease) received platinum retreatment after weekly PTX/5FU. All three patients with sensitive disease and three of ten patients with resistant disease revealed response to platinum retreatment. Overall objective response rate by platinum retreatment after weekly PTX/5-FU was 46.2% (6/13). CONCLUSIONS: Weekly PTX/5FU followed by platinum retreatment could be a useful treatment strategy for recurrent ovarian cancer patients. We need to establish the standard treatment strategy for recurrent ovarian cancer patients with a poor prognosis.

Acid-suppressive effects of rabeprazole: comparing 10mg and 20mg twice daily in Japanese Helicobacter pylori-negative and -positive CYP2C19 extensive metabolisers.

BACKGROUND: Rabeprazole 10mg b.i.d. is often administered as therapy for eradication of Helicobacter pylori (H. pylori) and is also proposed as therapy for refractory gastro-oesophageal reflux disease. However, there has not been a comprehensive assessment of its acid-suppressive effects. AIMS: To compare the acid-suppressive effects of rabeprazole 10mg b.i.d. with 20mg b.i.d. considering H. pylori status. SUBJECTS: Thirteen H. pylori-negative and eleven H. pylori-positive Japanese CYP2C19 extensive metabolisers (<35 years). METHODS: Intragastric pH was measured for 24h three times in a randomised manner; on day 7 of the repeated administration of rabeprazole 10mg b.i.d. or 20mg b.i.d., or a placebo. RESULTS: In median intragastric pH value and percent time of pH>3.0, >4.0, >5.0, >6.0, and >7.0 for 24h, no significant differences were observed between the two doses in either H. pylori-negative or H. pylori-positive subjects. At either dose, these parameters were significantly higher in H. pylori-positive subjects than in H. pylori-negative subjects. Nocturnal acid breakthrough occurred in seven and two of the thirteen H. pylori-negative subjects and one and two of the eleven H. pylori-positive subjects at each dose, respectively. CONCLUSIONS: The effects of rabeprazole 10mg b.i.d. were equal to those of 20mg b.i.d. in H. pylori-positive subjects; whereas in H. pylori-negative subjects, 20mg b.i.d. was superior for prevention of nocturnal acid breakthrough.

Recombinant human betacellulin promotes the neogenesis of beta-cells and ameliorates glucose intolerance in mice with diabetes induced by selective alloxan perfusion.

Betacellulin (BTC), a member of the epidermal growth factor family, is expressed predominantly in the human pancreas and induces the differentiation of a pancreatic acinar cell line (AR42J) into insulin-secreting cells, suggesting that BTC has a physiologically important role in the endocrine pancreas. In this study, we examined the in vivo effect of recombinant human BTC (rhBTC) on glucose intolerance and pancreatic morphology using a new mouse model with glucose intolerance induced by selective alloxan perfusion. RhBTC (1 microg/g body wt) or saline was injected subcutaneously every day from the day after alloxan treatment. The intraperitoneal glucose tolerance test revealed no difference between rhBTC-treated and rhBTC-untreated glucose-intolerant mice at 2-4 weeks. However, glucose tolerance was significantly improved and body weight was significantly increased in rhBTC-treated mice compared with untreated mice at 8 weeks. Islet-like cell clusters, consisting mainly of beta-cells, were increased in the pancreas and were localized in contact with the ductal lining cells and sometimes with acinar cells. In conclusion, administration of rhBTC improved glucose tolerance in this mouse model by increasing beta-cell volume, primarily through accelerated neogenesis from ductal lining cells.

Pancreatic biopsy as a procedure for detecting in situ autoimmune phenomena in type 1 diabetes: close correlation between serological markers and histological evidence of cellular autoimmunity.

To better understand the pathogenesis of type 1 diabetes, we have developed pancreatic biopsy under laparoscope for recent-onset type 1 diabetic patients. The patients included 29 acute-onset type 1 diabetic patients, 5 latent-onset type 1 diabetic patients, and 1 type 2 diabetic patient. Their median age was 28 years, and the duration of diabetes at the time of biopsy was approximately 3 months. In 31 of 35 patients, we could obtain the pancreas tissue by punching. No serious complications, such as heavy bleeding, peritonitis, or pancreatitis, have been experienced. Pneumoderma was observed in two patients, and abdominal dull pain had continued for 2 days in two patients. However, special treatment was not necessary for these complications. T-cell-predominant infiltration to islets (insulitis) and hyperexpression of major histocompatibility complex class I antigens on islet cells were the two major findings and were observed in 17 of 29 recent-onset type 1 diabetic patients. These findings could be regarded as evidence of immune attack against beta-cells, and their presence was closely correlated with the presence of either anti-GAD or anti-IA-2 antibodies (P = 0.02). In conclusion, pancreatic biopsy under laparoscope is a safe procedure without serious complications, according to our findings, for detecting in situ autoimmune phenomenon in recent-onset type 1 diabetic patients.

The involvement of polyamines in the proliferation of cultured retinal pigment epithelial cells.

PURPOSE: To investigate the involvement of polyamines (putrescine, spermidine, and spermine) in the proliferation of retinal pigment epithelium (RPE) using cultured bovine RPE cells. METHODS: The polyamine content and the activities of rate-limiting enzymes in polyamine biosynthesis (ornithine decarboxylase [ODC] and S-adenosylmethionine decarboxylase [SAMDC]) and in polyamine biodegradation (spermidine spermine N1-acetyltransferase [SAT]) were measured after proliferative stimulation by 10% fetal calf serum (FCS). DNA synthesis was determined by [3H]thymidine incorporation into the acid-insoluble fraction after the addition of an inhibitor of ODC (alpha-difluoromethylornithine [DFMO]) or SAMDC (methylglyoxal bis[guanylhydrazone] [MGBG]). The effects of exogenous polyamines on DNA synthesis after the additions of inhibitors also were determined. RESULTS: ODC and SAMDC activities were elevated after stimulation by FCS and reached their peaks 16 hours and 4 hours, respectively, after the addition of FCS. SAT activity was not increased. Polyamine content was increased significantly after stimulation by FCS. DFMO did not inhibit DNA synthesis induced by FCS, and only putrescine content was decreased significantly among polyamines. However, MGBG inhibited DNA synthesis in a dose-dependent manner, and the amounts of spermidine and spermine were decreased significantly. Exogenous polyamines, especially spermine, restored MGBG-induced inhibition of DNA synthesis. CONCLUSIONS: Polyamines are essential for the proliferation of cultured bovine RPE cells. These data suggest that, of the polyamines, spermine has the greatest effect on DNA synthesis although other polyamines can substitute for spermine at higher concentrations with similar results. As for polyamine metabolism in RPE proliferation, it is possible that SAMDC is the key enzyme rather than ODC.

Overexpression of the aldose reductase gene induces apoptosis in pancreatic beta-cells by causing a redox imbalance.

To determine the role of the polyol metabolizing pathway under hyperglycemic conditions, the effects of aldose reductase (AR) on the cellular functions of pancreatic beta-cells were examined. Stable transfectants of rat AR cDNA were obtained with a pancreatic beta-cell line, HIT, in which a negligible amount of AR was originally expressed. Overproduction of AR triggered DNA fragmentation, as judged with the TUNEL method and agarose gel electrophoresis. Morphological analysis by electron microscopy also clearly showed apoptosis of the AR-overexpressing HIT cells. Induction by interleukin-1beta of gene expression such as those of an inducible form of nitric oxide synthase (NOS-II) and Mn-superoxide dismutase (Mn-SOD), was much lower in the transfectants than in the control cells, while the expression of constitutively expressed genes such as those for Cu,Zn-superoxide dismutase and insulin was not changed. The susceptibility to interleukin-1beta stimulation of the expression of the NOS II and Mn-SOD genes was due to suppressed NF-kappaB activity, which is essential for the expression of these genes. In addition, the intracellular NADPH/NADP+ ratio was considerably lower in the AR-transfected cells than in control cells. Thus, the overexpression of AR in pancreatic beta-cells induced apoptosis that may be caused by a redox imbalance.

Acute retinal necrosis late in the second trimester.

PURPOSE: To report treatment of a patient with acute retinal necrosis during pregnancy. METHODS: A 24-year-old woman in her twenty-third week of gestation was diagnosed with acute retinal necrosis. A combination of acyclovir and interferon therapy was started at 25 weeks. Pars plana vitrectomy was performed during the 26th week of gestation. RESULTS: The necrotizing retina became gliotic within 3 weeks of surgery. The patient's visual acuity improved to LE, 20/40. A healthy baby was delivered at 39 weeks of gestation. CONCLUSION: Combination therapy of acyclovir and interferon followed by surgery partially restored the patient's vision without affecting fetal development.

Organ-specific expression of ß-glucuronidase activity driven by the Arabidopsis heat-shock promoter in heat-stressed transgenic Nicotiana plumbaginifolia.

The expression of the Arabidopsis heat-shock protein (HSP) 18.2 promoter ß-D-glucuronidase (GUS) chimera gene was studied in various organs of the transgenic Nicotiana plumbaginifolia during the recovery phase at normal temperatures (20-22  °C) following heat-shock (HS) treatment. The optimum HS temperature for GUS activity in the anthers, petals and capsules was 42  °C, but in immature seeds and the placentas of capsules it was 36  °C and 39  °C, respectively. No apparent GUS activity was observed in any organs except for dry seeds after HS at 45  °C, although the activity in dry seeds was apparent even after HS at 48  °C. After HS at 42  °C, GUS activity in the flower tissues was the highest before anthesis and declined thereafter.

Delayed recovery of ß-glucuronidase activity driven by an Arabidopsis heat shock promoter in heat-stressed transgenic Nicotiana plumbaginifolia.

The expression of the Arabidopsis heat shock protein (HSP) 18.2 promoter-ß-D-glucuronidase (GUS) chimera gene was investigated in transgenic Nicotiana plumbaginifolia plants during the recovery phase at normal temperatures (20-22  °C) after a heat shock (HS) treatment. GUS activity increased during the recovery phase after HS at 42  °C for 2 h, and maximal GUS activity was observed after 12 h at normal temperatures, at levels 50-100 times higher than the activity immediately after HS. After HS at 44  °C, little GUS activity was observed during the first 20-24 h at normal temperatures, but the activity increased gradually thereafter, to reach a maximum at 40-50 h. After HS at 45  °C, no GUS activity was observed throughout the experimental period. RT-PCR analysis showed that GUS mRNA remained for 10 h after a 2-h HS at 42  °C and for 40 h after a 2-h HS at 44  °C. These findings demonstrate that brief HS treatment, especially at a sublethal temperature, induces a long-term accumulation of HSP-GUS mRNA during the recovery phase.

Suppressed phenylalanine ammonia-lyase activity after heat shock in transgenic Nicotiana plumbaginifolia containing an Arabidopsis HSP18.2-parsley PAL2 chimera gene.

The activity of phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) after heat shock (HS) in leaves and buds of transgenic Nicotiana plumbaginifolia containing an Arabidopsis HSP18.2 promoter-parsley phenylalanine ammonia-lyase 2 (HSP18.2-PAL2) chimera gene was examined. Immediately after HS treatment at 44 degrees C for 5 h, the PAL activity in both transgenic and normal (untransformed) plants was 35-38% lower than that before HS. At normal temperature (25-26 degrees C), the PAL activity recovered within 5 h of ending the HS treatment in normal plants, but not until 12-24 h in transgenic plants containing the HSP18.2-PAL2 gene. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed the presence of parsley PAL2 mRNA in transgenic plants, which remained for 8-12 h following 5-h HS at 44 degrees C; the mRNA was not observed before HS. The content of chlorogenic acid (CGA; 3-caffeoylquinic acid) decreased drastically 8-12 h after HS in transgenic plants, but only slightly in normal plants. Thus, the decrease in PAL activity accompanied by expression of the parsley PAL2 gene after HS treatment corresponded to the decrease in CGA synthesis. These results might be attributed to post-transcriptional degradation of endogenous PAL mRNA triggered by transcription of the transgene.

Pathological changes in HPCD pigs with prednisolone induced recrudescence of pseudorabies virus.

In HPCD pigs inoculated with PRV, latent PRV could be reactivated in-vivo by the administration of large doses of prednisolone 3 months after the primary infection. In two pigs, virus shedding was without clinical signs of disease, whereas depression of circulating lymphocytes was prominent. Reactivation of PRV was also demonstrated by cultivation of the brain cortex on the 7th day and the mandibular lymph node on the 9th day after the prednisolone began treatment. Coincident with the virus isolation, characteristic lesions were observed in 2 pigs in the central nervous tissues and mandibular lymph nodes and these were composed of cell necrosis and eosinophilic intranuclear inclusion bodies. Cells containing the intranuclear inclusion bodies had immature and mature PRV particles. Results of the present study with HPCD pigs indicated that the lesions in the brain and lymph node accompanied by eosinophilic intranuclear inclusion bodies were pathogonomonic lesions induced by reactivation of PRV.

Digital fluorescein fundus angiography with DFC-512 and IMAGEnet systems.

Image processing with direct acquisition digital fundus fluorescein angiography using the OIS DFC-512 and Topcon IMAGEnet were assessed. Fundus diseases included central and branch retinal vein occlusion, diabetic retinopathy, age-related macular degeneration, central serous choroidopathy and angioid streaks. Analogue images were taken with Kodak Tri-X 35 mm film as well. Although the resolution of the original image was less than that of 35 mm film, it was improved by image processing, particularly contrast enhancement. One type of contrast enhancement, available in both systems, exaggerated lack of light at the peripheral area of the original image. This drawback was not seen with another type of enhancement processing, available in IMAGEnet. However, the latter produced an artificial hypofluorescent area around an intensely hyperfluorescent lesion and also exaggerated hyper- and hypo-fluorescent patches of the background. Nevertheless, these processing methods were particularly useful in case of media opacity like cataract, even when the 35 mm film picture is not clear. Other processing techniques like pseudocolor and relief, etc. were used just for different ways of presentation. The two systems are basically equivalent, except for contrast enhancement. There was no problem for diagnosis in both systems, but the artificial effects must be borne in mind when interpreting the image.

Computer assisted image analysis using the subtraction method in indocyanine green angiography.

The choroidal vessels are three dimensionally distributed and very complex in their patterns. They often appear to be overlaid in indocyanine green (ICG) angiograms so it is harder to analyze ICG angiography than fluorescein angiography. When an earlier frame is subtracted from a later frame in a sequence of angiograms, the fluorescence which has increased during the time between the two frames can theoretically be demonstrated. We applied computer-assisted image subtraction methods in selected clinical cases of directly acquired digital ICG angiography to demonstrate how this method works. We used software already installed in an IMAGEnet computer system (Topcon) for image subtraction. We applied the subtraction technique in 18 cases with various diseases. When two images with a time difference of several seconds were subtracted, filling of the choriocapillaris, the neovascularization or the pathological vessels could be observed. When the images had a time difference of several minutes, intrachoroidal dye leakage could be seen more clearly. This method is very helpful for analyzing pathological changes in ICG angiography in clinical cases, when two images are selected appropriately.

The relationship between gap-junctional intercellular communication and the proliferative activity of retinal pigment epithelial cells.

PURPOSE: To investigate the relationship between gap-junction mediated intercellular communication and the proliferative activity of retinal pigment epithelial cells in the retinal tear base. METHODS Retinal tears were created experimentally in white rabbits, and the changes of intercellular communication via gap junctions between retinal pigment epithelial cells in the retinal tear base were investigated using the dye-coupling method, which involves observing the spread of a fluorescent dye, Lucifer Yellow CH. In addition, the proliferative activity of these cells was investigated using an antibody for proliferating cell nuclear antigen and was compared with the changes in intercellular communication. RESULTS: Immediately after the creation of retinal tears, extensive spreading of Lucifer Yellow CH into adjacent cells was observed, which was markedly reduced 1 week later. After 1 month, the spreading of dye to adjacent cells was observed again. On the other hand, proliferative activity was enhanced at 1 week after retinal tear creation and was reduced again after 1 month. CONCLUSIONS: It is known that tumor cells with enhanced proliferative activity show decreased intercellular communication via gap junctions. The present study suggests a possible relationship between intercellular communication and the proliferative activity of retinal pigment epithelial cells in the base of retinal tears.

Indocyanine green angiograms of choroidal nevi. comparison between confocal and nonconfocal scanning laser ophthalmoscope and fundus video camera.

PURPOSE: A fundus video camera and a nonconfocal scanning laser ophthalmoscope (SLO) detect direct light and indirect light, whereas a confocal SLO detects mostly direct light. Differences in confocal and nonconfocal SLO images and fundus video camera images are most likely due to their different optical systems. These differences were examined in indocyanine green (ICG) angiograms of a choroidal nevus. METHODS: A confocal SLO, a nonconfocal SLO, and a high resolution digital fundus video camera were used to obtain ICG angiograms of pigmented choroidal nevi in 4 patients for 30 minutes following dye injection. RESULTS: All the angiograms showed a hypofluorescent region in the nevus until 10-14 minutes after dye injection, except in 1 patient in whom no hypofluorescent region was seen in an early confocal-SLO angiogram. From 20 minutes to 30 minutes postinjection, the hypofluorescent regions were still visible in all fundus video camera angiograms and nonconfocal SLO angiograms but not in confocal SLO angiograms. CONCLUSIONS: Early angiograms taken with the three angiography systems showed a similar appearance of the choroidal nevus. However, late ICG angiograms with a confocal SLO showed different images from those taken with a nonconfocal SLO or a fundus video camera. It is suggested that the angiography system and the aperture size of an SLO should be selected according to the aspect of the pigmented choroidal nevus that is of interest in late-phase ICG angiography.

Pathologic changes in pigs with prednisolone-induced recrudescence of herpesvirus infection.

In pigs inoculated with Aujeszky's disease virus (ADV), recrudescence of herpesvirus infection was induced by daily administration of 1,000 mg of prednisolone for 5 days at 2 (group A) or 5 (group B) months after the primary infection. At necropsy in group A pigs, ADV was recovered from nasal secretions 3 to 9 days after prednisolone treatment initiation and from the brain cortex 10 days after treatment initiation; ADV was not recovered from group B pigs. In pigs of both groups killed 10 days after treatment initiation, 2 types of characteristic lesions were found. One type was a nonsuppurative encephalitis that consisted of neuronal necrosis, neuronophagia, and formation of eosinophilic intranuclear inclusion bodies. The 2nd type had basophilic intranuclear inclusion bodies in the enlarged endothelial cells of the kidneys, liver, lungs, adrenal glands, and lymph node sinusoids. Cells containing intranuclear inclusion bodies had immature and mature herpesvirus particles. Therefore, the brain lesions containing the eosinophilic inclusion bodies were considered to be due to ADV. Basophilic inclusion bodies in the endothelial cells were due to porcine cytomegalovirus. These observations indicated that prednisolone treatment resulted in recrudescence of ADV and porcine cytomegalovirus infections.

Phagocytic and digestive activities of retinal pigment epithelium at experimentally produced retinal tears.

The present study examined the phagocytic and digestive activities of retinal pigment epithelial cells (RPE cells) at sites of experimentally produced retinal tears, using the phagocytosis of the latex particles by these cells as a measure of phagocytic activity, and their acid phosphatase activity as a measure of their digestive activity. Four days after retinal tear formation, acid phosphatase activity of RPE cells at the site of retinal tears was the same as that of the RPE cells in the surrounding normal retina. However, at one week, four weeks and six months after tear formation, acid phosphatase activity of RPE cells at the site of retinal tears was higher than that of the RPE cells in the surrounding normal retina. Four weeks after retinal tear formation, a latex particle solution was injected onto the retinal tear or subretinally. The RPE cells at the site of retinal tears phagocytized the latex particles earlier than did the RPE cells of the eye which was injected subretinally. At three hours after the injection, the RPE cells at the site of retinal tear had phagocytized more latex particles in their cytoplasm than RPE cells in normal retina. It can be concluded that RPE cells at the site of retinal tear had higher phagocytic and digestive activity than did RPE cells in normal retina.

Incising the thick retrolental fibrovascular tissue with a hooked sclerotome in persistent hyperplastic primary vitreous.

A technique for incising thick retrolental fibrovascular tissue and extensive cyclitic membrane is reported in a case of anterior persistent hyperplastic primary vitreous. A membranectomy was performed in a 1-month-old post-lensectomy baby via a limbal approach. A sclerotome tip was hooked to cut through an extremely thick fibrovascular tissue by rotating the sclerotome by its grip. Sutherland microscissors (Grieshaber, Switzerland) and a vitrectomy cutter were used for further membranectomy. The baby was followed-up until age 18 months. A total of 3 membranectomy sessions were required because of rapid cyclitic membrane formation, severe centripetal retraction of the membrane on the ciliary processes, and posterior synechia. Thorough membranectomy and cutting the iris edge maintained a clear pupillary area during the 13-month postoperative period. Extremely thick retrolental fibrovascular tissue is a challenging condition that can be dealt with by delicate instrumentation.