A randomised, placebo controlled clinical trial of the aldose reductase inhibitor CT-112 as management of corneal epithelial disorders in diabetic patients.

AIM: To evaluate the efficacy of topical aldose reductase inhibitor CT-112 (5-[3-ethoxy-4-pentyloxyphenyl]-2,4-thiazolidinedione) on corneal epithelial barrier function in diabetic patients. METHODS: This was a prospective, randomised, double masked placebo controlled study. 34 eyes of 34 diabetic patients were randomly assigned treatment with 0.25% eye drops of CT-112 (n = 22) or a placebo (n = 12) four times a day for 8 weeks. Corneal fluorescein staining and corneal sensation were examined before treatment as well as 4 and 8 weeks after administration. Corneal epithelial permeability to fluorescence was measured with an anterior fluorophotometer. RESULTS: Average scores of superficial punctate keratopathy and corneal sensitivity did not differ significantly between the two groups at any time. Whereas average fluorescein concentrations did not differ significantly for the CT-112 and placebo groups before treatment, they did differ significantly 4 and 8 weeks after treatment (4 weeks, p = 0.0327; 8 weeks, p = 0.0143). CONCLUSION: The topical aldose reductase inhibitor, CT-112 improves the corneal epithelial barrier function in diabetic patients.

Selective urinary excretion of phosphatidyl ethanolamine in patients with chronic glomerular diseases.

Urinary phospholipids and lipoproteins in chronic glomerular diseases were analyzed. The subjects used were 26 patients consisting of 14 with chronic glomerulonephritis and 12 with nephrotic syndrome. Nine healthy normals served as controls. Phospholipids were isolated by one-dimensional thin-layer chromatography (TLC) using an internal standard for quantification and partially by two-dimensional TLC and, furthermore, quantified by two different methods to ascertain the kinds of phospholipids. Urinary lipoproteins were isolated by density gradient ultracentrifugation and analyzed by electrophoresis. The urinary excretion of phosphatidyl ethanolamine (PE) was recognized exclusively in the patient group and that of phosphatidyl serine (PS) in most cases with nephrotic syndrome. The daily urinary PE excretion rate was closely correlated to the urinary albumin excretion rate. However, phosphatidyl choline (PC) and sphingomyelin (SPH), which are main phospholipids in serum and red blood cell membranes, in most cases were hardly detected in urine. These observations were confirmed by two-dimensional TLC using valuable spot tests for identification of phospholipids and also by the two different quantification methods. In density gradient ultracentrifugation, urinary lipoproteins did not form such peaks as seen in the profiles of serum lipoproteins. The presence of urinary lipoproteins in two nephrotic patients has been shown, but although the method used was not very sensitive, it was suggested that lipoproteins were hardly excreted into urine as the lipoprotein deficient fraction (LPDF) (d greater than 1.21 g/ml), in which albumin is predominant. PE was found mainly in LPDF of urine, although the amount of PE in urinary lipoproteins was very limited.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in the concentrations of urinary proteins after physical exercise.

The influence of physical exercise on the urinary excretion of proteins was examined in 17 male high school baseball players. Their urine was collected before and after exercise to determine the concentrations of total protein, albumin, beta 2-microglobulin and creatinine along with the activity of N-acetyl-beta-D-glucosaminidase (EC 3.2.1.30). Concentrations of total protein, albumin, beta 2-microglobulin and creatinine increased significantly (p less than 0.01) after exercise, while N-acetyl-beta-D-glucosaminidase activity did not increase. Similar results were obtained when the concentrations of these urinary components were calculated on the basis of a urinary density of 1.024, and when they were expressed relative to the amount of creatinine. Positive correlations were seen among total protein, albumin, beta 2-microglobulin and creatinine concentrations, but not between the beta 2-microglobulin concentration and N-acetyl-beta-D-glucosaminidase activity. Isoenzyme activities of N-acetyl-beta-D-glucosaminidase in the urine were determined by electrophoresis on cellulose acetate plates. After exercise, the A-form increased slightly, and the B-form decreased slightly, but these changes were not statistically significant.

Comparison of postoperative surface light scattering of different intraocular lenses.

AIMS: This retrospective study was conducted to compare postoperative surface scattering of four kinds of intraocular lens (IOL). METHODS: Sixty-seven eyes of 67 patients who had undergone cataract surgery were enrolled in this study. One of four IOLs was used in the patients; MA60BM in 17 patients (MA group), SA60AT in 17 patients (SA group), AR40 in 16 patents (AR group) and ClariFlex in 17 patients (CL group). Measurement of scattering from the anterior surface of the IOL was measured with area densitometry using a Scheimpflug camera (EAS-1000, Nidek, Aichi) for 3 years after the surgery. RESULTS: The density of IOL surface scattering increased starting 1 year after surgery and throughout the 3-year period in the MA group and starting at 6 months through 3 years in the SA group, whereas the density was stable in the AR and CL groups. The density of surface scattering in the MA and SA groups at 3 years after surgery was significantly higher than in the AR and CL groups. CONCLUSION: The surface scattering of MA60BM and SA60AT is higher than that of AR40 and ClariFlex. The grades of surface scattering differ among the manufacturers, even with the same acrylic material.

On the transport and cleavage of ova in the rats' and the rabbits' oviducts with IUD.

PIP: To investigate the possible mode of IUD action, 2 experiments on rats and rabbits were performed to measure time differences of ova passage through the oviduct. A silk thread was inserted into 1 uterine horn while the other served as a control. Superovulation was induced with HCG and the reproductive tract of each animal removed 12, 24, 48, or 72 hours later. In the experiment with rabbits artificial ova labeled with radioisotope were inserted into the fimbria of the Fallopian tubes and passage measured 12, 24, 40, or 56 hours later. In animals sacrificed after 24 to 48 hours, ova were detected in both the IUD and control oviducts but not in the uterus. At 72 hours approximately the same number of fertilized ova were found in each oviduct and in the uteri of both the control and IUD horns. The rabbit experiments also failed to show significant differences. It was concluded that an IUD does not influence speed of transport of the ova in rabbits or rats.^ieng

Antiseptic effects at injection sites.

To our knowledge, there are no published papers detailing antisepsis for injection sites. In view of this, the efficacies of povidone-iodine (PVP-I) ethanol solution and chlorhexidine (CH) ethanol, the agents most commonly used for antisepsis of the operative field, were compared. Before and after the injection site was disinfected with either of these antiseptics, specimens of indigenous bacteria on the skin were collected by the cylinder scrub method, and the bacteria reduction rate and the reduction factor (RF) were determined to evaluate the efficacy of antisepsis. The bacteria reduction rate and RF value obtained for PVP-I ethanol were 95.1 +/- 11.2 and 2.1 +/- 0.9% and those for CH ethanol were 93.5 +/- 9.3 and 1.8 +/- 0.9%. Since there were individual differences in cell count before antisepsis, no significant difference was seen in bactericidal activity. However, slightly more favorable results were obtained with PVP-I ethanol. Although it is impossible to eradicate completely the indigenous microbes with currently available methods, it is considered important for the prevention of infection of the injection site to decrease bacterial counts as much as possible.

Direct reduction of alcohols: highly chemoselective reducing system for secondary or tertiary alcohols using chlorodiphenylsilane with a catalytic amount of indium trichloride.

The direct reduction of alcohols using chlorodiphenylsilane as a hydride source in the presence of a catalytic amount of indium trichloride is described. Benzylic alcohols, secondary alcohols, and tertiary alcohols were effectively reduced to give the corresponding alkanes in high yields. A compound bearing both primary and secondary hydroxyl groups was reduced only at the secondary site to afford the primary alcohol after workup with Bu(4)NF. This system showed high chemoselectivity only for the hydroxyl group while not reducing other functional groups that are readily reduced by standard reducing systems. Thus alcohols bearing ester, chloro, bromo, or nitro groups, which are sensitive to LiAlH(4) or Zn/H(+), were selectively reduced only at the hydroxyl sites by the chlorodiphenylsilane/InCl(3) system. NMR studies revealed the reaction course. The hydrodiphenylsilyl ether is initially formed and then, with InCl(3) acting as a Lewis acid, forms an oxonium complex, which accelerates the desiloxylation with donation of the hydrogen to the carbon.

Motility of single one-headed kinesin molecules along microtubules.

The motility of single one-headed kinesin molecules (K351 and K340), which were truncated fragments of Drosophila two-headed kinesin, has been tested using total internal reflection fluorescence microscopy. One-headed kinesin fragments moved continuously along the microtubules. The maximum distance traveled until the fragments dissociated from the microtubules for both K351 and K340 was approximately 600 nm. This value is considerably larger than the space resolution of the measurement system (SD approximately 30 nm). Although the movements of the fragments fluctuated in forward and backward directions, statistical analysis showed that the average movements for both K340 and K351 were toward the plus end of the microtubules, i.e., forward direction. When BDTC (a 1.3-S subunit of Propionibacterium shermanii transcarboxylase, which binds weakly to a microtubule), was fused to the tail (C-terminus) of K351, its movement was enhanced, smooth, and unidirectional, similar to that of the two-headed kinesin fragment, K411. However, the travel distance and velocity of K351BDTC molecules were approximately 3-fold smaller than that of K411. These observations suggest that a single kinesin head has basal motility, but coordination between the two heads is necessary for stabilizing the basal motility for the normal level of kinesin processivity.

The effect of long-term use of intrauterine devices.

PIP: Of 10,223 Japanese women examined for uterine cancer between 1952 and 1967, 1333 were or had been wearing an IUD during that period; 1075 of the IUD users were studied to assess the results of long-term IUD use and its effect on fertility after removal. Results indicated that fertility after removal was normal depending on the age of the patient. The patients' ages ranged from 21 to 35 years. Closure for medical reasons, e.g., hemorrhage, pain, and unintended pregnancy, were most frequent during the 1st year of use; however, the IUD was removed in almost 1/2 of the patients after 5 years of use because of desire for pregnancy, physician's advice, or other personal reasons. Pregnancy rates before IUD insertion were 64.6/100 woman-years of use (total woman-months, 83,065); after IUD insertion, the rate was 1.05/100 woman-years of use (total woman-months, 37,642); and following removal, the rate was 23.43 (based on 449 cases). Vaginal cytology did not differ in Class 1, 2, and 3 smears between 1058 IUD users and 7226 non-IUD users.^ieng

[The development of newborn C 57 BL/KsJ-dbm mice produced from eggs fertilized in vitro].

The purpose of this study was to examine the development of newly born C57BL/KsJ-dbm mice produced from eggs fertilized in vitro. The embryos derived from fertilization in vitro (which was performed by using db/db eggs and adrenalectomized db/db (Adrex) spermatozoa,) were transferred to the oviduct of MRL/MpJ pseudopregnant recipients 30 hr after insemination. 376 of these embryos yielded 65 young. Weight gain and urine glucose, plasma glucose and insulin levels were measured in these young as well as in Adrex males. The young produced by fertilization in vitro showed hyperglycemia, hyperinsulinemia and obesity. The physiological abnormalities in these young were similar to those in db/db young produced by natural mating between heterozygote (db/+) males and females. Adrex males did not show hyperglycemia but did show hyperinsulinemia. These results indicate that in vitro fertilization and embryo transfer is an effective means of producing fetuses or newborns with an overt genotype in genetically diabetic obese (db) mice.

[Establishment of a new hereditary-cataract mouse (CSM) strain derived from SENCAR mouse].

A mouse representing a new hereditary cataract strain was found in a mouse colony and a new line was established strain CSM. These mice were investigated genetically, histologically and biochemically. The results suggested that this cataract was apparently inherited through two recessive autosomal genes. Histologically the denucleation process of lens fibers was abnormal and small vacuoles appeared in the equatorial region of the lens cortex at 12 days. Biochemically, insoluble protein and sodium increased in the lens with age.

Interleukin-6 overexpression cannot generate serious disorders in severe combined immunodeficiency mice.

C57BL/6 human interleukin-6 (IL-6) transgenic mice develop mesangial proliferative glomerulonephritis with massive IgG1 plasmacytosis and die of renal failure in early life. To test whether the IL-6 overexpression could cause development of mesangial proliferative glomerulonephritis without plasmacytosis or promote proliferation of immature B cells that have not undergone immunoglobulin gene rearrangement, the IL-6 transgene was introduced into mice with severe combined immunodeficiency (SCID). In the immunocompetent littermate IL-6 transgenic mice, there were various symptoms such as plasmacytosis, nephropathy, anemia, and thrombocytosis, accompanied by marked increases in serum IL-6 levels as they aged. All these mice died by 25 weeks of age. In contrast, the SCID-IL-6 transgenic mice had no such abnormalities, except certain hematological changes, although the transgene was expressed in various tissues. In these mice, the serum IL-6 levels were 10- to 15-fold higher than those in the nontransgenic mice, and they remained constant throughout their lives. Furthermore, there were no signs of lymphoid development. This study demonstrates that deregulation of IL-6 expression does not stimulate cell growth or differentiation of immature B cells, and thus does not result in plasmacytosis and age-related increases in IL-6 production, and also does not generate mesangial proliferative glomerulonephritis.

Induction of autoimmune disease by graft-versus-host reaction across MHC class II difference: modification of the lesions in IL-6 transgenic mice.

We examined the effect of IL-6 on the development of autoimmune diseases (primary biliary cirrhosis, Sjögren's syndrome) employing murine graft-versus-host reaction (GVHR) model with MHC class II disparity. For this purpose, we used IL-6 transgenic (B6.6) mice in which a high level of IL-6 was detected. C57Bl/6 (B6) spleen T cells were injected into B6.6 mated with B6.C-H-2bm12 (bm12) mutant mice ((bm12 x B6.6)F1) and GVHR with MHC class II disparity was induced. The transgenic hybrid mice with GVHR showed a larger spleen index and contained a higher serum level of IL-6 than those without GVHR. Autoimmune-like lesions in transgenic recipients became weakened compared with those in non-transgenic (bm12 x B6)F1 recipients. In contrast, levels of antimitochondrial antibodies in (bm12 x B6.6)F1 GVHR group were significantly higher than that of (bm12 x B6)F1 GVHR group. These results indicate that IL-6 excessively produced in vivo might regulate the progression of autoimmune diseases.

Immunologic abnormality in NZB/NZW F1 mice. Thymus-independent occurrence of B cell abnormality and requirement for T cells in the development of autoimmune disease, as evidenced by an analysis of the athymic nude individuals.

Both NZB nu/+ and NZW nu/+ mice were microbially clean by cesarean section. The (NZB x NZW)F1 hybrid (NZB/W) nu/nu mice and nu/+ littermates were then generated by mating of NZB nu/+ with NZW nu/+mice under specific pathogen-free conditions. The female NZB/W F1 nu/nu mice did not develop autoimmune kidney disease, whereas all of nu/+ female littermates mice exhibited proteinuria and died of renal failure with a 50% survival time of 35 wk. Namely, nude mice had no signs of proteinuria up to the time of their death caused by other diseases rather than glomerulonephritis, and their mean survival time was greater than 45 wk. Nude mice had also no anti-ssDNA antibody in their serum. However, splenic B cells of NZB/W nude mice exhibited hyper-responsiveness to both LPS and B151-TRF2, a T cell-derived polyclonal B cell-stimulation factor, and produced large numbers of Ig-secreting cells and anti-TNP plaque-forming cells as well as anti-ssDNA antibody comparable to the nu/+ littermate mice. Interestingly, thymus-engrafted NZB/W nude mice developed autoimmune disease exemplified by the induction of anti-ssDNA antibody and proteinuria at approximately the same time as their nu/+ littermates. These results indicate that the B cell hyper-responsiveness found in NZB/W mice is apparently determined by the T cell-independent process, and T cells are obligatorily required for the development of autoimmune disease in NZB/W mice.