The Clinical Outcome of Anatomical Reattachment of the TFCC to the Ulnar Fovea using an ECU Half-Slip and an Interference Screw.

We developed an original reattachment technique using a half-slip of the extensor carpi ulnaris (ECU) tendon with a very small titanium interference screw for chronic foveal avulsion of the TFCC. The clinical outcome of 66 wrists with foveal detachment of the TFCC treated by this procedure was examined.A distally based ECU half-slip was harvested, inserted into the TFCC, sutured to the remnant of the TFCC, and pulled out through a 2.5-mm bone tunnel at the centre of the fovea. The ECU half-slip was subsequently anchored to the ulnar fovea with a small titanium interference screw. We evaluated 66 wrists of 65 patients with a minimum follow-up of 1 year. Ulnar variance was neutral in 47 wrists, negative in 5 and positive in 14 wrists. Causes of injury were falls in 34 patients, traffic accidents in 12, sports activities in 9, labour in 2 and unknown in 8 patients. In the positive variance wrists, ulnar shortening was performed before the reattachment. The clinical outcome was evaluated using our original DRUJ evaluating system.Preoperatively, severe wrist pain was reported in 50 wrists and moderate pain in 16 wrists. Severe no-endpoint DRUJ instability was noted in 65 wrists, while 1 wrist demonstrated moderate DRUJ instability. Only 2 wrists had supination loss by 20 degrees. At the final follow-up, no pain was felt in 55 wrists, mild pain in 3 wrists, and 8 patients had moderate pain. One wrist exhibited a 30-degree loss of supination. The DRUJ was stable in 55 wrists, mildly unstable in 3, moderately unstable in 4 and severely unstable in 4 wrists. There were 50 excellent, 9 good, 3 fair and 4 poor results.The technique of anatomical reattachment of the TFCC to the ulnar fovea using an ECU half-slip tendon is effective for chronic foveal avulsion of the TFCC with severe DRUJ instability.

Postprandial changes in plasma concentrations of growth hormone and ghrelin around weaning in the goat.

We measured and compared plasma levels of GH and ghrelin in response to feeding in 4-week-old (milk replacer-fed) and 13-week-old (alfalfa hay cube-fed) goats, in order to elucidate whether or not the postprandial regulation of these hormone levels changes around weaning. Furthermore, we examined the effects of suckling from the dam or intravenous glucose administration on both hormone and insulin levels in kids. In 4-week-old goats, feeding of a milk replacer diet significantly increased plasma GH levels without changing level of ghrelin. In contrast, in 13-week-old goats, feeding of hay cubes did not change the levels of either ghrelin or GH. Suckling of milk directly from the dams significantly increased the levels of GH and insulin, but not ghrelin, in kids. Finally, intravenous injection of glucose (0.625 mmol/kg BW) did not cause any significant increase in the levels of GH or ghrelin, despite a significant increase in the levels of insulin and glucose. From these results, we conclude that the regulatory system of the somatotropic axis is altered by weaning or weaning-associated processes, and that ghrelin levels may not be involved in this alteration in young goats.

Deep lamellar keratoplasty by deep parenchyma detachment from the corneal limbs.

AIM: To improve the deep lamellar keratoplasty technique. METHOD: For the easy and reliable perfomance of deep lamellar keratoplasty (DLKP), detachment of Descemet's membrane through the corneal limber flap was improved. To expose Descemet's membrane, the parenchyma was detached by hydrodelamination through a sclerocorneal flap made in the corneal limbs. The parenchyma was removed after the pseudochamber between it and Descemet's membrane was maintained with viscoelastic material. The corneal graft was placed with a running suture. 22 eyes were treated. RESULTS: Complete exposure of Descemet's membrane was obtained in 20 of the 22 eyes (91%). The membrane was perforated in five of the 22 eyes (23%) during surgery, and two of the 22 eyes (9%) were converted to penetrating keratoplasty. These two eyes developed keratoconus after acute corneal hydrops. CONCLUSION: Compared with the conventional procedure, this new method provides easy, reliable exposure of Descemet's membrane.

Effects of long-chain fatty acids on cytosolic triacylglycerol accumulation and lipid droplet formation in primary cultured bovine mammary epithelial cells.

Mammary epithelial cells have recently been shown to express and secrete leptin into milk and to accumulate triacylglycerol (TAG) in cytosol. We examined the effects on the accumulation of cytosolic TAG of free fatty acid addition to the medium bathing bovine mammary epithelial cells (bMEC). Both saturated (palmitic and stearic) and unsaturated (oleic and linoleic) fatty acids stimulated the accumulation of TAG in a concentration-dependent manner from 50 to 400 microM and the expression of mRNA expression for CD36, which is involved in the uptake and secretion of long-chain fatty acids. However, leptin mRNA expression and lipid droplet formation were significantly increased only by the addition of unsaturated, but not saturated, fatty acids. Interestingly, both types of fatty acids stimulated alphas1-casein mRNA expression. These data suggest that the expression of leptin is related to droplet formation, whereas CD36 is related to cytosolic TAG accumulation, and that fatty acids or cytosolic TAG accumulation also have a role to accelerate differentiation of bMEC as shown by casein synthesis.

Suppressing effects of bisphenol A on the secretory function of ovine anterior pituitary cells.

We investigated the action of bisphenol A (BPA) on cellular GH release and content, cell number, GHmRNA expression, and concentrations of cellular cyclic AMP ([cAMP]c) and calcium ion ([Ca2+]c) in primary cultured ovine anterior pituitary cells. The following results were found: (1) BPA as well as nonylphenol (NP) at 10(-6) to 10(-3) M significantly and concentration-dependently suppressed basal and GHRH-stimulated GH release, and the cellular GH content, (2) BPA suppressed the cell number in a time- and concentration-dependent manner, (3) 10(-4)M BPA suppressed GHmRNA expression to 68% of control (BPA-free), and abolished GHRH (10(-8) M)-induced increases in [cAMP]c and [Ca2+]c. From these findings we conclude that BPA possesses a suppressing action on GH synthesis and release, and this suppressing action is probably related to impairment of cellular signal transduction systems in ovine anterior pituitary cells.

Rapid suppressing action of insulin-like growth factor-I (IGF-I) on GH release from anterior pituitary cells of goats.

Goat anterior pituitary cells were cultured to investigate the effects of insulin-like growth factor-I (IGF-I), insulin, and growth hormone (GH) on basal and GH-releasing hormone (GHRH)-induced GH release. Changes in cellular Ca2+ concentrations were also assessed to enable discussion of the cellular mechanisms of IGF-I. The cells were cultured for 48 h, and then stimulated with GHRH (10 nmol/l) for 30 min, with or without each test substance. In the control cells, IGF-I (10 and 100 ng/ml) significantly raised the basal, but did not change GHRH-induced GH release, resulting in the abolishment of GH release induced by GHRH in the presence of 100 ng/ml IGF-I. However, there was no significant effect of insulin (10, 100, and 1000 microU/ml) on basal and GHRH-induced GH release. In the cells cultured for 48 h with each test substance but stimulated for 30 min without the test substance, no significant change in the basal and GHRH-stimulated GH release was observed. Regardless of treatment, there was no significant effect on intra-cellular GH content. Analysis with a confocal laser microscope revealed that IGF-I (100 ng/ml) significantly increased the basal, but significantly reduced GHRH (10 nmol/l)-induced increase in cellular Ca2+ concentrations. From these findings we conclude that IGF-I exerts an acute suppressing action on the GHRH-induced GH release, which partly involves changes in cellular Ca2+ metabolism in goat somatotrophs.

Existence of GPR40 functioning in a human breast cancer cell line, MCF-7.

GPR40, which has recently been identified as a G-protein-coupled cell-surface receptor for long-chain fatty acids, was assessed in a human breast cancer cell line (MCF-7). We detected GPR40 mRNA by RT-PCR and found that oleate and linoleate, but not palmitate or stearate, caused an increase in cellular Ca(2+) concentrations, which was partially blocked by the pertussis toxin (PTX) treatment. We examined the expression of GPR40 mRNA by quantitative RT-PCR in the relation to cell number. It was significantly increased at the beginning and at the end of cell proliferation. These results indicate the possibility that GPR40 for long-chain fatty acids may be involved in cellular function such as cell proliferation, providing a new perspective for the action of long-chain fatty acids on mammary epithelial cells.

Effects of acetate and butyrate on the expression of leptin and short-form leptin receptor in bovine and rat anterior pituitary cells.

The effects of acetate and butyrate on leptin and leptin receptor (OB-R) expression in bovine and rat anterior pituitary were examined. In bovine tissues, leptin gene expression using RT-PCR was observed in fat and anterior pituitary but not in liver. Isolated anterior pituitary cells cultured in Dulbecco's modified Eagle's medium (DMEM) for 3 days were further cultured for 48 h in DMEM containing 10 mM acetate or butyrate or without any fatty acids as control. Western blot analysis revealed that the abundance of leptin protein was greater in the presence of acetate and butyrate than that for the control culture. Leptin abundance was increased in a dose- and time-dependent manner in bovine anterior pituitary cells. However, leptin expression in rat cells, of which the basal level was much greater than that in ovine cells, was significantly decreased by the culture with butyrate. In addition, we studied the effects of both fatty acids on OB-R mRNA expression using semi-quantitative RT-PCR. The results showed that butyrate significantly decreased the expression in both bovine and rat cells. These findings indicate that acetate and butyrate enhance leptin expression in bovine, but not in rat anterior pituitary cells while butyrate suppresses OB-Ra expression in both rat and bovine pituitaries.

Metabolic and lactational responses during recombinant bovine tumor necrosis factor-alpha treatment in lactating cows.

This study examined the effects of recombinant bovine tumor necrosis factor-alpha (rbTNF) administration on metabolic and hormonal responses and lactational performance in dairy cows. Twelve lactating Holstein cows were injected subcutaneously with rbTNF (2.5 microg per kg per d) or saline (3 ml per head per d) at 1200 h daily for 7 d (d 0-6) and used in a crossover design. The rbTNF treatment induced increases in plasma haptoglobin, nonesterified fatty acid, cortisol, and growth hormone levels compared with the control levels. The rbTNF-treated cows had lower triiodothyronine and insulin-like growth factor-1 concentrations than control cows. In a somatoliberin challenge on d 6, the somatotropin response to somatoliberin (0.25 microg/kg) was smaller in the rbTNF group than in the control. The rbTNF treatment also produced increases of the nitrite plus nitrate concentration in plasma and milk during the period between d 1 and 7. Milk yield was reduced by rbTNF administration from d 1 to 8. The percentage of milk fat was increased on d 1-7 by rbTNF treatment, but milk protein content in the rbTNF group was decreased on d 5 and 7 as compared with that in the control group. These results support the possibility that tumor necrosis factor-alpha is responsible for the changes in hormone secretion, milk production and composition, and inflammatory parameters observed during coliform mastitis.

Effects of nutritional conditions around weaning on carbonic anhydrase activity in the parotid gland and ruminal and abomasal epithelia of Holstein calves.

Thirty-two male Holstein calves were used to investigate the effects of nutritional conditions around weaning and aging on carbonic anhydrase (CA) activity in the parotid gland and epithelium from the rumen and abomasum. We fed calf starter and lucerne hay as well as milk replacer (group N) or fed milk replacer either with (group S) or without (group M) administration of short-chain fatty acids (SCFA) through polypropylene tubing into the forestomach until 13 weeks of age. The diets were fed at 1000 hours and 1600 hours, and SCFA were administrated after milk replacer feeding at 1600 hours. Slaughter and tissue sampling were carried out between 1300 hours and 1430 hours at 1, 3, 7, 13, and 18 weeks of age. Tissue samples from five adult (1.5-2.0 years-old) Holstein steers were obtained from a local abattoir. In group N, CA activity in the parotid gland gradually and significantly increased toward the adult value, whilst in the epithelium from the rumen and abomasum, adult values were reached at 3 and 7 weeks of age, respectively. At 13 weeks, the activity for group N was significantly higher than that for the other two groups in the parotid gland, but there was no significant difference in the epithelium from the rumen and abomasum. The concentration of the carbonic isozyme VI in the parotid gland also changed with age but, in contrast to CA activity, had not reached adult levels by 13 weeks of age. In groups M and S, parotid saliva did not show any change toward an alkaline pH or toward a reciprocal change in the concentrations between Cl(-) and HCO(3)(-), even at 13 weeks of age. From these results we conclude that a concentrate-hay based diet around weaning has a crucial role in CA development in the parotid gland, but not in the epithelium of the rumen and abomasum.

Childhood T-cell acute lymphoblastic leukemia with four distinct immunophenotypes representing different stages of T-cell development.

The authors report on a 14-year-old boy who developed T-cell acute lymphoblastic leukemia (FAB:L1) displaying 4 immunophenotypically distinct leukemic cell populations by 3-color immunofluorescence staining. Cytogenetic analysis at diagnosis showed 46,XY,add(4)(p16)[12]/46,XY[2]. A single rearrangement of the T-cell antigen receptor beta- and gamma-chain genes in these cells indicated monoclonality of the leukemic cells. These findings suggest that leukemic blast cells of monoclonal origin in this case were divided into 4 immunophenotypic populations, representing various stages of differentiation.

Scabronine G-methylester enhances secretion of neurotrophic factors mediated by an activation of protein kinase C-zeta.

Glial cells release neurotrophic factors that maintain neurons functionally. Previously, we have shown that the scabronines isolated from Sarcodon scabrosus enhanced the secretion of neurotrophic factors from 1321N1 human astrocytoma cells. In the present study, we examined the mechanism of newly synthesized scabronine G-methylester (ME)-induced secretion of neurotrophic factors from 1321N1 cells. The dramatic neuronal differentiation of rat pheochromocytoma cells (PC-12) was observed by scabronine G-ME-conditioned medium of 1321N1 cells. Scabronine G-ME increased the secretion of nerve growth factor (NGF) and interleukin-6 (IL-6) from 1321N1 cells with the enhancement of their mRNA expressions. Scabronine G-ME concentration-dependently inhibited the carbachol-induced inositol phosphate accumulation in 1321N1 cells, which was reversed by GF109203X, an inhibitor of protein kinase C (PKC) isoforms. Furthermore, GF109203X inhibited the scabronine G-ME-induced mRNA expressions of both NGF and IL-6 and the differentiation of PC-12 cells, showing that scabronine G-ME activated PKC. Although scabronine G-ME enhanced activities of neither conventional nor novel types of PKCs, it translocated PKC-zeta to membranes in intact cells and cell-free condition. Furthermore, recombinant PKC-zeta activity was also increased by scabronine G-ME, suggesting the involvement of PKC-zeta in the effect of scabronine G-ME. Concerning the downstream effectors of the PKC-zeta, scabronine G-ME translocated nuclear factor-kappaB to nucleus, and enhanced its transcriptional activity. In addition, scabronine G-ME caused the degradation of inhibitor of nuclear factor-kappaB concentration-dependently, which was inhibited by GF109203X. These results suggest that scabronine G-ME potentially enhances the secretion of neurotrophic factors from 1321N1 cells mediated via the activation of PKC-zeta.

Effects of adenosine 5'-triphosphate and growth hormone on cellular H+ transport and calcium ion concentrations in cloned bovine mammary epithelial cells.

The present experiment was carried out to investigate the effects of exogenous adenosine 5'-triphosphate (ATP) and growth hormone (GH) on cellular H(+) efflux rate (extracellular acidification rate) and Ca(2+) concentration ([Ca(2+)](c)) in cloned bovine mammary epithelial cells (bMEC) raised from the mammary gland of a 26-day-pregnant Holstein heifer. Perifusion of 2-day cultured cells with a medium containing ATP (10, 100 and 1000 micromol/l) for 30 min caused a significant and concentration-dependent increase in the cellular H(+) efflux rate. ATP application (100 micromol/l) caused a transient and large increase in [Ca(2+)](c) in all cells. In contrast, perifusion with a medium containing bovine GH at 10, 50 and 250 ng/ml for 30 min caused a significant decrease in the cellular H(+) efflux rate in a concentration-dependent manner. However, bovine GH application (50 ng/ml) caused a small decrease followed by an increase, in some cases, in [Ca(2+)](c). In bMEC treated with lactogenic hormones (1 microgram/l prolactin, 1 nmol/ml dexamethasone and 5 microgram/ml insulin) for 2 days, the increased H(+) efflux rate induced by ATP was significantly reduced, whereas the negative response induced by GH was inversely and significantly changed to the positive. Treatment of the cells with lactogenic hormones reduced the increase in [Ca(2+)](c) induced by ATP stimulation, while it enhanced the increase in [Ca(2+)](c) induced by GH stimulation. Application of ATP or GH did not cause any significant changes in [pH](c). Treatment with lactogenic hormones enhanced GH receptor (GHR) transcription that was determined by RT-PCR. From these results, we conclude that exogenous application of ATP and GH causes prompt and significant responses in H(+) transport and [Ca(2+)](c) that were significantly changed in the opposite direction by the treatment with lactogenic hormones. The lactogenic hormone treatment also enhanced GHR transcription, which may change post-receptor signal transduction systems for both agents in the bMEC.

Clonal evolution from trisomy into tetrasomy of chromosome 8 associated with the development of acute myeloid leukemia from myelodysplastic syndrome.

Tetrasomy 8, though rare, is usually associated with trisomy 8, a far more common chromosomal abnormality in acute myeloid leukemia (AML). Yet the clonal relationship between trisomy 8 and tetrasomy 8 in the cases with these chromosomal abnormalities has been unclear. Here, we report a case of a 17-year-old male, diagnosed as having a myelodysplastic syndrome (MDS). Chromosome analysis showed the presence of trisomy 8. Five years later, he developed overt AML exhibiting tetrasomy 8 only. After chemotherapy, the blast cells in the bone marrow decreased to 3.4%, and the karyotype showed trisomy 8 alone. Fluorescence in situ hybridization using a probe specific for chromosome 8 showed that the percentages of cells exhibiting 2/ 3 /4 signals were 7.8/89.2/2.0 at the MDS stage, 20.5/36.1/41.0 when overt AML developed and 24.0/72.1/2.4 after chemotherapy. These results suggested that tetrasomy 8 is derived from the AML clone, possibly evolved from the MDS clone with trisomy 8. To our knowledge, this is the first detailed case report of clonal evolution from trisomy 8 into tetrasomy 8 associated with the development of AML from MDS.

Differentiation of rat pheochromocytoma cells by fomitellic acids, specific DNA polymerase inhibitors.

Fomitellic acid (FA) A and B are specific inhibitors of DNA polymerase alpha and beta. They showed cytotoxicity against rat pheochromocytoma cells (PC-12 cells) in a concentration-dependent manner. However, after PC-12 cells were cultivated with low concentrations of FAs, the cells extended neurites in greater degree similar to the cells cultivated with nerve growth factor. Another DNA polymerase alpha inhibitor, aphidicolin, also induced neurite outgrowth. Furthermore, PC-12 cells were strongly immunostained with anti-alpha-tubulin or anti-tau antibody after the treatment with FAs. These results suggest that weak inhibition of DNA polymerase activity induces the neurite outgrowth in PC-12 cells.

Maitotoxin-induced nerve growth factor production accompanied by the activation of a voltage-insensitive Ca2+ channel in C6-BU-1 glioma cells.

1. The aim of the present study was to determine the effects of maitotoxin on nerve growth factor production and the Ca2+ influx in clonal rat glioma cells (C6-BU-1). 2. Maitotoxin (1 - 10 ng ml-1) induced a profound increase in 45Ca2+ influx in an extracellular Ca2+-dependent manner. However, high KCl had no effect at all. These effects were supported by the results from the analysis of intracellular Ca2+ concentration using fura 2. 3. The maitotoxin-induced 45Ca2+ influx was inhibited by inorganic Ca2+ antagonists, such as Mg2+, Mn2+ and Co2+. The inhibitory effect of Co2+ was antagonized by increasing the extracellular Ca2+ concentrations. 4. Maitotoxin (3 ng ml-1) as well as A-23187 (1microM) and dibutyryl cyclic AMP (0.5 mM) caused an acceleration of nerve growth factor (NGF) production in C6-BU-1 cells, as determined by NGF enzyme immunoassay. 5. Reverse transcription polymerase chain reaction (RT - PCR) analysis showed that maitotoxin (10 ng ml-1) enhanced the expression of NGF mRNA, which was abolished by the removal of extracellular Ca2+. A-23187 also accelerated its expression. 6. These results suggest that maitotoxin activates a voltage-insensitive Ca2+ channel and accelerates NGF production mediated through a Ca2+ signalling pathway in C6-BU-1 glioma cells.

Stimulation of neurotrophic factor secretion from 1321N1 human astrocytoma cells by novel diterpenoids, scabronines A and G.

Glial cells release neurotrophic factors that maintain neurons functionally. When rat pheochromocytoma cells (PC-12) were cultivated with the conditioned medium of human astrocytoma cells (1321N1) incubated with the new diterpenoids, scabronines A and G, isolated from Sarcodon scabrosus, they changed their morphology and there was neurite outgrowth. The scabronines increased the expression of mRNA for nerve growth factor (NGF), and the secretion of NGF from 1321N1 cells in a concentration-dependent manner. However, the enhanced neurite outgrowth produced by the conditioned media was slightly inhibited by NGF neutralizing antibody, and the concentration of NGF released in response to the scabronines was insufficient to cause differentiation. These results suggest that scabronines cause the secretion of other factors together with NGF from 1321N1 cells. The diterpenoids are useful drugs to clarify the mechanism of synthesis and secretion of neurotrophic factors.

[Clinical results of three types of intraocular lenses for small incision surgery].

We implanted three types of intraocular lens (IOL) in 30 eyes each by small-incision surgery. They were: foldable acryl IOL, foldable silicone IOL and polymethylmethacrylate (PMMA) IOL. We evaluated the following items during three years after surgery: visual acuity, astigmatism, glare disability, contrast sensitivity, tilt and decentering of IOL, corneal endothelial population, aqueous flare and aftercataract. Eyes foldable IOLs were more excellent than PMMA IOLs regarding visual acuity, astigmatism and aqueous flare which are indices for the early postoperative period. PMMA and acryl IOLs were more excellent than silicone IOLs regarding decentration of IOL and aftercataract which are indices for the late postoperative period.

A new factor derived from 1321N1 human astrocytoma cells causes differentiation of PC-12 cells mediated through mitogen-activated protein kinase cascade.

Glial cells play an important role in maintaining neural function. In the present study, we examined the effects of a factor derived from human astrocytoma cells (1321N1) on differentiation of rat pheochromocytoma cells (PC-12). The conditioned medium which had been used for culture of 1321N1 cells caused the differentiation of PC-12 cells, suggesting that 1321N1 cells release a neurotrophic factor. The factor was apparently distinct from well-known neurotrophic factors, such as nerve growth factor (NGF), since it was resistant to boiling and trypsin treatment. The molecular size of the factor was assumed to be below 1000 through dialysis and ultrafiltration experiments. Furthermore, PC-12 cells were differentiated synergistically by the combined addition of NGF and the conditioned medium of 1321N1 cells. Partially purified fraction of the factor by Sephadex G-15 gel filtration column caused the prolonged activation of mitogen-activated protein kinase (MAPK). The differentiation of PC-12 cells induced by the fraction or NGF disappeared after the treatment with PD98059, a specific inhibitor of MAPK kinase (MEK), suggesting the involvement of MAPK in the differentiation. These results suggest that the new low-molecular factor derived from glial cells causes differentiation of PC-12 cells mediated through an activation of MAPK.