Randomized, double-blind, crossover, placebo-controlled clinical trial to evaluate the effects of chicken hot water extract on insulin secretion.

OBJECTIVE: Essence of chicken (EOC), a hot water extract of chicken, is widely consumed in Southeast Asia as a beverage. EOC has an inhibitory effect on the elevation of blood glucose levels and a secretagogue effect on insulin. However, the mechanism by which EOC promotes insulin secretion is unknown. We aimed to verify the postprandial hyperglycemic inhibitory effect and the insulin secretory effect of EOC in healthy adults under appropriate placebo settings. In addition, we aimed to understand the mechanism underlying the insulin secretory effect of EOC. PATIENTS AND METHODS: Thirty-four healthy Japanese adults were fed 68 mL of EOC or control food, followed by 200 g of cooked rice. Blood glucose and plasma insulin levels were measured at 30, 45, 60, 90, and 120 min after the participants ate cooked rice. The trial had a randomized, double-blind, crossover, placebo-controlled design. RESULTS: The ingestion of EOC induced an increase in the maximum blood concentration (Cmax) of insulin and shortened the time required to reach the maximum blood concentration following rice consumption. Ingestion of the test beverage resulted in a significantly higher insulinogenic index than that obtained after ingestion of the control beverage. No side effects were observed in this study. Mechanistic experiments revealed that EOC stimulated significant (p < 0.05) secretion of GLP-1 from NCI-H716 human intestinal L cells at 0.1, 1, and 10 mg/mL. CONCLUSIONS: Consuming EOC when eating rice supports pancreatic function. Daily consumption of EOC could elevate the early-phase insulin response; therefore, it could prevent diabetes in Asians with low insulin secretion.

A role for phosphoinositide 3-kinase in bacterial invasion.

Listeria monocytogenes is a bacterial pathogen that invades cultured nonphagocytic cells. Inhibitors and a dominant negative mutation were used to demonstrate that efficient entry requires the phosphoinositide (PI) 3-kinase p85alpha-p110. Infection with L. monocytogenes caused rapid increases in cellular amounts of PI(3, 4)P2 and PI(3,4,5)P3, indicating that invading bacteria stimulated PI 3-kinase activity. This stimulation required the bacterial protein InlB, host cell tyrosine phosphorylation, and association of p85alpha with one or more tyrosine-phosphorylated proteins. This role for PI 3-kinase in bacterial entry may have parallels in some endocytic events.

Requirement of atypical protein kinase clambda for insulin stimulation of glucose uptake but not for Akt activation in 3T3-L1 adipocytes.

Phosphoinositide (PI) 3-kinase contributes to a wide variety of biological actions, including insulin stimulation of glucose transport in adipocytes. Both Akt (protein kinase B), a serine-threonine kinase with a pleckstrin homology domain, and atypical isoforms of protein kinase C (PKCzeta and PKClambda) have been implicated as downstream effectors of PI 3-kinase. Endogenous or transfected PKClambda in 3T3-L1 adipocytes or CHO cells has now been shown to be activated by insulin in a manner sensitive to inhibitors of PI 3-kinase (wortmannin and a dominant negative mutant of PI 3-kinase). Overexpression of kinase-deficient mutants of PKClambda (lambdaKD or lambdaDeltaNKD), achieved with the use of adenovirus-mediated gene transfer, resulted in inhibition of insulin activation of PKClambda, indicating that these mutants exert dominant negative effects. Insulin-stimulated glucose uptake and translocation of the glucose transporter GLUT4 to the plasma membrane, but not growth hormone- or hyperosmolarity-induced glucose uptake, were inhibited by lambdaKD or lambdaDeltaNKD in a dose-dependent manner. The maximal inhibition of insulin-induced glucose uptake achieved by the dominant negative mutants of PKClambda was approximately 50 to 60%. These mutants did not inhibit insulin-induced activation of Akt. A PKClambda mutant that lacks the pseudosubstrate domain (lambdaDeltaPD) exhibited markedly increased kinase activity relative to that of the wild-type enzyme, and expression of lambdaDeltaPD in quiescent 3T3-L1 adipocytes resulted in the stimulation of glucose uptake and translocation of GLUT4 but not in the activation of Akt. Furthermore, overexpression of an Akt mutant in which the phosphorylation sites targeted by growth factors are replaced by alanine resulted in inhibition of insulin-induced activation of Akt but not of PKClambda. These results suggest that insulin-elicited signals that pass through PI 3-kinase subsequently diverge into at least two independent pathways, an Akt pathway and a PKClambda pathway, and that the latter pathway contributes, at least in part, to insulin stimulation of glucose uptake in 3T3-L1 adipocytes.

Requirement for activation of the serine-threonine kinase Akt (protein kinase B) in insulin stimulation of protein synthesis but not of glucose transport.

A wide variety of biological activities including the major metabolic actions of insulin is regulated by phosphatidylinositol (PI) 3-kinase. However, the downstream effectors of the various signaling pathways that emanate from PI 3-kinase remain unclear. Akt (protein kinase B), a serine-threonine kinase with a pleckstrin homology domain, is thought to be one such downstream effector. A mutant Akt (Akt-AA) in which the phosphorylation sites (Thr308 and Ser473) targeted by growth factors are replaced by alanine has now been shown to lack protein kinase activity and, when overexpressed in CHO cells or 3T3-L1 adipocytes with the use of an adenovirus vector, to inhibit insulin-induced activation of endogenous Akt. Akt-AA thus acts in a dominant negative manner in intact cells. Insulin-stimulated protein synthesis, which is sensitive to wortmannin, a pharmacological inhibitor of PI 3-kinase, was abolished by overexpression of Akt-AA without an effect on amino acid transport into the cells, suggesting that Akt is required for insulin-stimulated protein synthesis. Insulin activation of p70 S6 kinase was inhibited by approximately 75% in CHO cells and approximately 30% in 3T3-L1 adipocytes, whereas insulin-induced activation of endogenous Akt was inhibited by 80 to 95%, by expression of Akt-AA. Thus, Akt activity appears to be required, at least in part, for insulin stimulation of p70 S6 kinase. However, insulin-stimulated glucose uptake in both CHO cells and 3T3-L1 adipocytes was not affected by overexpression of Akt-AA, suggesting that Akt is not required for this effect of insulin. These data indicate that Akt acts as a downstream effector in some, but not all, of the signaling pathways downstream of PI 3-kinase.

Phosphoinositide 3-kinase is required for insulin-induced but not for growth hormone- or hyperosmolarity-induced glucose uptake in 3T3-L1 adipocytes.

The(1) regulatory mechanism of glucose uptake in 3T3-L1 adipocytes was investigated with the use of recombinant adenovirus vectors encoding various dominant negative proteins. Infection with a virus encoding a mutant regulatory subunit of phosphoinositide (PI) 3-kinase that does not bind the 110-kDa catalytic subunit (delta p85) inhibited the insulin-induced increase in PI 3-kinase activity co-precipitated by antibodies to phosphotyrosine and glucose uptake in a virus dose-dependent manner. Overexpression of a dominant negative RAS mutant in which Asp57 is replaced with tyrosine (RAS57Y) or of a dominant negative SOS mutant that lacks guanine nucleotide exchange activity (delta SOS) abolished the insulin-induced increase in mitogen-activated protein kinase activity, but had no effect on PI 3-kinase activity or glucose uptake. Although GH and hyperosmolarity attributable to 300 mM sorbitol each promoted glucose uptake and translocation of glucose transporter (GLUT)4 to an extent comparable to that of insulin, these stimuli triggered little or no association of PI 3-kinase activity with tyrosine-phosphorylated proteins. Overexpression of delta p85 or treatment of cells with wortmannin, an inhibitor of PI 3-kinase activity, had no effect on glucose uptake or translocation of GLUT4 stimulated by GH or hyperosmolarity. Moreover, overexpression of delta SOS or RAC17N also did not affect the increase in glucose uptake induced by these stimuli. A serine/threonine kinase Akt, a constitutively active mutant of which was previously shown to stimulate glucose uptake, is activated by insulin, GH, and hyperosmolarity to approximately 4-fold, approximately 2.1-fold, and approximately 2.3-fold over basal level, respectively. These results suggest that insulin-induced but neither GH- or hyperosmolarity-induced glucose uptake is PI 3-kinase-dependent, and neither RAS nor RAC is required for glucose uptake induced by these stimuli in 3T3-L1 adipocytes.

Activation of translation initiation factor eIF2B by insulin requires phosphatidyl inositol 3-kinase.

Eukaryotic initiation factor eIF2B mediates a key regulatory step in peptide-chain initiation and is acutely activated by insulin, although, it is not clear how. Inhibitors of phosphatidylinositide 3-kinase blocked activation of eIF2B, although rapamycin, which inhibits the p70 S6 kinase pathway, did not. Furthermore, a dominant negative mutant of PI 3-kinase also prevented activation of eIF2B, while a Sos-mutant, which blocks MAP kinase activation, did not. The data demonstrate that a pathway distinct from MAP and p70 S6 kinases regulates eIF2B. Glycogen synthase kinase-3 (GSK-3) phosphorylates and inactivates eIF2B. In all cases, eIF2B and GSK-3 were regulated reciprocally. Dominant negative PI 3-kinase abolished the insulin-induced inhibition of GSK-3. These data strongly support the hypothesis that insulin activates eIF2B through a signalling pathway involving PI 3-kinase and inhibition of GSK-3.

Comparative study of vitreous oxygen tension in human and rabbit eyes.

For the first time, vitreous oxygen tension in the human vitreous body was measured using a polarographic oxygen electrode during vitreous surgery. The same method was used in a rabbit experimental model. In the human eye, the mean oxygen tensions of the anterior peripheral vitreous body, central vitreous body and posterior vitreous body were 16.7 +/- 3.7 mm Hg (mean +/- 1 SD), 15.9 +/- 2.8 mmHg and 19.9 +/- 4.8 mm Hg, respectively, while the preretinal oxygen tension of the detached retina was 30.0 +/- 4.8 mm Hg. In the rabbit eye, the mean oxygen tensions of the anterior peripheral vitreous body, central vitreous body and posterior vitreous body were 13.9 +/- 4.3 mm Hg, 16.0 +/- 3.5 mm Hg and 22.5 +/- 2.1 mm Hg, respectively. The preretinal oxygen tension of the attached retina in the rabbit eye was 39.5 +/- 3.4 mm Hg, while that of the detached retina decreased to 4.3 mm Hg. When comparing human with rabbit eyes, it appears that the vitreous oxygen tensions are similar, except over the detached retina, even though the ocular circulation differs. We believe that human retinal vasculature plays an important role in perfusing the detached retina with relatively high levels of oxygen.

Simultaneous pattern reversal ERG and VER recordings. Effect of stimulus field and central scotoma.

The effects of the sizes of the stimulus field and of an artificially created central scotoma on simultaneously recorded pattern-reversal electroretinogram (P-ERG) and pattern-reversal visual-evoked response (P-VER) were investigated. With an increase in the stimulus field from 4 degrees X 4 degrees to 12 degrees X 12 degrees, the amplitude of the P-ERG increased steadily. The amplitude of the P-VER also showed an increase up to a stimulus field of 6 degrees X 6 degrees or 8 degrees X 8 degrees, but showed no increase with further expansion of the stimulus field. A central scotoma, created by placing a square of black paper at the center of the 12 degrees X 12 degrees stimulus field, was increased from 4 degrees X 4 degrees to 10 degrees X 10 degrees by 2-degree increments. Amplitudes of both the P-ERG and the P-VER decreased with increasing central scotoma size. The P-VER decreased significantly with a 4 degrees X 4 degrees central scotoma. Although both the P-ERG and the P-VER were influenced by the overall stimulus field and the central scotoma, there was a distinct difference in their behavior. The P-VER showed saturation above a certain stimulus field size and, with a central scotoma of 4 degrees X 4 degrees, much more reduction than the P-ERG, suggesting that the P-VER depends more on the macular area than does the P-ERG. The P-ERG also exhibited a substantial macular dependency, which, however, was not as great as that of the P-VER. The greater macular dependency of the P-VER compared to the P-ERG, as observed in our study, reflects the larger anatomic representation of the macula at the higher visual level.

Electroretinographic findings in fellow eyes of patients with central retinal vein occlusion.

In the study of 50 patients with unilateral central retinal vein occlusion, abnormal electroretinograms were observed in the fellow eyes of 18 patients (36%). The abnormal electroretinograms were supernormal in 15 patients (30%) and negative (+) in the fellow eyes of 3 patients (6%). The amplitudes of a and b waves were significantly greater in the fellow eye than those in normal controls. During a 1- to 9-year follow-up, central retinal vein occlusion developed in three fellow eyes (6%). The abnormal electroretinograms in the fellow eyes reflected such factors as circulatory disturbance of the retina that might have led to central retinal vein occlusion in the affected eye.

Effects of prolonged oral cimetidine, ranitidine, and famotidine therapy on antipyrine elimination.

The effects of three histamine H2-receptor antagonists on the antipyrine metabolizing capacity of the liver were studied in 32 patients with gastrointestinal disorders and in three healthy volunteers. The drugs studied were 800 mg and 400 mg of cimetidine, 300 mg of ranitidine, and 40 mg of famotidine daily. Only cimetidine at 800 mg/day inhibited hepatic antipyrine metabolism, and the inhibitory effects disappeared rapidly with no accumulation.

Changes in the anterior chamber configuration after small-incision cataract surgery with posterior chamber intraocular lens implantation.

PURPOSE: To report quantitative changes in the anterior chamber configuration after small-incision cataract surgery with implantation of a posterior chamber intraocular lens by means of ultrasound biomicroscopy. METHODS: We examined the anterior chamber configuration of 20 eyes of 20 patients before and 3 months after small-incision cataract surgery (phacoemulsification and aspiration plus foldable intraocular lens implantation through a 3.0- to 4.0-mm self-sealing wound) by means of ultrasound biomicroscopy. The following variables were measured: the anterior chamber depth at the center of the cornea, the angle-opening distance 250 microns from the scleral spur (AOD250), the angle-opening distance 500 microns from the scleral spur (AOD500), and the trabecular-iris angle. RESULTS: The anterior chamber depth at the center of the cornea, AOD250, AOD500, and trabecular-iris angle increased significantly after surgery. The preoperative anterior chamber depth at the center of the cornea and trabecular-iris angle were negatively correlated with the differences between the postoperative and preoperative values (P < .01). The preoperative values of all variables examined were negatively correlated with the ratios of the postoperative value to the preoperative value (P < .002). CONCLUSIONS: The present results showed that small-incision cataract surgery significantly deepened the anterior chamber and widened its angle. The more shallow the preoperative anterior chamber was, the greater the postoperative change of the chamber was; and the more narrow the preoperative angle was, the greater the postoperative change of the angle was.

Electroretinogram b/a wave ratio improvement in central retinal vein obstruction.

The electroretinogram (ERG), especially the b/a wave ratio, is considered a good indicator of retinal ischaemia in central retinal vein obstruction (CRVO). Seven CRVO patients who showed b/a wave ratio improvement from < 1.0 [negative type (-) ERG] to > or = 1.0 and one from 1.07 to 1.53 were studied. Three mechanisms of change were observed: firstly, the b-wave amplitude increased without an a-wave amplitude decrease (group A, n = 2); secondly, the b-wave amplitude increased with an a-wave amplitude decrease (group B, n = 4); and, thirdly, both decreased, but the a-wave amplitude decreased more markedly (group C, n = 2). In group A, the visual acuities improved markedly. In group B, the visual acuities improved in two cases in which the b-wave amplitude reached the normal range; the visual acuities did not improve in two cases in which the b-wave amplitude did not reach the normal range. In group C, the visual acuities remained poor. The negative (-) ERG or significantly reduced b/a wave ratio is associated with ischaemic CRVO and did not occur because of the filtering effect of the haemorrhage, which may reduce the stimulus light for the ERG. Improvement of the reduced b/a wave ratio with an increased b-wave amplitude was accompanied by improvements in fundus appearance and visual acuity in CRVO. The results suggest that the retinal ischaemia in CRVO, as revealed by the ERG and fluorescein angiogram, may be reversible in some cases.

Electrophysiological studies of spontaneous retinal degeneration in celestial goldfish (chotengan).

Spontaneous retinal degeneration of Celestial goldfish was studied histologically and electrophysiologically. Retinal degeneration was divided into three stages. Until the age of 75 days (Stage 0) the retina showed normal histological development and the electroretinograms (ERGs) exhibited dominant b-waves similar to those of adult common goldfish. By the age of 105 days (Stage 1), when the eyeballs began to protrude laterally, histological changes were first observed in the retinal pigment epithelium and photoreceptor layer. The b-wave amplitude decreased to 52% of that of Stage 0 with a prolongation of the peak latency. By the age of 135 days (Stage 2), when eyeballs protruded further anterodorsally, the retinal degeneration extended to the entire retinal layers and no b-waves were detected. The deterioration of the b-wave amplitude was closely correlated to the histological findings of retinal degeneration. The Celestial goldfish is considered to be a new model of hereditary retinal degeneration.

Measurement of vitreous oxygen tension in human eyes.

The vitreous oxygen tension in the human vitreous body was measured using a polarographic oxygen electrode during vitreous surgery. To the best of the authors' knowledge, this is the first time these measurements have been taken in the human eye. The vitreous oxygen tensions in the anterior peripheral vitreous body, central vitreous body, and posterior vitreous body were 16.7 +/- 3.7 mmHg (mean +/- SD, n = 9), 15.9 +/- 2.8 mmHg and 19.9 +/- 4.8 mmHg, respectively. The preretinal oxygen tensions over the detached retina and detached macula were 30.0 +/- 4.8 mmHg (n = 5) and 15.3 +/- 2.1 mmHg (n = 3), respectively, with the latter significantly lower than the former (P less than 0.01). In one case, after inhalation of 100% oxygen the oxygen tension over the retinal scar resulting from cryoretinopexy in a previous retinal detachment surgery increased remarkably to 160 mmHg.

Multicenter genetic study of retinitis pigmentosa in Japan: I. Genetic heterogeneity in typical retinitis pigmentosa.

A nationwide, multicenter study of typical retinitis pigmentosa (RP) was carried out in collaboration with 18 hospitals throughout Japan to obtain current information for genetic counseling. We analyzed the genetic heterogeneity of RP based on the parental consanguinity of 434 probands registered during a 6-month period in 1990. A gradual decline in the frequency of consanguineous marriage was recognized among the normal parents of RP patients. The relative frequencies of inheritance patterns were estimated as: autosomal recessive, 25.2%; autosomal dominant, 16.9%; X-linked, 1.6%; and simplex, 56.3%. A comparison of these results with previous reports in Japan revealed a decline in the relative frequency of autosomal recessive cases and an increase in simplex cases. This suggests a decrease in the incidence of autosomal recessive retinitis pigmentosa in Japan, as well as the necessity for exhaustive investigations aimed at identifying inheritance patterns for RP patients seeking genetic counseling.

Multicenter genetic study of retinitis pigmentosa in Japan: II. Prevalence of autosomal recessive retinitis pigmentosa.

Retinitis pigmentosa (RP) is a group of genetically heterogeneous diseases with autosomal recessive (AR), autosomal dominant, and X-linked modes of inheritance. Autosomal recessive retinitis pigmentosa (ARRP) is the most common form in Japan. A genetic analysis was done to determine the prevalence of ARRP indirectly, to provide an estimation of changing trends in the overall prevalence of RP. Data on the frequency of consanguinity and marriage year of normal parents of 59 ARRP patients were obtained from a nationwide multicenter survey of typical retinitis pigmentosa conducted in 1990. The gene frequency of ARRP was 0.01145 (Dahlberg's formula). In 1990, the number of young symptomatic ARRP patients decreased, while the number of patients aged 40 years and older increased. The total number of symptomatic ARRP patients in 1990 was nearly 21% higher than in 1970. Despite a dramatic decline in consanguinity in recent decades in Japan, the number of ARRP patients has increased. This increase is attributed to greater longevity and overall population growth. Our results suggest that the total number of RP patients has not decreased, and may even have increased.

[A case of bilateral continuous central retinal artery occlusion].

BACKGROUND: We report a case of bilateral simultaneous central retinal artery occlusion (CRAO). The time lag was 21 hours. CASE: The patient was a 78-year-old woman. She had hypertension, arteriosclerosis, and bilateral thrombosis of the legs. Both eyes could only distinguish light and dark at the first medical examination. RESULTS: We started steroid and urokinase therapy and used a vasodilator of peripheral vessels. When the patient left the hospital, visual acuity had improved to the ability of counting fingers (0.01 leftward 30 cm on the left). CONCLUSIONS: Both visual field and color vision had improved slightly. We think this bilateral CRAO resulted from thrombosis of both central retinal arteries at almost the same time.

[Effects of endothelin on retinal blood vessels].

Endothelins (ETs), which are one of the most potent vasoconstrictors, were discovered in 1988. ETs are involved in the regulation of vascular smooth muscle and are thought to function like a local hormone. In this study, we measured changes in the caliber of retinal blood vessels caused by the intravitreal injection of ET 1 in rabbits, using fundus photography. The retinal artery showed an immediate vasoconstriction after the injection (0.1 ml) of ET 1 at 10(-6)M. However, ET 1 in lower concentrations (10(-7)M and 10(-8)M) caused vasodilations initially and subsequently vasoconstrictions. A dose-response relation was found. The blood vessel caliber reduced maximumly to 53% the value before injection. The vasoconstriction due to ET 1 may induce retinal ischemia and hypoxia. We speculate that ET 1 may be involved in the regulation of retinal blood vessels.

[Analysis of the long-term prognosis for conjunctival malignant melanomas in Japan].

PURPOSE: To investigate the long-term prognosis for primary conjunctival malignant melanomas in Japan. MATERIALS & METHODS: We conducted a survey of 61 cases which had been reported in a 38-year period (1959 to 1996). We gathered information regarding the survival of patients, the post-operative follow-up period, the causes of death, and recurrences. Answers were obtained segarding 51 cases (84%). Detailed progress was identified in 23 of these cases. The survival rates were calculated using the Kaplan-Meier method. RESULTS: The survival rates were 95.1% after 1 year, 72.9% after 3 years, and 53.4% after 5 years. These values are relatively low compared with those reported in Europe and the United States.

[Manometric study of the biliary sphincter zone: a comparison between the length of phasic activity and the narrow distal segment].

Motor activity of the sphincter of Oddi has been evaluated in 26 patients who underwent ERCP examination. Manometric recordings from the common bile duct and the sphincter of Oddi were performed with a single end-lumen perfused catheter. The number of successive 3 mm spaced black marks, observed during the pull-through from the first appearance of the proximal sphincter activity to that of duodenal pressure, was used to measure the length of the sphincter zone. In 14 of 26 patients, the length of phasic activity zone of the sphincter of Oddi was 9 mm. However, the narrow distal segment of the common bile duct was 15.1 +/- 3.6 mm in length. In same patients, the length of phasic activity zone was shorter than the anatomical one. The mean sphincter of Oddi pressure (SOP) at the distance of 6 mm from the papillary pore (77.8 +/- 22.1 mmHg) was greater than that of 9 mm (58.4 +/- 27.0 mmHg) and 12 mm (51.0 +/- 16.9 mmHg). These results suggest that the location of the orifice of catheter should be taken into consideration when comparing the SOP.