Effect of dietary mannanase-hydrolysed copra meal on growth performance and intestinal histology in broiler chickens.

We investigated mannanase-hydrolysed copra meal (MCM), which contains ß-1,4-mannobiose (MNB), for its capacity to improve growth performance and activate intestinal villus function. Seven-day-old chicks were separated into four flocks with an equal mean body weight and then fed a basal diet (control) or a diet supplemented with 0.02% or 0.1% MCM. After 7 weeks, the feed intake and body weight were determined and then used to calculate the feed efficiency (FE). Moreover, the intestinal segments were examined by light microscopy and scanning electron microscopy (SEM) for cellular and morphological changes in the villus. Although feed intake was not significantly different among the experimental groups, the body weight gain and FE were significantly higher in the 0.1% MCM group than in the control group (p < 0.05), while feed intake tended to be higher in the 0.02% and 0.1% MCM groups. The cellular area of the ileum was significantly higher in the 0.02% and 0.1% groups in relation to that in the control group (p < 0.05). Furthermore, the cellular area of the duodenum and the jejunum tended to be higher in the 0.02% and 0.1% MCM groups. For the correlation analysis, a significant correlation was observed between the dosage of MCM and the cell area of the duodenum, jejunum and ileum. Moreover, the number of mitotic cells was higher in the 0.1% MCM group. As shown by SEM, the cells at the villi tips were protuberant in appearance in the 0.02% and 0.1% MCM treatments when compared with the relatively flat cells of the control. On the duodenal villus surface of the 0.1% MCM group, some cells devoid of microvilli were observed, suggesting that the increased protuberance of these cells represents increased absorption activity. Although intestinal villus height and area did not significantly differ among groups, the levels of these parameters tended to increase in the experimental groups relative to the control. The present morphological findings reveal that MNB might be effective for activating intestinal absorptive function, and that the functional activation promotes the growth of the chickens.

Analysis of gut immune-modulating activity of beta-1,4-mannobiose using microarray and real-time reverse transcription polymerase chain reaction.

beta-1,4-Mannobiose (MNB) supplementation has been shown to prevent Salmonella Enteritidis infection in broilers by improving Salmonella Enteritidis clearance and increasing IgA production. This study examined in detail the gut immunomodulatory activity of MNB using microarray and real-time quantitative PCR analysis. One-day-old chicks were orally administered 0.1% (wt/wt) MNB 3 times a week for 28 d. Control birds received vehicle alone. Body weights and fecal IgA levels were monitored weekly. On d 28, spleen and bursa of Fabricius were removed and weights were recorded; samples of ileum, jejunum, cecum, spleen, thymus, and bursa of Fabricius were collected for histological examination; and ileum samples were collected for RNA extraction. No significant difference in BW or organ weights was observed between MNB-treated and untreated control birds, and no histological abnormalities were observed in any of the tissues examined. The MNB-treated chickens had significantly higher levels of fecal IgA over all 4 wk when compared with control birds. Microarray and reverse transcription PCR analysis revealed the upregulation of several genes involved in immune responses, including those involved in antigen recognition, processing and presentation (MHC class I and II), interferon-related genes, and genes involved in host defense. These results provide insight into the mechanism of action of dietary MNB in the intestine and confirm that MNB acts as a potent immune-modulating agent, exerting combined effects on the intestinal immune system.

Absorption of calcium, magnesium, phosphorus, iron and zinc in growing male rats fed diets containing either phytate-free soybean protein or soybean protein isolate or casein.

The effect of dietary phytate-free soybean protein (PFS) on intestinal mineral absorption and retention was examined in growing male rats using a three-day mineral balance technique. The rats were fed diets containing PFS, soybean protein isolate (SPI) or casein at a 20% level for 5 wk. Total calcium (Ca), magnesium (Mg), phosphorus (P), iron (Fe) and zinc (Zn) contents in diets were adjusted to 0.35, 0.05, 0.7, 0.0035 and 0.003%, respectively, by supplementation of the diet with their salts. Mineral absorption and retention ratios in rats fed the PFS diet were significantly higher than those in rats fed either the SPI or casein diet. These results suggest that PFS may be a promising dietary protein source for improving the mineral bioavailability in humans.

Inhibition of root elongation in microgravity by an applied electric field.

Roots grown in an applied electric field demonstrate a bidirectional curvature. To further understand the nature of this response and its implications for the regulation of differential growth, we applied an electric field to roots growing in microgravity. We found that growth rates of roots in microgravity were higher than growth rates of ground controls. Immediately upon application of the electric field, root elongation was inhibited. We interpret this result as an indication that, in the absence of a gravity stimulus, the sensitivity of the root to an applied electric stimulus is increased. Further space experiments are required to determine the extent to which this sensitivity is shifted. The implications of this result are discussed in relation to gravitropic signaling and the regulation of differential cell elongation in the root.

Effects of "body compression" on parameters related to ascites formation: therapeutic trial in cirrhotic patients.

Decreased effective circulating blood volume is an important factor in ascites formation in liver cirrhosis. We designed a "body compression" apparatus as a means to restore effective blood volume and investigated its effectiveness in reducing ascites formation in cirrhotics in terms of its effect on parameters of ascites formation noted below. The subjects, eight cirrhotics with ascites and eight cirrhotics without ascites were given spironolactone (50-75 mg/day) and furosemide (40-80 mg/day) while they received a diet containing 85 mEq of sodium per day. All four limbs and the lower abdomen were compressed with constant pressure [height (cm) divided by 13.6 mmHg] once, for 3h, using stroke rehabilitation splints, while patients lay supine. In cirrhotics both with and without ascites, urine volume, urinary sodium excretion, and creatinine clearance during the body compression were greater than values during control (non-compression) periods (urine volume, means 285 vs 169 ml/3h; P < 0.001, urinary sodium excretion 15.8 vs 9.5 mEq/3h; p < 0.001, creatinine clearance 74 vs 59 ml/min, P < 0.001, respectively). The increased basal plasma renin activity, angiotensin II, aldosterone, and norepinephrine levels in all cirrhotics were significantly decreased by the body compression. In another group of six cirrhotics who received no diuretics or albumin, repeat body compression alleviated ascites in three with well preserved renal function, but was ineffective in three with markedly impaired renal function. These results suggest that the improvement in renal function brought about by the body compression is attributable to an increase in effective circulating blood volume. This maneuver may be a useful complementary therapy in patients with cirrhotic ascites with well preserved renal function.

Inhibition of root elongation in microgravity by an applied electric field.

Roots grown in an applied electric field demonstrate a bidirectional curvature. To further understand the nature of this response and its implications for the regulation of differential growth, we applied an electric field to roots growing in microgravity. We found that growth rates of roots in microgravity were higher than growth rates of ground controls. Immediately upon application of the electric field, root elongation was inhibited. We interpret this result as an indication that, in the absence of a gravity stimulus, the sensitivity of the root to an applied electric stimulus is increased. Further space experiments are required to determine the extent to which this sensitivity is shifted. The implications of this result are discussed in relation to gravitropic signaling and the regulation of differential cell elongation in the root.

Tomographic distribution of acetylated histone H4 in plant chromosomes, nuclei and nucleoli.

Root tip cells of broad bean (Vicia faba L. cv. 'Wase soramame') and barley (Hordeum vulgare L. cv. 'Minorimugi') were immunostained with antibodies specific for acetylated histone H4. With an antiserum that recognizes histone H4 acetylated at lysine-5, the nucleolar organizing region (NOR) in mitotic chromosomes was strongly labeled in both species. The broad bean had two signals in the metaphase and telophase chromosome complements and four signals in the prophase and anaphase chromosome complements, while the barley had four signals in the metaphase and telophase chromosome complements and eight signals in the prophase and anaphase complements. Five different patterns of signals were observed at interphase: in type I only nucleoli were wholly stained; in type II perinucleolar knob-like signals and/or fiber-like signals emanated from the nucleus; in type III aggregate signals appeared in the nucleolus; in type IV many small dot-like signals were distributed throughout the nucleus, except nucleoli; and in type V string-like or some granule-like signals appeared in the nucleoli. Type II was very similar to previous results by in situ hybridization with sense rDNA probes. Type III was similar to the patterns of DNA synthesis recognized as chromatin domains by anti-BrdU antibodies. Type V was very similar to the results of in situ hybridization with pTa71, rDNA probes and the appearance of the dense fibrillar components of the nucleolus.

Absorption and metabolism of lipids in rats depend on fatty acid isomeric position.

This study determined the effect of dietary (dipalmitoyl) triacylglycerol (TAG) structure on the absorption and metabolism of fat in the presence or absence of cholesterol. Sprague-Dawley male rats weighing approximately 180 g were fed diets containing 10 g/100 g diet 1,3-dipalmitoyl-2-oleoyl-glycerol-rich oil (POP) or 1(3),2-dipalmitoyl-3(1)-oleoyl-glycerol-rich oil (PPO), either with or without cholesterol supplementation (0.5 g/100 g diet), for 2 wk. There were no differences in the growth of rats fed the various diets. The apparent absorption efficiency of dietary fat especially the apparent absorption efficiency of palmitic acid, was significantly lower in the POP groups than in the groups fed the PPO diets. The excretion of cholesterol in feces of rats fed the POP diet with cholesterol was significantly greater than those fed the PPO diet containing cholesterol. The excretion of total bile acids in the rats fed cholesterol was significantly greater than in those fed the diets without cholesterol. Liver cholesterol was significantly greater in rats fed cholesterol and was not affected by the TAG structure. Plasma cholesterol was significantly lower in the POP dietary groups than in the PPO groups. These results suggest that the absorption and metabolism of fat depend on the positional isomer of the fatty acid.

The effect of follicular fluid on intracellular free calcium levels in human spermatozoa.

In this study, we demonstrate the effect of follicular fluid (FF) on free cytosolic calcium ion ([Ca2+]i) in human spermatozoa, using a fluorescent indicator "fura 2," and we evaluate the factors that increase [Ca2+]i in FF. A significant and immediate [Ca2+]i increase (656.3 +/- 148.6 nM) was elicited by intact FF. The increase in [Ca2+]i elicited by lipid-stripped FF was only 50% (322 +/- 68.7 nM) of that elicited by intact FF. However, 82% of the [Ca2+]i increase elicited by intact FF was observed when lipid-stripped FF was supplemented with progesterone, but not when it was supplemented with estradiol. Also, progesterone at the concentrations of 1 ng/ml to 1 microgram/ml caused a significant increase in [Ca2+]i by itself, but estradiol produced a small effect, ranging from 3% to 9% of the effect produced by progesterone. These results indicate that progesterone plays a major role in the lipid fraction in FF to elicit the entry of calcium into human spermatozoa.

[Autologous blood transfusion in cardiac surgery over 70-year-old patients].

We discussed the problems for autologous blood transfusion particularly in the preoperative blood donation about the patients over 70-year-old who underwent cardiac surgery in contrast with the younger patients. Following results were obtained: 1) Because aged patients have the tendency of anemia before predonation, full storage of blood donation could not be successed. 2 of 8 over 70-year-old patients were able to do the preoperative phlebotomy, and their mean volume were 355 gram as whole blood. On the other hand, 96% patients in younger group were phlebotomized preoperatively, and their mean value of storaged blood volume were 757 gram. 2) As the examinations about the iron-related parameters, not all aged patients were iron deficiency status. Their reticulocyte counts were nearly equal level to the younger group and plasma concentration of erythropoietin were higher in the aged patients than that in the younger group. These results indicate that erythropoiesis in the bone marrow was deteriorated in the aged patients. 3) In aged patients, all of them were required homologous blood transfusion at their perioperative terms. We thought that they have had the anemia before preoperation and our tolerable, allowable level to the postoperative anemia was not lower in the aged patients as against to the younger group. 4) We performed the autologous plasma donation with the membranous plasma separator added to the whole blood donation. It was easy and safe method, and circulatory indices were not changed before and after plasma-separation even in the aged patients. These autologous plasma were usefully administered as the volume expander postoperatively.

Extent of pontine pyramidal tract Wallerian degeneration and outcome after supratentorial hemorrhagic stroke.

BACKGROUND AND PURPOSE: Pyramidal tract Wallerian degeneration has been detected on magnetic resonance imaging (MRI) as T2-weighted high-intensity areas. We analyzed the relation between the extent of brain stem Wallerian degeneration and activities of daily living (ADL) after supratentorial hemorrhagic stroke. METHODS: Twenty-six patients with supratentorial hemorrhage were examined on the coronal T2-weighted image of the pons 3 months or later after stroke, and the percentage of Wallerian degeneration in the pons was calculated. The patients were divided into three groups. In group A (n = 6), MR films were taken 3 to 6 months from the onset, and the ADL assessment was done within 2 months from the MRI. In group B (n = 11), MR films were taken 3 to 6 months from the onset, and the ADL assessment was done within 10 months from the MRI (mean, 15.5 months from the onset). In group C (n = 9), MR films were taken after 10 to 17 months (mean, 12.0 months) from the ictus, and the ADL assessment was done simultaneously. Barthel Index score was used for quantitative ADL assessment. RESULTS: All patients showed various degrees of pontine pyramidal tract Wallerian degeneration associated with capsular involvement by the hematoma. In group A, the percentage of degeneration did not correlate with the Barthel Index score (r = .2101, P = .6895). An inverse relation between percentage of degeneration and Barthel Index score was seen in groups B (r = .7354, P = .0099) and C (r = .888, P = .0014). In groups B and C, Wallerian degeneration was higher in patients with Barthel scores less than 60 (P = .005). CONCLUSIONS: The extent of pontine Wallerian degeneration on MRI 3 months or later after the stroke correlated with the patient's Barthel Index score 1 year after the stroke.

Six new triterpenoidal glycosides including two new sapogenols from Albizziae Cortex. V.

Six new triterpenoid glycosides called julibrosides A1-A4, B1 and C1 were isolated from Albizziae Cortex, the dried stem bark of Albizzia julibrissin Durazz. Their structures were determined based on spectral and chemical evidence. Julibrosides B1 and C1 had new sapogenols, designated julibrogenin B and C, respectively, while julibrosides A3 included N-acetyl-D-glucosamine as a sugar component.

[Effect of recombinant human erythropoietin on autologous blood pre-donation in open heart surgery].

We have used recombinant human erythropoietin (EPO) with an autologous blood predonation in open heart surgery looking forward to preventing patient's blood level of hemoglobin and quick recovery in post-operative period. In our results, patient's value of hemoglobin (Hb) and hematocrit (Ht) decreased due to autologous blood predonation. In group A (autologous blood predonation with EPO administration), however, predonated blood volume were larger than in group B (without EPO administration), decreased value of Hb and Ht were smaller than in group B. The counts of reticulocyte were higher in group A at the operative day. Among six cases phlebotomized with EPO administration, five cases required no homologous blood transfusion for their hospital course. Postoperative recovery of patient's Ht value were earlier in preoperative blood donation group. In particular, patients who administered EPO intravenously have showed fair recovery from anemia. EPO is very effective drug to prevent patients from the developing anemia as a complication of autologous blood predonation. We conclude that autologous blood predonation with EPO administration is beneficial method to reduce homologous blood requirement in open heart surgery.

Vasoactive intestinal peptide- and peptide histidine isoleucine amide-like immunoreactivity colocalize with vasopressin-like immunoreactivity in the canine hypothalamo-neurohypophysial neuronal system.

The distribution of vasoactive intestinal peptide (VIP) and peptide histidine isoleucine amide (PHI) was investigated in the canine hypothalamus by immunocytochemistry. VIP- and PHI-like immunoreactive neurons were detected in the magnocellular supraoptic and paraventricular nucleus. These magnocellular VIP- and PHI-producing neurons coexist with vasopressin-like immunoreactivity and send axons to the median eminence and neurohypophysis. These findings may serve as an anatomical basis for studying the function of VIP and PHI on pituitary hormone secretion.

Light and electron microscopic immunocytochemistry of neurotensin-like immunoreactive neurons in the rat hypothalamus.

Neurotensin-like immunoreactive neuronal perikarya, fibers and terminals in the rat hypothalamus, particularly in the arcuate nucleus, the paraventricular nucleus and the median eminence, were investigated by light and electron microscopic immunocytochemistry. The main distributional areas of immunoreactive neuronal perikarya were found to be the arcuate nucleus, the periventricular nucleus and the paraventricular nucleus by light microscopic immunocytochemistry. Immunoreactive neuronal perikarya showed a characteristic distributional pattern in the arcuate nucleus. In the paraventricular nucleus they were distributed in both the magnocellular and parvocellular portions. A large number of immunoreactive terminals were observed throughout the external layer of the median eminence, particularly its lateral portion. A moderate number of immunoreactive terminals were also observed in the internal layer of the median eminence. By electron microscopic immunocytochemistry immunoreactive neuronal perikarya both in the arcuate and paraventricular nuclei showed generally well-developed cell organelles such as mitochondria, r-ER, and Golgi complex. In addition, immunoreactive dense granules were dispersed throughout the perikarya. A large number of immunoreactive terminals containing immunoreactive dense granules, clear vesicles and mitochondria were observed in the vicinity of pericapillary spaces of the external layer of the median eminence. This observation strongly suggests that neurotensin-like immunoreactive substance is released into the portal capillaries.

Morphological survey of neurotensin-like immunoreactive neurons in the hypothalamus.

Neurotensin-like immunoreactive neuronal perikarya, fibers and terminals in in the rat hypothalamus were investigated by light and electron microscopic immunocytochemistry. Distributional density and pattern of these elements were clarified. Fine structure of immunoreactive neuronal perikarya with respect to development of cell organellae and immunoreactive dense granules was also elucidated. Features of immunoreactive processes, dendrites and preterminal axons were examined electron microscopically. In addition to the above findings by light and electron microscopic immunocytochemistry, we examined the coexistence of dopamine and neurotensin-like immunoreactive substances in these same neurons in the arcuate and periventricular nuclei. This was proved by the application of fluorescence histochemistry and immunocytochemistry on the same sections. Moreover, we speculated that the ascending noradrenergic neurons influence the neurotensin immunoreactive neurons in the paraventricular nucleus since a marked decrease in the number of neurotensin-like immunoreactive neuronal perikarya was observed after transection of ascending noradrenergic pathway.

Some cellular characteristics of somatostatin neurons and terminals in the periventricular nucleus of the rat hypothalamus and median eminence. Electron microscopic immunohistochemistry.

Somatostatin neuronal perikarya and their processes, presumably dendrites, in the periventricular nucleus of the rat hypothalamus and terminals in the median eminence were observed by electron microscopic immunohistochemistry. Neuronal perikarya and processes contained immunoreactive dense granules (100-120 nm in diameter) and other cellular components such as polysomes, rER membranes occasionally showed high electron density. Few axo-somatic terminals were found on the somatostatin neurons, but we could detect a number of preterminal axons on immunoreactive processes, presumably dendrites. Therefore, we considered that somatostatin neurons receive mainly neuronal input through axo-dendritic synapses rather than through axo-somatic ones. In the somatostatin terminals in the external layer of the median eminence immunoreactivity was completely restricted on the granules.

Increase by visual stimuli in turnover of 5-hydroxytryptamine in the superior colliculi of the rabbit.

1. Irregular light falshes were played on to one eye of dark adapted rabbits for periods of 20-80 min. The concentration of 5-hydroxyindol-3-ylacetic acid (5-HIAA) and of 5-hydroxytryptamine (5-HT) were estimated in left and right superior colliculi, thalami and hippocampi. 2. In rabbits exposed to such visual stimuli for 30-60 min, there was an increase in the 5-HIAA content of the colliculus contralateral to the stimulated retina which aberaged 17% (P = 0-02), but no rise was seen if the exposure was shortened to 20 or prolonged to 80 min. At no time was there a difference in 5-HIAA content between right and left thalamus or right and left hippocampus. 3. Stationary or strictly repetitive visual stimuli produced no difference between the 5-HIAA content of left and right superior colliculus. 4. No difference in 5-HT concentration between the two colliculi was found after any form of visual stimulation, nor did any changes occur in the other parts of the brain which were examined. 5. Irregular, prolonged visual stimualtion thus appears to activate tryptaminergic neurones terminating in the colliculi. The possibility is discussed that the 5-HT released at this site might act as a brake to neuronal activity under conditions when habituation to the stimuli is not yet complete.