Regulation of the lysophosphatidylserine and sphingosine 1-phosphate levels in autologous whole blood by the pre-storage leukocyte reduction.

BACKGROUND AND OBJECTIVES: The effect of leukoreduction and storage periods on the accumulation of bioactive lysophospholipids and substances in human autologous blood (AB units) has not been fully investigated. MATERIALS AND METHODS: The accumulation of bioactive lysophospholipids such as sphingosine 1-phosphate (S1P) and lysophosphatidylserine (LysoPS) in AB units during the storage was investigated. The time-dependent changes and the effect of the filtration in pre-storage leuckoreduction (LR) and unmodified samples derived from 46 AB units were analysed. Additionally, the changes of lysophospholipids and platelet releasate, namely ß-thromboglobulin (ß-TG), induced by exposure of whole blood (WB) or platelet-rich plasma (PRP) to the filter material were analysed. RESULTS: LysoPS, but not S1P levels, time-dependently and significantly increased in both unmodified and LR samples. LysoPS significantly decreased in LR compared with unmodified samples, whereas S1P increased in LR compared with unmodified samples. In addition, exposure of WB and/or PRP to the filter material in vitro resulted in increased levels of S1P, LysoPS and ß-TG. CONCLUSIONS: LR effectively reduced the accumulation of LysoPS in AB units. On the other hand, it increased concentrations of S1P due to platelet activation by exposure to the filter material. These suggest that increases of S1P levels in LR and LysoPS in the unmodified samples were mainly caused by the leukocytes and/or platelets and that LR was effective in inhibiting the accumulation of LysoPS.

Simulated Cuspal Deflection and Flexural Properties of Bulk-Fill and Conventional Flowable Resin Composites.

CLINICAL RELEVANCE: Some bulk-fill flowable resin composites produce less cuspal deflection than a conventional incrementally filled flowable resin composites. SUMMARY: Objective: This study investigated simulated cuspal deflection and flexural properties of bulk-fill and conventional flowable resin composites.Methods and Materials: Five bulk-fill and six conventional flowable resin composites were evaluated. Aluminium blocks with a mesio-occlusal-distal cavity were prepared and randomly divided into groups for each of the different measurement techniques and were further subdivided according to the type of flowable resin composite. The simulated cuspal deflection caused by the polymerization of resin composite within an aluminium block was measured using a highly accurate submicron digimatic micrometer or a confocal laser scanning microscope (CLSM). In addition, the flexural properties of tested resin composites were measured to investigate the relation between cuspal deflection and flexural properties, and the resin composites were observed using scanning electron microscopy.Results: Simulated cuspal deflection of some bulk-fill flowable resin composites was found to be significantly lower than or similar to those for conventional counterparts, regardless of the measurement method. There were statistically significant differences in flexural properties depending on the material, regardless of the type of flowable resin composite. Pearson correlation analysis did not reveal a statistically significant relationship between flexural properties and cuspal deflection.Conclusion: These results indicate that some bulk-fill flowable resin composites exhibit lower cuspal deflection with the bulk-filling technique than is shown by conventional flowable resin composites using the incremental filling technique. Simulated cuspal deflection can be measured using either a micrometer or CLSM, but this experiment failed to show a significant relationship between cuspal deflection and flexural properties of flowable resin composites.

Effect of Reduced Universal Adhesive Application Time on Enamel Bond Fatigue and Surface Morphology.

OBJECTIVE:: The purpose of this study was to evaluate the effect of reduced application times of universal adhesives on enamel bond fatigue and surface morphology of the treated enamel with constant force atomic force microscopy (AFM). METHODS:: Four universal adhesives-Adhese Universal (AU), Clearfil Universal Bond Quick (CU), G-Premio Bond (GP), and Scotchbond Universal Adhesive (SU)-were evaluated in a laboratory for their ability to adhesively bond resin composite to enamel. Shear bond strengths were initially determined using 15 specimens per test group for each adhesive. Shear fatigue strengths were then determined using 20 specimens per test group for each the adhesives. The fatigue specimens were loaded using a sine wave at a frequency of 20 Hz for 50,000 cycles or until failure occurred. AFM observations, surface Ra roughness measurements, and geometric surface area evaluations of enamel surface treated with the adhesive agents were also conducted. RESULTS:: A strong relationship was found between the initial shear bond strength and shear fatigue strength for enamel surface Ra roughness but not for geometric surface area. The initial shear bond strength and shear fatigue strength of CU and GP were not influenced by different application times, unlike those of AU and SU. While the surface area of enamel treated with the adhesive agents was not significantly influenced by different application times and type of adhesive, surface Ra roughness of the enamel in the AU and SU groups significantly increased with increasing application time, unlike CU and GP. CONCLUSIONS:: The results of this study suggest that universal adhesives, used with reduced application times, have adequate Ra surface roughness to provide sufficient resistance to enamel bond fatigue at application times from <1 second to 20 seconds, while the geometric surface area of adhesive-treated enamel did not show any significant changes at these different application times.

Application of the Self- Assembling Peptide P11-4 for Prevention of Acidic Erosion.

The aim of this study was to use ultrasonography to evaluate the effect of the self-assembling peptide P11-4 on acid erosion prevention. Curodont Repair (CR), which includes peptide P11-4, was used. Rectangular prisms of bovine enamel (4×1×1 mm) were immersed in pure orange juice for a period of 5 minutes six times per day for 28 days. These samples were divided into four groups of six specimens each and treated differently for an additional period of 28 days: 1) baseline group specimens were stored in artificial saliva; 2) CR group specimens were exposed to curodont without acid challenge; 3) NCRA (no curodont+acid challenge) specimens were treated with orange juice without curodont exposure; and 4) CRA (CR+acid challenge) specimens were treated with curodont before treatment with orange juice. The propagation time of longitudinal ultrasonic velocity (UV) was measured. Ultrastructural observation of each tested enamel surface was carried out using field-emission scanning electron microscopy (SEM). The UV data were analyzed using two-way analysis of variance with time and treatment as confounding factors. Post hoc pairwise tests among groups were performed using the Tukey honestly significant difference test. The average UV in intact bovine enamel for the baseline group ranged from 4,483 to 4,549 m/s and did not vary significantly within the test period. The average ultrasonic velocity (UV) in all samples decreased after the initial erosion. The UV in NCRA decreased further over time. Increased UVs were found for CR and CRA. For CR and CRA, there was no significant difference in UV at the end of the experiment from the initial value before erosion. In the results of SEM observation, the CR and CRA groups had similar morphologic features in that etching patterns were not clearly due to precipitation between the enamel rods. From the results of this in vitro study, it might be concluded that applying enamel matrix derivatives and self-assembling peptides on erosive lesions can improve remineralization.

Effect of active immunization against inhibin on hormonal concentrations and semen characteristics in Shiba bucks.

Active immunization against inhibin increased ovulation rate in females; in males, the effects of active immunization against inhibin on hormonal concentrations and sperm production need more investigation. To test the hypothesis that active immunization against inhibin increases FSH secretion and sperm output, the present study was undertaken to determine the effects of active immunization against inhibin on hormonal profile and sperm production in Shiba bucks. The bucks were actively immunized against inhibin alpha-subunit (immunized group, n=6) or Freund adjuvant (control group, n=5) four times, at 5-weeks intervals. Blood samples were collected twice-weekly and two successive ejaculates of semen were collected (with an artificial vagina) once-weekly. Plasma concentrations of FSH, LH and testosterone were measured by radioimmunoassay (RIA) and sperm motility characteristics were measured by computer-assisted sperm analysis (CASA). All inhibin-immunized bucks produced antibodies against inhibin. Relative to control bucks, in immunized bucks there were significant increases in plasma FSH concentrations and in sperm concentrations from 5 to 9 weeks and from 8 to 11 weeks, respectively, after primary immunization. However, plasma concentrations of LH and testosterone, semen volume, percentage of motile spermatozoa and motility parameters (straight-line velocity, curvilinear velocity and linearity index) were similar in both groups. In conclusion, active immunization against inhibin alpha-subunit increased FSH secretions and enhanced sperm production in bucks, whereas LH and testosterone concentrations, semen volume and sperm motility parameters were unaffected. Active immunization against inhibin could be used to improve fertility in Shiba bucks.

Ovarian and hormonal response of female goats to active immunization against inhibin.

This study was conducted to evaluate the effect of active immunization against inhibin on hormonal levels and the ovulation rate in goats. Ten adult Shiba goats (Capra hircus) in two groups were used in this study. The first group was injected with inhibin vaccine (immunized, n=5) and the second group was injected with Freund's adjuvant (control, n=5) followed by three booster injections at 4-week intervals. After the third booster injection, three consecutive periods of oestrus were induced using prostaglandin F(2alpha) at intervals of 11 days. Blood samples were collected at 2-6 h intervals and the ovaries were monitored using B-mode ultrasonography. All inhibin-immunized goats generated antibodies that bound (125)I-labelled bovine inhibin and their FSH concentrations were significantly higher than corresponding values in the control group. Also, inhibin-immunized goats had significantly higher preovulatory oestradiol-17beta (P<0.01) and higher concentrations of progesterone in the luteal phase (P<0.05). Immunization of goats against inhibin resulted in a significant (P<0.01) increase in ovulation rate (control: 1.7+/-0.3 vs immunized: 7.6+/-1.1). These results demonstrate that active immunization against inhibin enhances ovarian follicular development and ovulation rate by promoting an increase in pituitary FSH secretion. Therefore, immunization against inhibin may be a useful alternative to the conventional approach of superovulation in goats.

Influence of cefotaxime on renal function during the treatment of infections.

A study was conducted of the effects of cefotaxime, a third generation cephalosporin antibiotic, on the function of the kidney, using several indices of renal function including urinary concentrations of the renal enzyme N-acetyl-beta-D-glucosaminidase (NAG). In 6 patients with respiratory infections and normal renal function (group I), the urinary concentrations of NAG before and after the administration of cefotaxime 2 to 4g daily were 5.7 +/- 0.6 U/L and 5.5 +/- 0.9 U/L, respectively (NS). Similarly, 9 patients with chronic renal failure who were not undergoing haemodialysis showed pre- and post-treatment urinary NAG concentrations of 8.7 +/- 4.0 U/L and 6.6 +/- 1.7 U/L, respectively (NS), while the corresponding values in 12 renally impaired patients undergoing haemodialysis (group III) were 8.1 +/- 3.5 U/L and 8.9 +/- 3.8 U/L, respectively (NS). With regard to other parameters of renal function (serum creatinine, BUN, beta 2M, and creatinine clearance), no statistically significant differences were found between the values obtained before and after therapy with cefotaxime. Therefore, it was concluded that the influence of cefotaxime on renal function is slight, and that this antibiotic can be safely used to treat patients with infections in the presence of renal dysfunction.

Effect of intraperitoneal administration of heparin to patients on continuous ambulatory peritoneal dialysis (CAPD).

The effect of intraperitoneal administration of heparin to patients on continuous ambulatory peritoneal dialysis (CAPD) remains obscure. We examined 8 patients on CAPD to investigate its effect. When 2.5 U/ml or 5 U/ml of heparin was given intraperitoneally, t1/2 of heparin activity in the dialysate was 0.5 to 2 hrs, and 6 hrs after administration its activity was 0.5 U/ml and 1.4 U/ml respectively. Whole blood clotting time was hardly affected because the transfer of heparin to the plasma was minimal. The plasma antithrombin III (AT-III) levels were comparable to patients on hemodialysis, but the AT-III level in the dialysate was only 1.5% of those in the plasma. We conclude that the intraperitoneal administration of heparin at these doses is effective in preventing fibrin precipitation when intraperitoneal AT-III levels are expected to be relatively increased such as at the start of CAPD or in the presence of peritonitis.

Analysis of mandible of newly developing strain of Japanese white rabbit (Nib:JWNS).

In the present paper, we applied analysis of the mandible to a new strain of Japanese White rabbit being developed (Nib: JWNS), at 5th and 8th generations (F5 and F8), and evaluated the process of establishment, making comparison with the parent colonies (I, K and L), and other established JW colonies (A, E, O and R). The mandibles were measured at 12 sites and the data obtained were calculated by principal component analysis and discriminant analysis. As a result of principal component analysis, the parent colonies were found to be close to one another with intermediate size of the mandible and height of condyloid and angular processes, but in F5 and F8 the mandibles were slightly shorter in height and different from those in the parent colonies. On the other hand, results of discriminant analysis revealed that mandibles were discriminated correctly 100% in colonies A, O and R, and 90% in E and I. Colonies A, O, R, E and I were therefore regarded as established strains. In the developing colonies, the discriminant rate was 70% in F5 and 80% in F8. One mandible from F5 and two from F8 were wrongly classified to the parent colony L, which was the lowest discriminant rate among the colonies examined (61.5%). The results of both statistical analyses suggest that the JWNS are almost established at 8th generation as a new strain.

Study on the environmental background to aggravating factors for renal function related to pregnancy and delivery in IgA nephritis.

Various effects on renal function and factors which tend to cause aggravation in pregnancy and delivery in patients with renal diseases have been described previously. In the present study, the authors undertook personal and telephone inquiries to patients with renal insufficiency due to aggravated renal dysfunction occurring during the course of pregnancy and delivery, and to those showing a favorable course despite renal dysfunction before pregnancy. Sixty-four patients with IgA nephritis were investigated, covering 101 cases of pregnancy in them. The management of the pregnant women and their social and home (family/domestic) environments were taken into consideration, since these are important for giving guidance in their daily life to overcome various factors which influence the course of spontaneous pregnancy and delivery. The following findings were obtained. (1) In patients with hypertension, toxemia of pregnancy, abnormal delivery and abortion were frequently complicated. (2) Patients under greater mental and physical stresses during the course of pregnancy and after delivery showed a poorer prognosis for both mother and fetus. (3) Generally speaking, guidance should be given not to be overconfident or overdefensive in pregnancy. (4) Working women revealed a prognosis which was poorer than that of housewives.

[Phosphate kinetics in patients on chronic hemodialysis].

To investigate the phosphate kinetics in hemodialysis (HD), 8 patients in a stable condition, who were receiving HD three times a week for 4 hours per session, were investigated. Plasma phosphate was under 7 mg/dl, and residual renal function had almost disappeared. Dialysate containing phosphate was prepared by adding Na2HPO4 using a micro-infusion pump from the inlet of single pass dialysate in the individual dialysate delivery system. In the first week, Na2HPO4 was not added as the control period. In the second session of the second and third week, Na2HPO4 was added to give a phosphate concentration of 1.0 and 2.0 mmol/l in the dialysate, respectively. Total phosphate mass removal was 777 +/- 46.64 mg in the control period, 403 +/- 67.21 mg in the second week, and -230 +/- 214.8 mg in the third week. Total phosphate mass removal in the second and third week was significantly lower than that of the control period. Plasma phosphate concentration was significantly decreased after the HD compared with before the HD in the control and second week. There was no significant difference in plasma phosphate concentration between the period before HD and at 48 hours in the control and the second week. Plasma phosphate concentration before HD not only depended on phosphate mass removal by HD, but also on other factors. We suggest that dialysate containing phosphate might prevent excessive phosphate removal from non-extracellular compartments during HD.

Influence of 1 alpha-hydroxy vitamin D3 (0.25 micrograms/day) and calcium carbonate on patients with chronic renal failure at the predialytic stage.

The influence of a small amount of 1 alpha (OH)D3 on patients with chronic renal failure at the predialytic stage (CRFPS patients) receiving CaCO3 was investigated. 7 patients given CaCO3 (3 g/day) were administered 1 alpha (OH)D3 (0.25 micrograms/day) over a period of 12 weeks. 2 patients were eliminated from the study because of obvious deterioration of renal function. The others revealed no significant changes in levels of serum creatinine, adjusted calcium, phosphate, calcium x phosphate product, alkaline phosphatase, C-terminal parathyroid hormone, pH and bicarbonate. Our results indicated that even 0.25 micrograms/day 1 alpha (OH)D3 should not be prescribed to CRFPS patients given 3 g/day CaCO3. We recommend a little sole use of CaCO3 in CRFPS patients, paying attention to any exacerbation of renal function due to a rise in serum calcium concentration, when therapy for secondary hyperparathyroidism is required.

Treatment of secondary hyperparathyroidism in patients on maintenance hemodialysis.

The effects of CaCO3 were investigated in 7 patients undergoing maintenance hemodialysis who had been treated with 1 microgram/day 1 alpha (OH)D3 and 2 g/day Al(OH)3. CaCO3 (3 g/day) was administered instead of Al(OH)3. The levels of serum total calcium and ionized calcium were significantly increased, the level of C-terminal parathyroid hormone was appreciably decreased and the levels of serum aluminum in all patients were reduced. There were no significant changes in the levels of serum phosphate, calcium x phosphate product, alkaline phosphatase, calcitonin, magnesium and bicarbonate. It is concluded that 3 g/day CaCO3 is equivalent to 2 g/day Al(OH)3 in terms of its phosphate-binding effect, and the prescription of CaCO3 together with 1 alpha (OH)D3 ameliorates secondary hyperparathyroidism.

A case of primary systemic amyloidosis with AA protein.

We present here a case of young adult-onset primary systemic amyloidosis with AA protein. A 29-year-old male presenting with macrohematuria was transferred to our hospital because of aggravation of renal function. Amyloid protein was detected in the bladder, stomach and thyroid. The amyloid protein proved to be AA type by potassium permanganate histochemical analysis. Since secondary amyloidosis was excluded by the laboratory data and there was no family history of amyloidosis, a diagnosis of primary systemic amyloidosis with AA protein was reached. Although the patient was prescribed dimethyl sulphoxide, his renal function worsened with gastrocolic symptoms and a bleeding tendency. Hemodialysis (HD) was then initiated. After starting the HD, the patient's general condition recovered and subsequently the patient on treatment with maintenance HD was discharged.

Early morphological changes of tubules in rats with chronic renal failure.

We investigated the early changes of tubules and effect of the oral adsorbent, AST-120, on the early changes of tubules in rats with chronic renal failure. Sprague-Dawley rats were divided into two groups with and without AST-120, after 3/4 nephrectomy. Although there were no significant differences in levels of blood urea nitrogen, serum creatinine, creatinine clearance, inulin clearance, para-aminohippuric acid clearance and urinary N-acetyl-beta-D-glucosaminidase at week 8 between the two groups, the amount of 24-hour urinary protein excretion and the direct systolic blood pressure at week 8 were significantly decreased in the group with AST-120. Examinations by light microscopy at week 8 revealed that proteinaceous casts in the tubules, tubular dilatation and infiltration of monocytes into the interstitium in the group with AST-120 were less prominent than those in the group without AST-120. A significant difference in numbers of proteinaceous casts was noted at week 8 between the two groups. In rats with chronic renal failure at the early stage, it is concluded that the formation of proteinaceous casts, resulting in tubular damage, is increased and that AST-120 delays the occurrence of proteinaceous casts by delaying the increase in urinary protein excretion.

[The influence of correction of acidosis on plasma level of branched-chain amino acids in chronic hemodialysis patients].

Seven patients undergoing chronic hemodialysis three times a week and whose plasma bicarbonate concentration on predialysis was consistently under 18 mmol/l due to bicarbonate dialysis (BCD), were treated with BCD for 2 weeks, then switched to acetate-free biofiltration (AFB) for 8 weeks. In both periods, the same high flux dialyzer (AN69HF) was used. The treatment time, dialysate flow rate and blood flow rate were kept constant in each patient during both periods. Plasma bicarbonate concentration (HCO3-), serum urea nitrogen (SUN), serum creatinine (Cr) and plasma amino acids concentrations (AA) were measured before dialysis and KT/V was calculated on the 2nd days of the last week in both periods. HCO3- on AFB was significantly higher than that on BCD (16.4 +/- 0.9 vs 19.9 +/- 1.8 mmol/l; p < 0.05). SUN on AFB was significantly lower than that on BCD even though the dialysis schedule and dietary content were not changed (84.7 +/- 3.7 vs 76.6 +/- 3.8 mg/dl; p < 0.05). TP, Cr and KT/V were not significantly different. Plasma total amino acid concentration (TAA) and plasma essential amino acid concentration (EAA) were not significantly different in both periods. In contrast, plasma branched-chain amino acid concentrations (BCAA) on AFB were significantly higher than that on BCD (313.5 +/- 44.3 vs 390.3 +/- 50.7 mumol/l; p < 0.05). Plasma BCAA concentrations, valine (VAL), leucine (LEU) and isoleucine (ILE), were significantly higher on AFB than that on BCD, respectively (p < 0.05). These findings suggest that optimal correction of the metabolic acidosis in chronic hemodialysis patients by AFB leads to a significant increase in plasma BCAA concentration.

Effects of recombinant human erythropoietin (rHuEPO) on nutritional status of hemodialysis patients: investigation of direct anabolic effects of rHuEPO.

To investigate whether the nutritional improvement achieved by recombinant human erythropoietin (rHuEPO) treatment is the result of anemia correction with rHuEPO or the direct anabolic effects of rHuEPO per se, nutritional assessment was performed in 2 studies (study I and II) on hemodialysis (HD) patients. Nutritional assessment included blood biochemistry determinations, anthropometric measurements, daily protein intake (DPI) and dialysis efficiency. In study I, 5 HD patients who had not been given rHuEPO and had a hematocrit (Hct) of < or = 25%, were administered rHuEPO at the initial dose of 96.2 U/kgBW. Nutritional assessment of these patients was performed before rHuEPO treatment and every 4 weeks until the 24th week after rHuEPO treatment. In study II, the same nutritional assessment as in study I except for DPI, was performed in 2 groups with the same Hct level and dialysis regimen; an EPO group (n = 8) previously given rHuEPO (88.2 +/- 13.7 U/kgBW, 25.8 +/- 2.5 mos) and a non-EPO group (n = 8) not given rHuEPO. In study I, the mean Hct level was significantly increased 4 weeks after rHuEPO treatment (23.3 +/- 0.6 to 26.9 +/- 0.9%). However, the nutritional parameters and dialysis efficiency were nearly constant over 24 weeks, suggesting either the absence of a short-term direct anabolic effect of rHuEPO or masking of such an effect due to general condition improvement by anemia correction with rHuEPO. In study II, no significant differences in nutritional assessment were confirmed between the groups, suggesting that a long-term direct anabolic effect of rHuEPO may not exist and nutritional improvement may result from correction of anemia with rHuEPO.(ABSTRACT TRUNCATED AT 250 WORDS)

Three-quarters nephrectomy in rats as a model of early renal failure.

To produce a compatible model of early renal failure easily, we prepared three-quarters nephrectomized animal. 20 Sprague-Dawley rats were divided into the following two groups: 10 rats which received three-quarters nephrectomy (Nx group), assuming the weight of both kidneys to be equal, and another 10 rats which underwent sham operations (S group). The levels of creatinine clearance in Nx group and S group were 286.5 +/- 33.5 vs. 431.1 +/- 55.9 microliters/min/100 g BW (P less than 0.001) on week 2, and 233.0 +/- 16.7 vs. 562.3 +/- 62.9 microliters/min/100 g BW (P less than 0.001) on week 10, respectively. The indirect and direct systolic blood pressure (SBP) values of Nx group and S group at the 10th week were 154.5 +/- 5.5 vs. 131.1 +/- 3.6 mmHg (P less than 0.01), and 148.2 +/- 4.8 vs. 130.7 +/- 6.6 mmHg (P less than 0.01), respectively. Significant changes in the levels of urinary protein excretion, urinary sodium output, urinary epinephrine, urinary norepinephrine and plasma renin activity between both groups were recognized on the 10th week. The planar area of Nx group was significantly increased as compared to that of S group (11.0 +/- 0.3 vs. 7.2 +/- 0.1 x 10(-3) mm2, P less than 0.001). It is concluded that this model resembles early renal failure in humans because slow progression of renal dysfunction occurred with mild elevation of SBP, and that it is adequate for evaluating the influence of glomerular hypertrophy, resulting in glomerulosclerosis.