Effect of Whey-Derived Lactopeptide ß-Lactolin on Memory in Healthy Adults: An Integrated Analysis of Data from Randomized Controlled Trials.

CONTEXT: Epidemiological studies have shown that consumption of dairy products reduces the risk of dementia and cognitive decline in older individuals. Tryptophan-tyrosine-related ß-lactopeptides and their representative ß-lactolin of glycine-threonine-tryptophan-tyrosine tetra-peptide have been identified as agents in dairy products, which improve cognitive function as well as memory function via the activation of the dopaminergic system in a mouse model of amnesia. Previous clinical trials have shown that supplementation with ß-lactolin improves memory retrieval in healthy older adults. Specifically, ß-lactolin improved the scores in some neuropsychological tests. However, the effects of ß-lactolin on memory function have not been clarified. OBJECTIVES: The aim of this study was to evaluate the effect of ß-lactolin on memory function using statistical methods. DATA SOURCES: We searched the Web of Science, Cochrane Library, and JDream III until November 2021 to identify relevant randomized controlled trials for integrated analysis. DATA SYNTHESIS: Three randomized controlled trials evaluating the effect of ß-lactolin on memory in healthy adults were selected for the integrated analysis. The results showed that the score of cued recall among the neuropsychological tests in the ß-lactolin group was significantly higher than that in the placebo group (g=0.33; 95% CI: 0.10, 0.55). In addition, the total memory score was higher but this difference was not significant (g=0.17; 95% CI: -0.09, 0.43). CONCLUSIONS: Taken together, these results suggest that supplementation with ß-lactolin improves cued recall in healthy older adults.

Proposal of a simplified technique for staining bacterial spores without applying heat--successful modification of Moeller's method.

BACKGROUND AND AIMS: As the bacterial spores are difficult to stain, a number of staining techniques including their modifications have been proposed to date. Most of the conventional staining procedures unexceptionally contain the step of staining with steamed dye reagent in order to increase the stainability of the spores. We made an attempt to improve the conventional Moeller's methods for staining bacterial spores. METHODS: Spores of Bacillus species were stained with our modified Moeller's spore stain and evaluated for its staining properties. We investigated the stainability of both of the conventional and the modified Moeller's methods and the evaluation was made whether or not the step of steaming of Kinyoun's carbol-fuchsine dye reagent could be omitted by adding to aliquots of Tergitol 7, in place of the conventional dye solution steamed for some interval over hot blue flame of a Bunsen burner. RESULTS: We successfully omitted the heating step of steaming the Kinyoun's carbol fuchsine dye solution in the Moeller's method of bacterial spore stain, by the replacement of Kinyoun's carbol-fuchsine dye solution involving 2 drops of Tergitol 7, nonionic polyglycol ether surfactants type NP-7 (Sigma-Aldrich Japan, Tokyo, Japan) per 10 ml of Kinyoun's carbol-fuchsine dye solution. Bacillus spores stained pink to red and vegetative bacterial cells stained blue, although without applying any heating step during the whole course of staining processes including the fixation process. The novel staining method of our proposal resulted in far better satisfactory stainability in comparison with the conventional Moeller's method with the steaming dye solution. CONCLUSIONS: The modified spore stain without applying any heating step using the Kinyoun's carbol-fuchsine dye solution with an addition of Tergitol 7 aliquots was demonstrated to be reproducible and yielded consistent and satisfactory stainability. This simplified staining procedure is rapid to perform and found to be applicable to detect the bacterial spores in routine clinical microbiology laboratories.

Liver Failure From Ultra-Short Bowel Syndrome on the Intestinal Transplant Waiting List: A Retrospective Study.

BACKGROUND: Patients with intestinal failure (IF) are candidates for intestinal transplantation (ITx). In Japan, these patients have few opportunities to undergo cadaveric ITx because of low rates of organ donation. The donor criteria and recipient priority for ITx are still unknown. We reviewed our cases of IF to investigate which patients should be prioritized for ITx. METHODS: Patients with IF who were registered as candidates for cadaveric ITx between January 2010 and November 2015 in our institute were included in this retrospective study. Their data were gathered from their charts and analyzed. RESULTS: Five patients were included. Their primary diseases included total colon aganglionosis (n = 1), chronic idiopathic intestinal pseudo-obstruction syndrome (n = 2), superior mesenteric vein embolization (n = 1), and graft loss after ITx (n = 1). Two patients died of liver failure (LF) during the waiting period. The remaining three are now alive and waiting for transplantation. The lengths of the remaining intestine were more than 20 cm in living cases but less than 20 cm in fatal cases. In the fatal cases, they had several episodes of catheter-related blood stream infection, which caused LF and acute renal failure. CONCLUSIONS: We identified two patients with less than 20 cm residual small bowel who died after acute deterioration of liver function. Patients with ultra-short bowel could have a higher risk of LF. Therefore, they should be referred as soon as possible to a specialized hospital where ITx is a choice of treatment for IF.

Expression of a Synthetic Gene of CTDM by Transgenic Animals.

BACKGROUND: The purpose of this study was to produce molecules that can precisely regulate the complement and coagulation system and to assess the expression of such molecules in transgenic animals. METHODS: The CTDM gene, which is composed of the delta-1-99 amino acid (aa) C1-INH, EGF domain 4-6 of thrombomoduline (TM), short consensus repeat (SCR) 2-4 of DAF(CD55), and SCR 2-4 of MCP(CD46) was established. The codon usage for expression in mammals was adopted. The cDNA of CTDM was subcloned into the pCPI site (the human insulin promoter and a cytomegalovirus enhancer). pCPI-CTDM was transfected into pig endothelial cells (PEC). The expression of the molecule was clearly assessed by means of flow cytometry. RESULTS: BD3F1 female mice were induced to superovulate and were then crossed with BD3F1 males. Micro-injection and embryo transfer were performed by standard methods, thus generating transgenic mice that express CTDM. The mice carried the CTDM plasmid, as verified by PCR. Tissue expression levels in transgenic mouse lines generated with the constructs were follows: pancreas, 1.0; brain, 5.4; thymus, 0.3; heart, 0.2; lung, 1.2; liver, 0.1; kidney, 0.1; intestine, 0.4; and spleen, 1.6. A naive control mouse was also analyzed in the exact manner as for the transgenic mice. CONCLUSIONS: A synthetic CTDM gene with codon usage optimized to the mammalian system represents a critical factor in the development of transgenic animals.

HLA-G1, but Not HLA-G3, Suppresses Human Monocyte/Macrophage-mediated Swine Endothelial Cell Lysis.

The inhibitory function of HLA-G1, a class Ib molecule, on monocyte/macrophage-mediated cytotoxicity was examined. The expression of inhibitory receptors that interact with HLA-G, immunoglobulin-like transcript 2 (ILT2), ILT4, and KIR2DL4 (CD158d) on in vitro-generated macrophages obtained from peripheral blood mononuclear cells and the phorbol 12-myristate 13-acetate (PMA)-activated THP-1 cells were examined by flow cytometry. cDNAs of HLA-G1, HLA-G3, HLA-E, and human ß2-microglobulin were prepared, transfected into pig endothelial cells (PECs), and macrophage- and the THP-1 cell-mediated PEC cytolysis was then assessed. In vitro-generated macrophages expressed not only ILT2 and ILT4 but CD158d as well. The transgenic HLA-G1 on PEC indicated a significant suppression in macrophage-mediated cytotoxicity, which was equivalent to that of transgenic HLA-E. HLA-G1 was clearly expressed on the cell surface of PEC, whereas the levels of HLA-G3 were much lower and remained in the intracellular space. On the other hand, the PMA-activated THP-1 cell was less expressed these inhibitory molecules than in vitro-generated macrophages. Therefore, the HLA-G1 on PECs showed a significant but relatively smaller suppression to THP-1 cell-mediated cytotoxicity compared to in vitro-generated macrophages. These results indicate that by generating HLA-G1, but not HLA-G3, transgenic pigs can protect porcine grafts from monocyte/macrophage-mediated cytotoxicity.

Human HLA-Ev (147) Expression in Transgenic Animals.

BACKGROUND: In our previous study, we reported on the development of substituting S147C for HLA-E as a useful gene tool for xenotransplantation. In this study we exchanged the codon of HLA-Ev (147), checked its function, and established a line of transgenic mice. METHODS: A new construct, a codon exchanging human HLA-Ev (147) + IRES + human beta 2-microgloblin, was established. The construct was subcloned into pCXN2 (the chick beta-actin promoter and cytomegalovirus enhancer) vector. Natural killer cell- and macrophage-mediated cytotoxicities were performed using the established the pig endothelial cell (PEC) line with the new gene. Transgenic mice with it were next produced using a micro-injection method. RESULTS: The expression of the molecule on PECs was confirmed by the transfection of the plasmid. The established molecules on PECs functioned well in regulating natural killer cell-mediated cytotoxicity and macrophage-mediated cytotoxicity. We have also successfully generated several lines of transgenic mice with this plasmid. The expression of HLA-Ev (147) in each mouse organ was confirmed by assessing the mRNA. The chick beta-actin promoter and cytomegalovirus enhancer resulted in a relatively broad expression of the gene in each organ, and a strong expression in the cases of the heart and lung. CONCLUSION: A synthetic HLA-Ev (147) gene with a codon usage optimized to a mammalian system represents a critical factor in the development of transgenic animals for xenotransplantation.

Mitochondrial DNA replication in human T lymphocytes is regulated primarily at the H-strand termination site.

The most unique feature in the replication of mitochondrial DNA (mtDNA) is that most of the newly synthesized heavy strands (H-strands) terminate prematurely, resulting in the formation of displacement loop (D-loop) strands. Only the H-strand which proceeds past the termination site is a true nascent H-strand leading to the overall replication on a circular mtDNA molecule. The physiological significance of the D-loop formation has long been unclear. To examine the role of premature termination in mtDNA replication, we therefore developed a method for selectively measuring both the total amount of nascent H-strands and the amount of true nascent H-strands using ligation-mediated polymerase chain reaction, which, for the first time, enabled us to estimate the frequency of premature termination. The stimulation of cell proliferation with interleukin 2 and phytohemagglutinin in human peripheral T lymphocytes caused an increase in the net replication rate of mtDNA. In stimulated cells, in comparison to resting ones, the amount of true nascent H-strands increased approx. 2.6-fold while the total amount of nascent H-strands remained unchanged, indicating that premature termination decreased while the initiation of replication remained the same. Our findings thus demonstrate the first clear example that premature termination plays a primary role in the up-regulation of the net rate of mtDNA replication in human cells.

Effects of macrophage colony-stimulating factor (M-CSF) on the development, differentiation, and maturation of marginal metallophilic macrophages and marginal zone macrophages in the spleen of osteopetrosis (op) mutant mice lacking functional M-CSF activity.

Immunohistochemical techniques using an anti-mouse panmacrophage monoclonal antibody and anti-mouse monoclonal antibodies specific for marginal metallophilic macrophages or marginal zone macrophages were used to detect red pulp macrophages, marginal metallophilic macrophages, and marginal zone macrophages in the spleen of op/op mice. In the mutant mice, the red pulp macrophages were reduced to about 60% of those in the normal littermates and the marginal metallophilic macrophages and marginal zone macrophages were absent. After administration of recombinant human macrophage colony-stimulating factor (rhM-CSF), numbers of red pulp macrophages increased rapidly, reaching levels found in normal littermates 1 week later. In contrast, the marginal metallophilic macrophages as well as the marginal zone macrophages appeared slowly after rhM-CSF administration and their numbers were less than half of the baseline level of normal littermates even at 12 weeks of administration. The distribution of marginal metallophilic macrophages and marginal zone macrophages appearing after M-CSF administration was irregular in the spleen of the op/op mice. These splenic macrophage subpopulations differed in their responses to rhM-CSF, suggesting that distinct mechanisms may be involved in their development and differentiation. The splenic red pulp macrophages present in unmanipulated op/op mice are an M-CSF-independent macrophage population. Although the marginal metallophilic macrophages and marginal zone macrophages are thought to be M-CSF-dependent, their development and differentiation appear to be influenced by locally produced M-CSF or other cytokines.

Development, differentiation, and phenotypic heterogeneity of murine tissue macrophages.

In murine ontogeny, macrophage precursor cells develop in the yolk sac and fetal liver. Primitive macrophages also appear in the yolk sac, migrate to various tissues, and differentiate into several fetal macrophage populations. Because the development of the monocytic cell lineage is incomplete in the early stage of fetal hematopoiesis, primitive/fetal macrophages are considered to originate from granulocyte-macrophage colony forming cells or earlier macrophage precursors, bypassing the early monocytic cell series. In adult mice rendered severely monocytopenic by administration of strontium-89, resident macrophages are maintained by self-renewal. In contrast, administration of liposome-encapsulated dichloromethylene diphosphonate (clodronate) results in the elimination of various tissue macrophage populations. The repopulation of affected macrophages is dependent on the increase of precursors in the liver and spleen during the period of macrophage depletion. Such precursors reconstitute heterogeneous macrophage subpopulations. In mice homozygous for the osteopetrosis (op) mutation, the absence of macrophage colony-stimulating factor (M-CSF) activity results in a deficiency of monocytes and monocyte-derived macrophages. However, immature macrophages are present in various tissues. Administration of M-CSF to op/op mice induces the increased proliferative capacity and the morphological maturation of macrophages. However, the responses of individual tissue macrophage subpopulations to M-CSF are different. These results indicate that macrophage development, differentiation, and proliferation are regulated by the tissue microenvironment including the in situ production of macrophage growth factors in both fetal and adult life.

Expression of glial fibrillary acidic protein (GFAP) in peripheral nerve sheath tumors. A comparative study of immunoreactivity of GFAP, vimentin, S-100 protein, and neurofilament in 38 schwannomas and 18 neurofibromas.

Immunoreactivity of glial fibrillary acidic protein (GFAP) in 38 schwannomas and 18 neurofibromas was evaluated and compared with the reactivity of vimentin, S-100 protein, and neurofilament protein. All cases were positive for vimentin and S-100 protein. GFAP was positively stained in the neoplastic cells of 15 of 38 schwannomas (38%) and in two of 18 neurofibromas (11%). The extensively stained GFAP-positive tumors tended to be deeply situated in the body. The GFAP-positive cells were usually spindle-shaped and appeared preferentially in the perivascular region of hyalinized, thick blood vessels.

Selective 3-hydroxylation deficiency of lidocaine and its metabolite in Dark Agouti rats.

Selective and marked 3-hydroxylase deficiency of lidocaine and its N-deethylated metabolite, MEGX, were observed in male and female DA rats. These findings suggest that cytochrome P450 isozymes metabolizing debrisoquine may be involved in the 3-hydroxylations of lidocaine and MEGX in rats and humans.

Impairment of debrisoquine 4-hydroxylase and related monooxygenase activities in the rat following treatment with propranolol.

The effect of repetitive oral administration of propranolol on hepatic microsomal drug metabolizing enzyme activities in the rat was investigated. Propranolol ring (4-, 5- and 7-)hydroxylase activities were markedly decreased, but, interestingly, N-desisopropylase activity was increased after propranolol administration. A marked decrease in enzyme activity after propranolol pretreatment was also observed with debrisoquine 4-hydroxylation. In addition, a similar decrease was observed with imipramine 2-hydroxylation which co-segregates with debrisoquine/sparteine type polymorphic drug oxidation, but not with imipramine N-demethylation. These results suggest the selective impairment of debrisoquine 4-hydroxylase by propranolol pretreatment.

Participation of the CYP2D subfamily in lidocaine 3-hydroxylation and formation of a reactive metabolite covalently bound to liver microsomal protein in rats.

Lidocaine metabolism was investigated in rat liver microsomes and in a reconstituted system containing P450BTL, a cytochrome (P450) isozyme belonging to the CYP2D subfamily (Suzuki et al., Drug Metab Dispos 20: 367-373, 1992). P450BTL biotransformed lidocaine into 3-hydroxylidocaine (3-OH-LID) but not monoethylglycinexylidide and 2-methylhydroxylidocaine, in the reconstituted system including NADPH-P450 reductase and dilauroylphosphatidylcholine. An antibody against P450BTL inhibited microsomal lidocaine 3-hydroxylase activity by 97%. Thus, P450BTL and/or its immunorelated P450 isozyme(s) belonging to the CYP2D subfamily appear to be involved in lidocaine 3-hydroxylation. Furthermore, the antibody also suppressed the amounts of a lidocaine metabolite(s) bound to microsomal protein. These results suggest that the CYP2D subfamily biotransformed lidocaine into 3-OH-LID via an epoxy intermediate, which binds to microsomal macromolecules.

Another approach for aortic valve replacement through left thoracotomy.

We report a 54-year-old man with a history of esophagectomy and retrosternal esophagogastric anastomosis for esophageal cancer through right thoracotomy in whom cardiac failure developed due to aortic regurgitation. He underwent aortic valve replacement through a left thoracotomy with division of two great arteries and their strong traction toward the surgeon by stay sutures. He has been well for 3 years postoperatively in New York Heart Association class I.

Continuous subcutaneous infusion of buprenorphine for cancer pain control.

The continuous subcutaneous infusion of buprenorphine, a new approach to the relief of severe cancer pain, has been carried out using a portable infusion pump. The efficacy of this method was examined in 30 patients by visual analogue scale. An infusion rate of 4 micrograms/kg/day following intramuscular administration of 0.004 micrograms/kg gave satisfactory pain relief without serious complications. The minimum effective blood concentration was not detectable by high-performance liquid chromatography. Advantages of this therapy are its simplicity, applicability in many types of cancer, multiple sites of administration, and easier training on the part of health personnel.

The presence of siderosomes in synovioblasts (B cells) of chronic spontaneous hemarthrosis. Histologic, electron microscopic, and roentgenographic electron microanalysis studies of three cases.

The synovium of three patients with chronic spontaneous hemarthrosis was examined by light and electron microscopy and roentgenographic electron microanalysis. The synovium showed hyperplastic villous processes with proliferation of synovioblasts (B cells) and synovial macrophages (A cells). Erythrophagocytosis was observed in synovial macrophages but not in synovioblasts. Synovioblasts, however, had more numerous siderosomes than did synovial macrophages. Ferritin granules were identified in the cytoplasms of synovial macrophages and synovioblasts as well as in the intercellular space. It was noted that the longer the interval following the hemorrhagic episode, the fewer the siderosomes and erythrophagosomes in synovial macrophages. Therefore, it is likely that synovioblasts take up ferritin and/or inorganic iron released from synovial macrophages in chronic local iron overload.

The effects of an angiotensin-converting inhibitor (enalapril) on patients with mild cardiac failure--evaluating cardiac function based on the relationship between daily walking pace and heart rate.

BACKGROUND: Heart failure has been evaluated by several methods, the New York Heart Association (NYHA) classification of heart failure based on symptoms being used most frequently. However, the degree of heart failure assessed by these criteria does not always correlate with cardiac function in daily life. HYPOTHESIS: The aim of the study was to evaluate cardiac function based on the walking pace/heart rate (HR) relationship to assess the effects of enalapril, an angiotensin-converting enzyme inhibitor, in patients with mild to moderate cardiac function. METHODS: To evaluate cardiac function objectively, we developed a method using a pedometer to count the steps walked while simultaneously recording HR using a Holter electrocardiograph (ECG). Step-count walk rate (WR) was recorded on the magnetic tape of the Holter apparatus, and both HR and walking pace were calculated automatically by the Holter ECG analysis system. Data were determined every hour, and mean pace and HR were plotted along the x and y axes, respectively. The slope of HR x WR was calculated using the least squares method. The slope and the total number of steps were regarded as indicators of cardiac function and quality of life, respectively. We analyzed 36 subjects, consisting of 8 normal volunteers, 8 patients in New York Heart Association (NYHA) class I. 11 in class II, and 9 in class III chronic mild heart failure, during maximal exercise work load by bicycle ergometer; furthermore, fractional shortening of the left ventricle on echocardiogram was determined in 14 patients with chronic mild heart failure and was compared with the slope of HR x WR. Enalapril was administered at a daily dose of 2.5-10 mg for 1-24 months (mean 6 months) in 60 patients to evaluate the effects of this drug on these parameters. RESULTS: There was a significant inverse relationship between maximal work load and the HR x WR slope, and also between the fractional shortening and the slope, suggesting that the slope may reflect the severity of cardiac dysfunction. Furthermore, the slope decreased significantly from 1.8 +/- 1.26 before enalapril to 1.0 +/- 0.94 (mean +/- standard deviation) after drug administration, while the total number of steps increased significantly from 4842 +/- 3581 to 7804 +/- 4793. CONCLUSION: The slope of the graph relating step count and HR proved to be a good, objective indicator of cardiac function, and enalapril therapy improved this parameter.

Lymphocyte activation effect of (1-->6)-2,5-anhydro-D-glucitol and it derivatives with 3,4-di-O-methyl and sulfate groups.

(1-->6)-2,5-Anhydro-3,4-di-O-methyl-D-glucitol (2a) and (1-->6)-2,5-anhydro-D-glucitol (2c) and its sulfated derivative (2d) were synthesized and their biological activities were evaluated in regards to the effects on murine lymphocytes. The polymers showed different effects on the lymphocytes depending on the substituent groups. The sulfated polymer (2d) induced mitogenic activities, and specifically activated the CD4(-)CD8(-) subset of lymphocytes.

Hemodynamic and hemolytic features of the St. Jude Medical valve prostheses.

We performed valvular replacement in 86 cases (108 valves, 43 males, 43 females) from July 1978 to July 1981 with St. Jude Medical valves which utilize two discs made of pyrolytic carbon and employ a bileaflet central opening system. Ages ranged from 13 to 68 years (average 42.3). For all cases in this study, we performed anti-coagulant therapy. The incidence of thromboembolic complication was zero. With regard to postoperative clinical evaluation on valve function and chronic hemolysis, we compared the cases of St. Jude Medical valves with those of Starr-Edwards (S.E.) valves (aortic: Model 2320, mitral: Model 6400), Carpentier-Edwards (C.E.) valves and cases of open mitral commissurotomy. As for valve function such as left atrioventricular diastolic pressure gradient, mitral effective orifice area both at rest and on exercise, the St. Jude Medical valve yielded best results. Next was the C.E. and third was the S.E. The results of the St. Jude Medical valve group and those of the open mitral commissurotomy group were equivalent. In comparison with ball type cardiac valve prostheses and bioprostheses, the St. Jude Medical valve has excellent hemodynamic characteristic. Concerning hemolysis, the St. Jude Medical was below only the C.E., however the degree of hemolysis was so low that the St. Jude Medical valve holds great promise as central flow mechanical valve prostheses.

Effects of shosaikoto, an oriental herbal medicinal mixture, on age-induced amnesia in rats.

The possibility of naturally occurring amnesia was examined in aging Fischer F254 rats. Animals 110 weeks old showed a loss of learning and memory in a passive avoidance responses (PAR) failure test. When 120 mg/kg/day of Shosaikoto extract was placed in the diet after 72 weeks of age to 110 weeks, it improved age-induced PAR failure in both the two-compartment avoidance box test and the step-down test. The responses of 110-week-old Shosaikoto-treated rats on both PAR failures were almost the same as those of non-treated 6-week-old rats. In these Shosaikoto-treated rats, dopamine was increased and norepinephrine and vanillyl mandelic acid were decreased in brain relative to those in non-treated aging rats.