Research Note: Expression of IGF-1 and IGF-1 receptor proteins in skeletal muscle fiber types in chickens with hepatic fibrosis.

We investigated the expression of insulin-like growth factor 1 (IGF-1) and IGF-1 type 1 receptor (IGF-1R) in skeletal muscle fiber types in chickens with hepatic fibrosis induced by bile duct ligation (BDL). Eleven hens, approximately 104 weeks old, were randomly assigned to BDL (n = 4) and sham surgery (SHAM; n = 7) groups. In BDL hens, histopathology revealed marked bile duct proliferation and liver fibrosis. The cross-sectional area (CSA) of myofibers from both the pectoralis (PCT) muscles significantly decreased in the BDL group compared with the SHAM group (P < 0.01). In contrast, the CSA of myofibers from the femorotibialis lateralis (FTL) muscle did not decrease in the BDL group. Type I fibers were large, round, and hypertrophic. Elongated type IIA and IIB fibers were also present. For IGF-1 immunostaining, the immunoreaction intensity was higher in the PCT in the BDL group than the SHAM group. Within the BDL group, type I fibers from FTL had a stronger immunoreaction intensity than the type II fibers. For IGF-1R immunostaining, the intensity of the immunoreactions was similar within the PCT in the BDL group compared with the SHAM group. For FTL, type I fibers had stronger reactions to IGF-1R than type II fibers in the BDL group. These results suggest that type I fibers express both IGF-1 and IGF-1R and become hypertrophic in chickens with hepatic fibrosis.

Bioactivities generated from meat proteins by enzymatic hydrolysis and the Maillard reaction.

Bioactive peptides are released from meat proteins by enzymatic hydrolysis (i.e., gastrointestinal digestion, aging/storage, fermentation, and protease treatment). Such peptides attribute physiological functions to meat and meat products and are promising food ingredients for developing functional foods. Meat by-products (e.g., blood and collagen) are also good sources for generating bioactive peptides, since they are produced in large quantities and are rich in proteins. Although protein-derived bioactive peptides are attractive ingredients, their changes by the Maillard reaction during processing, cooking, and storage should be investigated. This article briefly reviews the production of bioactive peptides from meat and meat by-products. Such diverse peptides affects circulatory, nervous, alimentary, and immune systems. Then, the bioactivities of Maillard reaction products (MRPs) generated from protein hydrolysates are discussed. Special attention is paid to bioactivities of 2,5-dimethyl-4-hydroxy-3(2H)-furanone (DMHF) inhalation. As such activities, we have evaluated the impact of DMHF on blood pressure, moods, brainwaves, and dietary intake. Our efforts for understanding various aspects and implication of peptides and MRPs from meat proteins would open new avenues in the meat and food industry.

DMHF (2,5-dimethyl-4-hydroxy-3(2H)-furanone), a volatile food component with attractive sensory properties, brings physiological functions through inhalation.

2,5-Dimethyl-4-hydroxy-3(2H)-furanone (DMHF) is an aroma compound found in various foods, and used widely in the flavor and perfume industry. Dilute DMHF solutions exhibit a strawberry-like flavor while DMHF concentrates have a caramel-like aroma. DMHF is an important flavor compound contributing to the sensory properties of various natural products and thermally processed foods. DMHF is generated by the Maillard reaction during cooking and processing and affects the palatability of foods. Although Maillard reaction products (e.g., melanoidins) have physiologically positive effects, effects of odors generated from by this reaction are relatively unknown. This chapter initially overviewed the Maillard reaction and the generation of volatile compounds. Then, properties of DMHF, which is an attractive volatile food component, is discussed. We focused particularly on bioactivities of DMHF inhalation in our previous studies.

Bioactive Properties of Maillard Reaction Products Generated From Food Protein-derived Peptides.

Food protein-derived peptides are promising food ingredients for developing functional foods, since various bioactive peptides are released from food proteins. The Maillard reaction, which plays an important role in most processed foods, generates various chemical components during processing. Although changes of amino acids or proteins and reduced sugars by the Maillard reaction have been studied extensively, such changes of peptides by the Maillard reaction are still not resolved enough. Since food protein-derived peptides are widely utilized in many processed foods, it deserves concern and research on the changes of peptides by the Maillard reaction in foods during processing or storage. This chapter initially overviewed food protein-derived bioactive peptides. Then, Maillard reaction products generated from peptides are discussed. We focused particularly on their bioactivities.

Egg yolk protein and egg yolk phosvitin inhibit calcium, magnesium, and iron absorptions in rats.

Egg yolk decreases the absorption of iron. The effects of egg yolk protein and egg yolk phosvitin on the absorption of calcium, magnesium, and iron were investigated by in vivo studies. Male Wistar rats were fed purified diets containing casein, soy protein, or egg yolk protein for 14 d. The apparent absorptions of calcium, magnesium, and iron in the rats fed the yolk protein-based diet were lower than those in rats fed the casein- and soy protein-based diets. The apparent phosphorus absorption and the apparent protein digestibility in the yolk protein group were lower than those in the casein and soy protein groups. In the feces of the yolk protein group, serine comprised more than 30% of the amino acids. The addition of egg yolk phosvitin to the casein diets at levels of 1% and 2% (w/w) produced effects on calcium and magnesium absorptions similar to those produced by the diet containing yolk protein. The tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) pattern suggested that phosphopeptide fragments having molecular masses of 28, 22, and 15 kDa were evident in the contents of the small intestine of the rats fed phosvitin diets. These results indicate that yolk protein, when compared with casein and soy protein, decreases calcium and magnesium absorption via the resistance of phosvitin to proteolytic action.

Advanced bladder cancer in a young female: a case report.

We present a rare case of advanced bladder cancer in a young female. A 27-year-old woman had the chief complaint of proteinuria. A clinical examination revealed a papillary, broad-based bladder tumor with a clinical stage of T3, N3, M0. Preoperatively, 3 courses of neoadjuvant chemotherapy with methotrexate, adriamycin, and cisplatin were performed, and proved to be effective. Radical cystectomy was done and the histopathologically it was diagnosis as Grade 2 transitional cell carcinoma, which did not show any p53 gene mutation. The patient's postoperative clinical course was uneventful, and she remained disease free for 27 months. Bladder carcinoma in patients under 30 years of age tends to have a early stage and a low grade. However, the above described 27-year-old female patient demonstrated the advanced stage bladder tumor. Therefore, it should be kept in mind to accurately evaluate young patients with transitional cell carcinoma of the bladder and not to rule out the possibility of advanced disease even though a patient is young.

Peptide inhibitors for angiotensin I-converting enzyme from enzymatic hydrolysates of porcine skeletal muscle proteins.

Inhibitors of angiotensin I-converting enzyme (ACE) have been shown to have antihypertensive effects and have been utilized for pharmaceuticals and physiologically functional foods. In the present study, efforts were directed to find ACE inhibitory activities derived from muscle proteins. Porcine skeletal muscle proteins were hydrolyzed by eight proteases, and the inhibitory activities of the hydrolysates toward ACE were measured. Among the digests of the water-insoluble protein fraction prepared from muscle, thermolysin digest demonstrated the highest activity. Also, among hydrolysates of porcine myosin produced by the same enzymes, thermolysin digest showed the most potent inhibitory activity. Two ACE inhibitory peptides were purified from thermolysin digest of myosin. The sequences of these inhibitory peptides, named myopentapeptides A and B, were Met-Asn-Pro-Pro-Lys and Ile-Thr-Thr-Asn-Pro. These sequences were found in the primary structure of the myosin heavy chain. The concentrations of the peptides showing 50% inhibition values (IC(50)) of ACE were 945.5 and 549.0 µM, respectively. Also, six tripeptides, Met-Asn-Pro, Asn-Pro-Pro, Pro-Pro-Lys, Ile-Thr-Thr, Thr-Thr-Asn, and Thr-Asn-Pro, which have parts of the sequences of the myopentapeptides, demonstrated activity. Their IC(50) values were 66.6, 290.5, >1000, 678.2, 672.7, and 207.4 µM, respectively.

UV-induced Lactobacillus gasseri mutants resisting sodium chloride and sodium nitrite for meat fermentation.

Lactobacillus gasseri, one of the predominant lactobacilli in human intestinal tracts, is utilized for probiotics and dairy starter cultures. However, since L. gasseri is relatively sensitive to sodium chloride and sodium nitrite (essential compounds for meat products), it is difficult to utilize this species for conventional fermented meat products. In this study, efforts were directed to generate mutants of L. gasseri resisting sodium chloride and sodium nitrite. UV irradiation of the strain of L. gasseri JCM1131(T) generated several mutants resisting these compounds. A mutant strain 1131-M8 demonstrated satisfactory growth in meat containing 3.3% sodium chloride and 200 ppm sodium nitrite. Although proteins extracted from the cell surface of 1131-M8 were slightly different from those of the original strain, other biochemical characteristics of both strains were indistinguishable. These results suggest that the L. gasseri mutant obtained in this study could be utilized as a starter culture to develop probiotic meat products.

Effect of intestinal Lactobacillus starter cultures on the behaviour of Staphylococcus aureus in fermented sausage.

The effects of Lactobacillus strains isolated from intestinal tracts for starter cultures of fermented sausage on the growth rate and enterotoxin production of Staphylococcus aureus were studied at two fermentation temperatures of 20 degrees C and 35 degrees C. Initial inoculated populations in the sausage batter were approx. 10(4) cfu/g for S. aureus and 10(7) cfu/g for the Lactobacillus strain as a starter culture. Samples of sausage were taken during fermentation and analyzed for pH and microbial populations. In control lots without inoculation of Lactobacillus strains, staphylococcal enterotoxin was detected during fermentation at each temperature. Of three intestinal Lactobacillus strains, L. rhamnosus FERM P-15120 and L. paracasei subsp. paracasei FERM P-15121 inhibited the growth and enterotoxin production of S. aureus in sausages during fermentation at both temperatures, although L. acidophilus FERM P-15119 could not satisfactorily suppress them. The effect of the two selected strains in meat fermentation (i.e., fermentation time, acid production, inhibition of S. aureus) was the same as that of a commercial L. sake starter culture for fermented sausage. These results suggest the intestinal Lactobacillus strains selected in this study could be utilized as a starter culture to produce new fermented meat products that are microbiologically safe.

Salivacin 140, a novel bacteriocin from Lactobacillus salivarius subsp. salicinius T140 active against pathogenic bacteria.

Fifteen of 353 environmental isolates of lactic acid bacteria consistently showed activity against Listeria monocytogenes, Streptococcus mutans, Actinomyces viscosus, and/or Propionibacterium acnes. Strain T140, isolated from the surface of Japanese pampas grass leaves and identified as Lactobacillus salivarius subsp. salicinius, also had activity against several Lactobacillus species, Staphylococcus aureus and Yersinia enterocolitica. Since the antagonistic factor(s) produced by T140 was sensitive to a proteolytic enzyme, it was concluded that a bacteriocin (named salivacin 140) was involved in the inhibition activity. Strain T140 required a high initial pH (7.5-8.5) in agar plates for bacteriocin production.

Contribution of the glycolytic pathway to enzymatic metmyoglobin reduction in myocytes.

The metmyoglobin (oxidized form of myoglobin) reducing enzyme system was examined in rat myocytes. An inhibitor of the glycolytic pathway (2-deoxy-D-glucose) inhibited the reduction of metmyoglobin in myocytes. An inhibitor of the citric acid cycle (malonic acid), did not inhibit this reaction. From the results of this study along with those of our previous work, we proposed that the glycolytic pathway may have a significant role in the enzymatic metmyoglobin reduction, presumably by its supply of cytoplasmic NADH for the NADH-cytochrome b5 reductase system, in muscle.

Localization of metmyoglobin-reducing enzyme (NADH-cytochrome b(5) reductase) system components in bovine skeletal muscle.

Localization of metmyoglobin-reducing enzyme system components (NADH-cytochrome b(5) reductase, cytochrome b(5) outer mitochondrial membrane cytochrome b) was demonstrated in bovine skeletal muscle by immunohistochemical techniques. Both NADH-cytochrome b(5) reductase and OM cytochrome b were located in red fibers. However, localization of cytochrome b(5) did not show a definite difference between fiber types. Using an immunoblotting technique. NADH-cytochrome b(5) reductase was found predominantly in the mitochondrial fraction, but it was also detected at lower levels in the microsomal fraction. OM cytochrome b was found predominantly in the mitochondrial fraction, but cytochrome b(5) was detected only in the microsomal fraction. The results from this study, along with previous work about the localization of myoglobin in muscle, suggest that NADH-cytochrome b(5) reductase reduces metmyoglobin by using OM cytochrome b at the mitochondrial surface and, in part, by using cytochrome b(5) at the sarcoplasmic reticulum.

[Statistic studies on bacteria isolated from urinary tract infections (report 6: isolation rate and drug sensitivity from 1988 through 1989)].

We examined the bacteria isolated from urine of patients in our department from 1988 though 1989. A total of 809 strains were isolated from the outpatients. They were 241 strains of gram-positive bacteria (29.8%). E. coli (32.0%) was isolated most frequently, followed by Enterococcus spp. (9.9%), Proteus group (7.4%), S. epidermidis (6.2%), P. aeruginosa (5.1%) and others. A total of 412 strains were isolated from the inpatients. They consisted of 173 strains of gram-positive bacteria (42.0%). Enterococcus spp. (17.0%) was isolated most frequently, followed by P. aeruginosa (12.6%), S. epidermidis (9.7%), E. coli (7.3%), S. aureus (6.6%) and others. Fewer P. aeruginosa, S. marcescens and Proteus group were isolated from the outpatients and fewer S. marcescens, E. coli and K. pneumoniae from inpatients than previously. More S. aureus were isolated abruptly from the inpatients and more E. coli were isolated from the outpatients than previously. E. cloacae, S. epidermidis, S. aureus and Corynebacterium spp. all isolated from inpatients showed lower antimicrobial sensitivity than those isolated from the outpatients.

[Preoperative collection of autologous blood with recombinant human erythropoietin therapy].

The major problems in liquid storage of donated blood are the limitation of blood volume preserved and the time consumed. We studied on the usefulness of utilization of recombinant human erythropoietin (rH-EPO) for preoperative collection of autologous blood. Patients were divided into 2 groups: Group 1 (12 patients) received 3000 unit of rH-EPO intravenously every day. Group 2 (10 patients) was not treated with rH-EPO. All the patients were given ferrous sulfate 100 mg of iron orally every day. As a rule, 400 ml blood was donated two or three times a week for rH-EPO treated group, while, for control group 400 ml blood was collected once a week. Some patients did not donate 400 ml because of their critical condition. The blood volume preserved became 780.9 +/- 284.1 ml in Group 1, and 910.8 +/- 263.9 ml in Group 2. However, the number of days until operation were significantly shorter in Group 1 (8.7 +/- 3.6 days) than in Group 2 (26.2 +/- 11.2 days). Hemoglobin and Hematocrit levels recovered promptly in rH-EPO treated group. In control group, 2 patients complained of nausea and vomiting caused by severe anemia. On the other hand, no one with rH-EPO therapy did not show any side effects and complications. Our study indicates that the administration of rH-EPO enables us to collect an adequate volume of autologous blood preoperatively in a shorter period of time.

Characterization of bacteriocins from Enterococcus faecium with activity against Listeria monocytogenes.

Laboratory cultures and environmental isolates of bacteria were screened for antagonism towards Listeria monocytogenes using an agar spot test. Seven of the 163 strains that were tested, one Streptococcus bovis, one Enterococcus casseliflavus, two E. avium and three E. faecium, consistently displayed antilisterial activity. Cell-free, pH-neutralized supernatants prepared from the three E. faecium strains (JBL1061, JBL1083 and JBL1351) exhibited strong antilisterial activity against L. monocytogenes, and were subjected to more detailed analyses. The antagonistic factors produced by these three strains were sensitive to chloroform and several proteolytic enzymes, resistant to heat (121 degrees C, 20 min), and stable over a wide pH range (3.0-10.0). Moreover, they were listericidal without causing cell lysis. These data suggest that a bacteriocin(s) is involved in the inhibition of L. monocytogenes by E. faecium JBL1061, JBL1083 and JBL1351.

Lectin-like activity of Lactobacillus acidophilus strain JCM 1026.

The lectin-like activity of the Lactobacillus acidophilus strain JCM 1026 was studied by hemagglutination and hemagglutination-inhibition assays. L. acidophilus strain JCM 1026 was found to hemagglutinate human and animal erythrocytes. Neuraminidase-treatment of human-type O erythrocytes enhanced the activity. Treatment of the bacterial cells with proteinase K reduced hemagglutinating activity significantly. Although several mono- and disaccharides did not inhibit hemagglutination, several different glycoproteins did. These data indicate that a proteinaceous lectin-like component(s) recognizing carbohydrate-containing molecules is located on the cell surface of L. acidophilus.

[Contralateral ureteral metastasis from renal cell carcinoma: a case report].

A case of contralateral ureteral metastasis from renal cell carcinoma is reported. A 52-year-old man underwent left nephrectomy for renal cell carcinoma of November 20, 1986. He was clinically asymptomatic for 4 years 8 months after the operation. He was admitted on August 14, 1991, because of right loin pain. Right retrograde pyelography and percutaneous pyelography showed a filling defect in the right ureter at the level of L3. After the right ureter was explored, the tumor lesion of ureter was resected and end to end anastomosis of the ureter was performed. Histopathologic examination showed a metastatic clear cell carcinoma consistent with a renal primary. The contralateral ureteral metastasis from renal cell carcinoma is very rare and only 15 cases have been reported previously.

Genomic analysis of Pediococcus starter cultures used to control Listeria monocytogenes in turkey summer sausage.

The pulsed-field technique of clamped homogeneous electric field electrophoresis was employed to characterize and size genomic DNA of three pediocin-producing (Ped+) and two non-pediocin-producing (Ped-) strains of Pediococcus acidilactici. Comparison of genomic fingerprints obtained by digestion with the low-frequency-cleavage endonuclease AscI revealed identical restriction profiles for four of the five strains analyzed. Summation of results for 10 individually sized AscI fragments estimated the genome length to be 1,861 kb for the four strains (H, PAC1.0, PO2, and JBL1350) with identical fingerprints. Genomic analysis of the pediocin-sensitive, plasmid-free strain P. acidilactici LB42 with the unique fingerprint revealed nine AscI fragments and a genome length of about 2,133 kb. Ped- (JBL1350) and Ped+ (JBL1095) starter cultures (one each) were used to separately prepare turkey summer sausage coinoculated with a four-strain Listeria monocytogenes mixture (ca. 10(5) CFU/g). The starter cultures produced equivalent amounts of acid during fermentation, but counts of L. monocytogenes were reduced to a greater extent in the presence of the Ped+ starter culture (3.4 log10 unit decrease) than in the presence of the Ped- starter culture (0.9 log10 unit decrease). Although no listeriae were recovered from sausages following the cook/shower, appreciable pediocin activity was recovered from sausages prepared with the Ped+ strain for at least 60 days during storage at 4 degrees C. The results of this study revealed genomic similarities among pediococcal starter cultures and established that pediocins produced during fermentation provide an additional measure of safety against listerial proliferation in turkey summer sausage.

[Clinical statistics on operations for inpatients at Department of Urology, Tokai University Hospital 1975-1989].

Operations on inpatients during the 15-year period from 1975 to 1989 were analyzed statistically. A total of 3,791 operations were performed at our Department of Urology, including 717 operations on the kidney, 316 operations on the ureter, 583 operations on the bladder, 811 operations on the prostate, 136 operations on the urethra, 1,097 operations on the scrotum, 43 operations on the adrenal gland, 19 operations on the retroperitoneal space and 69 operations on other organs.

Immunofluorescence microscopic studies on distribution of Lactobacillus kefiranofaciens and Lactobacillus kefir in kefir grains.

Immunofluorescence staining was applied to observe distribution of two Lactobacillus species in kefir grains with cauliflower floret forms. Kefiran-producing, encapsulated Lactobacillus kefiranofaciens was located all over the grain and increased toward the center, while Lactobacillus kefir populated only a small region at the surface layers.