Bifidobacterium breve alters immune function and ameliorates DSS-induced inflammation in weanling rats.

BACKGROUND: Bifidobacterium breve M-16V (M16V) is a probiotic bacterial strain with a long tradition of use in neonatal intensive care units in some countries. Previous study showed that the effects of M16V administration on gene expression were greater during the weaning period than in the neonatal period and were greater in the colon than in the small intestine and spleen, suggesting that M16V has anti-inflammatory effects. In this study, we evaluated the effects of inflammation during the weaning period and the effects of M16V on normal and inflammatory conditions. METHODS: From postnatal day (PD) 21 to 34, weanling rats were administered of 2.5 × 10(9) of M16V daily, and colitis was induced by administration of 2% dextran sulfate sodium from PD28 to 35. Colitis severity, immune function, and microbiota were investigated. RESULTS: Colitis caused a reduction in body weight gain, colon shortening, poor nutritional status, anemia, changes in blood and spleen lymphocyte populations, spleen T-cell malfunctions, and alterations in colon microbiota. M16V administration improved some but not all of the changes induced by colitis. CONCLUSION: M16V could suppress inflammation and, therefore, can be considered a safe strain to use not only during the neonatal period but also the weaning period.

Long-term prognosis of hearing loss in patients with unilateral Ménière's disease.

CONCLUSION: The finding of deteriorated hearing loss at the initial visit at middle to high frequencies is a factor of poor hearing prognosis in Ménière's disease. Early intervention with instructions for lifestyle changes may lead to good outcomes in hearing. OBJECTIVE: An attempt was made to examine long-term changes in hearing loss in unilateral Ménière's disease and factors associated with prognosis of hearing loss retrospectively. METHODS: Based on their last hearing level of the affected ear, 36 patients were subdivided into two groups: the poor prognosis of hearing (PPH) group and the good prognosis of hearing (GPH) group. RESULTS: In the PPH group, the hearing levels at the initial visit at middle and high frequencies were significantly worse than those in the GPH group. Moreover, the hearing loss progressed during the first 2 years of the disease, and stayed flat to approximately 50 dB at the later stage. Conversely, the hearing loss at the onset in the GPH group showed no further progression over the first 2 years, and remained constant to approximately 35 dB at the later stage. In addition, the mean intervals from the onset to the initial visit in the PPH group were significantly longer than those in the GPH group.

Inhibition of intestinal polyp growth by oral ingestion of bovine lactoferrin and immune cells in the large intestine.

Studies using animal models have demonstrated that ingestion of bovine lactoferrin (bLF) inhibits carcinogenesis in the colon and other organs of experimental animals. As a result of these studies, a blinded, randomized, controlled clinical trial was conducted in the National Cancer Center Hospital, Tokyo, Japan to determine whether ingestion of bLF had an effect on the growth of colorectal polyps in humans. Patients with colorectal polyps ≤5 mm diameter and likely to be adenomas ingested 0, 1.5, or 3.0 g bLF daily for 1 year. Ingestion of 3.0 g bLF suppressed the growth of colorectal polyps and increased the level of serum human lactoferrin in trial participants 63 years old or younger. The purpose of the present study was to investigate correlations between immune parameters and changes in polyp size. Trial participants with regressing polyps had increased NK cell activity, increased serum hLF levels (indicating increased neutrophil activity), and increased numbers of CD4+ cells in the polyps. These findings are consistent with a correlation between higher immune activity and suppression of colorectal polyps. In addition, participants with regressing polyps had lower numbers of PMNs and increased numbers of S100A8+ cells in the polyps, consistent with a correlation between lower inflammatory potential in the colon and suppression of colorectal polyps. Trial participants ingesting bLF had increased serum hLF levels, a possible increase in systemic NK cell activity, and increased numbers of CD4+ and CD161+ cells in the polyps. Taken together, our findings suggest that bLF suppressed colorectal polyps by enhancing immune responsiveness.

Time-dependent expression profiles of microRNAs and mRNAs in rat milk whey.

Functional RNAs, such as microRNA (miRNA) and mRNA, are present in milk, but their roles are unknown. To clarify the roles of milk RNAs, further studies using experimental animals such as rats are needed. However, it is unclear whether rat milk also contains functional RNAs and what their time dependent expression profiles are. Thus, we prepared total RNA from whey isolated from rat milk collected on days 2, 9, and 16 postpartum and analyzed using microarrays and quantitative PCR. The concentration of RNA in colostrum whey (day 2) was markedly higher than that in mature milk whey (days 9 and 16). Microarray analysis detected 161 miRNAs and 10,948 mRNA transcripts. Most of the miRNAs and mRNA transcripts were common to all tested milks. Finally, we selected some immune- and development-related miRNAs and mRNAs, and analysed them by quantitative PCR (in equal sample volumes) to determine their time-dependent changes in expression in detail. Some were significantly more highly expressed in colostrum whey than in mature milk whey, but some were expressed equally. And mRNA expression levels of some cytokines and hormones did not reflect the protein levels. It is still unknown whether RNAs in milk play biological roles in neonates. However, our data will help guide future in vivo studies using experimental animals such as rats.

Risk factors for poor outcome of a single Epley maneuver and residual positional vertigo in patients with benign paroxysmal positional vertigo.

CONCLUSION: The findings suggest that in patients with posterior semicircular canal (PSCC) benign paroxysmal positional vertigo (P-BPPV), head trauma and prolonged bedrest, but not inner ear disease, are risk factors for poor outcome of a single Epley maneuver and persistent residual positional vertigo. OBJECTIVES: We first examined the efficacy of a single Epley maneuver and then assessed the time course in remission of residual positional vertigo in patients with idiopathic P-BPPV and secondary P-BPPV. METHODS: A total of 157 patients with idiopathic P-BPPV and 40 patients with secondary P-BPPV (secondary to head trauma in 8 patients, to prolonged bedrest in 14 patients, and to inner ear disease in 18 patients) were treated with a single Epley maneuver. RESULTS: The negative rates of the Dix-Hallpike test on day 7 after a single Epley maneuver in both patients with P-BPPV secondary to head trauma (25%) and those with prolonged bedrest (36%) were significantly lower than that (73%) in patients with idiopathic P-BPPV. Additionally, the remission of residual positional vertigo in the former groups of patients was significantly delayed in comparison with that of the latter group. However, there were no significant differences in the efficacy of a single Epley maneuver and persistent residual positional vertigo between idiopathic P-BPPV and P-BPPV secondary to inner ear disease.

Purification of RNA from milk whey.

MicroRNAs (miRNAs) are small regulatory RNA molecules that modulate specific target mRNAs and play very important roles in physiological processes. They were recently detected in body fluids such as blood, urine, saliva, and milk. These body fluid miRNAs have been studied thoroughly as potential diagnostic biomarkers. However, there have been few studies of milk miRNAs, and their roles are not clearly understood. Milk is the only nutritional source for newborn infants, and bovine milk is used widely as a dairy product. Thus, it is important to study milk miRNAs. In general, body fluid RNA concentrations are extremely low and of diverse existence types. In this chapter, we compare two silica membrane column-based RNA purification kits, and also compare RNA obtained directly from whey with that isolated from whey-derived exosomes.

Energy requirement assessed by doubly-labeled water method in patients with advanced amyotrophic lateral sclerosis managed by tracheotomy positive pressure ventilation.

This study aimed to clarify the energy requirement in patients with amyotrophic lateral sclerosis (ALS) undergoing tracheostomy positive pressure ventilation with tracheostomy. Total energy expenditure (TEE) was measured in 10 hospitalized bedridden ALS patients using the doubly-labeled water (DLW) method. The mean TEE/day and TEE/fat- free mass estimated by DLW method were 934 ± 201 kcal/day and 34.8 ± 5.5 kcal/kg/day, respectively. The mean TEE/resting metabolic rate (RMR) was 0.85 when RMR was estimated by the Harris-Benedict equation, 0.91 by Dietary Reference Intake (DRI), and 0.97 by Ganpule's equation using fat-free mass (FFM). The ratios of TEE to measured RMR were 1.05, 1.15 and 1.23 in three patients. In conclusion, multiplying measured RMR by 1.1 to 1.2 is considered to be appropriate to estimate energy need. However, because it is difficult to measure RMR directly in a clinical setting, an appropriate equation for estimating RMR for ALS patient should be developed.

Effects of sleep position on time course in remission of positional vertigo in patients with benign paroxysmal positional vertigo.

CONCLUSION: The findings suggest that it is easy for otoconial debris dislodged from the utricle to fall into the posterior semicircular canal (PSCC) or the horizontal semicircular canal (HSCC) of the undermost ear during sleep, but not to exit from the uppermost ear in patients with benign paroxysmal positional vertigo (BPPV). OBJECTIVE: The aims of the present study were two-fold. (1) To examine the association between the preferred side of head-lying during sleep and the side of the affected ear in patients with both posterior canal BPPV (P-BPPV) and horizontal canal BPPV (H-BPPV). (2) To see whether that position affects the time course in remission of their positional vertigo. METHODS: One hundred and sixteen patients with P-BPPV and 40 patients with H-BPPV who showed a habitual preference for right or left side sleeping position were included in this study. RESULTS: The side of the affected ear was significantly associated with the head-lying side during sleep in patients with P-BPPV and was closely but not significantly associated with it in patients with H-BPPV. However, the head-lying side during sleep did not affect the remission rate of their positional vertigo.

Influences of casein hydrolysate ingestion on cerebral activity, autonomic nerve activity, and anxiety.

This study examined the influences of the oral ingestion of casein hydrolysate from bovine milk at rest physiologically and psychologically. Eleven male university students were given a casein hydrolysate drink (H) or a maltitol drink as a control (C) in a crossover study. Just before and one hour after ingestion of each drink, the total-hemoglobin (tHb) concentrations at ten points of the prefrontal cortex to evaluate cerebral activity, and heart rate variability (HRV) to evaluate autonomic nerve activity through spectral analysis were measured as physiological indicators. The Japanese version of the State--Trait Anxiety Inventory--state anxiety (STAI-s) score was also used, as a psychological indicator. In comparison between H and C ingestion, a significant difference is observed only in tHb concentrations at one of ten points. At this point, the change in tHb concentration was lower after H ingestion compared to C ingestion. And in comparison between before and after ingestion of each drink, a significant increase in tHb concentration at two points after C ingestion, a significant increase in parasympathetic activity and decrease in sympathetic activity after H ingestion, and a significant decrease in STAI-s score in H ingestion were observed. These results suggest that ingestion of the casein hydrolysate may keep prefrontal cortex activity stable while maltitol ingestion partially increases the activity. Moreover, there is a possibility that casein hydrolysate might decrease sympathetic activity, increase parasympathetic activity, and lower anxiety. We conclude that the bovine milk casein hydrolysate may have more relaxing effects than maltitol.

microRNA as a new immune-regulatory agent in breast milk.

BACKGROUND: Breast milk is a complex liquid that provides nutrition to the infant and facilitates the maturation of the infant's immune system. Recent studies indicated that microRNA (miRNA) exists in human body fluid. Because miRNAs are known to regulate various immune systems, we hypothesized that human breast milk contains miRNAs that may be important for the development of the infant's immune system. FINDINGS: We profiled miRNA expression in human breast milk and detected high expression levels of immune-related miRNAs in the first 6 months of lactation. Furthermore, these miRNA molecules are stable even in very acidic conditions, indicating that breast milk allows dietary intake of miRNAs by infants. CONCLUSIONS: Our findings provide new insight into how breast milk can modulate the development of the infant's immune system. This study suggests the transfer of genetic material as miRNA from human to human occurs by means other than through sexual reproduction.

Is glycemic index of food a feasible predictor of appetite, hunger, and satiety?

This review assesses the feasibility of using glycemic index (GI) as a predictor of appetite, hunger and satiety by surveying published human intervention studies. We also discuss the relationship between GI and two appetite/satiety control hormones, leptin and ghrelin. Ingestion of high-GI food increased hunger and lowered satiety in short-term human intervention studies. This effect may be attributed to the rapid decline in blood glucose level following a hyperinsulinemic response caused by a sharp and transient increase in blood glucose level that occurs after the ingestion of high-GI food, which is defined as the glucostatic theory. However, appetite, hunger and satiety after the ingestion of foods with varying GI were inconsistent among long-term human intervention studies. From the few relevant long-term studies available, we selected two recent well-designed examples for analysis, but they failed to elicit clear differences in glycemic and insulinemic responses between high- and low-GI meals (consisting of a combination of different foods or key carbohydrate-rich foods incorporated into habitual diets). One of the reasons that these studies could not predict glycemic response to mixed meals is presumably that the GI of each particular food was not reflected in that of the mixed meals as a whole. Thus, it is difficult to conclude that the GI values of foods or mixed meals are a valid long-term predictor for appetite, hunger and satiety. Both insulin and insulin-mediated glucose uptake and metabolism in adipose tissue affect blood leptin concentration and its diurnal pattern. Circulating ghrelin level is suppressed by carbohydrate-rich meals, presumably via glycemia and insulinemia. Accordingly, low-GI foods may not necessarily increase satiety or suppress appetite and/or hunger because of the lack of insulin-mediated leptin stimulation and ghrelin suppression. However, insulin-mediated leptin stimulation and ghrelin suppression per se is not consistent among studies; thus we were not able to identify a clear relationship among GI, satietogenic leptin, and appetitic ghrelin.

Effects of hypergravity on histamine H1 receptor mRNA expression in hypothalamus and brainstem of rats: implications for development of motion sickness.

CONCLUSION: The study findings suggest that histamine was released from the axon terminals in the hypothalamus and brainstem and the released histamine activated post-synaptic H1 receptors there, resulting in the development of motion sickness. OBJECTIVES: We first examined which subtype of post-synaptic histaminergic receptor was responsible for the development of motion sickness. We then examined whether H1 receptors were up-regulated in various areas of the rat brain after 2 G hypergravity load, because the stimulation of H1 receptor was reported to up-regulate the level of H1 receptor protein expression through augmentation of H1 receptor mRNA expression. MATERIALS AND METHODS: For this purpose, we used an animal model of motion sickness, using pica (eating non-nutritive substances such as kaolin), as a behavioral index in rats. RESULTS: After 2 G hypergravity load, rats ate a significant amount of kaolin, indicating that they suffered from motion sickness. The hypergravity-induced kaolin intake was suppressed by mepyramine, but not by terfinadine or zolantizine. This finding indicates that cerebral post-synaptic H1 but not H2 or peripheral H1 receptors play an important role in the development of motion sickness. The expression of H1 receptor mRNA was up-regulated in the hypothalamus and brainstem, but not in the cerebral cortex after 2 G hypergravity load in rats.

3D analysis of benign positional nystagmus due to cupulolithiasis in posterior semicircular canal.

CONCLUSIONS: The characteristic of both the vertical-torsional positional nystagmus with long time constant and its disappearance at the neutral head position could diagnose cupulolithiasis in posterior semicircular canal (PSCC) in the eight patients with the PSCC type of benign paroxysmal positional vertigo (P-BPPV). OBJECTIVE: The aim of the study was to diagnose cupulolithiasis in patients with P-BPPV. PATIENTS AND METHODS: We used three-dimensional rotation axis analysis of nystagmus of the vertical-torsional positional nystagmus in 111 patients with P-BPPV and evaluated its time constant. We then examined whether the vertical-torsional positional nystagmus with long time constant disappeared at the neutral head position where the axis of the heavy cupula of the affected PSCC is aligned with gravity. RESULTS: The first parameter showed a wide variation that could be divided into two groups: one lasting more than 40 s in 8 patients and another below 20 s in 103 patients. Since the time constant of the positional nystagmus induced by cupulolithiasis was much longer than that induced by canalolithiasis, this finding suggests that cupulolithiasis in the PSCC induced the vertical-torsional positional nystagmus with a long time constant in the group of eight patients. The vertical-torsional positional nystagmus disappeared in these patients at the neutral head position, where the axis of the cupula of affected PSCC aligned with gravity.

Lack of chronic oral toxicity of chemopreventive bovine lactoferrin in F344/DuCrj rats.

Studies were undertaken to determine whether bovine lactoferrin (bLF) and related compounds, shown to prevent carcinogenesis in the colon and other organs in rats, have any toxic effects in long-term feeding studies. In experiment I, male F344/DuCrj rats received a basal diet containing 0.2% bLF for 40 weeks. No adverse findings were noted, furthermore, serum triglyceride level was significantly decreased to 72% of the control level, suggesting preventive effects against the metabolic syndrome. In experiment II, male and female F344/DuCrj rats were fed a basal diet containing 0.02, 0.2, 2.0 and 5.0% bLF, 2.0% bLF hydrolysate (bLF-H) or 0.1% lactoferricin (LFcin), a peptide derived from bLF, for 60 weeks in males and 65 weeks in females. No toxicological effects, including carcinogenicity, were evident in either sex. The results of the studies provide subjective support for safety of clinical studies of bLF for supplement use.

Differential diagnosis of true and pseudo-bilateral benign positional nystagmus.

CONCLUSIONS: Two patients showing two rotational axes of their positional nystagmus had canalolithiasis in bilateral posterior semicircular canals (PSCCs), leading to the diagnosis of true bilateral benign paroxysmal positional nystagmus (P-BPPN). Another 18 patients had a single rotation axis of their positional nystagmus with short time constant (TC) on one side and long TC on the other. Since canalolithiasis in the uppermost PSCC may be transient cupulolithiasis, evoking positional nystagmus with long TC, the diagnosis of the latter patients should be pseudo-bilateral P-BPPN. OBJECTIVE: To differentiate the true bilateral posterior canal type of P-BPPN from pseudo-bilateral P-BPPN. PATIENTS AND METHODS: The rotational axis and TC of positional nystagmus were three-dimensionally analyzed in 20 patients showing geotropic torsional nystagmus on the left and right Dix-Hallpike (D-H) maneuvers. RESULTS: Two patients showed two rotational axes of their positional nystagmus, which were perpendicular to the plane of the PSCCs. There were no differences in TCs of their positional nystagmus between bilateral D-Hs. Another 18 patients showed a single rotational axis of their positional nystagmus, which was perpendicular to the plane of either the left or right PSCC. TCs of their positional nystagmus were short on one side and long on the other.

Changes in slow phase eye velocity and time constant of positional nystagmus at transform from cupulolithiasis to canalolithiasis.

Changes in slow phase eye velocity (SPEV) and time constant (TC) of benign paroxysmal positional nystagmus in horizontal canal type were examined at transitional period from cupulolithiasis (apogeotropic nystagmus) into canalolithiasis (geotropic nystagmus) in two patients. SPEV and TC of positional nystagmus were tri-dimensionally analyzed. The first patient showed an apogeotropic nystagmus. Head rotation to the left in supine position induced a right-beating nystagmus with an initial SPEVof 15.3 degrees/s and a TC of 133 s. The nystagmus then gradually declined with a TC of 31.3 s after reaching a maximum SPEV of 28.8 degrees/s. After the nystagmus disappeared, he showed a geotropic nystagmus. The second patient showed a left-beating nystagmus with an initial SPEV of 2.5 degrees/s and a TC of 141 s when his head was rotated to the right in supine position. The nystagmus then gradually declined with a TC of 8.05 s after reaching a maximum SPEV of 16.7 degrees/s. After the nystagmus disappeared, he showed a geotropic nystagmus. The present findings suggested that in both patients, at the period of an increase of SPEV of the positional nystagmus with the shortening of its TC, cupulolithiasis transformed into canalolithiasis.

Natural history of benign paroxysmal positional vertigo and efficacy of Epley and Lempert maneuvers.

We assessed the efficacy of Epley maneuver in patients with posterior canal benign paroxysmal positional vertigo (P-BPPV) and Lempert maneuver in patients with horizontal canal BPPV (H-BPPV). In patients with P-BPPV, positional vertigo in patients treated by Epley maneuver was significantly resolved more quickly than that in untreated patients. But in patients with H-BPPV, there were no significant differences of time course in remission of positional vertigo between untreated patients and patients treated by Lempert maneuver. Among the untreated patients, the positional vertigo in patients with H-BPPV was significantly resolved more quickly than that in patients with P-BPPV. Epley maneuver was effective for the treatment of patients with P-BPPV, whereas the efficacy of Lempert maneuver for the treatment of patients with H-BPPV was limited. The natural courses in remission of positional vertigo in untreated patients with H-BPPV showed significantly faster resolution than that in patients with P-BPPV.

Prevention of carcinogenesis and cancer metastasis by bovine lactoferrin.

Increasing attention is being paid to chemopreventive agents for individuals at high risk of cancer. We have concentrated on bovine lactoferrin (bLF), an 80 kDa iron-binding glycoprotein known to have anti-microbial and immunoprotective effects. Lactoferrin is particularly abundant in colostrum, and is also present in tears, saliva and seminal and uterine secretions. However, only little is known regarding its influence on carcinogenesis. We have shown preventive effects of bLF and its fragment peptide, lactoferricin (bLFcin), consisting of a 25 amino acid sequence without iron binding capacity, on chemically-induced colon carcinogenesis in the rat and transplanted carcinoma cell metastasis in the mouse. The mechanisms are wide-spectrum, including elevation of caspase-1 and IL-18 in the small intestine, enhancement of the cell killing activity of cytotoxic T and natural killer (NK) cells, and anti-inflammatory and anti-angiogenic effects. It also inhibits the induction of liver CYP1A2, a carcinogen activating enzyme, and induces apoptosis in the colon epithelium of carcinogen treated rats. Thus, bLF possesses multi-functional potential to suppress carcinogenesis and is a good candidate for practical application in humans.

Microarray analysis of stress-related gene expression in patients with Ménière's disease.

We developed an original microarray carrying 1,467 cDNAs of stress-related genes for the assessment of stress responses. In this study, we used microarray analysis to assess the stress-related gene expression profiles in peripheral leukocytes in 2 patients with definite Ménière's disease. In the attack and active phases, mRNA expression levels of 57 genes and 163 genes were either up-regulated more than twofold or down-regulated by less than half in patient 1 and patient 2, respectively. Patient 1 had sporadic episodes of vertigo attack, while patient 2 had an intractable course with frequent vertigo attacks, suggesting that the magnitude of changes in gene expression is correlated with the severity of the disorder in Ménière's disease. The expression of a total of 26 genes commonly changed in both patients in the attack and active phases and returned to the baseline levels in the remission phase, suggesting the involvement of the distinct group of stress-related genes in the development of vertigo attacks in Ménière's disease. We then examined the effects of caloric stimulation on the stress-related gene expression profiles in peripheral leukocytes in 5 healthy volunteers. Although unilateral caloric stimulation with cold water caused acute vertigo with nystagmus, the expression profiles of stress-related genes did not significantly change after this experiment. This finding indicated that the up- or down-regulated genes in the attack and active phases in patients with Ménière's disease are not secondary to vertigo or vertigo-associated anxiety. All these findings suggested that the distinct group of stress-related genes contributed to the development of vertigo attacks of Ménière's disease and that stress-related gene expression profiles in peripheral leukocytes can be a predictive and therapeutic tool for episodic vertigo attacks in patients with Ménière's disease.

[Incidence of vertigo and dizziness disorders at a university hospital].

Subjects were 626 patients reporting vertigo or dizziness seen at the University Hospital Department of Otolaryngology from April 2001 to September 2003. Patients were diagnosed based on diagnostic criteria prescribed by the Japan Society for Equilibrium Research. The most common peripheral vestibular disorder was benign paroxysmal positional vertigo (32%), followed by Meniere's disease (12%). All peripheral vestibular disorders accounted for 65%. Central vestibular disorder accounted for 7%, of which space-occupying lesions in the posterior fossa accounted for 1.0% and cerebral infarction 1.9%. Dizziness due to orthostatic hypotension accounted for 4.0%. Among past reports on clinical statistics of vertigo, the incidence of vertigo and dizziness disorders differed greatly, but our research and the past 2 reports based on diagnostic criteria prescribed by the Japan Society for Equilibrium Research showed almost the same incidence, i.e., BPPV of 30-40%, Meniere's disease of 7-10%, other peripheral vestibular disorders of 15-20%, and central vestibular disorder of 6-8%. Unified diagnostic criteria are thus important in the statistical analysis of vertigo disorders.