Effects of Lactobacillus plantarum Strain OLL2712 Culture Conditions on the Anti-inflammatory Activities for Murine Immune Cells and Obese and Type 2 Diabetic Mice.

Studies on the health-promoting effects of lactic acid bacteria (LAB) are numerous, but few provide examples of the relationship between LAB function and culture conditions. We verified the effect of differences in culture conditions on Lactobacillus plantarum OLL2712 functionality; this strain exhibits anti-inflammatory activity and preventive effects against metabolic disorders. We measured interleukin-10 (IL-10) and IL-12 production in murine immune cells treated with OLL2712 cells prepared under various culture conditions. The results showed that the IL-10-inducing activities of OLL2712 cells on murine immune cells differed dramatically between OLL2712 groups at different culture phases and using different culture medium components, temperatures, and neutralizing pHs. In particular, exponential-phase cells had much more IL-10-inducing activity than stationary-phase cells. We confirmed that the Toll-like receptor 2 (TLR2) stimulation activity of OLL2712 cells depended on culture conditions in conjunction with IL-10-inducing activity. We also demonstrated functional differences by culture phases in vivo; OLL2712 cells at exponential phase had more anti-inflammatory activity and anti-metabolic-disorder effects on obese and diabetic mice than those by their stationary-phase counterparts. These results suggest that culture conditions affect the functionality of anti-inflammatory LAB.IMPORTANCE While previous studies demonstrated that culture conditions affected the immunomodulatory properties of lactic acid bacteria (LAB), few have comprehensively investigated the relationship between culture conditions and LAB functionality. In this study, we demonstrated several culture conditions of Lactobacillus plantarum OLL2712 for higher anti-inflammatory activity. We also showed that culture conditions concretely influenced the health-promoting functions of OLL2712 in vivo, particularly against metabolic disorders. Further, we characterized a novel mechanism by which changing LAB culture conditions affected immunomodulatory properties. Our results suggest that culture condition optimization is important for the production of LAB with anti-inflammatory activity.

Influenza 2014-2015 among pregnant Japanese women: primiparous vs multiparous women.

This study was performed to determine whether multiparous pregnant women are prone to influenza. A questionnaire survey was conducted at 19 centres located throughout Japan, targeting all 6,694 postpartum women within 7 days after birth before leaving the hospital. All women gave birth during the study period between March 1, 2015, and July 31, 2015. Data regarding vaccination and influenza infection in or after October 2014, age, previous experience of childbirth, and number and ages of cohabitants were collected. Seventy-eight percent (n = 51,97) of women given questionnaires responded. Of these, 2,661 (51 %) and 364 (7.0 %) women reported having been vaccinated and having contracted influenza respectively. Multiparous women had a higher risk of influenza regardless of vaccination status (8.9 % [121/1362] vs 5.7 % [74/1299], relative risk [95 % confidence interval], 1.80 [1.36 to 2.38] for vaccinated and 9.3 % [112/1198] vs 4.3 % [57/1328], 2.18 [1.60 to 2.97] for unvaccinated women) compared to primiparous women. The risk of influenza increased with increasing number of cohabitants: 4.8 % (100/2089), 7.5 %, (121/1618), 9.0 %, (71/785), and 10.4 % (58/557) for women with 1, 2, 3, and ≥4 cohabitants respectively. Family size is a risk factor for influenza infection in pregnancy.

Identification of a Lactobacillus plantarum strain that ameliorates chronic inflammation and metabolic disorders in obese and type 2 diabetic mice.

In this study, we identified a strain of lactic acid bacteria (LAB) that induces high levels of IL-10 production by immune cells, and evaluated the ability of the strain to suppress chronic inflammation and ameliorate metabolic disorders in in vitro and in vivo models. Among a collection of LAB strains, Lactobacillus plantarum strain OLL2712 (OLL2712) induced the highest levels of IL-10 production in mouse-derived dendritic cells and peritoneal macrophages. The anti-inflammatory effects of this strain were evaluated using a co-culture system comprising RAW 264.7 and 3T3-L1 cells. We also administered heat-killed OLL2712 to obese and type 2 diabetic KKAy mice for 3 wk to evaluate the in vivo effects of the strain. The OLL2712 significantly decreased the production of proinflammatory cytokines in vitro. Likewise, the administration of OLL2712 significantly suppressed proinflammatory cytokine levels in both the visceral adipose tissue and the serum of KKAy mice, and reduced serum triglyceride concentrations. The strain also alleviated oxidative stress and adrenaline levels in the serum of KKAy mice. On the other hand, Lactobacillus gasseri strain MEP222804 (a moderate IL-10 inducer) did not ameliorate the systemic inflammation and hyperlipidemia in KKAy mice. Our results suggest that treatment with strong IL-10-inducing LAB has the potential to ameliorate metabolic disorders by suppressing chronic inflammation in the host animal.

A double-blind randomized controlled trial of patient-controlled epidural analgesia with or without a background infusion following initial spinal analgesia for labor pain.

BACKGROUND: Patient-controlled epidural analgesia (PCEA) combined with spinal analgesia is an option for pain relief in labor. However, the effect of a CBI on the analgesic requirements of laboring women is still debated. This double-blind study investigated the effect of CBI with PCEA following spinal analgesia on the local anesthetic requirements of parturients during labor. METHODS: Sixty-six nulliparous women were randomly assigned to a standard PCEA protocol (5-mL demand bolus, 10-min lockout) with or without a CBI of 6 mL/h. The epidural solution consisted of 0.1% ropivacaine with fentanyl 2 microg/mL. Labor analgesia was initiated in both groups with intrathecal bupivacaine 2.5 mg plus fentanyl 25 microg. The number of demands per hour and the hourly dose of ropivacaine were calculated for both groups. RESULTS: The median [range] number of analgesic boluses per hour in the PCEA group that were demanded: 2.4 [0.8-12.2] and delivered: 1.6 [0.8-2.6], were significantly greater than those in the PCEA+CBI group: 0.7 [0.4-4.2] and 0.6 [0.2-1.3] (P 0.05). However, the hourly ropivacaine dose in the PCEA group (7.9 [3.9-13.2] mg/h) was not significantly different from that in the PCEA+CBI group (8.4 [6.0-12.5] mg/h). CONCLUSION: In laboring nulliparous patients provided initial labor analgesia with spinal anesthesia, the use of a continuous background infusion decreases PCEA demand dosing, but not the total hourly amount of ropivacaine and fentanyl used.

Alu-related 5q35 microdeletions in Sotos syndrome.

Haploinsufficiency of the NSD1 gene due to 5q35 microdeletions or intragenic mutations causes Sotos syndrome (SoS). In 46 of the 49 Japanese deletion cases, common deletion breakpoints were located at two flanking low copy repeats (LCRs), implying that non-allelic homologous recombination (NAHR) between LCRs is the major mechanism for the common deletion. In the other three cases of atypical deletions, the mechanism(s) of deletions remains unanswered. We characterized the atypical microdeletions using fluorescence in situ hybridization (FISH), quantitative real-time polymerase chain reaction (qPCR), and Southern blot hybridization. All the deletion breakpoints in the three cases were successfully determined at the nucleotide level. Two deletions are 1.07 Mb (SoS19) and 1.23 Mb (SoS109) in size, and another consisted of two deletions with sizes of 28 kb and 0.72 Mb, intervened by an intact 29-kb segment (SoS44). All deletions were smaller than a typical 1.9-Mb common deletion. Alu elements were identified in both deletion breakpoints in SoS19, one of deletion breakpoints in SoS109, and both deletion breakpoints of the proximal 28-kb deletion in SoS44. Alu-mediated NAHR is strongly suggested at least in two of atypical deletions. Finally, qPCR is a very useful method to determine deletion breakpoints even in repeat-related regions.

Systemic circulatory parameters. comparison between patients with normal tension glaucoma and normal subjects using ambulatory monitoring.

PURPOSE: To compare circadian changes of systemic circulation in patients with normal tension glaucoma (NTG) and normal subjects. METHODS: Forty-three patients with NTG and 226 normal subjects were enrolled in this study. Circulatory parameters, including blood pressure (BP) and pulse rate (PR), were measured in all subjects for 49 hours using an ambulatory monitoring system. In addition to a comparison between NTG patients and normal controls, the same parameters were compared between NTG patients who had progressive field defects and NTG patients who had stable field defects. RESULTS: The BP in NTG patients was significantly higher than in normal subjects. The nocturnal dip of BP in NTG patients was similar to the dip in normal subjects. However, the BP dip in NTG patients showing progressive visual field defects was significantly smaller than in patients with NTG showing stable visual field defects. Blood pressure fluctuation in sleep in the "progressive" patients was significantly greater than in the "stable" patients. Patients with NTG whose random BP was in a normal range showed a higher BP than normal subjects. The dip in PR in NTG patients was significantly lower than in normal subjects. CONCLUSIONS: An insufficient physiological nocturnal BP dip or a greater nocturnal fluctuation in BP may disturb the microcirculation of, and/or may directly damage, the optic nerve, resulting in increasing field loss in NTG.

Effect of epidural epinephrine infusion with bupivacaine on labor pain and mother-fetus outcome in humans.

BACKGROUND AND OBJECTIVES: Epinephrine is used with local anesthetics to prolong the duration of epidural analgesia and decrease the peak plasma concentrations of local anesthetics. In the practice of obstetric anesthesia, the utero-placental and fetal effects of epinephrine are controversial issues. We designed a prospective, randomized, and double-blind study to examine the effects of epinephrine infusion on the quality of analgesia and uterine or umbilical blood flows with Doppler ultrasound, as well as the duration of the first or the second stage of labor, and fetal outcome. METHODS: Twenty-eight parturients received continuous epidural bupivacaine 0.25% (4 mL/h) combined either with epinephrine (20 microg/h) (n = 13) or without epinephrine (n = 15) for analgesia during labor. If patients requested additional analgesia, an additional bolus of 1% or 1.5% lidocaine (6 to 10 mL) was administered. RESULTS: The total amount of additional lidocaine was greater in the plain bupivacaine group (130 [0, 280] mg; median [25th, 75th percentile] with P < .05) than in the epinephrine group (0 [0, 60] mg). Epinephrine infusion did not alter the resistance of the uterine and umbilical arteries as measured by resistance index. The duration of the first or second stages of labor did not significantly differ in the 2 groups. Epinephrine infusion did not change the fetal heart rate or the blood gas data in the umbilical artery. CONCLUSIONS: A low-dose epidural infusion of epinephrine decreased anesthetic requirements.

Changes in pulmonary histology and exfoliated bronchoalveolar cells induced by in vivo introduction of the tumor necrosis factor-alpha gene.

An inflammatory cytokine, tumor necrosis factor (TNF)-alpha, has been implicated in the pathogenesis of inflammatory lung diseases such as interstitial pneumonia (IP). To clarify the role of the inflammatory cytokine in the pathogenesis of lung inflammation, we introduced a murine TNF-alpha gene into murine lungs by the hemagglutinating virus of Japan (HVJ)-liposome method. Seven days after the TNF-alpha gene introduction resulted in marked cellular infiltration of alveoli, and mild histological change was observed 28 days after the gene introduction. Electron microscopic analysis revealed minimal deposition of collagen fibrils. Analysis of the BAL revealed that the total cell number was markedly increased 3 and 7 days after the gene introduction, and more than 90% of the cells were macrophages. The increase in the cell number was returned to below the normal level 28 days after the gene introduction. During the development of IP, TNF-alpha may regulate pathologic change of the pulmonary interstitium and alveolar cells.

In vivo gene transfer of an extracellular domain of platelet-derived growth factor beta receptor by the HVJ-liposome method ameliorates bleomycin-induced pulmonary fibrosis.

A number of investigators have reported augmented expression of PDGF in lungs with idiopathic pulmonary fibrosis (IPF) or with other types of pulmonary fibrosis. To accomplish such a regulation of PDGF activity, we constructed an expression plasmid of the extracellular domain of PDGF receptor beta chain (XR), which lacks intracellular tyrosine kinase domain and transmembrane portions, and estimated the therapeutic effects of XR gene transfer through the trachea on bleomycin-induced lung fibrosis of C57BL/6 mice using the hemagglutinating virus of Japan(HVJ)-liposome method. The XR gene transfer ameliorated the increases in the wet weight and hydroxyproline content and the histopathologic changes of the lung induced by bleomycin. These findings suggest that PDGF plays a crucial role in the pathogenesis of pulmonary fibrosis, and that XR gene transfer using the HVJ-liposome method may limit the progression of pulmonary fibrosis.

Introduction of plasmid DNA and oligonucleotides into lung epithelial cells by the hemagglutinating virus of Japan (HVJ)-liposome method.

The hemagglutinating virus of Japan (HVJ) fused with liposomes provides a unique transfection vehicle with characteristics of both virus vector and liposome. Here we investigate the efficiency and safety of the HVJ-liposome technique in delivering foreign genes and oligonucleotides into the lung of the Wistar rat. A plasmid vector containing the Escherichia coli beta-galactosidase (beta-gal) gene and the chicken beta-actin promoter was transfected via the trachea using the HVJ-liposome method. Cytochemical staining showed expression of exogenous beta-gal activity in airway epithelial cells, alveolar macrophages, and alveolar type II cells. This activity persisted at least 28 days after administration of the genes. FITC-labeled oligonucleotides also were introduced into the same types of lung cells as those expressing beta-gal. After instillation of HVJ-liposome, anti-HVJ antibodies were detected in the sera of the rats, but even after repeated administration of HVJ-liposome, no marked histopathologic change was observed while exogenous beta-gal expression was detected in pulmonary cells.

[Diffuse alveolar hemorrhage after allogeneic bone marrow transplantation].

A 34-year-old woman with chronic myeloid leukemia underwent allogeneic bone marrow transplantation (BMT) after receiving high-dose chemotherapy and total body irradiation. She experienced progressively dry cough 51 days after BMT, and chest X-ray films showed patchy infiltrations in the lower fields of both lungs on the 66th day after BMT. The symptoms of cough, fever, and hypoxemia worsened. The patchy infiltrations continued to spread and fuse. Diffuse alveolar hemorrhage (DAH) was diagnosed on the basis of high-resolution CT and bronchoalveolar lavage findings. Treatment with high-dose methyl prednisolone pulse therapy, antibiotics, and haptoglobin resolved the patient's DAH symptoms. DAH was thought to be secondary to thrombotic microangiopathy. The majority of patients who experience DAH after BMT eventually die. The remission observed in our case was rare, and illustrated that steroid therapy can be effective for DAH after BMT.

Interleukin-6 secreting phaeochromocytoma associated with clinical markers of inflammation.

Phaeochromocytomas have been shown to produce not only catecholamines but other neuropeptides and hormones, with a variety of clinical manifestations. We report a 70-year-old female patient with phaeochromocytoma exhibiting sustained hypertension, low-grade fever, thrombocytosis, and elevated levels of plasma fibrinogen and C-reactive protein. Serum interleukin (IL)-6 levels were significantly elevated, whereas serum IL-1 alpha and IL-1 beta were not detectable. After surgical removal of the tumour, hypertension and low-grade fever disappeared, and the laboratory finding including serum IL-6 concentrations became normal. Immunohistochemical study of the tumour showed positive staining for IL-6. Culture of the resected tumour revealed the production of large amounts of IL-6. It is suggested that IL-6 secreted by the tumour was responsible for some of the clinical manifestations in this patient.

[Effects of in vivo instillation of genes coding for cytokines on pulmonary fibrosis].

Interstitial pneumonia is characterized by alveolitis that results in interstitial fibrosis. To study the role of humoral factors in the pathogenesis of interstitial fibrosis, we introduced expression vectors into Wistar rats via the trachea, to cause local overexpression of these humoral factors in the lung. Genes for human interleukin (IL)-6 and for the IL-6 receptor caused lymphocytic alveolitis without marked proliferation of fibroblasts. In contrast, overexpression of the genes for human transforming growth factor (TGF)-beta 1 and for human platelet-derived growth factor (PDGF)-B caused only mild cellular infiltration in the alveoli. However, both caused marked proliferation of fibroblasts and deposition of collagen fibrils. Introducing an expression vector that coded for a mutant form of the PDGF beta receptor that lacks its cytoplasmic domain markedly alleviated the pathohistologic changes caused by bleomycin in murine lungs. These findings show that TGF- and PDGF-B may be closely related to fibrosis in the lung, and that artificial regulation of them may be effective for treatment of lung fibrosis.

Lung cancer producing interleukin-6.

A patient with complaints of high fever and left shoulder pain was found to have a large mass in the left upper lobe on chest roentgenogram. Laboratory evaluation revealed marked thrombocytosis, hypoalbuminemia, and increased serum concentrations of CRP, fibrinogen and interleukin-6 (IL-6). A transcutaneous biopsy specimen revealed large cell carcinoma. Tumor production of IL-6 was confirmed by immunohistochemical staining with an anti-human IL-6 monoclonal antibody (MH60).

[Study on human eye movements induced by prolonged eye closure and lid opening--report 2. Closing and opening of eyelids].

The eye movement during prolonged eye closure and the following eye opening has not been studied precisely. In the present study we evaluated the eye movement during prolonged eye closure and the following eye opening using a magnetic search oil method. In a bright room, the exact moment of recognizing a visual target during eye opening could not be determined. Therefore, other conditions were used to elucidate the precise relationship between the presence and lack of visual input during eye opening. Eye movement during eye opening was accomplished in three phases. The first phase had rapid movements like visually guided saccades toward the imaged target. The second phase had slow movements and drift toward the imaged target, and the final eye position was reached within 3 degrees from the target from 5 different gaze directions. The third phase was final visually evoked corrective saccades and the subjects found the target. We discuss the origin of these eye movements.

[Study on human eye movements induced by blinks--report 1. Three kinds of blinking].

The eye movement associated with blinking was studied with a magnetic search coil method. Lid movements and eye movements associated with spontaneous blinks, voluntary blinks, and reflex blinks induced by puffs of air or flashes of light were analysed. Eye movements associated with spontaneous blinks and voluntary blinks had initial downward and adducted movements followed by upward and abducted movements of the eyeball. Reflex blinks initially showed the same downward and adducted movements, but upward and abducted movements were not constantly obtained in given subjects. The velocity-amplitude ratio was equal among the three kinds of blinks in both horizontal and vertical movements. The velocity-amplitude ratio of the eye movements associated with blinks and visually-guided saccadic eye movements was compared. The horizontal and vertical eye movements associated with blinks were slower than those of saccadic eye movements in the same subject. Therefore, the eye movements associated with the blinks may have a different neural mechanism in the brain from that of the saccadic eye movement system.

Antitumor activity of SPM VIII, a derivative of the nucleoside antibiotic spicamycin, against human tumor xenografts.

The antitumor activity of spicamycin analogue SPM VIII against human stomach, breast, lung, colon and esophageal cancers was compared to that of mitomycin C (MMC) in the human tumor-nude mice xenograft model. Comparative studies of SPM VIII given i.v. at 6 mg/kg/day daily for 5 days and MMC given i.v. at 6.7 mg/kg on day 1 revealed that the antitumor spectrum of SPM VIII showed a different pattern from that of MMC and that SPM VIII caused tumor mass reductions in more tumors than did MMC in colon cancers (4/12 versus 1/11). In addition to this study, a comparative study of SPM VIII given i.v. at 12 mg/kg/day 8 times at 3- or 4-day intervals and 5'-deoxy-5-fluorouridine (5'-DFUR) given po at 185 mg/kg/day 5 days per week for 4 weeks showed that SPM VIII had the highest effect on SC-9 human stomach cancer and COL-1 human colon cancer among the 3 compounds, resulting in a significant reduction of tumor mass. Although other pharmacological studies are in progress, these results suggest that SPM VIII might be a novel antitumor compound effective for human cancers including cancer of the digestive organs.

Antitumor activity of a spicamycin derivative, KRN5500, and its active metabolite in tumor cells.

KRN5500, (6-[4-Deoxy-4-(2E,4E)-tetradecadienoylglycyl]amino-L-glycero - beta-L-mannoheptopyranosyl]amino-9H-purine), was semi-synthesized in an attempt to increase the therapeutic effects of spicamycin analogues. The present study evaluated the antitumor activity of KRN5500 against murine tumors and human tumor xenografts. KRN5500 prolonged the survival of P388 leukemia- and B16 melanoma-bearing mice but was marginally effective on colon adenocarcinoma 26. The antitumor activity of KRN5500 (4 mg/kg/day x 5, IV) against xenografts of 10 human stomach, 14 colon and 2 esophageal cancers was evaluated with two parameters: the tumor growth inhibition rate (TGIR) and the tumor mass reduction by comparison with mitomycin C (MMC, 6.7 mg/kg/day x 1,IV). KRN5500 showed a marked efficacy in the human tumor xenograft model. The overall response rate of 26 cancers to KRN5500, evaluated by TGIR, was approximately equal to their response rate to MMC (72% vs. 73%). However, more tumors were reduced by KRN5500 than by MMC (52% vs. 39%). It is notable that the response rates of 14 colon cancers to KRN5500 were significantly higher than those to MMC, both in TGIR (69% vs. 58%) and in tumor mass reduction (46% vs. 23%). Among the tumors sensitive to KRN5500, COL-1 showed a marked response (TGIR 93%) and a significant reduction in tumor mass (0.22-fold the starting volume). In the mode of action, KRN5500 was found to show an inhibitory effect on protein synthesis in P388 cells (IC50 1.5 microM). However, KRN5500 was ineffective even at 170 microM in inhibition of protein synthesis in rabbit reticulocyte lysates.(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro and ex vivo free radical scavenging activities of carazostatin, carbazomycin B and their derivatives.

Free radical scavenging activities of various carbazole compounds, carazostatin, carbazomycin B and their chemically modified derivatives were studied in vitro and ex vivo. Among these compounds, carazostatin, which was isolated as a free radical scavenger from the culture of Streptomyces chromofuscus, showed the most potent inhibitory activity against lipid peroxidation of rat brain homogenate in vitro. Carbazomycin B, a known antimicrobial antibiotic, also exhibited strong activity in this system. Although O-modified derivatives of carazostatin and carbazomycin B retained considerable activity, N,O-dimethyl derivatives did not suppress the peroxidation. On the other hand, the results from the ex vivo evaluation of these carbazoles in the lipid peroxidation system of mouse blood plasma showed that the original compounds as well as their O-modified derivatives had a strong inhibitory activity upon oral administration to mice. These findings suggest that these natural carbazoles and their effective derivatives can protect tissues from the peroxidative damage due to generation of free radicals.