Effect of phospholipid acyl chain modulation on vitamin E incorporation into pulmonary artery endothelial cell membranes

Incorporation of vitamin E (α-tocopherol) was measured in total membranes of pulmonary artery endothelial cells (PAEC) following treatment with eight synthetic phosphatidylethanolamines (PE) (Palmitoyloleoyl, 16:0–18:1 PE₁; distearoyl, 18:0–18:0 PE₂; dioleoyl, 18:1–18:1 PE₃; stearoyl- linoleoyl, 18:0–18:2 PE₄; dilinoleoyl, 18:2–18:2 PE₅; stearoyl-arachiod-nyl, 18:0–20:4 PE₆; diarachidonyl, 20:4–20:4 PE₇; and stearoyl-docosahexenoyl, 18:0–22:6 PE₈). Endogenous PE content of native membranes was 0.88 ± 0.01 nmol/mg protein. Incorporation of PE irrespective of fatty acid content significantly (P < 0.02) increased the PE content of total membranes. Vitamin E incorporation in control membranes was 63 ± 9 nmol/mg protein. Incorporations of vitamin E in PE₁- to PE₇-treated cells were significantly (P < 0.05) increased compared to controls and were comparable to each other. Vitamin E incorporation into PE₈-treated cells was threefold greater (P < 0.001) thatn controls and twofold greater (P < 0.001) than PE₁- to PE₇-treated cells. Increased PE content results in increased vitamin E incorporation into PAEC membranes irrespective of the fatty acids present on the acyl chain, and maximal incorporation of vitamin E in PE₈-treated cells may relate to the increased carbon chain length rather than to the degree of unsaturation at the sn₂ position. © 1993 Wiley-Liss, Inc.     

Effect of thyroid hormone on concentrations of plasma calcitonin in broiler chicks

The purpose of these studies was to determine the effect of thyroidectomy (Tx), and thyroid hormone (T₃/T₄) treatment on concentrations of plasma CT in chicks. In addition, the turnover of CT in Tx- and T₃/T₄-treated chicks was estimated using a novel nonradioactive salmon CT preparation. One-week-old broiler chicks (Gallus domesticus) (n=75) were divided into three groups. Group I was sham-injected daily (i.m. saline), Group II was injected with 50 μg/day of T₃/T₄ while Group III was injected with the goitrogen, methimazole, (150 mg/kg BW per day) for 8 weeks. Chicks (8–9 weeks old) were implanted with catheters in the brachial wing vein and administered ruthenium-labeled salmon CT. Blood samples were collected at 30 s, 1, 2, 4, 8, 20 min, and 3 h after injection. Results showed that concentrations of plasma CT were decreased in T₃/T₄-injected birds. There was no significant effect of methimazole on circulating concentrations of plasma CT. The half-life of CT was significantly increased (P<0.05) in both T₃/T₄-injected (n=6; 1.34±0.16 min) and goitrogen-treated birds (n=2; 5.81±2.83 min) compared to controls (n=7; 54±3 s) The results demonstrate that changes in concentrations of plasma thyroid hormones can significantly affect concentrations of plasma CT.     

Determination of major UDP-glucuronosyltransferase enzymes and their genotypes responsible for 20-HETE glucuronidation

The compound 20-HETE is involved in numerous physiological functions, including blood pressure and platelet aggregation. Glucuronidation of 20-HETE by UDP-glucuronosyltransferases (UGTs) is thought to be a primary pathway of 20-HETE elimination in humans. The present study identified major UGT enzymes responsible for 20-HETE glucuronidation and investigated their genetic influence on the glucuronidation reaction using human livers (n = 44). Twelve recombinant UGTs were screened to identify major contributors to 20-HETE glucuronidation. Based on these results, UGT2B7, UGT1A9, and UGT1A3 exhibited as major contributors to 20-HETE glucuronidation. The K_m values of 20-HETE glucuronidation by UGT1A3, UGT1A9, and UGT2B7 were 78.4, 22.2, and 14.8 μM, respectively, while V_max values were 1.33, 1.78, and 1.62 nmol/min/mg protein, respectively. Protein expression levels and genetic variants of UGT1A3, UGT1A9, and UGT2B7 were analyzed in human livers using Western blotting and genotyping, respectively. Glucuronidation of 20-HETE was significantly correlated with the protein levels of UGT2B7 (r² = 0.33, P < 0.001) and UGT1A9 (r² = 0.31, P < 0.001), but not UGT1A3 (r² = 0.02, P > 0.05). A correlation between genotype and 20-HETE glucuronidation revealed that UGT2B7 802C>T, UGT1A9 −118T₉>T₁₀, and UGT1A9 1399T>C significantly altered 20-HETE glucuronide formation (P < 0.05–0.001). Increased levels of 20-HETE comprise a risk factor for cardiovascular diseases, and the present data may increase our understanding of 20-HETE metabolism and cardiovascular complications.     

Skeletal ontogeny of the vertebral column and of the fins in shi drum Umbrina cirrosa (Teleostei: Perciformes: Sciaenidae), a new candidate species for aquaculture

The osteological development of the vertebral column and fins in shi drum Umbrina cirrosa was studied in order to improve knowledge for its introduction in Mediterranean aquaculture. The osteological development was studied in 171 individuals, of total length (L_T) from 2·7 to 30·2 mm that were reared under the mesocosm technique. Vertebral ontogeny starts at 3·4 and 4·0 mm L_T, with the formation of the first cartilaginous neural and haemal arches, and spines, respectively, and is completed with the full attainment of epicentrals (12·5 mm L_T). The formation of vertebral centra occurs between 4·1 and 7·4 mm L_T. Pectoral supports are the first fin elements to develop (3·0 mm L_T), followed by those of the caudal fin (3·8 mm L_T), pelvic fin (3·9 mm L_T) and finally by those of the dorsal and anal fins (4·5 mm L_T). The caudal fin is the first to develop fin rays and attain the full count of principal fin rays (4·5–6·8 mm L_T), but the last to be fully completed with the formation of procurrent fin rays (6·9–17·5 mm L_T). The next fins starting to present rays are the dorsal (5·3 mm L_T) and the pectoral fins (5·6 mm L_T), while the anal and pelvic fins are the last (5·7 mm L_T). Following the caudal principal fin rays (6·8 mm L_T), the dorsal, anal (6·9 mm L_T), pelvic (7·4 mm L_T) and pectoral fins (9·8 mm L_T) are the next with fully completed ray counts. Aggregation of qualitative changes, such as the appearance of cartilages, the beginning and the complement of the ossification process and the full complement of elements in U. cirrosa were measured as cumulative frequency counts. These measurements reveal three ontogenetic intervals: one very developmentally active period during early life stages (from 3 to 5·9 mm L_T), a second slower developmental period (from 6·0 to 8·9 mm L_T) and finally a period of ontogeny more focused on structure refinement up to metamorphosis and settlement (>9·0 mm L_T).     

Early kit mortality and growth in farmed mink are affected by litter size rather than nest climate

We investigated the effects of nest box climate on early mink kit mortality and growth. We hypothesised that litters in warm nest boxes experience less hypothermia-induced mortality and higher growth rates during the 1st week of life. This study included data from 749, 1-year-old breeding dams with access to nesting materials. Kits were weighed on days 1 and 7, dead kits were collected daily from birth until day 7 after birth, and nest climate was measured continuously from days 1 to 6. We tested the influences of the following daily temperature (T) and humidity (H) parameters on the number of live-born kit deaths and kit growth: T_mean, T_min, T_max, T_var (fluctuation) and H_mean. The nest microclimate experienced by the kits was buffered against the ambient climate, with higher temperatures and reduced climate fluctuation. Most (77.0%) live-born kit deaths in the 1st week occurred on days 0 and 1. Seven of 15 climate parameters on days 1 to 3 had significant effects on live-born kit mortality. However, conflicting effects among days, marginal effects and late effects indicated that climate was not the primary cause of kit mortality. Five of 30 climate parameters had significant effects on kit growth. Few and conflicting effects indicated that the climate effect on growth was negligible. One exception was that large nest temperature fluctuations on day 1 were associated with reduced deaths of live-born kit (P<0.001) and increased kit growth (P=0.003). Litter size affected kit vitality; larger total litter size at birth was associated with greater risks of kit death (P<0.001) and reduced growth (P<0.001). The number of living kits in litters had the opposite effect, as kits in large liveborn litters had a reduced risk of death (P<0.001) and those with large mean litter size on days 1 to 7 had increased growth (P=0.026). Nest box temperature had little effect on early kit survival and growth, which could be due to dams’ additional maternal behaviour. Therefore, we cannot confirm that temperature is the primary reason for kit mortality, under the conditions of plenty straw access for maternal nest building. Instead, prenatal and/or parturient litter size is the primary factor influencing early kit vitality. The results indicate that the focus should be on litter size and dam welfare around the times of gestation and birth to increase early kit survival in farmed mink.     

Sphingomonas gimensis sp. nov., a novel Gram-negative bacterium isolated from abandoned lead–zinc ore mine

A novel bacterial strain designated 9PNM-6^T was isolated from an abandoned lead–zinc ore mine site in Meizhou, Guangdong Province, China. The isolate was found to be Gram-negative, rod-shaped, orange-pigmented, strictly aerobic, oxidase- and catalase-positive. Growth occurred at 0–4 % NaCl (w/v, optimum, 0 %), at pH 6.0–8.0 (optimum, pH 7.0) and at 15–32 °C (optimum, 28–30 °C). Phylogenetic analysis based on 16S rRNA gene sequence similarities showed that strain 9PNM-6^T belongs to the genus Sphingomonas, with the highest sequence similarities with Sphingomonas jejuensis NBRC 107775^T (99.7 %), Sphingomonas koreensis KCTC 2882^T (95.1 %) and Sphingomonas dokdonesis KCTC 12541^T (95.1 %). The chemotaxonomic characteristics of strain 9PNM-6^T were consistent with those of the genus Sphingomonas. The predominant respiratory quinone was identified as ubiquinone Q-10, the major polyamine as sym-homospermidine, and the major cellular fatty acids as C_18:1 ω7c, C_16:0, C_16:1 ω7c and/or C_16:1 ω6c and C_14:0 2-OH. The major polar lipids are sphingoglycolipid, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatideylcholine, an unidentified phospholipid and four unidentified aminolipids. The genomic DNA G+C content of strain 9PNM-6^T was determined to be 69.2 ± 0.6 mol%. Based on comparative analyses of morphological, physiological and chemotaxonomic data, and levels of DNA–DNA relatedness values, strain 9PNM-6^T is considered to represent a novel species of the genus Sphingomonas, for which the name Sphingomonas gimensis sp. nov. (Type strain 9PNM-6^T = GIMCC 1.655^T = CGMCC 1.12671^T = DSM 27569^T) is proposed.     

<Emphasis Type="Italic">Spirosoma daeguensis</Emphasis> sp. nov., isolated from beach soil

A Gram-stain-negative, non-motile, non-spore-forming, rod-shaped, aerobic bacterium, designated 15J9-6^T, was isolated from beach soil on Jeju Island, South Korea. Strain 15J9-6^T, grew at 10–30°C (optimum growth at 25°C) and pH 7–8 (optimum growth at pH 7) on R2A, NA, and TSA agar. Phylogenetically, the strain was closely related to members of the genus Spirosoma (92.3–90.1% 16S rRNA gene sequence similarities) and showed highest sequence similarity to Spirosoma panaciterrae DSM 21099^T (92.3%). The G+C content of the genomic DNA of strain 15J9-6^T was 45.7 mol%. The strain contained phosphatidylethanolamine, two unidentified aminophospholipids, an unidentified phospholipid, and an unidentified lipid as the major polar lipids; menaquinone MK-7 as the predominant respiratory quinone and summed feature 3 (C_16:1 ω6c/C_16:1 ω7c; 30.1%), C_16:1 ω5c (23.1%), iso C_15:0 (13.3%), and C_16:0 (8.4%) as the major fatty acids which supported the affiliation of strain 15J9-6^T to the genus Spirosoma. The results of physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain 15J9-6^T from recognized Spirosoma species. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain 15J9-6^T represents a novel species of the genus Spirosoma, for which the name Spirosoma daeguensis sp. nov. is proposed. The type strain is 15J9-6^T (=KCTC 52036^T =JCM 31995T)     

THE EFFECT OF ESTRADIOL ON THE CELL KINETICS IN THE UTERINE AND CERVICAL EPITHELIUM OF NEONATAL MICE

The effect of estradiol-17β on the length of the various phases of the cell cycle was studied in the neonatal mouse uterine, and cervical epithelium. A double labelling method was used, and in addition labelled mitoses were counted. In the uterus proper, estradiol shortens the length of the total cell cycle, T_c, from 17-9 hr to 15-7 hr, and the duration of S phase, T_s, from 6–7 to 5-1 hr 6 hr after estradiol treatment. 12 hr after estradiol treatment, T_c is shortened to 7-4 hr and T_s to 4–5 hr. The shortening of T_c at 12 hr is mainly due to an effect on T_G1, which is shortened from 8–55 hr in untreated animals to 1–8 hr in estradiol treated animals. The T_c of cervix epithelium cells in untreated animals was found to be 21-8 hr. After treating the mice for 6 hr with estradiol the T_c was now increased to 47 hr and further to 61 -2 hr following 12 hr treatment with the hormone. T_s increases from 8-3 hr to 15-2 hr following 6 hr estradiol treatment, and to 15-4 hr after 12 hr treatment. The effect is most pronounced in T_G1, which is lengthened from 10–95 hr in untreated animals to 28-1 hr and 43 hr, respectively, in animals treated for either 6 or 12 hr with estradiol.     

Effect of ovine TSH, thiourea, ovine prolactin and bovine growth hormone on plasma thyroxine and tri-iodothyronine levels in rainbow trout,Salmo gairdneri

Summary A single injection of ovine TSH (1.1 g or 2.2 g/g body weight) effected a significant (P<0.01) increase in plasma T₄ levels in rainbow trout within 9 h and 1 h respectively. In fish given the higher dose the T₄ levels were still significantly higher (P<0.01) than in the controls 24 h after the injection. Ovine TSH had no significant effect on plasma T₃ levels. Plasma T₃/T₄ ratios in fish given the higher TSH dose were significantly lower than the controls at 3 h (P<0.01), 6 h (P=0.01), 9 h and 24 h (P<0.05) while in fish given the lower dose they were significantly lower than in controls at 3 h (P<0.01), 6 h (P=0.01) and 9 h (P<0.05).T₄ levels in TSH-injected fish were significantly higher (P<0.05) than in controls after 3 and 11 injections but were not significantly different after 16 injections. Plasma T₃ and T₃/T₄ ratios did not differ significantly in fish given multiple injections of either TSH or saline.Plasma T₄ levels in fish treated with thiourea were significantly lower (Pº0.01) than in comparable controls after 6, 9 and 15 days and plasma T₃ levels were lower (P<0.05) after 15 days treatment. T₃/T₄ ratios did not differ significantly at any time period.A single injection of bovine STH (10 g/g body weight) effect a significant rise in plasma T₄ levels 9 h (P=0.05) and 24 h (P<0.01) after the injection while ovine prolactin in a similar dose was without effect. Plasma T₃ levels in saline injected fish showed a steady decline over the 24 h following the injection whereas levels in prolactin and STH injected animals tended to rise. T₃ levels in the saline-injected fish were significantly higher (P<0.01) than in prolactin and STH injected animals 1 h after the injection but were significantly lower (P<0.01) than in the hormone-treated fish 24 h after the injection. There were no significant differences in T₃/T₄ ratios in any of the groups. Multiple injections of STH effected a significant increase (P<0.05) in plasma T₄ compared with the controls; prolactin had no significant effect. There was no significant differences in plasma T₃ concentration or T₃/T₄ ratios in any of the groups.     

Retinoic acid can enhance the stimulation by thyroid hormone of heme oxygenase activity in the liver of thyroidectomized rats

The effects of retinoic acid (RA) (50 μg/100 g body wt. per day) on hepativ heme oxygenase activty, δ-aminolevulinate synthase (ALAS) activity and on cytochrome P-450 content were determined in thyroidectomized rats treated with T₃ (10 μg/100 g body wt. per day) or diluent. RA, when administered for 3 days, failed to influence significantly the activity of either heme oxygenase or ALAS, however, the retinoid depleted hepatic cytochrome P-450 content by 17% (P < 0.01) and microsomal heme content by 47% (P < 0.001). T₃ administration enhanced heme oxygenase activity by 72% (P < 0.001) and ALAS activity by 251% (P < 0.001) above levels in diluent treated controls and depleted cytochrome P-450 levels by 55% (P < 0.001) and heme levels by 75% (P < 0.001). When RA and T₃ were administered together, the retinoid markedly enhanced the T₃ stimulation of heme oxygenase activity; 173% above controls (P < 0.001), and 61% above T₃ alone (P < 0.001). However, RA failed to influence the effect of T₃ on ALAS activity or cytochrome P-450 depletion. The results indicate that RA can influence the levels of hepatic cytochrome P-450 and can modulate the stimulation of heme oxygenase activity by thyroid hormone in vivo.     

Validation and Comparison of the Therapeutic Efficacy of Boron Neutron Capture Therapy Mediated By Boron-Rich Liposomes in Multiple Murine Tumor Models

Boron neutron capture therapy (BNCT) was performed at the University of Missouri Research Reactor in mice bearing CT26 colon carcinoma flank tumors and the results were compared with previously performed studies with mice bearing EMT6 breast cancer flank tumors. Mice were implanted with CT26 tumors subcutaneously in the caudal flank and were given two separate tail vein injections of unilamellar liposomes composed of cholesterol, 1,2-distearoyl-sn-glycer-3-phosphocholine, and K[nido-7-CH₃(CH₂)₁₅–7,8-C₂B₉H₁₁] in the lipid bilayer and encapsulated Na₃[1-(2`-B₁₀H₉)-2-NH₃B₁₀H₈] within the liposomal core. Mice were irradiated 30 hours after the second injection in a thermal neutron beam for various lengths of time. The tumor size was monitored daily for 72 days. Despite relatively lower tumor boron concentrations, as compared to EMT6 tumors, a 45 minute neutron irradiation BNCT resulted in complete resolution of the tumors in 50% of treated mice, 50% of which never recurred. Median time to tumor volume tripling was 38 days in BNCT treated mice, 17 days in neutron-irradiated mice given no boron compounds, and 4 days in untreated controls. Tumor response in mice with CT26 colon carcinoma was markedly more pronounced than in previous reports of mice with EMT6 tumors, a difference which increased with dose. The slope of the dose response curve of CT26 colon carcinoma tumors is 1.05 times tumor growth delay per Gy compared to 0.09 times tumor growth delay per Gy for EMT6 tumors, indicating that inherent radiosensitivity of tumors plays a role in boron neutron capture therapy and should be considered in the development of clinical applications of BNCT in animals and man.     

Thyroid hormones are necessary for the metamorphosis of tarpon Megalops cyprinoides leptocephali

This study investigates the effects of thyroxine (T₄), triiodothyronine (T₃) and thiourea (TU) on the metamorphosis of tarpon Megalops cryprinoides leptocephali. TU is an anti-thyroid hormone drug that inhibits the production of T₄ and T₃ in the thyroid tissue. Fully grown tarpons leptocephali were collected at the river mouth and, in the laboratory, were immediately treated with 100 ppb T₄, 10 ppb T_3, or 300 ppm TU. The appropriate concentrations were validated in a preliminary dose response experiment. Morphological and physiological characteristics that indicate metamorphic processes were measured every 2 days. T₄ and T₃ slightly speeded up the metamorphosis of tarpons compared with the control group. The experimental treatments produced accelerated reductions in length, increases in head/body ratio, swimbladder development, and loss of body water and sodium. In contrast, TU treatment caused metamorphic stasis with complete inhibition of metamorphosis between days 6 and 8. Thyroid hormone treatment stimulated fast otolith growth while TU treatment stopped otolith growth between days 6 and 9. Leptocephali in T₄, T₃ and control groups completed metamorphosis in 10-14 days, but TU-treated tarpons remained in the metamorphic leptocephalus stage more than 22 days. In addition, the inhibition of leptocephalus metamorphosis by 300 ppm TU can be reversed in the presence of 10 ppb T₃. These results indicate that thyroid hormones are involved in regulating the metamorphosis of leptocephali.     

Rapid accumulation of adoptively transferred CD8+ T cells at the tumor site is associated with long-term control of SV40 T antigen-induced tumors

We previously established a model to study CD8⁺ T cell (T_CD8)-based adoptive immunotherapy of cancer using line SV11 mice that develop choroid plexus tumors in the brain due to transgenic expression of Simian Virus 40 large T antigen (Tag). These mice are tolerant to the three dominant T_CD8-recognized Tag epitopes I, II/III and IV. However, adoptive transfer of spleen cells from naïve C57BL/6 (B6) mice prolongs SV11 survival following T_CD8 priming against the endogenous Tag epitope IV. In addition, survival of SV11 mice is dramatically increased following transfer of lymphocytes from Tag-immune B6 mice. In the current study, we compared the kinetics and magnitude of Tag-specific T_CD8 accumulation at the tumor site following adoptive transfer with a high dose of either Tag-immune or naïve donor cells or decreasing doses of Tag-immune lymphocytes. Following adoptive transfer of Tag-immune cells, epitope I- and IV-specific T_CD8 accumulated to high levels in the brain of SV11 mice, peaking at 5–7 days, while epitope IV-specific T_CD8 derived from naïve donors required three weeks to achieve peak levels. A similar delay in the peak of epitope IV-specific T_CD8 accumulation was observed when tenfold fewer Tag-immune donor cells were administered, reducing control of tumor progression. These results suggest that efficient and prolonged control of established autochthonous tumors is associated with high-level early accumulation of adoptively transferred T cells. We also provide evidence that although multiple specificities are represented in the Tag immune donor lymphocytes, epitope IV-specific donor T_CD8 play a predominant role in control of tumor growth.     

Age and growth of Siberian sculpin (Cottus poecilopus) and young brown trout (Salmo trutta) in a subalpine Norwegian river

Age, growth and density of Siberian sculpin (Cottus poecilopus) and young brown trout (Salmo trutta) within two sections of River Atna; above Lake Atnsjøen [Section 1 at altitudes between 739 and 715 m] and below Lake Atnsjøen [Section 2 at altitudes between 430 and 370 m] was studied during a 6-year period (1986–91). The water temperature was considerably lower in Section 1 than in Section 2, as the number of days with a water temperature above 10?°C (T _{D ≥ 10?° C} ) from spring to August 1 ranged between 2–26 and 26–52 days, respectively. Juvenile brown trout (age 0+) attained a significantly smaller body size in Section 1 than in Section 2; mean length ±SD was 35 ± 8 mm (ranged 27–46) and 43 ± 7 mm (range 38–46), respectively. In Section 2, there was a highly positive correlation between the body length of 0+ brown trout and mean water temperature in June (p<0.005), and also to some extent in Section 1 (p=0.11). Individuals of age 1+ did not exhibit any such difference, while fish in age group 2+ were larger in Section 1 than in Section 2. By using the number of days with a water temperature between the range 5–10?°C (T _{D ≥ 5 ? 10?° C} ) as test variables, we found a highly positive correlation between the August 1 body length of 0+ brown trout and T _{D ≥ 9}?°C from spring to August 1 in Section 2 (p<0.05), as opposed to T _{D ≥ 7}?°C for trout in Section 1 (p=0.11). Young Siberian sculpin (age 0+ and 1+) also exhibited slower growth in Section 1 than in Section 2, but this was not the case among older specimens. In the year with the lowest temperature measured (1987), no 0+ Siberian sculpin were caught in any of the two sections, indicating that low temperature affects their survival. Both species exhibited large spatial and temporal variation in density. Thus, data on abundance and growth sampled on one occasion at one site can not be regarded as representative for these two fish populations.     

<Emphasis Type="Italic">Halomonas heilongjiangensis</Emphasis> sp. nov., a novel moderately halophilic bacterium isolated from saline and alkaline soil

A moderately halophilic bacterium, designated strain 9-2^T, was isolated from saline and alkaline soil collected in Lindian county, Heilongjiang province, China. The strain was observed to be strictly aerobic, Gram-negative, rod-shaped, oxidase-positive, catalase-positive and motile. It was found to require NaCl for growth and to grow at NaCl concentrations of 0.5–14 % (w/v) (optimum, 7–10 %, w/v), at temperatures of 10–45 °C (optimum 25–30 °C) and at pH 5.0–10.0 (optimum pH 8.0). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 9-2^T is a member of the genus Halomonas and is closely related to Halomonas desiderata DSM 9502^T (96.68 %), Halomonas campaniensis DSM 1293^T (96.46 %), Halomonas ventosae DSM 15911^T (96.27 %) and Halomonas kenyensis DSM 17331^T (96.27 %). The DNA–DNA hybridization value was 38.9 ± 0.66 % between the novel isolate 9-2^T and H. desiderata DSM 9502^T. The predominant ubiquinones were identified as Q9 (75.1 %) and Q8 (24.9 %). The major fatty acids were identified as C_16:0 (22.0 %), Summed feature 8 (C_18:1 ω6c/C_18:1 ω7c, 19.6 %), Summed feature 3 (C_16:1 ω6c/C_16:1 ω7c, 12.6 %), C_12:0 3-OH (12.0 %) and C_10:0 (11.7 %). The DNA G+C content was determined to be 69.7 mol%. On the basis of the evidence presented in this study, strain 9-2^T is considered to represent a novel species of the genus Halomonas, for which the name Halomonas heilongjiangensis sp. nov. is proposed. The type strain is 9-2^T (=DSM 26881^T = CGMCC 1.12467^T).     

Functional validation of the genetic architecture of Salmonella Enteritidis persistence in 129S6 mice

The Gram-negative bacteria, Salmonella, cause a broad spectrum of clinical diseases in humans, ranging from asymptomatic carriage to life-threatening sepsis. We have designed an experimental model to study the contribution of genetic factors to the persistence of Salmonella Enteritidis during the late phase of infection in 129S6/SvEvTac and C57BL/6J mice. C57BL/6J mice cleared the bacteria from their reticuloendothelial system within a period of 42 days, whereas the 129S6 mice still presented a high bacterial load. Using this model, we have identified ten Salmonella Enteritidis susceptibility loci (Ses1, Ses1.1, and Ses3–Ses10) associated with bacterial persistence in target organs of 129S6/SvEvTac mice using a two-locus epistasis QTL linkage mapping approach. Significant statistical interactions were detected between Ses1 on chromosome 1 and Ses5 on chromosome 7 and between Ses1 and Ses4 on chromosome X. In this study, we functionally validated the genetic architecture of Salmonella persistence in 129S6 mice using single- (129S6.B6-Ses1.2 that combines Ses1 and Ses1.1 loci, 129S6.B6-Ses4, and 129S6.B6-Ses5) and double-congenic mice (129S6.B6-Ses1.2/Ses4 and 129S6.B6-Ses1.2/Ses5). These experiments demonstrate functional interactions between Ses1.2 and Ses4 or Ses5 that improve Salmonella Enteritidis clearance, validating the critical role played by gene–gene interactions in the contribution to bacterial clearance heritability. Improved bacterial clearance in double-congenic mice could be explained by the impact of Ses4 and Ses5 in combination with Ses1.2 on T_H polarization since a T_H2 bias (decreased Ifng and increased Il4 mRNA levels and reduced IgG2a immunoglobulins in the serum) was observed in 129S6.B6-Ses1.2/Ses5 mice and a T_H17 (high Il17 expression) bias in 129S6.B6-Ses1.2/Ses4.     

Affective pictures processing is reflected by an increased long-distance EEG connectivity

Analysis of affective picture processing by means of EEG has invaded the literature. The methodology of event-related EEG coherence is one of the essential methods used to analyze functional connectivity. The aims of the present study are to find out the long range EEG connectivity changes in perception of different affective pictures and analyze gender differences in these long range connected networks. EEGs of 28 healthy subjects (14 female) were recorded at 32 locations. The participants passively viewed emotional pictures (IAPS, unpleasant, pleasant, neutral). The long-distance intra-hemispheric event-related coherence was analyzed for delta (1–3.5 Hz), theta (4–7.5 Hz), and alpha (8–13 Hz) frequency ranges for F₃–T₇, F₄–T₈, F₃–TP₇, F₄–TP₈, F₃–P₃, F₄–P₄, F₃–O₁, F₄–O₂, C₃–O₁, C₄–O₂ electrode pairs. Unpleasant pictures elicited significantly higher delta coherence values than neutral pictures (p < 0.05), over fronto-parietal, fronto-occipital, and centro-occipital electrode pairs. Furthermore, unpleasant pictures elicited higher theta coherence values than pleasant (p < 0.05) and neutral pictures (p < 0.05). The present study showed that female subjects had higher delta (p < 0.05) and theta (p < 0.05) coherence values than male subjects. This difference was observed more for emotional pictures than for neutral pictures. This study showed that the brain connectivity was higher during emotional pictures than neutral pictures. Females had higher connectivity between different parts of the brain than males during emotional processes. According to these results, we may comment that increased valence and arousal caused increased brain activity. It seems that not just single sources but functional networks were also activated during perception of emotional pictures.     

Prediction of enteric methane emissions from Holstein dairy cows fed various forage sources

Milk fatty acid (FA) profile has been previously used as a predictor of enteric CH₄output in dairy cows fed diets supplemented with plant oils, which can potentially impact ruminal fermentation. The objective of this study was to investigate the relationships between milk FA and enteric CH₄ emissions in lactating dairy cows fed different types of forages in the context of commonly fed diets. A total of 81 observations from three separate 3×3 Latin square design (32-day periods) experiments including a total of 27 lactating cows (96±27 days in milk; mean±SD) were used. Dietary forages were included at 60% of ration dry matter and were as follows: (1) 100% corn silage, (2) 100% alfalfa silage, (3) 100% barley silage, (4) 100% timothy silage, (5) 50 : 50 mix of corn and alfalfa silages, (6) 50 : 50 mix of barley and corn silages and (7) 50 : 50 mix of timothy and alfalfa silages. Enteric CH₄output was measured using respiration chambers during 3 consecutive days. Milk was sampled during the last 7 days of each period and analyzed for components and FA profile. Test variables included dry matter intake (DMI; kg/day), NDF (%), ether extract (%), milk yield (kg/day), milk components (%) and individual milk FA (% of total FA). Candidate multivariate models were obtained using the Least Absolute Shrinkage and Selection Operator and Least-Angle Regression methods based on the Schwarz Bayesian Criterion. Data were then fitted into a random regression using the MIXED procedure including the random effects of cow, period and study. A positive correlation was observed between CH₄ and DMI (r=0.59,P<0.001), whereas negative associations were observed between CH₄ and cis9-17:1 (r=−0.58, P<0.001), and trans8, cis13-18:2 (r=−0.51,P<0.001). Three different candidate models were selected and the best fit candidate model predicted CH₄ with a coefficient of determination of 0.84 after correction for cow, period and study effects and was: CH₄ (g/day)=319.7−57.4×15:0−13.8×cis9-17:1−39.5×trans10-18:1−59.9×cis11-18:1−253.1×trans8, cis12-18:2−642.7×trans8, cis13-18:2−195.7×trans11, cis15-18:2+16.5×DMI. Overall and linear prediction biases of all models were not significant (P>0.19). Milk FA profile and DMI can be used to predict CH₄emissions in dairy cows across a wide range of dietary forage sources     

Interleukin 6 Accelerates Mortality by Promoting the Progression of the Systemic Lupus Erythematosus-Like Disease of BXSB.Yaa Mice

IL6 is a multifunctional cytokine that drives terminal B cell differentiation and secretion of immunoglobulins. IL6 also cooperates with IL21 to promote differentiation of CD4⁺ T follicular helper cells (T_FH). Elevated serum levels of IL6 correlate with disease flares in patients with systemic lupus erythematosus (SLE). We previously reported that IL21 produced by T_FH plays a critical role in the development of the SLE-like disease of BXSB.Yaa mice. To examine the possible contributions of IL6 to disease, we compared disease parameters in IL6-deficient and IL6-competent BXSB.Yaa mice. We report that survival of IL6-deficient BXSB.Yaa mice was significantly prolonged in association with significant reductions in a variety of autoimmune manifestations. Moreover, B cells stimulated by co-engagement of TLR7 and B cell receptor (BCR) produced high levels of IL6 that was further augmented by stimulation with Type I interferon (IFN1). Importantly, the frequencies of T_FH and serum levels of IL21 were significantly reduced in IL6-deficient mice. These findings suggest that high-level production of IL6 by B cells induced by integrated signaling from the IFN1 receptor, TLR7 and BCR promotes the differentiation of IL21-secreting T_FH in a signaling sequence that drives the lethal autoimmune disease of BXSB.Yaa mice.     

Helicobacter pylori Infection Induces Anemia,Depletes Serum Iron Storage,and Alters Local Iron-Related and Adult Brain Gene Expression in Male INS-GAS Mice

Iron deficiency anemia (IDA) affects > 500 million people worldwide, and is linked to impaired cognitive development and function in children. Helicobacter pylori, a class 1 carcinogen, infects about half of the world’s population, thus creating a high likelihood of overlapping risk. This study determined the effect of H. pylori infection on iron homeostasis in INS-GAS mice. Two replicates of INS-GAS/FVB male mice (n = 9-12/group) were dosed with H. pylori (Hp) strain SS1 or sham dosed at 6–9 weeks of age, and were necropsied at 27–29 weeks of age. Hematologic and serum iron parameters were evaluated, as was gene expression in gastric and brain tissues. Serum ferritin was lower in Hp SS1-infected mice than uninfected mice (p < 0.0001). Infected mice had a lower red blood cell count (p<0.0001), hematocrit (p < 0.001), and hemoglobin concentration (p <0.0001) than uninfected mice. Relative expression of gastric hepcidin antimicrobial peptide (Hamp) was downregulated in mice infected with Hp SS1 compared to sham-dosed controls (p<0.001). Expression of bone morphogenic protein 4 (Bmp4), a growth factor upstream of hepcidin, was downregulated in gastric tissue of Hp SS1-infected mice (p<0.001). Hp SS1-infected mice had downregulated brain expression of tyrosine hydroxylase (Th) (p = 0.02). Expression of iron-responsive genes involved in myelination (myelin basic protein (Mbp) and proteolipid protein 2 (Plp2)) was downregulated in infected mice (p = 0.001 and p = 0.02). Expression of synaptic plasticity markers (brain derived neurotrophic factor 3 (Bdnf3), Psd95 (a membrane associated guanylate kinase), and insulin-like growth factor 1 (Igf1)) was also downregulated in Hp SS1-infected mice (p = 0.09, p = 0.04, p = 0.02 respectively). Infection of male INS-GAS mice with Hp SS1, without concurrent dietary iron deficiency, depleted serum ferritin, deregulated gastric and hepatic expression of iron regulatory genes, and altered iron-dependent neural processes. The use of Hp SS1-infected INS-GAS mice will be an appropriate animal model for further study of the effects of concurrent H. pylori infection and anemia on iron homeostasis and adult iron-dependent brain gene expression.