Fusobacterium nucleatum-induced imbalance in microbiome-derived butyric acid levels promotes the occurrence and development of colorectal cancer

BACKGROUND Colorectal cancer(CRC)ranks among the most prevalent malignant tumors globally.Recent reports suggest that Fusobacterium nucleatum(F.nucleatum)contributes to the initiation,progression,and prognosis of CRC.Butyrate,a short-chain fatty acid derived from the bacterial fermentation of soluble dietary fiber,is known to inhibit various cancers.This study is designed to explore whether F.nucleatum influences the onset and progression of CRC by impacting the intestinal metabolite butyric acid.AIM To investigate the mechanism by which F.nucleatum affects CRC occurrence and development.METHODS Alterations in the gut microbiota of BALB/c mice were observed following the oral administration of F.nucleatum.Additionally,DLD-1 and HCT116 cell lines were exposed to sodium butyrate(NaB)and F.nucleatum in vitro to examine the effects on proliferative proteins and mitochondrial function.RESULTS Our research indicates that the prevalence of F.nucleatum in fecal samples from CRC patients is significantly greater than in healthy counterparts,while the prevalence of butyrate-producing bacteria is notably lower.In mice colonized with F.nucleatum,the population of butyrate-producing bacteria decreased,resulting in altered levels of butyric acid,a key intestinal metabolite of butyrate.Exposure to NaB can impair mitochondrial morphology and diminish mitochondrial membrane potential in DLD-1 and HCT116 CRC cells.Consequently,this leads to modulated production of adenosine triphosphate and reactive oxygen species,thereby inhibiting cancer cell prolif-eration.Additionally,NaB triggers the adenosine monophosphate-activated protein kinase(AMPK)signaling pathway,blocks the cell cycle in HCT116 and DLD-1 cells,and curtails the proliferation of CRC cells.The combined presence of F.nucleatum and NaB attenuated the effects of the latter.By employing small interfering RNA to suppress AMPK,it was demonstrated that AMPK is essential for NaB’s inhibition of CRC cell proliferation.CONCLUSION F.nucleatum can promote cancer progression through its inhibitory effect on butyric acid,via the AMPK signaling pathway.

Cortical and subcortical gamma amino acid butyric acid deficits in anxiety and stress disorders:Clinical implications

Anxiety and stress disorders are a major public health issue. However, their pathophysiology is still unclear. The gamma amino acid butyric acid(GABA) neurochemical system has been strongly implicated in their pathogenesis and treatment by numerous preclinical and clinical studies, the most recent of which have been highlighted and critical review in this paper. Changes in cortical GABA appear related to normal personality styles and responses to stress. While there is accumulating animal and human neuroimaging evidence of cortical and subcortical GABA deficits across a number of anxiety conditions, a clear pattern of findings in specific brain regions for a given disorder is yet to emerge. Neuropsychiatric conditions with anxiety as a clinical feature may have GABA deficits as an underlying feature. Different classes of anxiolytic therapies support GABA function, and this may be an area in which newer GABA neuroimaging techniques could soon offer more personalized therapy. Novel GABAergic pharmacotherapies in development offer potential improvements over current therapies in reducing sedative and physiologic dependency effects, while offering rapid anxiolysis.

Seed Priming with Beta-Amino Butyric Acid Improves Abiotic Stress Tolerance in Rice Seedlings

We studied the influence of seed priming with beta-amino butyric acid（BABA） on the growth, physiological and biochemical parameters of seedlings with varied abiotic stress tolerance, which were raised and grown under unstressed and stressed（NaCl/PEG-6000） conditions. Under stressed conditions, the growth of rice seedlings was less when compared to control plants. After BABA priming, the seedling growth increased both under unstressed and stressed conditions as compared to the respective controls. BABA priming of rice seeds caused increase in the photosynthetic pigment content of the leaves, modified the chlorophyll a fluorescence related parameters and also enhanced the photosystem activities of seedlings when compared to their respective non-primed controls. BABA priming also caused increased mitochondrial activities of the rice seedlings. Moreover, BABA priming significantly reduced malondialdehyde content in the seedlings and also resulted in accumulation of proline especially in the NaCl tolerant variety Vyttila 6. BABA seed priming also enhanced the activity of nitrate reductase enzyme and activities of antioxidant enzymes like guaiacol peroxidase and superoxide dismutase. The presence of BABA was detected by high performance thin layer chromatography analysis in the rice seeds whereas in the seedlings it was not detected. Thus, it can be inferred that the seed priming effect of BABA mainly occurred within the seeds, which was further carried to the seedlings. It is concluded that BABA priming of seeds improved the drought and salinity stress tolerance of all the three rice varieties and it was significantly evident in the drought tolerant variety Vaisakh and NaCl tolerant variety Vyttila 6, when compared to the stress sensitive variety Neeraja.

Batch and fed-batch production of butyric acid by Clostridium butyricum ZJUCB

The production of butyric acid by Clostridium butyricum ZJUCB at various pH values was investigated. In order to study the effect of pH on cell growth, butyric acid biosynthesis and reducing sugar consumption, different cultivation pH values ranging from 6.0 to 7.5 were evaluated in 5-L bioreactor. In controlled pH batch fermentation, the optimum pH for cell growth and butyric acid production was 6.5 with a cell yield of 3.65 g/L and butyric acid yield of 12.25 g/L. Based on these results, this study then compared batch and fed-batch fermentation of butyric acid production at pH 6.5. Maximum value (16.74 g/L) of butyric acid concentration was obtained in fed-batch fermentation compared to 12.25 g/L in batch fermentation. It was concluded that culti- vation under fed-batch fermentation mode could enhance butyric acid production significantly (P<0.01) by C. butyricum ZJUCB.

Efficient Separation of Butyric Acid by an Aqueous Two-phase System with Calcium Chloride

One of the bottlenecks for bioproduction of butyric acid as bulk chemical is the difficulty in separating butyric acid from the fermentation broth,compared with the petroleum-based chemical synthesis method.In the present work,a novel separation methodology was developed based on an aqueous two-phase system with inor-ganic salts.Calcium chloride was screened out for effective separation of butyric acid from butyric acid-water-salt systems.Within appropriate concentration range of butyric acid and salt,butyric acid was enriched in the upper phase and most of calcium ions remained in the lower phase.This"salting out"effect is very efficient to separate butyric acid from the simulated butyrate fermentation broth,which consists of butyric acid and acetic acid with concentration ratio of 4︰1,so that the final ratio of butyric acid/acetic acid in the upper phase is improved to 9.87. The aqueous two-phase system was used to separate butyric acid from the actual fermentation broth with satisfac-tory result.

Differential Effects of Butyric Acid on Mouse Skin Tumorigenesis

We studied the effects of butyric acid (BA) on mouse skin tumorigenesis using chronicanimal bioassays. Topical application of BA immediately after each treatment with 12-0-te-t radecanoyl phorbol-13-acetate (TPA) promoter' inhi bi ted skint umors. The effect was depe n -dent on the dose of BA applied. BA showed no marked inhibitory effect on either skin tumorinitiation or complete tumorigenesis induced by dimet hyl benzant hracene (DMBA ). Si nce t u -mor promotion reportedly involves epigenetic events whereas tumor initiation or complete tu-morigenesis takes place through genetic pathways, it is Possible that BA exerts its antitumori-genic effects mainly by altering the epigenetic events responsible for tumor promotion. The re-sults of the study could further be used to study the mechanism of action and modification ofantitumorigenic effects of BA in combination with other substances

Effect of Indole 3-Butyric Acid and Media Type on Adventitious Root Formation in Sheanut Tree (<i>Vitellaria paradoxa</i>C. F. Gaertn.) Stem Cuttings

Shea nuts play an important role in food security for rural folks within sub-Sahara Africa, serving as the main source of income for many people living in Northern Ghana. Unfortunately, the full economic potential of the Sheanut tree has not been fully realized due to the difficulty involved in its domestication. This difficulty in vegetatively propagating sheanut trees has greatly hindered its cultivation and the realization of its true economic potential. Two experiments were conducted to investigate the effects of rooting media and varying indole 3-butyric acid (IBA) concentrations on adventitious root formation in cuttings taken from coppiced sheanut trees. Results indicated that 3000 ppm produced significantly (p 0.05) better rooting (57.5%) than 5000 ppm (30%), 7000 ppm (45.0%) and the control (7.5%). Although the levels of soluble sugars (SS) and total free phenols (TFP) in the cutting were significantly (p 0.05) higher at the end of the experiment (after IBA treatment) compared to the start (prior to IBA treatment), the SS and TFP trends observed did not clearly explain the rooting differences found between the IBA levels investigated. Callus formation was significantly (p 0.05) higher (35.0%) in the control (no IBA). Generally, callus formation decreased with increasing IBA concentration. In the rooting media experiment, rooting was significantly (p 0.05) higher in the rice husk medium (35.0%) compared to that in the palm fiber (18.3%), saw dust (14.1%) and top soil (16.7%) media.

Liquid-Liquid Equilibria of Water + Butyric Acid + Nonanol Ternary System

Liquid-liquid equilibrium (LLE) data for the water + butyric acid + nonanol system have been determined experimentally at the temperatures of 298.15 K, 308.15 K and 318.15 K. Tie-line compositions were correlated by Othmer-Tobias method. The universal quasichemical functional group activity coefficient (UNIFAC) and modified UNIFAC methods were used to predict the phase equilibrium in the system using the interaction parameters between CH3, CH2, COOH, OH and H2O functional groups. Distribution coefficients and separation factors were evaluated for the immiscibility region.

Endogenous nitric oxide and hydrogen peroxide detection in indole-3-butyric acid-induced adventitious root formation in Camellia sinensis

Nitric oxide（NO）and hydrogen peroxide（H2O2）are essential signaling molecules with key roles in auxin induced adventitious root formation in many plants.However,whether they are the sole determinants for adventitious root formation is worth further study.In this study,endogenous NO and H2O2 were monitored in tea cutting with or without indole-3-butyric acid（IBA）treatment by using the fluorescent probes diaminofluorescein diacetate（DAF-2DA）and 2＇,7＇-dichlorodihydrofluorescein diacetate（DCF-DA）,respectively.The overproduction of NO and H2O2 was detected in the rooting parts of tea cuttings treated with or without IBA.But little NO and H2O2 was detected before the initiation phase of tea cuttings even with IBA treatment indicating that they might be not directly induced by IBA.Further carbon and nitrogen analysis found that the overproduction of NO and H2O2 were coincident with the consumption of soluble sugars and the assimilation of nitrogen.These results suggest that rooting phases should be taken into consideration with the hypothesis that auxin induces adventitious root formation via NO-and H2O2-dependent pathways and sink establishment might be a prerequisite for NO and H2O2 mediated adventitious root formation.

Rooting of Stem Cuttings with Different Indole 3 Butyric Acid (IBA) Treatments and Development of Micropropagation Protocol for <i>Piper betle</i>L. Node Culture

The present study, conducted during 2016 and 2017 seasons, aimed to investigate the effect of IBA on rooting of Piper betle L. stem cuttings (softwood and semi-hardwood). The experiment was undertaken in misting house field 2 UPM using the sand media to determine the adventitious roots initiation and development using the histological method. The cuttings were treated with different IBA concentrations (0, 500, 1000, 1500 and 2000 mg/L). The nodes explants were used in the development of a protocol for in vitro propagation of P. betle L., with different concentrations of Clorox with different times of immersion (20% Clorox 10 minutes, 30% Clorox 10 minutes, 20% Clorox 20 minutes, and 30% 20 minutes). In multiplication of the plantlets, Murashige and Skoog (MS) medium with different concentrations of BAP (0, 0.5, 1.0, 2.0 mg/L) were used to investigate the rooting of the explants. The results indicated that the types of the cuttings were different in the rooting capacity and the length of the roots. Moreover, it was found that in comparison with the control treatment, by a rise in the concentrations of the IBA, there was a significant upsurge in the rooting percentage, the root diameter, and the number of the roots. The results indicated that the types of cutting with 1000, 1500 and 2000 mg/L IBA perform better in the root percentage (100%) in the semi hardwood cuttings. The best results, however, were 2000 mg/L IBA in the semi hardwood cuttings, with the number of the roots to be 35.05, and the fresh weight of the roots to be 3.94 g, the dry weight of the roots to be 0.33 g, the length of the roots to be 391.88 cm, the roots diameter to be 1.21 mm, the surface area of the roots to be 121.83 cm2, and the root volume to be 2.99 cm3. Nonetheless, the optimal concentration of Clorox with the time immersion was 20% with the 20-minute immersion time, which produced a shoot induction percentage of 30% dead explants and a mean number of 70.00 shoots per explant and the optimal concentration of benzylaminopurine (BAP) at 1.0 mg/L. It is of note that a shoot induction percentage of 22.29% and a mean number of 4.1% number of auxiliary bud per treatment. P. betle shoots in MS medium without PGR MS (0.0) yielded a good rooting.

Growth inhibitory effect of 4-phenyl butyric acid on human gastric cancer cells is associated with cell cycle arrest

AIM: To investigate the growth effects of 4-phenyl butyric acid (PBA) on human gastric carcinoma cells and their mechanisms. METHODS: Moderately-differentiated human gastric carcinoma SGC-7901 and lowly-differentiated MGC-803 cells were treated with 5, 10, 20, 40, and 60 μmol/L PBA for 1-4 d. Cell proliferation was detected using the MTT colorimetric assay. Cell cycle distributions were examined using flow cytometry.RESULTS: The proliferation of gastric carcinoma cells was inhibited by PBA in a doseand time-dependent fashion. Flow cytometry showed that SGC-7901 cells treated with low concentrations of PBA were arrested at the G0/G1 phase, whereas cells treated with high concentrations of PBA were arrested at the G2/M phase. Although MGC-803 cells treated with low concentrations of PBA were also arrested at the G0/G1 phase, cells treated with high concentrations of PBA were arrested at the S phase. CONCLUSION: The growth inhibitory effect of PBA on gastric cancer cells is associated with alteration of the cell cycle. For moderately-differentiated gastric cancer cells, the cell cycle was arrested at the G0/G1 and G2/M phases. For lowly-differentiated gastric cancer cells, the cell cycle was arrested at the G0/G1 and S phases.

Quantitative analysis of butyric acid-induced nuclear ultrastructural alterations in cells of human lung giant cell carcinoma in vitro

The effects of butyric acid(BA)on the nuclear ultrastructure of humanlung giant cell carcinoma(Strain PLA-801 D)were observed with digital imageprocessing.It was found that the length of the nuclear circumference of the tu-mor cells incubated with 2mmol of BA was approximately equal to that of thecontrol whereas the nuclear area was increased by 1.4times,which implies thatthe nuclear profile tends to become more regular after BA treatment.In addition,the optical density of the nuclei of the experimental group decreased significantlyas compared with that of the control,which indicates that the chromatin in thenuclei was decreased by BA.It was concluded on the basis of the findings thatBA may have a biological effect of reverse-transformation on the malignant cells.

Effect of Indole Butyric Acid on the Transportation of Stored Calcium in Malus hupehensis Rhed. Seedling

Calcium （Ca） plays an important role in the metabolism of higher plants. Recently, research on Ca^2＋ in plants has been focused especially at the cellular and molecular levels. Uptake, transport, and distribution are also very important for Ca to accomplish its function at the whole-plant level. In this experiment, one-year-old apple seedlings （M. hupehensis Rehd.） were investigated to determine the distribution of stored Ca, the different forms of Ca, and Ca^2＋-ATPase activity after treatment with indole butyric acid （IBA）. The results showed that the total Ca measured in mature leaves and Ca^2＋- ATPase activity in tender leaves were higher compared with those in the control （CK）. Calcium nitrate and calcium chloride （ALe-Ca） and calcium phosphate and calcium carbonate （HAC-Ca） decreased in both mature leaves and shoots, whereas water-soluble calcium （H2O-Ca）, calcium pectate （NaCl-Ca）, and calcium oxalate （HCl-Ca） increased. The percentage of active calcium, calcium pectate, and water-soluble calcium increased, whereas the percentage of calcium phosphate and calcium carbonate decreased. When treated with IBA, calcium fractions and percentage of the different forms of Ca was enhanced in 40 part per million （ppm） IBA compared with 20 ppm IBA and water. The results indicated that IBA increased the percentage of both active calcium （NaCl-Ca and H2O-Ca） in tender shoots and boosted the transportation of stored Ca in plants. IBA promoted Ca^2＋-ATPase activity and Ca^2＋ uptake in tender shoots of M. hupehensis. It can improve the total Ca contents and the relative percentage of Ca.

Dietary <i>γ</i>-Aminobutyric Acid Shortens the Life Span of Stroke-Prone Spontaneously Hypertensive Rats

Dietary γ-amino butyric acid (GABA) has been suggested to decrease systolic blood pressure. This study aimed to ex-amine the effects of dietary GABA on the life span of stroke-prone spontaneously hypertensive rats (SHRSPs). In this study, life span was determined for SHRSPs provided 1% NaCl solution or 0.01% GABA in 1% NaCl solution as drinking water. The life span of the GABA-fed group (76.3 ± 1.65 days) was significantly shorter than that of the control group (81.6 ± 0.88 days). The results of this study may not be applicable to humans. Future studies will be necessary to elucidate the mechanism underlying this phenomenon.

Induced systemic resistance to Meloidogyne spp by <i>β</i>,amino butyric acid in tomato

β,amino butyric acid (BABA) induced resistance against?Meloigogyne?spp in tomato. Significantly (p?= 0.05) less, 41.11 second stage juveniles (J2) enter the roots of treated than, 116.66 J2 in untreated control plants. Root fresh and dry weight (Rfw, Rdw) were 2.87 and 0.12 g in treated compared with 4.78 and 0.30 g in nematode infected control plants respectively, 30 d after nematode inoculation. Sfw and Sdw were 8.62 and 0.60 g compared with 3.94 and 0.22 g in control plants. Foliage spray at 40 and 20 mM of BABA was more effective than 10 and 5 mM treatments. The former two concentrations recorded the lowest, average gall index, 2.11 compared with 3.33, 4.11, and 5 for the latter two concentrations and nematode infected control respectively. Results also indicated that treatments with BABA prior to nematode inoculation were superior in inducing resistance toMeloidogyne?spp over treatment at the time or after nematode inoculation.

响应面分析法优化一株窖泥源酪丁酸梭菌（Clostridium tyrobutyricum）RL1菌株产丁酸发酵培养基

从某浓香型白酒窖泥中筛选出1 株高产丁酸的酪丁酸梭菌(Clostridium tyrobutyricum)RL1 菌株,采用响应面分析法对其产丁酸培养基进行优化研究。首先采用Plackett-Burman 设计对基础培养基中9 个主要因素进行筛选,确定出对丁酸产量影响显著的3 个因素,依次为葡萄糖、碳酸钙和蛋白胨。对上述显著因素利用最陡爬坡试验逼近最大产酸响应区域,最后通过Box-Behnken 设计及响应面分析确定最佳培养基总成分组成为:葡萄糖85.29 g/L,酵母粉7.5 g/L,蛋白胨9.37 g/L,氯化钠7.5 g/L,乙酸钠4.5 g/L,磷酸氢二钾2.5 g/L,七水合硫酸镁0.3 g/L,硫酸铵1.5 g/L,七水合硫酸亚铁0.015 g/L,硫代乙醇酸钠0.5 g/L,碳酸钙11.6 g/L。优化后培养基的丁酸浓度为22.96 g/L,较优化前的14.23 g/L提高了61.35 %。

The specificity of ten non-digestible carbohydrates to enhance butyrate-producing bacteria and butyrate production in vitro fermentation

Butyrate and butyrate-producing bacteria are important indicators of gut microbial metabolism in human health.Ten non-digestible carbohydrates(NDCs),including inulin,fructooligosaccharide(FOS),oatsβ-glucans(OGS),oatsβ-glucan oligosaccharides(OGOS),Astragalus polysaccharides(APS),Astragalus oligosaccharides(AOS),xanthan gum oligosaccharides(XGOS),gellan gum oligosaccharides(GGOS),curdlan oligosaccharides(COS),and pullulan oligosaccharides(POS)were used to investigate NDC specifi city in modulating butyrate-producing bacteria and butyrate production in 48 h in vitro fermentation studies in combination with fecal inocula from 7 healthy donors and 11 patients with type 2 diabetes(T2D).We observed that the amount of these ten NDCs utilized depended on NDC structure and inter-individual gut microbial differences.XGOS and GGOS fermentations signifi cantly increased butyrate-producing bacteria(especially f_Lachnospiraceae)and butyric acid production.Furthermore,XGOS and GGOS fermentations showed a better ability to consistently modulate gut microbiota composition and metabolic properties between individuals of healthy donors or T2D patients when compared to inulin,FOS,APS,AOS,OGS,OGOS,COS and POS fermentation.This research indicated that xanthan gum and gellan gum oligosaccharides have strong specifi city to enhance butyrate-producing bacteria and butyrate production.

0.01%28-高芸苔素内酯·吲哚丁酸对寒地水稻生长及产量的影响

为明确0.01%28-高芸苔素内酯·吲哚丁酸对寒地水稻生长调节作用,采用随机区组试验设计,研究不同施药量对寒地水稻生长及产量的影响。结果表明,于分蘖期施用0.01%28-高芸苔素内酯·吲哚丁酸颗粒剂,能明显促进水稻分蘖,缩短始穗期至齐穗期天数,促进干物质积累,增强抗倒伏能力,提高水稻产量,对水稻生长安全。药剂施用量为22.5 kg/hm^(2)时,水稻穗粒数、结实率、千粒重最大,产量最高,是最佳施用量。

抑郁症患者治疗前后认知功能与前扣带回-氨基丁酸水平的关联

目的:探讨抑郁症患者治疗前后认知功能与前扣带回-氨基丁酸(GABA)水平的相关关系。方法:选取31例符合美国精神障碍诊断与统计手册第5版(DSM-5)抑郁症诊断标准的未服药患者和33例正常对照,患者人组后给予单一选择性5-羟色胺再摄取抑制剂治疗8周。采用认知功能成套测验(MCCB)评估认知功能,氢质子磁共振波谱成像技术检测前扣带回的GABA水平。结果:抑郁症组基线期前扣带回GABA相对于水的浓度(GABA+/W)低于对照组(P<0.05),治疗后升高(P<0.05);基线期抑郁症组前扣带回GABA+/W与言语学习和记忆以及视觉记忆和学习存在负相关(r=-0.40,P<0.05;r=-0.42,P<0.05);抑郁症组治疗前后前扣带回GABA+/W的差值与工作记忆评分的差值及推理和问题解决能力评分的差值存在正相关(r=0.58,P<0.05;r=0.66,P<0.05)。结论:抑郁症患者认知功能障碍可能与其抑郁、焦虑程度不相关,认知功能的改善可能与前扣带回GABA水平的升高有关。

外源IBA对无花果扦插苗抗氧化特性及IAA生物合成途径的影响

【目的】探讨适宜浓度吲哚丁酸(IBA)对无花果插穗生根萌芽、抗氧化性及生长素生物合成途径相关基因表达的影响,为其应用于无花果的育种、繁殖、推广和种植提供理论依据。【方法】以‘波姬红’无花果品种硬枝为插穗,观测不同质量浓度IBA(0,30,45,60,90 mg/L)处理下插穗生根性状、抗氧化特性,并对45 mg/L IBA处理及对照组的扦插枝条中段的腋芽进行转录组分析。【结果】(1)无花果插穗萌芽率和生根率在45 mg/L IBA处理时达到最大值,并与其他处理和对照差异显著。(2)随IBA浓度增加,插穗SOD和CAT活性先下降后上升,并均在45,60 mg/L IBA处理下显著低于对照,而POD活性无显著变化;各浓度IBA处理插穗中MDA和H2O2含量均显著高于对照,且45 mg/L IBA处理MDA显著低于其余处理。(3)45 mg/L IBA处理及对照组中共存在6 879个差异表达基因,KEGG富集显示有10个差异途径,GO富集分析表明生物学过程和分子功能为主要的生物学途径;与CAT、SOD相关的基因集中富集在过氧化物酶体通路上,POD相关基因则富集在苯丙烷生物合成通路中;IAA生物合成途径中代谢相关基因FcGH3显著上调表达,与信号转导相关基因FcAUX1、FcARG7和FcARF等显著下调表达。【结论】外源IBA处理会导致无花果插穗抗氧化酶和IAA生物合成途径中相关基因表达的差异变化,增强插穗抗逆性,促进插穗生根、萌芽、成苗,并以外源45 mg/L IBA促进效果最好。