Identification of Osmophilic Aspergillus Isolated from Rice and Their Radio-sensitivity

Osmophilic Aspergillus responsible for spoilage of rice, corn, milo and wheat have been isolated and identified. Fifteen strains were classified as members of the Aspergillus glaucus group, and were subdivided into A. ruber, A. repens, A. mangini, A. chevalieri and A. montevidensis. Nine strains were classified as members of the A. restrictus group, and were subdivided into A. gracilis, A. vitricolae and A. casiellus. The other 7 strains were classified as A. versicolor in the A. versicolor group, A. sulphureus in the A. ochraceus group, and A. niveus in the A. flavipes group.

All of dose-survival curves obtained with the conidia of 10 strains showed the sigmoidal type having the D₁₀ values between 18 and 30 krad. The survival curves obtained with the ascospores of A. glaucus group also showed the sigmoidal type having the D₁₀ values of 54 krad. Radio-sensitivity of the dry conidia was similar to that of the dry ascospores, having D₁₀ values between 50 to 58 krad.     

意大利蜜蜂产浆性状10个微卫星位点的遗传分析

我国培育成功的世界上蜂王浆产量最高的蜂种（Ea）是从20世纪30年代引进我国的意大利蜜蜂（Eb）中选育的。采用10个微卫星位点对蜂王浆高产蜜蜂、原种意大利蜜蜂（Ee）和本地意大利蜜蜂进行研究,以探明由于人工选择和地理隔离造成的其分子进化上的一些特征。结果,10个微卫星位点在3个蜂种中共扩增到96个等位基因,其中有48个等位基因是不同的,表明10个微卫星位点在3个蜂种中的高度多态性,而且由于人工选择和地理隔离已造成3蜂种在遗传基础上的一些分化。3蜂种的多态信息含量（PIC）分别为0．57、0．50、0．57,杂合度分别为0．60、0．57、0．61,二者均无显著差异。遗传距离的分析结果显示,Ee和Eb（0．14）、Eb和Ea（0．16）之间的遗传距离较近,而Ea和Ee（0.25）之间的遗传距离较远。等位基因频率的分析结果表明,6个位点的7个等位基因的频率（A29的159bp、A24的100bp和104bp、A7的110bp、A43的126bp、A14的221bp和A113的221bp）以Ee、Eb、Ea的顺序递增,Ea的这7个等位基因频率分别显著高于Ee和Eb。同时4个位点的4个等位基因的频率（A24的106bp、A43的140bp;A113的215bp和A14的219bp）以Ea、Eb、Ee的Ⅲ页序下降,Ea在这4个位点的频率分别显著低于Eb和Ee。这些位点的等位基因可能与蜂王浆产量有关。     

Stimulation of cell growth in the U-937 human myeloid cell line by honey royal jelly protein

Royal jelly was fractionated by ion-exchange chromatography and a protein (DIII protein) that had growth stimulating activity to the U-937 human myeloid cell line was obtained. The molecular weight of the DIII protein was 58 kDa on SDS-PAGE. The growth stimulating activity of the DIII protein was shown to be relatively heat and pH stable.     

不同人工代花粉对蜂群群势和生产性能的影响

早春将群势、蜂王年龄和质量基本一致的25群意大利蜜蜂Apis melliferaL.,随机分为5个处理,分别饲喂纯花粉和4种不同的人工代用花粉,观测它们对蜂群群势和生产性能的影响。结果表明,含有添加剂的各代用花粉组与纯花粉组的蜂群的蜂子数量差异均不显著,但均显著高于纯豆粕组(P<0.01);各组产蜜量随着纯花粉所占比例的减少而呈现递减的趋势,但饲喂含有60%花粉+40%豆粕+添加剂1组与纯花粉组没有明显的差异;含有添加剂的各代用花粉组的王台接受率均显著高于纯花粉组(P<0.01);饲喂含有添加剂的各代用花粉组与饲喂纯花粉组相比,王浆产量及蜂王浆中的10-HDA、水分、蛋白质的含量及酸度均没有差异。     

蜂王浆蛋白生物学功能的研究

蜂王浆是哺育蜂咽下腺与上颚腺分泌的供3日龄以内蜜蜂幼虫和蜂王食用的浆状物质,具有多种生物活性。王浆含有丰富的蛋白质,通过直接分离纯化王浆中的蛋白质分子,或通过王浆蛋白基因克隆表达以获得表达产物,进而研究王浆蛋白的功能。研究发现王浆中含有多种具有特定功能的蛋白质,如MRJP3具有免疫调节作用,Jelleine-I-IV及Royalisin具有抗菌作用,MRJP1具有抗肿瘤功能及可能的蜜蜂行为调节功能,57kDa蛋白及Apisimin的促细胞生长功能,57kDa蛋白具有抗疲劳功能等。随着王浆蛋白组分分离、基因克隆表达及其功能研究的深入,王浆蛋白将被应用到生物医药、细胞培养、组织工程等更多的研究领域。     

Some Aspects of the Enzyme Activities Involved in Coenzyme A Biosynthesis in Various Microorganisms

The distribution of the enzyme activities relating to CoA biosynthesis from pantothenic acid in various microorganisms and the effect of CoA on these activities are described.

High activities of partial reactions involved in CoA biosynthesis were surveyed in various type culture strains involving bacteria, actinomycetes, lactic acid bacteria, molds, and yeasts. Generally, higher activities were found in bacteria. CoA inhibited the phosphorylation of pantothenic acid, and resulted in a decrease of CoA production in all the CoA producing strains, while only a little inhibition by CoA was observed in the other reactions, and CoA production from pantothenic acid 4′-phosphate by Brevibacterium ammoniagenes IFO 12071 was not repressed even in the presence of 4mm of CoA. Extracellular excretion of the enzymes of CoA biosynthesis was observed when cells were in contact with sodium lauryl sulfate. Degrading activity against CoA and that against AMP were relatively lower in CoA producing strains when compared with those in other strains. It was confirmed that Brown’s route of CoA biosynthesis operates in Brevibacterium ammoniagenes IFO 12071.     

Antiapoptotic role of major royal jelly protein 8 of honeybee (Apis mellifera) venom

The dominant protein components of honeybee royal jelly (RJ) are major royal jelly proteins (MRJPs), which exhibit various biological properties. However, the biological basis of why bee venom contains MRJPs and what role MRJPs play in bee venom remains to be elucidated. This study reports the antiapoptotic role of MRJP 8 of Apis mellifera venom (AmMRJP 8) in melittin-treated mammalian cells. Recombinant AmMRJP 8 reduced caspase-3 activity and melittin-induced cell apoptosis. Additionally, recombinant AmMRJP 8 decreased the production levels of H₂O₂ and proinflammatory molecules. These results indicate that MRJP in bee venom plays a role in cell protection in bee venom-induced inflammatory responses.     

Antioxidant capacity of major royal jelly proteins of honeybee (Apis mellifera) royal jelly

Major royal jelly proteins (MRJPs) of honeybee royal jelly (RJ) exhibit antimicrobial and antioxidant activities. Although MRJPs of Apis mellifera RJ (AmMRJPs) responsible for antibacterial activity have been identified, AmMRJPs with antioxidant effects remain to be elucidated. Here we identified and compared the antioxidant activities of purified recombinant AmMRJPs 1–7, which are expressed in baculovirus-infected insect cells. Antioxidant assays of recombinant AmMRJPs 1–7 against H₂O₂ revealed that AmMRJPs reduce caspase-3 activity and oxidative stress-induced cell apoptosis and lead to increased cell viability. Consistent with these results, AmMRJPs 1–7 exhibit 1,1-diphenyl-2-picrylhydrazyl radical-scavenging activity and protect against oxidative DNA damage. These results indicate that AmMRJPs play a role as antioxidants in A. mellifera RJ.     

Identification of major royal jelly proteins in the brain of the honeybee Apis mellifera

The consumption of royal jelly (RJ) determines the differences between castes and behavioral development in the honeybee Apis mellifera. However, it is not known whether the proteins of RJ are related to these differences, or which proteins are responsible for the changes. To understand the functions of RJ proteins that are present in other tissues of the bee, in addition to hypopharyngeal gland, we used a polyclonal antibody anti-MRJP1 to investigate the presence of this protein in nervous system of honeybee. This study showed the presence of three polypeptides (p57, p70 and p128) in specific tissues of bee brain. Mushroom body, optic lobe and antennal lobe neuropils all contained proteins recognized by anti-MRJP1. Proteomic analysis showed that the three polypeptides are correlated with proteins of the MRJP family. p57 is correlated with MRJP1, p70 with MRJP3, while p128 may be an oligomeric form or a new polypeptide. Immunostaining of the brain and hypopharyngeal gland revealed differential expression of MRJPs in various brain regions and in different honeybee castes and subcastes. The identification and localization of these MRJPs contribute to the elucidation of the biological roles of this protein family.     

蜜蜂蜂王浆主蛋白(MRJPs)的研究进展

蜜蜂的蜂王浆主蛋白具有为蜂王和幼虫提供营养、影响蜂群社会行为及调节个体生理机能等作用,作为蜂王浆的主要成分对其他机体也可产生多方面的生物学功能。因此,近年来蜂王浆主蛋白的相关研究备受关注。本文针对蜂王浆主蛋白的发现、种类、功能、系统进化及其基因表达情况进行了系统综述。     

Ameliorative Effects of Propolis and Royal Jelly against CCl4-Induced Hepatotoxicity and Nephrotoxicity in Wistar Rats

Carbon tetrachloride (CCl₄) is known to have hepatotoxic and nephrotoxic effects. During the two-month CCl₄ exposure of Wistar rats, propolis extract (PE) and royal jelly (RJ) were added in order to test the potential protective effect against hepato-renal injury. Ketonuria, proteinuria, high creatinine and urea levels are the result of CCl₄-induced nephrotoxicity. Severe disorders of hematological indicators indicate anemia; high values of leukocytes indicate inflammatory condition. Cytogenetic impairments in hepatocytes, aggregation of platelets, and hypoproteinemia indicate severe liver impairment. Results suggest a more significant protective role of RJ compared to PE. Both extracts regulated proteinuria, ketonuria, hypoproteinemia and reduced platelet aggregation in the hepatic circulation. The increase in the number of erythrocytes (RBC) suggest protective effects against anemia; the decrease in the number of leukocytes can be linked to anti-inflammatory effects. PE and RJ have a beneficial effect against hepato-renal injury, anemia and anti-inflammatory conditions caused by CCl₄.     

Thiol Accumulation and Cysteine Desulfhydrase Activity in H2S-Fumigated Leaves and Leaf Homogenates of Cucurbit Plants

Fumigation of both, cucurbit plants and cucurbit leaf homogenateswith hydrogen sulfide (H₂S) resulted in an increase in solublethiol, mainly glutathione and cysteine. In leaf homogenatesthis increase was counteracted or prevented by the additionat 1 mM of inhibitors of pyridoxalphosphate dependent enzymesor of products of the cysteine desulfhydrase reaction. Thesecompounds inhibited cysteine desulfhydrase activity, but didnot severely affect O-acetylserine sulfhydrylase activity atthis concentration. These results provide circumstantial evidencethat cysteine desulfhydrase in its reverse reaction, but notO-acetylserine sulfhydrylase participates in the assimilationof atmospheric H₂S. (Received February 4, 1991; Accepted April 30, 1991)     

饲喂胡萝卜和蜂王浆对黄粉虫繁殖力的影响

采用多因素对比饲养试验,设计不同黄粉虫TemibriomolitorL.发育时期分别补饲胡萝卜、不同浓度的蜂王浆,研究胡萝卜和蜂王浆对黄粉虫繁殖力的影响。结果表明:各处理组比对照组羽化齐一,羽化率增高,产卵速度加快,总产卵量增多,良卵率、孵化率增高。幼虫期和成虫期都补饲2%的蜂王浆,可使产卵量极显著高于对照97.79%、孵化率极显著地高于对照8.90%;而只在成虫期补充2%的蜂王浆,也可使产卵量极显著高于对照90.65%、有效地提高孵化率6.66%。     

The evolution of food-producing glands in eusocial bees (Apoidea,Hymenoptera)1

A food-producing role for cephalic exocrine glands has arisen independently in both taxa of highly eusocial bees, Apis and Meliponini. With several exceptions, there is little evidence that food is produced by glands of solitary bees or by most bees at lower levels of sociality. We suggest that this association with sociality is due to four adaptive features of these glands: (1) food from the glands allows feces from queens and larvae to have a small volume, (2) the queen's fecundity can be increased, (3) nutrient recovery via cannibalism can be facilitated, and (4) rearing of emergency replacement queens is accelerated. Acceleration of the rearing of other castes and of queens in the normal process of colony fission is not clearly an advantage ascribed to these glands. Trophic eggs produced by meliponine colony workers are analogous to the secretions from food-producing glands in Meliponini and Apis workers.     

Quantification of Major Royal Jelly Protein 1 in Fresh Royal Jelly by Indirect Enzyme-Linked Immunosorbent Assay

Rabbit polyclonal antibody produced by a major royal jelly protein 1 (MRJP1) specific peptide reacted only with a MRJP1. Indirect ELISA with the antibody revealed a MRJP1 level of 4.12–4.67 g/100 g in different company's royal jelly, which almost agreed with that of a hexametric form of MRJP1 (apisin) measured by high performance liquid chromatography. These results suggest that MRJP1 exists mainly as apisin in royal jelly.     

Conservation of nutrients in larval tissue by cannibalizing honey bees

ABSTRACT. ¹⁴C-phenylalanine appeared in royal jelly secreted by worker honey bees ( Apis mellifera , L.) directly after eating ¹⁴-phenyl-alanine labelled larvae. The amount of labelled phenylalanine in royal jelly was associated directly with the volume of jelly produced. Loss of radio-label from the workers, in the form of O₂¹⁴C, increased when royal jelly secretion ended. These patterns show that conservation of nutrients in cannibalized tissue is enhanced when the cannibal bees are producing royal jelly. Specific activity of radiolabel in worker bee hypopharyngeal glands was higher than in thoracic muscle, showing selective movement within the bees of a nutrient acquired by cannibalism.     

乳腺癌干细胞和乳腺癌

肿瘤干细胞既包含干细胞的特性也包含肿瘤细胞的特性。乳腺癌起源于乳腺癌干细胞的说法能够合理地解释乳腺癌的不均一性及其治疗后的复发,这些变异的干细胞可能作为肿瘤预防策略的靶标。而且,由于乳腺癌干细胞能够抵抗辐射治疗和化学治疗,所以要想更好地治疗乳腺癌就需要寻找针对这些干细胞的靶标。我们综述了乳腺癌干细胞的发现、富集和分离、相关的信号途径,以及在乳腺癌治疗中的应用。     

Immunological Characterization of Honey Proteins and Identification of MRJP 1 as an IgE-Binding Protein

We encountered a fourth case of honey allergy in Japan. We characterized and identified the IgE-binding proteins in honey using the serum of a honey-allergenic patient. Immunoblot analysis revealed that IgE in the patient serum specifically bound to four proteins in each honey sample. At least three of these IgE-binding proteins were N-linked glycoproteins. To identify the 60-kDa IgE-binding protein in dandelion honey, the N-terminal sequences of the fragmented protein were analyzed, revealing the protein to be major royal jelly protein 1 (MRJP 1). Three IgE-binding proteins removed of N-linked oligosaccharide showed a large reduction in IgE-binding activity as compared with the intact protein. This suggests that the carbohydrates in the IgE-binding proteins are a major epitope for patient IgE.     

Histone demethylase KDM7A is required for stem cell maintenance and apoptosis inhibition in breast cancer

Histone demethylase KDM7A regulates neuronal differentiation and development in mammals. In this study, we found that KDM7A was also required for breast cancer stem cells (BCSCs) maintenance. Silencing KDM7A significantly reduced the BCSCs population and mamosphere formation in vitro, and inhibited breast tumor growth in vivo. Restoring KDM7A expression rescued the defect in stem cell maintenance. Our mechanism analysis suggested that KDM7A upregulated the stemness-associated factors KLF4 and c-MYC for BCSCs maintenance. In addition, KDM7A knockdown promoted apoptosis through decreasing BCL2 expression and BAD phosphorylation in breast cancer (BrCa). Furthermore, restoring KDM7A and BCL2 expression rescued apoptosis inhibition in breast cancer, suggesting that KDM7A inhibited apoptosis by upregulating the BCL2 level in breast cancer. In conclusion, KDM7A promotes cancer stem cell maintenance and apoptosis inhibition in breast cancer.     

Antimicrobial activity of major royal jelly protein 8 and 9 of honeybee (Apis mellifera) venom

Honeybee venom is a complex mixture of toxic components, including major royal jelly protein (MRJP) 8 and 9. MRJP 8 and MRJP 9 are allergens, and MRJP 8 reduces melittin-induced cell apoptosis. However, their functional roles are poorly understood, and their antimicrobial activities have not been determined. In this study, the antimicrobial role of MRJP 8 and MRJP 9 of honeybee (Apis mellifera) venom (AmMRJP 8 and AmMRJP 9) was demonstrated. The presence of AmMRJP 8 and AmMRJP 9 in the secreted venom was observed using antibodies against recombinant AmMRJP 8 and AmMRJP 9 produced in baculovirus-infected insect cells. Recombinant AmMRJP 8 and AmMRJP 9 exhibited an inhibitory activity against microbial serine proteases. Consistent with their inhibitory activity, they induced structural damage by binding to microbial surfaces, resulting in a broad-spectrum antimicrobial activity against bacteria and fungi. They had little effect on hemolysis. Therefore, AmMRJP 8 and AmMRJP 9 could function as antimicrobial agents in honeybee venom.