Food fat gets distributed in the blood and therefore may be removed from the blood. Fat removal is simple and efficient using the blood circulation through the blood fat removal column described herein. The resulting body weight after 20 blood fat removal procedures might be 230 lbs. The patient has reached her body size fitting the suits purchased before the weight accumulation
{"title":"Body Fat Removal","authors":"Michael V. Tyurin","doi":"10.55124/ijde.v1i1.150","DOIUrl":"https://doi.org/10.55124/ijde.v1i1.150","url":null,"abstract":"Food fat gets distributed in the blood and therefore may be removed from the blood. Fat removal is simple and efficient using the blood circulation through the blood fat removal column described herein. The resulting body weight after 20 blood fat removal procedures might be 230 lbs. The patient has reached her body size fitting the suits purchased before the weight accumulation","PeriodicalId":163207,"journal":{"name":"International Journal of Endocrinology: Diabetes and Metabolism","volume":"10 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115486026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
We have detected the changes in the molecular biologic properties of the lascobacilus strain Lactibacillus buchneri NRRLB1837 after its storage in the lyophilized state using some patented lyophilization media. The change noticed was the 3 orders of magnitude decrease of the natural competence for the genetic transformation capability of said lac-tobacillus strain. Using noted difference we have transformed L. buchneri NRRLB1837 with the largest plasmid of Lactobacillus casei MT205, possessing strong antibacterial activity against gram-positive, gram-negative bacteria and Candida albicans. The trans formants became resistant to Erythromycin (30 mcg/ml) and also had strong antibacterial activity, just like L. casei MT205, which was extractable from the concentrated by the evaporation cultural fluid of the respective transformants, The role of the lyophilization medium on the molecular biological properties of the lactobacilli stored lyophilized is dis-cussed. Introduction We have noticed about three orders in magnitude difference in the natural transfroma-tion capability in the strain of Lactobacillus buchneri NRRLB1837 depending on the par-ticular conditions of its lyophilization. Said difference is esssential, since sometimes the natural capability to uptake the foreign plasmid DNA gets below the threshold of de-tection and may not be noticed. We have detected, that for the strain of Lactobacillus buchneri NRRLB1837 such activity is the highest, if the lyophilization was performed using standard lyophilization medium comprising only sucrose and gelatin, but not the 0.1 M Trtis (pH8.0) and 10% adonit [1]. Routinely, deep freezers (-80oC) are used for the prolonged storage of the microbial cultures under the normal room temperatures. However, most of the shipments are performed with the microbial cultures, lyophilized for the long-term storage. We have noticed significant difference in the natural transforma-tion capability of Lactobacillus buchneri NRRLB1837 depending on the composition of the medium for its freeze drying. In particular, we noticed significant decrease of the natural genetic transformation capability in Lactobacillus buchneri NRRLB1837 depend-ing on the compositon of the medium for its freeze drying. The decrease of that capabil-ity by three orders of magnitude was observed for the freeze dryed culture of said lacto- bacilli freeze dryed in our patented medium for the freeze drying [1]. The core of our patent was the neutralization of the organic acids produced enormously by the intestinal lactobacilli by the 0.1 M Tris buffer (pH8.0) and the lyophilization viability was substan-tially (by 90%) increased when we added some cryoprotective sugar up to 10 vol % to said 0.1 M Tris buffer (autoclaved at 120oC for 30 min). Today we have faced the fact, that the natural competence of the plasmid DNA uptake reported by us earlier [2] was accompanied with the decreased frequency of the genetic transfer for the plasmid DNA
bucheri NRRLB1837，该质粒立即提供了4 kb、8 kb、12 kb、24 kb大小的质粒DNA分子，而不是用于干酪乳杆菌MT205转化质粒的原始大小(约120 kb[4])。讨论了冻干培养基组成对冻干后保存的乳酸菌分子生物学特性的影响。材料与方法在Lactobacillus MRS Medium[4]培养基中培养乳酸菌，维持其生长、培养基、选择性抗生素。对于平板，将1.25%的琼脂加入MRS培养基中，然后在120℃下高压灭菌30分钟。为了选择带有质粒DNA pCB20的转化子，我们在MRS琼脂中分别加入红霉素(Em)和林可霉素(Lm)，各20mcg/ml，然后进行高压灭菌并冷却至45℃，然后倒入100 mm培养皿[6]。将抗生素粉末按100 mg /支无菌加入无菌Eppendorf管[7]中，并按1 ml /支向无菌Eppendorf管中加入无菌ddH2O。在层流柜[8]中干燥所倒的培养皿，接种按所述[3]制备的天然转化菌。为了在培养皿表面传播变形菌，只使用无菌塑料环。为便于计算转化效率，只使用自然转化样品的七、六、五十进制稀释倍数。为了制备所述自然变压器样品的十进制稀释液，我们使用了db[10]。DB组成:0.1 M Tris, 0.15 M NaCl, 1.0 M尿素，10 mM CaCl2, 0.1 M一水柠檬酸，5 g牛血清白蛋白，1.0 g半胱氨酸-盐酸(pH 6.00)。用高压灭菌法在1atm下灭菌30min，用900微升[10]将DB分散到无菌的1.5 ml埃彭多夫管中。将得到的0.1 ml自然转化样品转移到含有900微升DB(所述样品的第二次稀释)的第二个Eppendorf管中，以此类推，以便对志愿者的粪便进行电镀十倍稀释。稀释粪便的步骤如下。将自然转化样品第一次稀释后的100微升等分液[11]加入到第二个1.5ml的Eppendorf管中，加入900微升DB等，分别进行第二次、第三次、第四次、第五次、第六次、第七次等稀释。从最后一次稀释受体细胞/自然转化体开始接种，以节省塑料微生物循环的用量。受体细胞的浓度在上述实验的五次重复中确定(为了适当的实验统计目的)。为此目的，如上所述制备了受体细胞的十进制稀释液，另外添加了受体细胞的第七、第八和第九稀释液。从最后一次稀释受体细胞开始进行接种，以节省使用的微生物循环量。如所述重复冻干。为了对我们的数据进行适当的统计分析，我们将布氏乳杆菌NRRLB 1837的培养物在俄罗斯科学院的收藏中保存了五次，我们重复了冻干过程，然后将相同的乳酸菌培养物冻干，因为它写在我们的专利中。该菌株从冻干状态中恢复，并用于实验，以揭示在俄罗斯科学院条件下冻干后自然转化能力的频率，并根据我们的专利# 1652336[1]。遗传转化前凝胶中提取的最大质粒DNA(约120 kb)的布氏乳杆菌NRRLB1837转化72h培养液的浓度，在100 ml MRS肉液中，37℃下培养72h;培养液在真空蒸发器[16]中进行真空蒸发。结果在5个独立实验中确定了受体细胞的浓度，对数据进行了适当的统计分析。所以，在我们所有的实验中，浓度从9 × 109到7 × 109不等。利用标准质粒DNA pCB20对布氏乳杆菌NRRLB1837进行遗传转化的自然转化效率评价用标准质粒DNA pCB20转染布氏乳杆菌NRRLB1837的受体细胞，发现了布氏乳杆菌NRRLB1837的emlmr抗性转化子，每用于该实验的受体细胞数量的emlmr抗性转化子的频率为9x10-5(数据为5个独立的遗传转化实验，以进行适当的统计评估)。L.细胞的遗传转化。
{"title":"Natural Competence for Transformation in Lactobacili","authors":"M. Tyurin","doi":"10.55124/ijde.v1i1.119","DOIUrl":"https://doi.org/10.55124/ijde.v1i1.119","url":null,"abstract":"We have detected the changes in the molecular biologic properties of the lascobacilus strain Lactibacillus buchneri NRRLB1837 after its storage in the lyophilized state using some patented lyophilization media. The change noticed was the 3 orders of magnitude decrease of the natural competence for the genetic transformation capability of said lac-tobacillus strain. Using noted difference we have transformed L. buchneri NRRLB1837 with the largest plasmid of Lactobacillus casei MT205, possessing strong antibacterial activity against gram-positive, gram-negative bacteria and Candida albicans. The trans formants became resistant to Erythromycin (30 mcg/ml) and also had strong antibacterial activity, just like L. casei MT205, which was extractable from the concentrated by the evaporation cultural fluid of the respective transformants, The role of the lyophilization medium on the molecular biological properties of the lactobacilli stored lyophilized is dis-cussed. \u0000Introduction \u0000We have noticed about three orders in magnitude difference in the natural transfroma-tion capability in the strain of Lactobacillus buchneri NRRLB1837 depending on the par-ticular conditions of its lyophilization. Said difference is esssential, since sometimes the natural capability to uptake the foreign plasmid DNA gets below the threshold of de-tection and may not be noticed. We have detected, that for the strain of Lactobacillus buchneri NRRLB1837 such activity is the highest, if the lyophilization was performed using standard lyophilization medium comprising only sucrose and gelatin, but not the 0.1 M Trtis (pH8.0) and 10% adonit [1]. Routinely, deep freezers (-80oC) are used for the prolonged storage of the microbial cultures under the normal room temperatures. However, most of the shipments are performed with the microbial cultures, lyophilized for the long-term storage. We have noticed significant difference in the natural transforma-tion capability of Lactobacillus buchneri NRRLB1837 depending on the composition of the medium for its freeze drying. In particular, we noticed significant decrease of the natural genetic transformation capability in Lactobacillus buchneri NRRLB1837 depend-ing on the compositon of the medium for its freeze drying. The decrease of that capabil-ity by three orders of magnitude was observed for the freeze dryed culture of said lacto- \u0000 \u0000bacilli freeze dryed in our patented medium for the freeze drying [1]. The core of our patent was the neutralization of the organic acids produced enormously by the intestinal lactobacilli by the 0.1 M Tris buffer (pH8.0) and the lyophilization viability was substan-tially (by 90%) increased when we added some cryoprotective sugar up to 10 vol % to said 0.1 M Tris buffer (autoclaved at 120oC for 30 min). Today we have faced the fact, that the natural competence of the plasmid DNA uptake reported by us earlier [2] was accompanied with the decreased frequency of the genetic transfer for the plasmid DNA ","PeriodicalId":163207,"journal":{"name":"International Journal of Endocrinology: Diabetes and Metabolism","volume":"1202 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124793818","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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