The role of routine post-natal abdominal ultrasound for newborns in a resource-poor setting: a longitudinal study

<p>Abstract</p> <p>Background-</p> <p>Neonatal abdominal ultrasound is usually performed in Nigeria to investigate neonatal symptoms rather than as a follow up to evaluate fetal abnormalities which were detected on prenatal ultrasound. The role of routine obstetric ultrasonography in the monitoring of pregnancy and identification of fetal malformations has partly contributed to lowering of fetal mortality rates. In Nigeria which has a high maternal and fetal mortality rate, many pregnant women do not have ante-natal care and not infrequently, women also deliver their babies at home and only bring the newborns to the clinics for immunization. Even when performed, most routine obstetric scans are not targeted towards the detection of fetal abnormalities.</p> <p>The aim of the present study is to evaluate the benefit of routinely performing abdominal scans on newborns with a view to detecting possible abnormalities which may have been missed ante-natally.</p> <p>Methods-</p> <p>This was a longitudinal study of 202 consecutive, apparently normal newborns. Routine clinical examination and abdominal ultrasound scans were performed on the babies by their mother's bedside, before discharge. Neonates with abnormal initial scans had follow-up scans.</p> <p>Results-</p> <p>There were 108 males and 94 females. There were 12 (5.9%) abnormal scans seen in five male and seven female neonates. Eleven of the twelve abnormalities were in the kidneys, six on the left and five on the right. Three of the four major renal anomalies- absent kidney, ectopic/pelvic kidney and two cases of severe hydronephrosis were however on the left side. There was one suprarenal abnormality on the right suspected to be a possible infected adrenal haemorrage. Nine of the abnormal cases reported for follow- up and of these, two cases had persistent severe abnormalities.</p> <p>Conclusions-</p> <p>This study demonstrated a 5.9% incidence of genito urinary anomalies on routine neonatal abdominal ultrasound in this small population. Routine obstetric USS is very useful but inadequate availability of skilled personnel and cost implications create great challenges in poor resource settings like Nigeria. However, awareness should be created so that parents who can afford such investigations can make informed decisions.</p

Membrane-Associated Heparan Sulfate Proteoglycan Is a Receptor for Adeno-Associated Virus Type 2 Virions

The human parvovirus adeno-associated virus (AAV) infects a broad range of cell types, including human, nonhuman primate, canine, murine, and avian. Although little is known about the initial events of virus infection, AAV is currently being developed as a vector for human gene therapy. Using defined mutant CHO cell lines and standard biochemical assays, we demonstrate that heparan sulfate proteoglycans mediate both AAV attachment to and infection of target cells. Competition experiments using heparin, a soluble receptor analog, demonstrated dose-dependent inhibition of AAV attachment and infection. Enzymatic removal of heparan but not chondroitin sulfate moieties from the cell surface greatly reduced AAV attachment and infectivity. Finally, mutant cell lines that do not produce heparan sulfate proteoglycans were significantly impaired for both AAV binding and infection. This is the first report that proteoglycan has a role in cellular attachment of a parvovirus. Together, these results demonstrate that membrane-associated heparan sulfate proteoglycan serves as the viral receptor for AAV type 2, and provide an explanation for the broad host range of AAV. Identification of heparan sulfate proteoglycan as a viral receptor should facilitate development of new reagents for virus purification and provide critical information on the use of AAV as a gene therapy vector

Isolation, Identification and Biochemical Characterization of Bioactive Peptides from Algae Protein Waste

Chlorella vulgaris has been the most popular edible microalgae without side effects. Algae essence is an industrial product derived from water extracts of microalgae, and high molecular weight algae protein is a by-product of algae essence production. More than 500 tons of algae essence byproduct produced in Taiwan every year, and re-processed into low economical-value animal feed. However, this algae essence byproduct might become an important protein source for the selection of novel ACE inhibitory peptides, antioxidative peptides and anticancer peptides by enzymatic hydrolysis. In this study, we screened bioactive peptides from algae protein digested with commercial enzymes. A bioactive hendeca-peptide was isolated from the pepsin hydrolysate of algae protein, and Edman degradation revealed its amino acid sequence to be Val-Glu-Cys-Tyr-Gly-Pro-Asn-Arg-Pro-Gln-Phe, the biofuctions were researched as below: 1.Researches on angiotensin I-converting enzyme (ACE) inhibitory activity The hendeca-peptide with IC50 value of 29.6 uM against ACE suggested a potent amount of ACE inhibitory activity compared with other peptides from the microalgae protein hydrolysates, which have a reported range between 11.4 and 315.3 uM. Inhibitory kinetics revealed a non-competitive binding mode. In addition, the hendeca-peptide completely retained its ACE inhibitory activity in a pH range of 2-10, temperatures of 20-100oC, as well as treated by a gastrointestinal enzyme in vitro, thus indicating its heat-, pH- and gastrointestinal enzyme stability. The combination of the biochemical properties of this isolated hendeca-peptide and a cheap algae protein resource make this process as an attractive alternative for producing high value product for blood pressure regulation as well as water and fluid balance. 2.Researches on antioxidative activity Algae protein was hydrolyzed using pepsin, and a potent antioxidative peptide of Val-Glu-Cys-Tyr-Gly-Pro-Asn-Arg-Pro-Gln-Phe was separated and isolated. The peptide could efficiently quench a variety of free radicals, including hydroxyl radical, superoxide radical, peroxyl radical, DPPH radical and ABTS radicals, and performed more efficiently than that observed for BHT, Trolox and peptides from marine protein sources in most cases. The peptide also has significant protective effects on DNA and prevents cellular damage caused by hydroxyl radicals. These results suggest that inexpensive algae protein could be a new alternative to produce antioxidative peptides. 3. Researches on AGS inhibitory activity The peptide fraction isolated from pepsin hydrolysate of algae protein was found to arrest the cell cycle and caused apoptosis in human gastric carcinoma cell lines (AGS); however no cytotoxicity was observed in human lung fibroblasts cell lines (WI-38) in vitro. The peptide fraction has a molecular weight of below 3 KD. The peptide fraction also revealed stronger antioxidative activity toward peroxyl radicals and LDL than natural antioxidant Trolox. A hendeca-peptide with an IC50 value against AGS cells of 256.4 uM was isolated from the peptide fraction and its amino acid sequence was Val-Glu-Cys-Tyr-Gly-Pro-Asn-Arg-Pro-Gln- Phe. The special amino acid sequence and structure of the peptide was important for anticancer effect compared to the synthetic peptide fragments. These results demonstrate that inexpensive algae protein could be a new alternative to produce anticancer peptides.微球藻為普遍可食用藻類，對人體並不具有任何不良的副作用。海藻精是利用水萃取微球藻所得之市售產品，而高分子量的海藻蛋白質則是伴隨海藻精生產製程中所產生的副產物。在臺灣每年有超過500噸的海藻精副產物因此而產生，此蛋白質大部分被重新乾燥製成低經濟價值的動物飼料販售及使用。因此海藻精副產物或許可藉由酵素水解而作為一個新的血管收縮轉換酵素抑制胜肽、抗氧化胜肽和抗癌胜肽的重要蛋白質來源。在本論文的研究中，我們利用商業化酵素水解海藻蛋白質並從中篩選具有生物活性的胜肽。結果顯示，可從海藻蛋白質之胃蛋白酶水解物中分離出一含有11個胺基酸的單一胜肽，此胜肽經Edman降解定序方法得知其胺基酸序列為Val-Glu-Cys-Tyr-Gly-Pro-Asn-Arg-Pro-Gln-Phe，其生理功能研究如下所示： 一、抑制血管收縮轉換酵素活性之研究 該胜肽於細胞體外試驗結果顯示其能有效抑制血管收縮轉換酵素之活性(IC50的抑制劑量為29.6 uM) ，此11胜肽相較於文獻中直接從海藻蛋白質水解物所分離出的胜肽 (IC50的抑制劑量範圍為11.4-315.3 uM) 是相當具有潛力的。在酵素抑制動力學的研究中，此胜肽對血管收縮轉換酵素抑制效果屬於非競爭型的抑制效應；除此之外，此11胜肽具有pH值穩定性 (pH 2—pH10)、熱穩定性 (40—100℃) 及腸胃酵素耐受性。因此由11胜肽的生化特性的結果顯示，低價值的海藻蛋白質可能是生產一個新的調節血壓的膳食補充品的選擇。 二、抗氧化活性之研究 從藻類蛋白質的胃蛋白酶水解物分離純化所得之Val-Glu-Cys-Tyr-Gly-Pro-Asn-Arg-Pro-Gln-Phe之胜肽，其細胞體外試驗結果顯示能夠有效的清除各種自由基，包括氫氧自由基、超氧自由基、過氧化自由基、DPPH自由基以及ABTS自由基；並且其清除自由基的效果較人工合成之抗氧化劑BHT、維生素E、維生素C和其它從海洋蛋白質來源所分離的胜肽為佳。此胜肽對氫氧自由基造成細胞及DNA的氧化傷害亦具有明顯的保護效果，並且對人類肺纖維細胞株 (WI-38細胞) 不具有細胞毐性等特性。本研究結果顯示，吾人可利用低價海藻蛋白質生產抗氧化胜肽。 三、抑制胃癌細胞活性之研究 藻類蛋白質的胃蛋白酶水解物經初歩的樹脂純化所得之胜肽區分物，在細胞體外試驗結果顯示此胜肽區分物可抑制胃腺癌細胞株 (AGS)的細胞週期及誘導其進行細胞自然凋亡作用，但對人類肺纖維細胞株 (WI-38細胞)不具有細胞毐性。在其它生化特性之研究方面，本研究所得之胜肽區分物大部分由小於3 kDa的小分子胜肽所組成且其清除peroxyl自由基及抑制低密度脂蛋白的氧化的效果較天然的抗氧化劑維生素E為佳。我們亦從此胜肽混合物中分離出Val-Glu-Cys-Tyr-Gly-Pro-Asn-Arg-Pro-Gln-Phe的單一胜肽，此胜肽於體外抑制AGS細胞效果之IC50劑量為256.4 uM﹔並且其特別的胺基酸組成和結構對抗癌效應是重要的。由以上之結果顯示，低價值的海藻蛋白質可作為新的生產抗癌胜肽的蛋白質之可能來源。Figure list of this dissertation…………………...….…….vi Table list of this dissertation……………………………...xi List of Abbreviation and Full name…………………………xiii Abstract (in Chinese)……………………………………………I Abstract (in English)…………………………………...……...IV 1. Introduction………………………………………….………….1 2. Literature Review 2.1. Peptides with angiotensin I-converting enzyme (ACE) inhibitive activity………………………………….……………..8 2.2. Peptides with antioxidative activity………………....11 2.3. Peptides with anticancer activity………………………14 3. Materials and Methods 3.1. Materials…………………………………………….…..……34 3.2. Preparation of enzymatic hydrolysate………………….35 3.3. Purification of bioactivity peptides from algae protein 3.3.1. Ammonium sulfate fractionation…………….………….36 3.3.2. Gel filtration chromatography………………….…….36 3.3.3. Ion exchange chromatography…………….…………….37 3.3.4. Reverse-phase high-performance chromatography (RP-HPLC)…………..…………...…………...…………37 3.4. Measurement of ACE inhibitory activity…...………..38 3.5. Determination of the inhibition pattern on ACE ……38 3.6. Trolox equivalent antioxidant capacity (TEAC) assay-39 3.7. 1. 1-diphenyl-2-picrylhydrazyl (DPPH) radicals scavenging activity assay………………………………………..39 3.8. Hydroxyl radicals scavenging ability assa…………….40 3.9. Superoxide radicals scavenging ability assay………..41 3.10. Oxygen radical absorbance capacity (ORAC) assay……….....42 3.11. Antioxidative activity toward copper-mediated oxidation of human low-density lipoprotein (LDL)…………...…………...42 3.12. Protection effect on oxidation-induced DNA damage………...43 3.13. Protection effect on oxidation-induced cell damage……..……43 3.14. Cytotoxicity of human gastric carcinoma AGS cell…………..44 3.15. Cell cycle kinetics analysis ……………………….…………..44 3.16. Molecular weight distribution analysis…………….……...…..45 3.17. Cytotoxicity of human normal lung WI-38 cells ………...…...45 3.18. Determination of amino acid sequence…………...…………...46 3.19. Stability of temperature and gastrointestinal enzyme ………...46 3.20. Statistical analysis ……………………………………...…......47 4. Results and discussion 4.1. ACE inhibitory activity of the peptides 4.1.1. Preparation of ACE inhibitory peptides …………….......48 4.1.2. Purification of ACE inhibitory peptides …………….......48 4.1.3. Determination of amino acid sequence ………….….......50 4.1.4. Structure-activity correlation studies …………................51 4.1.5. Stability of the hendeca-peptide…………........................52 4.1.6. Determination of the inhibition pattern on ACE………...54 4.1.7. Conclusion of the ACE inhibitory peptides.......................55 4.2. Antioxidant properties of the peptides 4.2.1. Preparation of antioxidant peptides………………….......64 4.2.2. Purification of antioxidative peptide ………………........65 4.2.3. Antioxidative characterization 4.2.3.1. ABTS radicals scavenging activity…………......67 4.2.3.2. DPPH radicals scavenging activity………..........67 4.2.3.3. Hydroxyl radicals scavenging activity……….....68 4.2.3.4. Superoxide radicals scavenging activity………..69 4.2.3.5. Peroxyl radicals scavenging activity………........71 4.2.3.6. Protection effect on oxidation-induced DNA damage……….....................................................73 4.2.3.7.Protection effect on oxidation-induced cell damage………………………………………….74 4.2.4. Conclusion of antioxidative peptide..................................75 4.3. AGS inhibitory activity of the peptides 4.3.1. Preparation and purification of AGS inhibitory peptides ………………………………………………...……..…84 4.3.2. Cytotoxicity activity against AGS cells ……………...…85 4.3.3. Effect on AGS cell cycle……………………………...…86 4.3.4. Antioxidant characterization 4.3.4.1. ABTS radicals scavenging activity…………..…88 4.3.4.2. Peroxy radicals scavenging activity……..……...88 4.3.4.3. Protection against LDL oxidation…..………......89 4.3.5. Molecular weight distribution………...............................90 4.3.6. Cytotoxicity activity of the hendeca-peptide against AGS Cells.…….........................................................................91 4.3.7. Conclusion of the anticancer peptide.............93 V. Future prospects .............................................................................101 VI. References.......................................................................................102 VII. Appendix 7.1. I. Chuan Sheih, Tony J. Fang, Tung-Kung Wu, 2009. Isolation and characterisation of a novel angiotensin I-converting enzyme (ACE) inhibitory peptide from the algae protein waste. Food Chemistry 115:279-284. 7.2. I-Chuan Sheih, Tung-Kung Wu, Tony J. Fang, 2009. Antioxidant properties of a new antioxidative peptide from algae protein waste hydrolysate in different oxidation systems. Bioresource Technology 100: 3419-3425

Isolation and characterisation of a novel angiotensin I-converting enzyme (ACE) inhibitory peptide from the algae protein waste

A hendeca-peptide with angiotensin I-converting enzyme (ACE) inhibitory activity was isolated from the pepsin hydrolysate of algae protein waste, a mass-produced industrial by-product of an algae essence from microalgae, Chlorella vulgaris. Edman degradation revealed its amino acid sequence to be Val-Glu-Cys-Tyr-Gly-Pro-Asn-Ai-g-Pro-Gln-Phe. Inhibitory kinetics revealed a non-competitive binding made with IC(50) Value against ACE of 29.6 mu M, suggesting a potent amount of ACE inhibitory activity compared with other peptides from the microalgae protein hydrolysates which have a reported range between 11.4 and 315.3 mu M. In addition, the purified hendeca-peptide completely retained its ACE inhibitory activity at a pH range of 2-10, temperatures of 40-100 degrees C, as well as after treatments in vitro by a gastrointestinal enzyme, thus indicating its heat- and pH-stability. The combination of the biochemical properties of this isolated hendeca-peptide and a cheap algae protein resource make an attractive alternative for producing a high value product for blood pressure regulation as well as water and fluid balance. (C) 2008 Elsevier Ltd. All rights reserved

AN ISOLATED PEPTIDE AND THE PREPARATION PROCESSES AND APPLICATIONS THEREOF

本發明揭露一種分離自一小球藻屬藻類(algae ofChlorellagenus)的肽，它具有血管緊縮素I-轉化酵素(angiotensin I-converting enzyme)抑制活性、抗氧化活性、抗發炎活性以及抑制癌細胞增生的活性。本發明亦揭露該肽的製備方法以及應用

Effects of fermentation on antioxidant properties and phytochemical composition of soy germ

Traditional soy-fermented foods, such as miso, douche, natto, and tempeh have been widely used as a dietary supplement in Asian countries, and numerous reports on their phenolics and antioxidant activities have been published. Soy germ contains 10-fold higher phenolics than whole soybean, hence using soy germ as fermentation substrate will be more efficient than whole soybean

Purification and Properties of a Novel Phenolic Antioxidant from Radix astragali Fermented by Aspergillus oryzae M29

The Chinese herb Radix astragalus (RA) has been widely used as a dietary supplement in Asia, and there are numerous reports on its bioactivities. However, there are no reports to date regarding the use of Aspergillus spp. in the culture medium of the RA plant for the production of phenolic antioxidants. In this study, utilizing the fungus Aspergillus to ferment the native RA has successfully resulted in a significant increase in the phenolic contents of RA, and the fermented RA also revealed much better antioxidant activity toward 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals, hydroxyl radical, superoxide radical and peroxyl radical than those of unfermented RA. Among these phenolics, a potent novel antioxidant was isolated and identified as 3,4-di(4'-hydroxyphenyl) isobutyric acid with a molecular weight of 272, by ESI-MS (electrospray ionization mass), (1)H NMR (nuclear magnetic resonance), (13)C NMR, DEPT (distortionless enhancement by polarization transfer)-NMR, HMQC (heteronuclear multiple quantum coherence), and HMBC (heteronuclear multiple bond correlation) spectra. These data demonstrated that the solid-state bioprocessing strategy could be an innovative approach to enhance the antioxidant activity of RA

The global role, impact, and limitations of Community Health Workers (CHWs) in breast cancer screening: a scoping review and recommendations to promote health equity for all

Introduction: Innovative interventions are needed to address the growing burden of breast cancer globally, especially among vulnerable patient populations. Given the success of Community Health Workers (CHWs) in addressing communicable diseases and non-communicable diseases, this scoping review will investigate the roles and impacts of CHWs in breast cancer screening programs. This paper also seeks to determine the effectiveness and feasibility of these programs, with particular attention paid to differences between CHW-led interventions in low- and middle-income countries (LMICs) and high-income countries (HICs).Methods: A scoping review was performed using six databases with dates ranging from 1978 to 2019. Comprehensive definitions and search terms were established for ‘Community Health Workers’ and ‘breast cancer screening’, and studies were extracted using the World Bank definition of LMIC. Screening and data extraction were protocolized using multiple independent reviewers. Chi-square test of independence was used for statistical analysis of the incidence of themes in HICs and LMICs.Results: Of the 1,551 papers screened, 33 were included based on inclusion and exclusion criteria. Study locations included the United States (n=27), Bangladesh (n=1), Peru (n=1), Malawi (n=2), Rwanda (n=1), and South Africa (n=1). Three primary roles for CHWs in breast cancer screening were identified: education (n=30), direct assistance or performance of breast cancer screening (n=7), and navigational services (n=6). In these roles, CHWs improved rates of breast cancer screening (n=23) and overall community member knowledge (n=21). Two studies performed cost-analyses of CHW-led interventions.Conclusion: This review extends our understanding of CHW effectiveness to breast cancer screening. It illustrates how CHW involvement in screening programs can have a significant impact in LMICs and HICs, and highlights the three CHW roles of education, direct performance of screening, and navigational services that emerge as useful pillars around which governments and NGOs can design effective programs in this area