Non-enzymatic glycation of proteins: From diabetes to cancer

Incubation of proteins with glucose leads to their non-enzymatic glycation and formation of Amadori products known as an early glycation product. Oxidative cleavage of Amadori products is considered as a major route to advanced glycation endproducts (AGEs) formation in vivo. Non-enzymatic glycation of proteins or Maillard reaction is increased in diabetes mellitus due to hyperglycemia and leads to several complications such as blindness, heart disease, nerve damage, and kidney failure. The early and advanced glycation products are accumulated in plasma and tissues of diabetic patients and cause production of autoantibodies against corresponding products. The advanced glycation products are also associated with other diseases like cancer. This review summarizes current knowledge of these stage specific glycated products as common and early diagnostic biomarkers for the associated diseases and the complications with the aim of a novel therapeutic target for the diseases.     

Specialty products made from goat milk

Although it may not be important in certain parts of the world, the contribution of goat milk to the economic and nutritional wellbeing of humanity is undeniable in many developing countries, especially in the Mediterranean, Middle East, Eastern Europe and South American countries. Goat milk has played a very important role in health and nutrition of young and elderly. Goat milk has also been known for its beneficial and therapeutic effects on the people who have cow milk allergy. These nutritional, health and therapeutic benefits enlighten the potentials and values of goat milk and its specialty products. The chemical characteristics of goat milk can be used to manufacture a wide variety of products, including fluid beverage products (low fat, fortified, or flavored) and UHT (ultra high temperature) milk, fermented products such as cheese, buttermilk or yogurt, frozen products such as ice cream or frozen yogurt, butter, condensed/dried products, sweets and candies. In addition, other specialty products such as hair, skin care and cosmetic products made from goat milk recently have gained a further attention. Nevertheless, high quality products can only be produced from good quality goat milk. The quality milk should have the potential to tolerate technological treatment and be transformed into a product that satisfies the expectations of consumers, in terms of nutritional, hygienic and sensory attributes. Taste is the main criteria used by consumers to make decisions to purchase and consume goat milk and its products. Typical goat taste is considered as a quality component in certain goat cheese products. Farmers can produce more value-added products for the economic sustainability of their business and the dairy goat industry in general.     

Glycation products in infant formulas: chemical,analytical and physiological aspects

Infant formulas are milk-based products, which are adapted to the composition of human milk. To ensure microbiological safety and long shelf life, infant formulas usually undergo rigid heat treatment. As a consequence of the special composition and the heat regimen, infant formulas are more prone to thermally induced degradation reactions than regular milk products. Degradation reactions observed during milk processing comprise lactosylation yielding the Amadori product lactulosyllysine, the formation of advanced glycation end products (AGEs), and protein-free sugar degradation products, as well as protein or lipid oxidation. Several methods have been developed to estimate the heat impact applied during the manufacturing of infant formulas, including indirect methods such as fluorescence analysis as well as the analysis of defined reaction products. Most studies confirm a higher degree of damage in infant formulas compared to regular milk products. Differences between various types of infant formulas, such as liquid, powdered or hypoallergenic formulas depend on the analyzed markers and brands. A considerable portion of protein degradation products in infant formulas can be avoided when process parameters and the quality of the ingredients are carefully controlled. The nutritional consequences of thermal degradation products in infant formulas are largely unknown.     

生态产品价值实现率评价方法——以丽水市为例

生态产品价值实现是指在维持生态系统稳定性和完整性的前提下,通过合理开发利用生态产品,将其生态价值转化为经济效益的过程。生态产品价值实现机制包含了促进生态产品价值实现的政策、市场和技术机制。生态产品通过各种途径实现的经济价值总和称为生态产品价值实现量。生态产品价值实现量与生态产品总值(GEP)的比值为生态产品价值实现率。评估生态产品价值实现率是评判生态产品价值实现状况的基础,是评估生态产品价值实现机制是否有效运转的重要前提。提出生态产品价值实现率的概念和核算方法,以浙江省丽水市为例,在核算GEP的基础上,评估生态产品价值实现量与实现率来分析丽水市生态产品价值实现状况、问题及影响因素,提出提高生态产品价值实现率的对策建议。研究表明,丽水市2019年GEP为4110.21亿元,生态产品价值实现量为1017.49亿元,生态产品价值实现率为24.76%。丽水生态产品价值实现模式主要有市场交易和政府补偿两种模式。市场交易模式贡献了95.84%的生态产品价值实现量,是目前丽水市最有效的生态产品价值实现模式,但存在价值实现效率不均衡、对于缺乏市场或是市场机制不成熟的生态产品不适用等问题。政府补偿模式贡...     

Bioanalytical strategy used in development of pharmacokinetic (PK) methods that support biosimilar programs

The development of biosimilar products is expected to grow rapidly over the next five years as a large number of approved biologics reach patent expiry. The pathway to regulatory approval requires that similarity of the biosimilar to the reference product be demonstrated through physiochemical and structural characterization, as well as within in vivo studies that compare the safety and efficacy profiles of the products. To support nonclinical and clinical studies pharmacokinetic (PK) assays are required to measure the biosimilar and reference products with comparable precision and accuracy. The most optimal approach is to develop a single PK assay, using a single analytical standard, for quantitative measurement of the biosimilar and reference products in serum matrix. Use of a single PK assay for quantification of multiple products requires a scientifically sound testing strategy to evaluate bioanalytical comparability of the test products within the method, and provide a solid data package to support the conclusions. To meet these objectives, a comprehensive approach with scientific rigor was applied to the development and characterization of PK assays that are used in support of biosimilar programs. Herein we describe the bioanalytical strategy and testing paradigm that has been used across several programs to determine bioanalytical comparability of the biosimilar and reference products. Data from one program is presented, with statistical results demonstrating the biosimilar and reference products were bioanalytically equivalent within the method. The cumulative work has established a framework for future biosimilar PK assay development.     

Kinetic modeling of the recA protein promoted renaturation of complementary DNA strands

Quantitative agarose gel assays reveal that the recA protein promoted renaturation of complementary DNA strands (phi X DNA) proceeds in two stages. The first stage results in the formation of unit-length duplex DNA as well as a distribution of other products ("initial products"). In the second stage, the initial products are converted to complex multipaired DNA structures ("network DNA"). In the presence of ATP, the initial products are formed within 2 min and are then rapidly converted to network DNA. In the absence of ATP, the initial products are formed nearly as fast as with ATP present, but they are converted to network DNA at a much lower rate. The time-dependent formation of initial products and network DNA from complementary single strands for both the ATP-stimulated and ATP-independent reactions can be modeled by using a simple two-step sequential kinetic scheme. This model indicates that the primary effect of ATP in the recA protein promoted renaturation reaction is not on the initial pairing step (which leads to the formation of initial products) but rather is to increase the rate at which subsequent pairing events can occur.     

Bioanalytical strategy used in development of pharmacokinetic (PK) methods that support biosimilar programs

The development of biosimilar products is expected to grow rapidly over the next five years as a large number of approved biologics reach patent expiry. The pathway to regulatory approval requires that similarity of the biosimilar to the reference product be demonstrated through physiochemical and structural characterization, as well as within in vivo studies that compare the safety and efficacy profiles of the products. To support nonclinical and clinical studies pharmacokinetic (PK) assays are required to measure the biosimilar and reference products with comparable precision and accuracy. The most optimal approach is to develop a single PK assay, using a single analytical standard, for quantitative measurement of the biosimilar and reference products in serum matrix. Use of a single PK assay for quantification of multiple products requires a scientifically sound testing strategy to evaluate bioanalytical comparability of the test products within the method, and provide a solid data package to support the conclusions. To meet these objectives, a comprehensive approach with scientific rigor was applied to the development and characterization of PK assays that are used in support of biosimilar programs. Herein we describe the bioanalytical strategy and testing paradigm that has been used across several programs to determine bioanalytical comparability of the biosimilar and reference products. Data from one program is presented, with statistical results demonstrating the biosimilar and reference products were bioanalytically equivalent within the method. The cumulative work has established a framework for future biosimilar PK assay development.     

Summary of the diverse situation of similar biotherapeutic products in the selected countries (August 2010)

The WHO guidelines on evaluating similar biotherapeutic products (SBPs) were adopted by the Expert Committee on Biological Standardization in 2009. The fundamental messages of the guidelines are that a) generic approach is not suitable for licensing SBPs, b) only products that have been subjected to a comparability exercise and show similarity to the reference biotherapeutic product (RBP) in terms of their quality, safety and efficacy are defined as SBPs, and c) the products that are not shown to be similar to the originator products as indicated in the guidelines should neither be described as "similar" nor called SBPs. In view of these, the products which have not been subjected to a head to head comparison with the RBP should be referred to as another term, e.g. 'non-innovator' therapeutic products. In order to review the current situation in each country, a survey was planned in line with the implementation workshop of the guidelines in August 2010. The results show that the diversity of regulatory framework for licensing SBPs and the ambiguous use of the terms, 'similar' or 'generic', present considerable challenges for the future use of SBPs.     

WHO informal consultation on regulatory evaluation of therapeutic biological medicinal products held at WHO Headquarters, Geneva, 19-20 April 2007.

This report reflects the discussion and conclusions of an informal consultation held on 19-20 April 2007 at the World Health Organization concerning the regulatory evaluation of therapeutic biological medicinal products. The objectives of this meeting were to discuss the current status of so-called "similar" biological medicinal products (biosimilars) and to review regulatory pathways and challenges in evaluating the quality, safety and efficacy of these products. Biosimilars are products that are subject to licensing with a reduced data package due to a proven 'similarity' to the licensed reference product. The meeting was attended by experts in biotherapeutics from regulatory agencies, industry and academia representing 16 countries worldwide. Dr. Elwyn Griffiths (Canada) acted as Chairman and Dr. James Robertson (UK) was the Rapporteur. The meeting strongly focused on the usage of biosimilars and the current regulatory situation in many different countries. The application of International Nonproprietary Names (INN) to biosimilars, their potential immunogenicity, and WHO international standards and reference materials were also discussed, alongside presentations from the innovator and generic manufacturing industries. The consultation recognized the importance of the terminology as well as a definition of biosimilars for future considerations of these products. However, achieving a global consensus on the terminology for these new challenging products was not attempted at the Consultation, and it was decided that a future WHO working group should act on this issue as a next step. For purposes of this meeting report only, the term 'biosimilars' is temporarily used to refer to this category of products. It became clear that biotherapeutics authorized on the basis of a reduced data package are available and being used in some countries, with more appearing on the market. The existence of divergent approaches to the regulatory oversight of biosimilars in different countries revealed a need for defining regulatory expectations for these products at the global level. While many countries are following the guideline developed within the EU for quality aspects, discrepancies remain regarding the non-clinical and clinical studies of these products. The Consultation recommended that the WHO should develop a guideline in this area in order to provide a framework for the development of regulatory pathways for these products worldwide. For this purpose, agreement on the scope, definition and terminology of these products was deemed necessary. The interchangeability and substitution of products were also flagged as areas in need of harmonization. A WHO working group should be established to develop a guideline that would promote global consensus on the regulation of biosimilars, assist in their registration and enhance the availability of safe and effective biosimilar products worldwide.     

DIFFERENTIAL THRESHOLD AND PSYCHOPHYSICAL POWER FUNCTION OF SWEETNESS SENSATION: APPLIED PSYCHOPHYSICS AND PROSPECT THEORY ON FORMULATING BAKING PRODUCTS

The main purpose of this project was to develop the optimal sweetness sensation framework for baking products. Phase 1 was designed to measure consumers' differential thresholds of sweetness sensation when the sweetener contents of various sweetener‐leveled products were increased and lowered by using the constant stimuli method. Results indicated that the consumers' differential thresholds increased when the initial sweetener levels of products increased. Phase 2 was designed to establish the psychophysical power function of sweetener‐content changes and consumers' psychological sensation of products with different sweetener levels by adopting the magnitude estimation method. Regardless of the initial sweetener levels of products, the results showed that consumers perceived the products as less sweet immediately after the sweetener contents of products dropped by even a small amount; however, consumers perceived products as sweeter only when products dramatically increased in contents of sweetener.     

国家公园生态产品价值实现的机制及模式——以神农架国家公园为例

探究生态产品价值实现的机制及模式是国家公园加快生态文明建设、推动"绿水青山"向"金山银山"转化的关键。构建了国家公园社会-经济-自然复合生态系统,分析国家公园生态产品特征并划分类别,以神农架国家公园试点为例,探究了国家公园生态产品价值的实现路径与机制,建立了国家公园生态产品价值实现模式。结果表明:(1)国家公园复合生态系统由社会、经济和自然生态系统构成,形成一个相互影响、相互关联的整体。(2)国家公园生态产品具有生物生产性、资源稀缺性、人类收益性、地域整体性、政策依赖性、利益平衡性等特征,识别与分类生态产品有助于复合生态系统的平稳运行与健康发展,进而实现生态产品价值。(3)国家公园生态产品总体分为公共性、准公共性和经营性三类,并由政府主导、社区主导、市场主导与多主体复合型路径组成的价值实现路径体系完成价值实现。(4)国家公园生态产品价值实现机制由生态保护机制、经济发展机制、社区进步机制和协调保障机制四部分构成,服务于生态保护、产业发展和社区振兴三个目标。(5)国家公园生态产品价值实现过程中存在一定问题,以生态保护为首要任务、旅游发展为实现路径、社区振兴为核心目标,构建神农架国家公园"生态保护-旅游发展-社区振兴"共生模式,并进一步提出生态产品价值实现的生态旅游模式,以协调处理神农架国家公园复合生态系统结构内的矛盾,实现国家公园生态产品价值的最大化。综上所述,丰富了生态产品价值实现理论,并为国家公园生态产品价值实现模式提供了重要的科学依据与决策参考。国家公园"生态保护-旅游发展-社区振兴"的共生模式以及国家公园生态产品实现的生态旅游模式,将为国家公园生态产品价值的实现提供更加全面与有效的路径与模式。     

Microbial Oxidation of the Isoprenoid Alkane Pristane

Microorganisms capable of utilizing pristane (2,6,10,14-tetramethylpentadecane) as a sole source of carbon and energy were isolated from soil specimens. A strain BPM 1613, tentatively classified in the genus Nocardia, accumulated several oxidation products of pristane in the culture fluid. Silica gel chromatography of the ethyl ether extract from the culture fluid yielded pristanol and pristanic acid as major products and pristyl pristanate and pristyl aldehyde as minor products. The confirmations on the estimated structures of these oxidation products are described in detail.     

WHO/KFDA joint workshop on implementing WHO guidelines on evaluating similar biotherapeutic products, Seoul, Republic of Korea 24-26 August, 2010

In August 2010, the World Health Organization and the Korea Food & Drug Administration jointly organized the first implementation workshop of WHO guidelines on evaluating similar biotherapeutic products (SBPs) at the global level. The objective of the Workshop was to facilitate implementation of the newly adopted WHO Guidelines into the practice of national regulatory authorities (NRAs). WHO Guidelines were recognized by the workshop participants as a tool for harmonizing regulatory requirements worldwide. By reviewing and practicing several case studies, better understanding and consensus on the principles of clinical trial designs were reached. However, variations in terms of the national requirements for quality, safety and efficacy of these products revealed diversity in the regulatory expectations in different countries and regions. In addition, lack of terminology for the products developed as copy products (so called "me too" products) with a partial comparability to an RBP, led to a great diversity in evaluating as well as naming these products. The workshop participants proposed the following actions: a) NRAs should make efforts to build their capacities for regulation of SBPs; b) WHO should revise WHO Guidelines for assuring the quality of products prepared by recombinant DNA technology (WHO TRS 814) and continue monitoring progress with the implementation of the Guidelines on evaluating SBPs. Publication of the outcome of the Workshop was recognized as another action that WHO should coordinate.     

The regulatory framework for similar biotherapeutic products in Cuba

Biopharmaceuticals make up a significant proportion of medicinal products used for the treatment of diseases such as cancer, arthritis, cardiac dysfunctions and AIDS. Access to therapies based on the use of these products has been limited as a result of the high marketing costs. Cuba has a biopharmaceutical industry with great potential for innovation, capable of developing new products and to produce others, like the biosimilars destined to fulfill the needs of its National Health System. The Center for State Control on the Quality of Drugs (CECMED) the Cuban NRA, is facing the challenge of regulating the approval of biosimilar products manufactured locally. Consequently, CECMED has issued a position paper establishing the basic principles for regulation of these products and a specific guideline on this was elaborated.     

Immunochemical evidence for the presence of advanced glycation end products in human lens proteins and its positive correlation with aging.

Prolonged incubation of protein with reducing sugar proceeds through a series of reactions involving early stage products to the advanced glycation end products with fluorescence, brown color, and cross-linking. Known collectively as the Maillard reaction, these changes have been suggested as factors in diabetic complications and the aging process. The early stage products have been demonstrated in vivo, but evidence for the presence in vivo of the advanced glycation end products has been limited. We sought to provide immunochemical evidence by the preparation and use of polyclonal and monoclonal antibodies to these end products (Horiuchi, S., Araki, N., and Morino, Y. (1991) J. Biol. Chem. 266, 7329-7332) as probes to identify and quantitate such compounds in human lens crystallins. Neither of the antibodies reacted with extracts from infant lenses, but fractions from adult lenses showed a significant reactivity, correlating with lens age. Our findings provide the first immunochemical evidence that human lens crystallins contain advanced glycation end products and that these products increase with tissue age.     

Importance of microbial natural products and the need to revitalize their discovery

Microbes are the leading producers of useful natural products. Natural products from microbes and plants make excellent drugs. Significant portions of the microbial genomes are devoted to production of these useful secondary metabolites. A single microbe can make a number of secondary metabolites, as high as 50 compounds. The most useful products include antibiotics, anticancer agents, immunosuppressants, but products for many other applications, e.g., antivirals, anthelmintics, enzyme inhibitors, nutraceuticals, polymers, surfactants, bioherbicides, and vaccines have been commercialized. Unfortunately, due to the decrease in natural product discovery efforts, drug discovery has decreased in the past 20 years. The reasons include excessive costs for clinical trials, too short a window before the products become generics, difficulty in discovery of antibiotics against resistant organisms, and short treatment times by patients for products such as antibiotics. Despite these difficulties, technology to discover new drugs has advanced, e.g., combinatorial chemistry of natural product scaffolds, discoveries in biodiversity, genome mining, and systems biology. Of great help would be government extension of the time before products become generic.     

Use of Symbiosis Products from Integrated Pulp and Paper and Carbon Steel Mills: Legal Status and Environmental Burdens

This study assesses the policy/legal status of both multistream residues and potential secondary products (“symbiosis products”) and whether there could be environmental benefits associated with the utilization of residues from integrated pulp and paper and carbon steel mills as raw materials for such secondary products. Waste‐related European Union (EU) and Finnish policy and legal instruments were reviewed to identify potential constraints for, and suggested next steps in, the development of potential process industry residue‐based symbiosis products. The products were soil amendment pellets, low‐grade concrete, and mine filler. A global warming potential (GWP) assessment and an exergy analysis were applied to these potential symbiosis products. Some indicative GWP calculations of greenhouse gas emissions associating similar and/or analogous products based on virgin primary raw materials, more energy‐intensive processes, and the alternative treatment of these residues as wastes are also presented. This study addresses GWP, exergy, and legal aspects in a holistic manner to determine the potential environmental benefits of secondary products within the EU legal framework. The GWP assessment and exergy analysis indicate that the utilization of multistream residues causes very low environmental burdens in terms of GWP. The utilization option can have potential environmental benefits in terms of GWP through process replacement and avoided landfilling and waste treatment impacts, as well as potentially through emission reductions from product replacement if suitable and safe applications can be identified. Waste regulation does not define the legal requirements under which utilizing residues in such novel concepts as introduced in this study would be possible, nor how waste status could be removed and product‐based legislation be applied to the potential products instead.     

Relationship between availability indices and plant uptake of nitrogen and phosphorus from organic products

A better knowledge of the plant-availability of nitrogen (N) and phosphorus (P) in organic products may help to improve the efficient use of these products as fertilizers. In the present study, availability indices for N and P of nine widely differing organic products obtained by different fractionation methods were compared with the plant uptake of N and P from these products. The fractionation methods included CaCl₂ extraction, thermal fractionation (heating of organic products), and pepsin extraction, for N, and extraction with diluted sulphuric acid, P-Bray-I, P-Olsen, and extraction using an iron oxide coated filter paper, for P. The results of pot experiments with ryegrass using a double-pot technique (Janssen, 1990) over 62 (N experiment) and 93 days (P experiment) were used as reference for plant-availabe N and P. The 0.01 M CaCl₂ extractable inorganic N reasonably predicted plant-available N only in organic products with a high inorganic N fraction. Thermal fractionation and pepsin extraction provided a reasonable index for mineralizable N in organic products having a high fraction of mineralizable N. Of the P fractionation methods, the extraction using iron oxide coated filter paper was the best indicator of plant-available P in the products.     

Screening of natural products in the laboratory and the field for control of pollen beetles

Pollen beetles, Meligethes spp. (Coleoptera: Nitidulidae), are among the most damaging pests of oilseed rape (Brassica napus L.). Increasing populations of pyrethroid‐resistant pollen beetles coupled with the prohibition of synthetic pest control products in organic farming means that other direct control methods are needed. A laboratory study was therefore conducted to evaluate the effect of natural products, combinations of natural products and additives as well as natural and synthetic insecticides on mortality of pollen beetles. In addition, field trials under both integrated and organic production were conducted to evaluate the efficacy of six natural products, nine combinations of natural products, two synthetic insecticides (integrated‐production trials only), two natural insecticides and two additives to reduce pollen beetles. The laboratory trial, using both a curative and preventive approach, showed that two of the eight natural products (lavender oil and stone meal) caused a mortality of over 98% within the first day of application. The other natural products were only partially effective compared with the untreated control. In the field trials, the natural products reduced the number of pollen beetles 1 day after application compared with the untreated control in both trial years and production systems. Promising substances were stone meal, Silico‐Sec and liquid manure. Nevertheless, the efficacy was not consistent, and no effect on oilseed rape yield was observed. In spite of this, these products have the potential to control pollen beetles under field conditions with comparable effects to synthetic insecticides. Further research should therefore focus on timing and frequency of applications as well as on the formulation of the natural products to increase the persistency of the products under field conditions.