Diet Screening for Immunoregulatory Milk via MicroRNA Profiling
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Solution Overview
Problem
Current methods for screening diets that enhance the immunoregulatory action of milk are limited, as they primarily focus on proteins and overlook the potential immunomodulatory effects of microRNAs present in breast milk.
Innovation Solution
A method involving the comparison of microRNA profiles in milk before and after ingestion of a diet or substance to identify increases in specific immunoregulatory microRNAs, such as miR-16, miR-17-5p, and miR-146a, which are then used to determine the immunostimulating action of the diet or substance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If current screening methods focus only on proteins in milk, then the screening process is simple and straightforward, but the immunomodulatory effects of microRNAs are overlooked and the screening comprehensiveness deteriorates
Solution Approach 1:
The screening method is segmented into two independent detection streams: one for proteins and one for microRNAs. Each stream can be performed separately using appropriate techniques (protein electrophoresis/Western blot for proteins, RT-PCR or microarray for microRNAs), allowing the simplicity of protein-only screening to be maintained while adding comprehensive microRNA detection capability.
Solution Approach 2:
The screening system is designed to detect multiple types of immunomodulatory substances (proteins and microRNAs) using a unified workflow framework. The same basic procedure (milk collection, sample preparation, data analysis) applies to both protein and microRNA detection, making the enhanced comprehensive method as efficient as the simpler protein-only approach.
2Loss of information
If microRNA profiling is added to the screening process, then the detection comprehensiveness improves, but the device complexity and method complexity increase
Solution Approach 1:
Standardized RNA extraction kits and pre-prepared reagents serve as intermediaries between the milk sample and the microRNA detection system. These intermediaries simplify the complex RNA extraction and cDNA synthesis steps, reducing the technical expertise required and making the enhanced screening method more accessible without compromising detection comprehensiveness.
Solution Approach 2:
The method uses parameter changes in the form of specific primers and probes designed for different microRNA targets. By changing only the sequence-specific parameters (primers, probes) rather than the entire detection system, the same complex apparatus can screen for multiple different microRNAs, reducing overall device complexity while maintaining detection comprehensiveness.
3Reliability
If multiple microRNA types are detected simultaneously, then the screening accuracy and reliability improve, but the measurement precision requirements and difficulty of detecting and measuring increase
Solution Approach 1:
The method creates copies of microRNA molecules through reverse transcription to generate cDNA, and subsequently through PCR amplification to create multiple copies of target sequences. This copying process amplifies the signal from scarce microRNA molecules, making them easier to detect and measure accurately while maintaining the reliability of simultaneous multi-microRNA detection.
Solution Approach 2:
The detection system incorporates internal controls and reference microRNAs that provide feedback on the quality and quantity of RNA extraction and amplification. This feedback mechanism allows for normalization of data and correction of variations, improving measurement precision and reducing the difficulty of accurate quantification across multiple microRNA targets.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for the identification of diets or substances that increase immunoregulatory microRNAs in milk, providing a novel means to enhance the immunostimulating action of breast milk, thereby promoting infant immunity.
Implementation Method 1
it binds to a 3'-untranslated region of target mRNAs in a sequence-complementary manner to suppress translation of the target mRNAs
Implementation Method 2
miRNAs which function in cells as translation regulatory molecules are present in a lipid bilayer called exosome, and are secreted out of the cells. Since it has also been confirmed that secreted miRNAs are incorporated into other cells
Data Source
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AI summary
A method for screening for a foodstuff providing production of milk having an immunoregulatory action, a novel foodstuff having an immunoregulatory action, and a method for producing it are provided. A diet or substance that increases or decreases an amount of microRNA present in milk of a mammal is identified by using correlation of microRNA profiles in the milk and a diet ingested by the mammal or a substance contained in the diet as an index to screen for a diet or a substance providing production of breast milk having an immunoregulatory action.