Processing Technologies for Bee Products: An Overview of Recent Developments and Perspectives

Increased demand for a more balanced, healthy, and safe diet has accelerated studies on natural bee products (including honey, bee bread, bee collected pollen royal jelly, propolis, beeswax, and bee venom) over the past decade. Advanced food processing techniques, such as ultrasonication and microwave and infrared (IR) irradiation, either has gained popularity as alternatives or combined with conventional processing techniques for diverse applications in apiculture products at laboratory or industrial scale. The processing techniques used for each bee products have comprehensively summarized in this review, including drying (traditional drying, infrared drying, microwave-assisted traditional drying or vacuum drying, and low temperature high velocity-assisted fluidized bed drying), storage, extraction, isolation, and identification; the assessment methods related to the quality control of bee products are also fully mentioned. The different processing techniques applied in bee products aim to provide more healthy active ingredients largely and effectively. Furthermore, improved the product quality with a shorter processing time and reduced operational cost are achieved using conventional or emerging processing techniques. This review will increase the positive ratings of the combined new processing techniques according to the needs of the bee products. The importance of the models for process optimization on a large scale is also emphasized in the future.

Glycan Profile Analysis of Engineered Trastuzumab with Rationally Added Glycosylation Sequons Presents Significantly Increased Glycan Complexity

Protein aggregation constitutes a recurring complication in the manufacture and clinical use of therapeutic monoclonal antibodies (mAb) and mAb derivatives. Antibody aggregates can reduce production yield, cause immunogenic reactions, decrease the shelf-life of the pharmaceutical product and impair the capacity of the antibody monomer to bind to its cognate antigen. A common strategy to tackle protein aggregation involves the identification of surface-exposed aggregation-prone regions (APR) for replacement through protein engineering. It was shown that the insertion of N-glycosylation sequons on amino acids proximal to an aggregation-prone region can increase the physical stability of the protein by shielding the APR, thus preventing self-association of antibody monomers. We recently implemented this approach in the Fab region of full-size adalimumab and demonstrated that the thermodynamic stability of the Fab domain increases upon N-glycosite addition. Previous experimental data reported for this technique have lacked appropriate confirmation of glycan occupancy and structural characterization of the ensuing glycan profile. Herein, we mutated previously identified candidate positions on the Fab domain of Trastuzumab and employed tandem mass spectrometry to confirm attachment and obtain a detailed N-glycosylation profile of the mutants. The Trastuzumab glycomutants displayed a glycan profile with significantly higher structural heterogeneity compared to the HEK Trastuzumab antibody, which contains a single N-glycosylation site per heavy chain located in the CH2 domain of the Fc region. These findings suggest that Fab N-glycosites have higher accessibility to enzymes responsible for glycan maturation. Further, we have studied effects on additional glycosylation on protein stability via accelerated studies by following protein folding and aggregation propensities and observed that additional glycosylation indeed enhances physical stability and prevent protein aggregation. Our findings shed light into mAb glycobiology and potential implications in the application of this technique for the development of “biobetter” antibodies.

Lab on a Chip: Bioreactors miniaturization for rapid optimization of biomedical processes and its impact on SARS-CoV-2 diagnosis

Lab on a Chip (LoC) as part of Microbioreactors (MBRs) constitute an emergent technology to carry out micro-bioprocesses based on microfluidics research. In this review, the usefulness of LoCs is exposed since its inception, demonstrating that it is a multidisciplinary research field, gathering different science branches to develop this technology. As a result, a beneficial point of advancement is reached, producing useful consumables for humanity. Some of the described LoCs throughout this work are also used to detect infectious diseases caused by bacteria or viruses, allowing accelerated studies on emerging or high-impact diseases, such as COVID-19. Here are also displayed with an updated panorama, different strategies to improve the use, applications in the biomedical field, and spread of these devices aimed at their availability to solve social problems.

Depression and Pain: use of antidepressant

Background: Emotional disorders are common comorbid affectations that exacerbate the severity and persistence of chronic pain. Specifically, depressive symptoms can lead to an excessive duration and intensity of pain. The use of antidepressant drugs is associated with pain reduction. The recent development of animal models has accelerated studies focusing on the underlying mechanisms of chronic pain and depression comorbidity. Aim: This review provides an overview of the comorbid relationship of chronic pain and depression, the clinical and pre-clinical studies performed on the neurobiological aspects of pain and depression, and the use of antidepressants as analgesics. Method: A systematic search of literature databases was conducted according to the pre-defined criteria. The authors independently conducted a focused analysis of the full-text articles. Results: Studies suggest that pain and depression are highly-intertwined and may co-exacerbate physical and psychological symptoms. One important biochemical basis for pain and depression focuses on the serotonergic and norepinephrine system, which have been shown to play an important role in this comorbidity. Brain structures that codify pain are also involved in mood. It is evident that using serotonergic and norepinephrine antidepressants are strategies commonly employed to mitigate pain. Conclusion: Literature indicates that pain and depression impact each other and play a prominent role in the development and maintenance of other chronic symptoms. Antidepressants continue to be a major therapeutic tool for managing chronic pain. Tricyclic antidepressants (TCAs) are more effective in reducing pain than selective serotonin reuptake inhibitors (SSRIs) and serotonin-noradrenaline reuptake inhibitors (SNRIs).

Formulation and Pharmacological Studies of Leaves of Moringa (Moringa oleifera), a Novel Hepatoprotection in Oral Drug Formulations

BACKGROUND: Moringa oleifera, Moringaceae, is a tree that is native to South East Asia. Various parts of this tree are commonly used in traditional medicine to treat inflammation, hepatitis, gastric ulcer, and other ailments. AIM: M. oleifera leaves extract was formulated into stable suspensions, characterized, and then evaluated for hepatoprotection activity against isoniazid. MATERIALS AND METHODS: The leaves were extracted using cold maceration, and suspensions of extract were prepared using sodium carboxymethyl cellulose (Na-CMC) as suspension agent at various concentrations (0.1, 0.5, and 1.0%). The formulations were analyzed by their appearance, color, odor, and taste. Density, pH, viscosity, re-dispersibility test, and sedimentation volume were observed. The stability of oral suspensions was analyzed in accelerated studies (5°C ± 2°C and 35°C ± 2°C for 12 h for 7 cycles) to find stable formulation, while the hepatoprotection activity was analyzed using an in vivo isoniazid-induced model. RESULTS: The appearance, color, odor, and taste of the suspensions were shown to be characteristic of the extract. Na-CMC at concentration 0.5% showed good physical properties. Stable suspension at dose 400 mg/kg BW per oral for 28 days exhibited a significant (p < 0.05) decrease in the serum glutamate oxaloacetate transaminase and serum glutamate pyruvate transaminase. CONCLUSION: Suspension containing M. oleifera leaves extract at 50 mg/5 mL was successfully obtained and showed physical properties that were appropriate and characteristic of this dosage form, suitable for hepatoprotection (400 mg/kg BW), making this an alternative to tablets.

Production of voice onset time (VOT) by senior Polish learners of English

Although research on foreign language learning among seniors has recently accelerated, studies on L2 phonology in this age group remain scarce. Seniors may be at a great disadvantage when it comes to learning the sounds of a foreign language because age of onset has been shown to correlate negatively with ultimate attainment especially for phonology (Piske et al. 2001). However, this is all the more reason to attempt a better understanding of the mechanisms of senior. This paper offers an attempt at shedding light on how senior learners with an age of L2 onset above 60 produce voiced and voiceless L2 word-initial stops. Twenty L1 Polish senior learners of English were asked to read a list of words containing word-initial voiced and voiceless plosives in their L2 English at A2+/B1 level according to CEFR. The results show that the senior Polish learners of English produce an in-between category for the English stops (with VOT longer than for Polish, but shorter than native English). The senior learners also experienced L1 drift, but mostly in the voiceless L1 Polish stops.

METTL4 catalyzes m6Am methylation in U2 snRNA to regulate pre-mRNA splicing

N 6-methylation of 2′-O-methyladenosine (Am) in RNA occurs in eukaryotic cells to generate N6,2′-O-dimethyladenosine (m6Am). Identification of the methyltransferase responsible for m6Am catalysis has accelerated studies on the function of m6Am in RNA processing. While m6Am is generally found in the first transcribed nucleotide of mRNAs, the modification is also found internally within U2 snRNA. However, the writer required for catalyzing internal m6Am formation had remained elusive. By sequencing transcriptome-wide RNA methylation at single-base-resolution, we identified human METTL4 as the writer that directly methylates Am at U2 snRNA position 30 into m6Am. We found that METTL4 localizes to the nucleus and its conserved methyltransferase catalytic site is required for U2 snRNA methylation. By sequencing human cells with overexpressed Mettl4, we determined METTL4’s in vivo target RNA motif specificity. In the absence of Mettl4 in human cells, U2 snRNA lacks m6Am thereby affecting a subset of splicing events that exhibit specific features such as 3′ splice-site weakness and an increase in exon inclusion. These findings suggest that METTL4 methylation of U2 snRNA regulates splicing of specific pre-mRNA transcripts.

METTL4 catalyzes m6Am methylation in U2 snRNA to regulate pre-mRNA splicing

N6-methylation of 2’-O-methyladenosine (Am) in RNA occurs in eukaryotic cells to generate N6,2’-O-dimethyladenosine (m6Am). Identification of the methyltransferase responsible for m6Am catalysis has accelerated studies on the function of m6Am in RNA processing. While m6Am is generally found in the first transcribed nucleotide of mRNAs, the modification is also found internally within U2 snRNA. However, the writer required for catalyzing internal m6Am formation had remained elusive. By sequencing transcriptome-wide RNA methylation at single-base-resolution, we identified human METTL4 as the writer that directly methylates Am at U2 snRNA position 30 into m6Am. We found that METTL4 localizes to the nucleus and its conserved methyltransferase catalytic site is required for U2 snRNA methylation. By sequencing human cells with overexpressed Mettl4, we determined METTL4’s in vivo target RNA motif specificity. In the absence of Mettl4 in human cells, U2 snRNA lacks m6Am thereby affecting a subset of splicing events that exhibit specific features such as overall 3’ splice-site weakness with certain motif positions more affected than others. This study establishes that METTL4 methylation of U2 snRNA regulates splicing of specific pre-mRNA transcripts.

Machine Learning Methods in Precision Medicine Targeting Epigenetic Diseases

Background: On a tide of big data, machine learning is coming to its day. Referring to huge amounts of epigenetic data coming from biological experiments and clinic, machine learning can help in detecting epigenetic features in genome, finding correlations between phenotypes and modifications in histone or genes, accelerating the screen of lead compounds targeting epigenetics diseases and many other aspects around the study on epigenetics, which consequently realizes the hope of precision medicine. Methods: In this minireview, we will focus on reviewing the fundamentals and applications of machine learning methods which are regularly used in epigenetics filed and explain their features. Their advantages and disadvantages will also be discussed. Results: Machine learning algorithms have accelerated studies in precision medicine targeting epigenetics diseases. Conclusion: In order to make full use of machine learning algorithms, one should get familiar with the pros and cons of them, which will benefit from big data by choosing the most suitable method(s).