High Protein Research Articles

Structure-Based Design and Development of Phosphine Oxides as a Novel Chemotype for Antibiotics that Dysregulate Bacterial ClpP Proteases.

A series of arylsulfones and heteroarylsulfones have previously been demonstrated to dysregulate the conserved bacterial ClpP protease, causing the unspecific degradation of essential cellular housekeeping proteins and ultimately resulting in cell death. A cocrystal structure of a 2-β-sulfonylamide analog, ACP1-06, with Escherichia coli ClpP showed that its 2-pyridyl sulfonyl substituent adopts two orientations in the binding site related through a sulfone bond rotation. From this, a new bis-aryl phosphine oxide scaffold, designated as ACP6, was designed based on a "conformation merging" approach of the dual orientation of the ACP1-06 sulfone. One analog, ACP6-12, exhibited over a 10-fold increase in activity over the parent ACP1-06 compound, and a cocrystal X-ray structure with ClpP confirmed its predicted binding conformation. This allowed for a comparative analysis of how different ligand classes bind to the hydrophobic binding site. The study highlights the successful application of structure-based rational design of novel phosphine oxide-based antibiotics.

Crosstalk between ubiquitination and translation in neurodevelopmental disorders

Ubiquitination is one of the most conserved post-translational modifications and together with mRNA translation contributes to cellular protein homeostasis (proteostasis). Temporal and spatial regulation of proteostasis is particularly important during synaptic plasticity, when translation of specific mRNAs requires tight regulation. Mutations in genes encoding regulators of mRNA translation and in ubiquitin ligases have been associated with several neurodevelopmental disorders. RNA metabolism and translation are regulated by RNA-binding proteins, critical for the spatial and temporal control of translation in neurons. Several ubiquitin ligases also regulate RNA-dependent mechanisms in neurons, with numerous ubiquitination events described in splicing factors and ribosomal proteins. Here we will explore how ubiquitination regulates translation in neurons, from RNA biogenesis to alternative splicing and how dysregulation of ubiquitin signaling can be the underlying cause of pathology in neurodevelopmental disorders, such as Fragile X syndrome. Finally we propose that targeting ubiquitin signaling is an attractive novel therapeutic strategy for neurodevelopmental disorders where mRNA translation and ubiquitin signaling are disrupted.

PELATIHAN PEMBUATAN SATE LILIT IKAN BANDENG SEBAGAI UPAYA MENINGKATKAN KETERAMPILAN DAN STATUS GIZI BAGI MASYARAKAT

Food is essential for human survival. The government has to provide and distribute food which is affordable and safe for the people to consume. The society in Muara Gembong District, Bekasi Regency, mainly makes a living from fish farming, agriculture, and trade. A vivid example of food availability is the utilization of local resources through 'food modification'. One popular culinary product is satay, which is usually made from meat. However, in this particular training, we used a local raw material, namely milkfish, as the main ingredient for making sate lilit (a type of satay). Milkfish contains high protein, namely around 20-24%; rich with micro and macro minerals; as well as has a soft, chewy, and savory texture. The results of the training, based on the results of the pre-test and post-test, showed an n-gain score of 0.6. This number signifies that the making of sate lilit was medium. Furthermore, in an effort to follow up on the aforementioned result, we hope that SMAN 1 Muara Gembong can become a part of the mentoring groups of the study program.

Effects of vitamin D supplementation on diabetic foot ulcer healing: a meta-analysis.

To systematically review the effect of vitamin D supplementation on diabetic foot ulcer (DFU) healing. The PubMed, Web of Science, Science direct, Ebsco host, CNKI, WanFang, VIP, and CBM databases were electronically searched to collect randomized controlled trials (RCTs) on the impact of vitamin D supplementation on DFUs from inception to 19 November 2022. Two researchers independently screened the literature, extracted the data and assessed the risk of bias of the included studies. Meta-analysis was then performed by using RevMan 5.3 software. A total of seven studies involving 580 patients were included. The results of meta-analysis showed that compared with control group, the wound healing efficiency rate (RR = 1.42, 95%CI 1.03 to 1.95, P= 0.03) and wound reduction rate (MD = 13.11, 95%CI 4.65 to 21.56, P < 0.01) of the experimental group were higher; the change values of the wound area (MD = -3.29, 95%CI -4.89 to 1.70, P < 0.01) and 25 (OH) D (MD = 9.63, 95%CI 6.96 to 12.31, P < 0.01) were larger. Supplementation of vitamin D on DFU patients can improve glucose metabolism and insulin indexes: hemoglobin A1c (MD = -0.44, 95%CI -0.62 to -0.26, P < 0.01), fasting insulin (MD = -3.75, 95%CI -5.83 to -1.67, P < 0.01), HOMA - β (MD = -5.14, 95%CI -8.74 to -1.54, P < 0.01), and quantitative insulin sensitivity check index (MD = 0.02, 95%CI 0.01 to 0.02, P < 0.01). It can also improve inflammation and oxidative stress markers: high sensitivity C-reactive protein (MD = -0.83, 95%CI -1.06 to -0.59, P < 0.01), erythrocyte sedimentation rate (MD = -15.74, 95%CI -21.78 to -9.71, P<0.01), nitric oxide (MD = 1.81, 95%CI 0.07 to 3.55, P= 0.04), and malondialdehyde (MD = -0.43, 95%CI -0.61 to -0.24, P<0.01). There was no statistically significant difference in changes of fasting plasma glucose, homeostasis model of assessment-insulin resistance, total antioxidant capacity, glutathione, very low density lipoprotein cholesterol, low density lipoprotein cholesterol, and high density lipoprotein cholesterol (P>0.05). The current evidence suggests that vitamin D supplementation can significantly promote DFU healing by lowering blood sugar and alleviating inflammation and oxidative stress. Key messages What is already known on this topic Diabetic foot ulcer (DFU) is a major complication of diabetes mellitus, with high morbidity, mortality and resource utilization. Vitamin D has the effect of lowering blood sugar, improving insulin sensitivity, and increasing anti-inflammatory response. Clinical research on vitamin D supplementation for the treatment of DFU is increasing, but due to the lack of combing and integration, the actual efficacy of vitamin D in patients is unclear. What this study adds This meta-analysis has shown that vitamin D supplementation can significantly promote DFU healing by lowering blood glucose and alleviating inflammation and oxidative stress. How this study might affect research, practice or policy This study preliminarily found the effectiveness of vitamin D supplementation on the healing of DFU, which can provide a reference for the treatment of DFU by medical staff.

CHARACTERISTICS OF JUWET FRUIT YOGHURT WITH THE ADDITION OF SKIM MILK AND LACTIC ACID BACTERIA STARTER

In this study, the manufacture of juwet fruit yoghurt was studied with different treatments of adding skim milk and lactic acid bacteria starter. Yogurt is a processed milk product made through bacterial fermentation. Yogurt is known as a good source of protein, calcium, and probiotics, which can support digestive health and the immune system. Juwet fruit has a high antioxidant and vitamin C content so that juwet fruit has interesting potential to be made into yoghurt. Skim milk is used to utilize high lactose and protein so that the growth of lactic acid bacteria can grow optimally. Lactic acid bacteria are used for preservation, providing texture, and adding flavor to yoghurt drinks. The purpose of this study was to determine the addition of skim milk and lactic acid bacteria starter on the physicochemical, microbiological, and organoleptic characteristics of juwet yoghurt. This study used a Completely Randomized Design (CRD) factorial pattern consisting of two factors. Factor 1 addition of skim milk (5%, 7%, 9%). Factor 2 addition of starter (5%, 7%, 9%). Each treatment was repeated three times. The data obtained were analyzed using analysis of variance (ANOVA). If there is a significant difference between treatments, it is continued with the Duncan test (DMRT) and organoleptic test using the Scoring test with 25 trained panelists. The treatment of adding 9% skim milk and 9% starter was the best treatment that produced juwet yoghurt with a protein content of 2.1% ± 0.036, total titratable acid 2.1% ± 0.1, total soluble solids 22.7ºBrix ± 1.53, total lactic acid bacteria 10.5 log CFU/ml ± 0.023, pH 3.7 ± 0.058, antioxidants 34.0% ± 0.117, viscosity 6.8% ± 0.058, Vitamin C 5.8 ± 0.3, and organoleptic tests of color 2.88 (rather like), taste 3.24 (rather like), aroma 2.68 (rather like), texture 3.76 (like).

Effect of Differences in Protein Levels and Length of Fermentation Time on Physicochemical Characteristics of Caspian Sea Yoghurt Green Tea

Caspian sea inoculum has been widely used for yogurt production, but it is rarely used in making green tea powder (GTP) yoghurt using milk with different protein content. This study aims to determine the changes in the characteristics of GTP yoghurt with Caspian sea inoculum using low fat UHT milk (SLF) or full cream UHT milk (SFC) during the fermentation period at 0 and 16 hours. The results showed that use of different types of UHT milk affected physicochemical characteristics. Fermentation for 16 hours caused a decrease in the pH of the sample to pH 4.3-4.4, but it was negatively correlated with an increase in the acidity. In addition, fermentation resulted increase the value of L* (brightness). The SLF sample has a greener color than the SFC sample. The viscosity of GTP yoghurt increased significantly at 16 hours in SFC and SLF samples. After 16 hours fermentation, the TPC became 4.15 mg GAE/g and 4.45 mg GAE/g with IC50 values ​​of 37.51 g/mL and 23.95 g/mL in SLF and SFC samples. The high protein content in UHT milk affected the high acidity, viscosity, and syneresis values ​​resulting a decrease in the TPC of GTP yoghurt.

Cellular prion protein acts as mediator of amyloid beta uptake by caveolin-1 causing cellular dysfunctions in vitro and in vivo.

Cellular prion protein (PrPC) was implicated in amyloid beta (Aβ)-induced toxicity in Alzheimer's disease (AD), but the precise molecular mechanisms involved in this process areunclear. Double transgenic mice were generated by crossing Prnp knockout (KO) with 5xFAD mice, and light-sheet microscopy was used for whole brain tissue analyses. PrPC-overexpressing cells were developed for in vitro studies, and microscopy was used to assess co-localization of proteins of interest. Surface-plasmon resonance (SPR) was used to investigate protein-binding characteristics. In vivo, PrPClevels correlated with reduced lifespan and cognitive and motor function, and its ablation disconnected behavior deficits from Aβ levels. Light-sheet microscopy showed that PrPCinfluenced Aβ-plaque burden but not the distribution of those plaques. Interestingly, caveolin-1 (Cav-1) KO neurons significantly reduced intracellular Aβ-oligomer (Aβo) uptake when compared to wild-type neurons. The findings shed new light on the relevance of intracellular Aβo, suggesting that PrPCand Cav-1 modulate intracellular Aβ levels andthe Aβ-plaque load. PrPC expression adversely affects lifespan and behavior in 5xFAD mice. PrPC increases Aβ1-40 and Aβ1-42 levels and Aβ-plaque load in 5xFAD mice. Cav-1 interacts with both PrPC and Aβ peptides. Knocking out Cav-1 leads to a significant reduction in intracellular Aβ levels.

Simple methodology to score micropattern quality and effectiveness.

Micropatterns (MPs) are widely used as a powerful tool to control cell morphology and phenotype. However, methods for determining the effectiveness of how well cells are controlled by the shape of MPs have been inconsistently used and studies rarely report on this topic, indicating lack of standardization. We introduce an evaluation score that quantitatively assesses the MP fabrication quality and effectiveness, which can be broadly used in conjunction with all currently available MP design types. This score uses four simple and quick steps: (i) scoring MP and (ii) background fabrication quality, (iii) defining the type(s) of MP of interest, and (iv) assigning so-called efficiency descriptors describing cell behavior. These steps are based on visual inspection and quick categorization of various aspects of MP fabrication quality and cell behaviour, presented in illustrations and microscopy image examples intended to serve as a reference "atlas". To illustrate the advantage of using this score, we determined differences in cell morphology and F-actin intensity between scored vs. non-scored cells. These measurements, which could be different in other studies, were chosen because both are understood as markers of cell phenotype and function. We combined intensity-calibrated immunofluorescence microscopy and image-based single cell protein analysis. Importantly, significant differences in cell morphology and cytoskeletal protein content between scored vs. non-scored cells were noted: the unconditional inclusion of all experimental read-outs (i.e., all MP data regardless of MP quality and effectiveness) into the final results significantly misjudged the experimental readouts vs. only including experimental read-outs of quality-controlled and effective MPs, identified by scoring. Specifically, non-scoring underestimated the F-actin intensity per cell and quantitative cellular morphometric descriptors circularity and solidity and overestimated aspect ratio. Scoring improved the precision of cellular readouts, advocating the use of a MP quality and efficiency score as a quantitative decision-supporting tool in deciding whether or not particular MPs should be used for experiments, saving time and money. This simple scoring methodology can be used for improving MP fabrication, comparing results across studies, benefiting basic science studies and potential future clinical use of MPs by introducing standardization.

Anti-inflammatory diet reduces risk of metabolic dysfunction-associated fatty liver disease among US adults: a nationwide survey

Background While dietary intervention was an important public health strategy for the prevention and intervention of metabolic dysfunction-associated fatty liver disease (MAFLD), the effect of diet-induced inflammation on MAFLD has not been studied in detail. Therefore, we aimed to analyze the relationship between dietary inflammatory index (DII) and MAFLD. Methods This study included data from the National Health and Nutrition Examination Survey 2017–2018. MAFLD was diagnosed based on the presence of hepatic steatosis, as determined by transient elastography, along with evidence of either overweight/obesity, type 2 diabetes mellitus, or metabolic dysfunction. DII was calculated using 27 dietary components collected through 24-hour dietary recall questionnaire. Weighted logistic regression was used to analyze the relationship between DII and MAFLD and its main components in three different models. Subgroup analyses were performed by age, sex, and alcohol use. Results A total of 1991 participants were included, and the MAFLD group had higher DII scores. After adjusting for age, sex, race, physical activity, smoking status, and alcohol use, the highest quartile of DII was associated with increased risk of MAFLD (OR:2.90, 95% CIs: 1.46, 5.75). Overweight/obesity, central obesity, low high density lipoprotein cholesterol (HDL-C) and high C-reactive protein (CRP) also shared the same characteristics in the main components of MAFLD. Results were consistent across subgroups (age, sex, and alcohol use). Conclusions A higher DII diet was positively associated with the risk of MAFLD in American adults, particularly as related to overweight/obesity, central obesity, high CRP level, and low HDL-C level.

Engineering artificial non-coding RNAs for targeted protein degradation.

Targeted protein degradation has become a notable drug development strategy, but its application has been limited by the dependence on protein-based chimeras with restricted genetic manipulation capabilities. The use of long non-coding RNAs (lncRNAs) has emerged as a viable alternative, offering interactions with cellular proteins to modulate pathways and enhance degradation capabilities. Here we introduce a strategy employing artificial lncRNAs (alncRNAs) for precise targeted protein degradation. By integrating RNA aptamers and sequences from the lncRNA HOTAIR, our alncRNAs specifically target and facilitate the ubiquitination and degradation of oncogenic transcription factors and tumor-related proteins, such as c-MYC, NF-κB, ETS-1, KRAS and EGFR. These alncRNAs show potential in reducing malignant phenotypes in cells, both in vitro and in vivo, offering advantages in efficiency, adaptability and versatility. This research enhances knowledge of lncRNA-driven protein degradation and presents an effective method for targeted therapies.

Intrinsic Disorder and Other Malleable Arsenals of Evolved Protein Multifunctionality.

Microscopic evolution at the functional biomolecular level is an ongoing process. Leveraging functional and high-throughput assays, along with computational data mining, has led to a remarkable expansion of our understanding of multifunctional protein (and gene) families over the past few decades. Various molecular and intermolecular mechanisms are now known that collectively meet the cumulative multifunctional demands in higher organisms along an evolutionary path. This multitasking ability is attributed to a certain degree of intrinsic or adapted flexibility at the structure-function level. Evolutionary diversification of structure-function relationships in proteins highlights the functional importance of intrinsically disordered proteins/regions (IDPs/IDRs) which are highly dynamic biological soft matter. Multifunctionality is favorably supported by the fluid-like shapes of IDPs/IDRs, enabling them to undergo disorder-to-order transitions upon binding to different molecular partners. Other new malleable members of the protein superfamily, such as those involved in fold-switching, also undergo structural transitions. This new insight diverges from all traditional notions of functional singularity in enzyme classes and emphasizes a far more complex, multi-layered diversification of protein functionality. However, a thorough review in this line, focusing on flexibility and function-driven structural transitions related to evolved multifunctionality in proteins, is currently missing. This review attempts to address this gap while broadening the scope of multifunctionality beyond single protein sequences. It argues that protein intrinsic disorder is likely the most striking mechanism for expressing multifunctionality in proteins. A phenomenological analogy has also been drawn to illustrate the increasingly complex nature of modern digital life, driven by the need for multitasking, particularly involving media.

SHC1 serves as a prognostic and immunological biomarker in clear cell renal cell carcinoma: a comprehensive bioinformatics and experimental analysis.

SHC1 plays a crucial regulatory role in various tumors, but its significance in predicting prognosis and immune response in clear cell renal cell carcinoma (ccRCC) is yet to be determined. In this study, we conducted a bioinformatics analysis of SHC1 expression, prognosis, and immunological functions in ccRCC using multiple databases. The association between SHC1 and immune infiltration, immune escape, and immunotherapy in ccRCC was systematically established. In addition, we validated our results by western blot of tumor and adjacent-tumor samples from nine ccRCC patients, as well as three renal carcinoma cell lines compared to a normal renal cell line. Our analysis revealed that the mRNA expression level of SHC1 in ccRCC tissues is significantly higher than that in normal tissues. Consistently, western blot experiment showed ccRCC tissues and cell lines exhibit higher protein levels that normal tissues and cell lines. Importantly, patients with low expression of SHC1 demonstrated a higher survival rate, indicating that SHC1 could serve as an independent prognostic factor for predicting survival in ccRCC. Additionally, high expression of SHC1 was associated with increased severe immune cell infiltration, enhanced immune escape, and higher immunotherapy scores. Hence, SHC1 emerges as a novel and easily detectable biomarker for predicting clinical outcomes, immune escape, and immunotherapy response in patients with ccRCC.

Alternating binding and p97-mediated dissociation of SDS22 and I3 recycles active PP1 between holophosphatases

Protein phosphatase-1 catalytic subunit (PP1) joins diverse targeting subunits to form holophosphatases that regulate many cellular processes. Newly synthesized PP1 is known to be transiently sequestered in an inhibitory complex with Suppressor-of-Dis2-number-2 (SDS22) and Inhibitor-3 (I3), which is disassembled by the ATPases Associated with diverse cellular Activities plus (AAA+) protein p97. Here, we show that the SDS22-PP1-I3 complex also acts as a thermodynamic sink for mature PP1 and that cycles of SDS22-PP1-I3 formation and p97-driven disassembly regulate PP1 function and subunit exchange beyond PP1 biogenesis. Förster Resonance energy transfer (FRET) analysis of labeled proteins in vitro revealed that in the p97-mediated disassembly step, both SDS22 and I3 dissociate concomitantly, releasing PP1. In presence of a targeting subunit, for instance Growth Arrest and DNA Damage-inducible protein 34 (GADD34), liberated PP1 formed an active holophosphatase that dephosphorylated its substrate, eukaryotic translation initiation factor 2 alpha (eIF2α). Inhibition of p97 results in displacement of the GADD34 targeting subunit by rebinding of PP1 to SDS22 and I3 indicating that the SDS22-PP1-I3 complex is thermodynamically favored. Likewise, p97 inhibition in cells causes rapid sequestration of PP1 by free SDS22 and I3 at the expense of other subunits. This suggests that PP1 exists in a steady state maintained by spontaneous SDS22-PP1-I3 formation and adenosine triphosphate (ATP) hydrolysis, p97-driven disassembly that recycles active PP1 between different holophosphatase complexes to warrant a dynamic holophosphatase landscape.

PH-Controlled Chemoselective Rapid Azo-Coupling Reaction (CRACR) Enables Global Profiling of Serotonylation Proteome in Cancer Cells.

Serotonylation has been identified as a novel protein posttranslational modification for decades, where an isopeptide bond is formed between the glutamine residue and serotonin through transamination. Transglutaminase 2 (also known as TGM2 or TGase2) was proven to act as the main "writer" enzyme for this PTM, and a number of key regulatory proteins (including small GTPases, fibronectin, fibrinogen, serotonin transporter, and histone H3) have been characterized as the substrates of serotonylation. However, due to the lack of pan-specific antibodies for serotonylated glutamine, the precise enrichment and proteomic profiling of serotonylation still remain challenging. In our previous research, we developed an aryldiazonium probe to specifically label protein serotonylation in a bioorthogonal manner, which depended on a pH-controlled chemoselective rapid azo-coupling reaction. Here, we report the application of a photoactive aryldiazonium-biotin probe for the global profiling of serotonylation proteome in cancer cells. Thus, over 1,000 serotonylated proteins were identified from HCT 116 cells, many of which are highly related to carcinogenesis. Moreover, a number of modification sites of these serotonylated proteins were determined, attributed to the successful application of our chemical proteomic approach. Overall, these findings provided new insights into the significant association between cellular protein serotonylation and cancer development, further suggesting that to target TGM2-mediated monoaminylation may serve as a promising strategy for cancer therapeutics.

Morphology of the Digestive Tube of the Amazonian Freshwater Stingray Potamotrygon wallacei (Elasmobranchii: Potamotrygonidae): A Stereological Approach.

This work aimed to describe and quantify the tissue components of the digestive tube of the neotropical freshwater stingray, Potamotrygon wallacei. For this, conventional histology and stereological methods were used to estimate tissue volume. The volumes of the four fundamental layers and the tissue components in the stomach (cardiac and pyloric) and spiral intestine were also estimated. In the cardiac stomach, the mucosa layer occupies 44.7% of the total volume of the organ wall. The gastric glands are the main components, and these structures alone represent 49.7% of this layer. This large number of gastric glands suggests a high potential for processing food items with a high protein content. The stereological methods were sensitive enough to show a reduction in the volume of the gastric glands from the cardiac region toward the pyloric region. Gastric glands are absent in the pyloric region of the stomach. However, the muscularis becomes thicker towards the pyloric region. The increase in smooth muscle thickness is due to the thickening of the inner muscular layer. This suggests that the role of the pyloric stomach may be related to the mixing of the chyme and assisting its passage to the spiral intestine. In the spiral intestine, data on the volume of the mucosa layer (and epithelial lining) suggest that the spiral valve has a large absorptive area. In several respects, the morphology of the digestive tube of P. wallacei is similar to that of other batoids. However, its slight morphological variations may be related to the habitat specificity of this species.

Kadar Protein, Kadar Albumin, dan Tingkat Kesukaan Brownies Mocaf Ikan Gabus

Background: Brownies are generally made from wheat flour, but can be made with local food mocaf flour. Mocaf protein is lower than wheat flour, wheat flour has 8-13% protein while mocaf has 1.2% protein content. Objectives: The purpose of the study was to determine the addition of snakehead fish to protein levels, albumin levels, and preference test for mocaf brownies.The addition of snakehead fish to mocaf brownies aims to increase protein levels. Snakehead fish contains high protein and albumin,protein of 16.2g/100g and albumin content of 6.224g/100g. Preference test is researched to find out brownies are acceptable to consumers. Method: The study used the Complete Randomized Design experimental method with the additionof snakehead fish 0%;7.3%;9.6%;11.7%, repetition 6 times. Protein content testing is carried out by the kjeldahl method, albumin levels by the biuret method and preference levels by organoleptic tests. Data analysis of protein levels and albumin levels using the ANOVA one-way test, and analysis of favorability data using the Friedman Test test then proceed to the Wilcoxon test. Results: The test results of protein levels and albumin levels get a value of p &gt;0.05, namely the addition of snakehead fish does not have a significant effect. The results of the favorability leveltest on color, aroma, taste get a value of p &lt; 0.05 that the addition of snakehead fish has a significant effect and the texture of p &gt; 0.05 is not significant. Test results with the addition of snakehead fish with the highest protein content and albumin content treatment 11.7%, the highest color test at 7.3% and 9.6%, the highest aroma test at 9.6%, the highest taste and texture test at 7.3%. Keywords : Albumin Levels, Mocaf Brownies, Preference Levels, Protein Levels, Snakehead Fish

Heparan sulfate proteoglycans in pathological protein aggregation and brain functionality

Proteoglycans are highly specialized proteins with specific interactive modules in their core proteins, allowing them to engage with a diverse range of cellular and structural proteins, contributing to many varied roles in cell processes. This review examines the roles of heparan sulfate (HS) proteoglycans (HSPGs) in the assembly of pathological protein aggregates in brain tissues, which negatively impact cognitive brain function. While HSPGs play roles in normal brain development, they also contribute to the abnormal accumulation and persistence of protein aggregates in Alzheimer&amp;rsquo;s and Parkinson&amp;rsquo;s diseases (AD and PD), prolonging their neurotoxic effects. HSPG-mediated effects on the innate immune system may also contribute to the development of neuroinflammation and pathogenesis of neurodegenerative changes in the central nervous system. These pathological developments lead to impaired cognition, permanent memory loss in dementia and AD, and an inability of the brain to exert normal neuromuscular control, affecting motor functions and body movement in PD. A deeper understanding of the properties of HS in these degenerative processes is essential for the development of therapeutic measures to treat these conditions. Significant improvements in neuroimaging, next-generation glycosaminoglycan analytical techniques, and functional glycomics hold great promise for elucidating the complexities of HS structure and function. These advancements may significantly aid in the development of HS biotherapeutics to treat these debilitating conditions, which are increasingly impacting the aging global population. While a number of promising therapeutic candidates have already emerged, further research is required to optimize their biotherapeutic applications in this challenging area of pathobiology, bioregulation, and the recovery of neural function.

The role of ubiquitin-conjugating enzyme in the process of spermatogenesis.

The ubiquitination is crucial for controlling cellular homeostasis and protein modification, in which ubiquitin-conjugating enzyme (E2) acts as the central player in the ubiquitination system. Ubiquitin-conjugating enzymes, which have special domains that catalyse substrates, have sequence discrepancies and modulate various pathophysiological processes in different cells of multiple organisms. E2s take part in the mitosis of primordial germ cells, meiosis of spermatocytes and the formation of mature haploid spermatids to maintain normal male fertility. In this review, we summarize the various types of E2s and their functions during distinct stages of spermatogenesis.

The cellular paraspeckle component SFPQ associates with the viral processivity factor ORF59 during lytic replication of Kaposi's Sarcoma-associated herpesvirus (KSHV).

Kaposi's sarcoma-associated herpesvirus (KSHV) relies on many cellular proteins to complete replication and generate new virions. Paraspeckle nuclear bodies consisting of core ribonucleoproteins splicing factor proline/glutamine-rich (SFPQ), Non-POU domain-containing octamer-binding protein (NONO), and paraspeckle protein component 1 (PSPC1) along with the long non-coding RNA NEAT1, form a complex that has been speculated to play an important role in viral replication. Paraspeckle bodies are multifunctional and involved in various processes including gene expression, mRNA splicing, and anti-viral defenses. To better understand the role of SFPQ during KSHV replication, we performed SFPQ immunoprecipitation followed by mass spectrometry from KSHV-infected cells. Proteomic analysis showed that during lytic reactivation, SFPQ associates with viral proteins, including ORF10, ORF59, and ORF61. These results are consistent with a previously reported ORF59 proteomics assay identifying SFPQ. To test if the association between ORF59 and SFPQ is important for replication, we first identified the region of ORF59 that associates with SFPQ using a series of 50 amino acid deletion mutants of ORF59 in the KSHV BACmid system. By performing co-immunoprecipitations, we identified the region spanning amino acids 101-150 of ORF59 as the association domain with SFPQ. Using this information, we generated a dominant negative polypeptide of ORF59 encompassing amino acids 101-150, that disrupted the association between SFPQ and full-length ORF59, and decreased virus production. Interestingly, when we tested other human herpesvirus processivity factors (EBV BMRF1, HSV-1 UL42, and HCMV UL44) by transfection of each expression plasmid followed by co-immunoprecipitation, we found a conserved association with SFPQ. These are limited studies that remain to be done in the context of infection but suggest a potential association of SFPQ with processivity factors across multiple herpesviruses.

The Effect of the Species Source of Muscle and/or Digestive Enzymes on the Utilization of Fish Protein Hydrolysates as a Dietary Protein Source in First Feed for Larval Walleye (Sander vitreus)

Fish protein hydrolysates used in larval diets have been prepared from a variety of fish species, with different enzymes used to hydrolyze the protein. This study’s objectives were to determine the effect of the dietary inclusion of fish muscle hydrolysates obtained from species-specific muscle/enzymes—versus hydrolysates produced from muscle/enzymes of a different species—on the growth performance, survival, skeletal development, intestinal peptide uptake, and muscle-free amino acid (FAA) composition of larval Walleye (Sander vitreus). Eight protein products were obtained for this study, comprising an unhydrolyzed and hydrolyzed product from each combination of muscle/enzymes from Walleye and Nile tilapia (Oreochromis niloticus). Four diets were produced, and the dietary protein was provided in a 50/50 ratio of unhydrolyzed and hydrolyzed protein from the respective muscle/enzyme combination. Four groups were fed one of the corresponding formulated diets, and two groups of larvae, fed a commercial starter diet and Artemia, respectively, served as reference groups. Larval Walleye fed the diet containing protein produced with the species-specific muscle and enzymes had a significantly higher weight after the study—30% higher than any other group. A significant interaction effect between muscle and enzyme sources on the growth of Walleye larvae was observed. The species-specific combination also led to a significant increase in postprandial FAA and indispensable amino acid concentrations in muscle. No significant differences were observed between the hydrolysate-fed groups in survival, deformity occurrence, or peptide uptake. Each hydrolysate-based diet significantly reduced skeletal deformities and survival compared to the commercial diet. The results of this study suggest that species-specific muscles and enzymes produce a more optimal dietary protein source for larval fish than non-species-specific products. Further research should focus on improving the physical properties of the formulated diets to reduce possible leaching of hydrolyzed protein and improve the survival of fish larvae.