High Protein Research Articles

A multiscale functional map of somatic mutations in cancer integrating protein structure and network topology.

A major goal of cancer biology is to understand the mechanisms underlying tumorigenesis driven by somatically acquired mutations. Two distinct types of computational methodologies have emerged: one focuses on analyzing clustering of mutations within protein sequences and 3D structures, while the other characterizes mutations by leveraging the topology of protein-protein interaction network. Their insights are largely non-overlapping, offering complementary strengths. Here, we established a unified, end-to-end 3D structurally-informed protein interaction network propagation framework, NetFlow3D, that systematically maps the multiscale mechanistic effects of somatic mutations in cancer. The establishment of NetFlow3D hinges upon the Human Protein Structurome, a comprehensive repository we compiled that incorporates the 3D structures of every single protein as well as the binding interfaces of all known protein interactions in humans. NetFlow3D leverages the Structurome to integrate information across atomic, residue, protein and network levels: It conducts 3D clustering of mutations across atomic and residue levels on protein structures to identify potential driver mutations. It then anisotropically propagates their impacts across the protein interaction network, with propagation guided by the specific 3D structural interfaces involved, to identify significantly interconnected network "modules", thereby uncovering key biological processes underlying disease etiology. Applied to 1,038,899 somatic protein-altering mutations in 9,946 TCGA tumors across 33 cancer types, NetFlow3D identified 1,4444 significant 3D clusters throughout the Human Protein Structurome, of which ~55% would not have been found if using only experimentally-determined structures. It then identified 26 significantly interconnected modules that encompass ~8-fold more proteins than applying standard network analyses. NetFlow3D and our pan-cancer results can be accessed from http://netflow3d.yulab.org.

Recombinant expression and characterization of the endochitinase Chit36-TA from Trichoderma asperellum in Komagataella phaffii for chitin degradation of black soldier fly exuviae.

The natural polymer chitin is an abundant source for valuableN-acetylchitooligosaccharides and N-acetylglucosamine applicable in several industries. The endochitinase Chit36-TA from Trichoderma asperellum was recombinantly expressed in Komagataella phaffii for the enzymatic degradation of chitin from unused insect exuviae into N-acetylchitooligosaccharides. Chit36-TA was purified by Ni-NTA affinity chromatography and subsequently biochemically characterized. After deglycosylation, the endochitinase had a molecular weight of 36kDa. The optimum pH for Chit36-TA was 4.5. The temperature maximum of Chit36-TA was determined to be 50°C, while it maintained > 93% activity up to 60°C. The chitinase was thermostable up to 45°C and exhibited ~ 50% activity after a 15min incubation at 57°C. Chit36-TA had a maximum specific enzyme activity of 50 nkat/mg with a Km value of 289µM with 4-methylumbelliferyl-N,N',N″-triacetyl-β-chitotrioside as substrate. Most tested cations, organic solvents and reagents were well-tolerated by the endochitinase, except for SDS (1mM), Cu2+ (10mM) and Mn2+ (10mM), which had stronger inhibitory effects with residual activities of 3, 41 and 28%, respectively. With a degree of hydrolysis of 32% applying colloidal shrimp chitin (1% (w/v)) and 12% on insect larvae (1% (w/v)) after 24h, the endochitinase was found to be suitable for the conversion of colloidal chitin as well as chitin from black soldier fly larvae into water-soluble N-acetylchitooligosaccharides. To prove scalability, a bioreactor process was developed in which a 55-fold higher enzyme activity of 49µkat/l and a tenfold higher protein expression of 1258mg/l were achieved.

Syntaxin-6 delays prion protein fibril formation and prolongs presence of toxic aggregation intermediates.

Prions replicate via the autocatalytic conversion of cellular prion protein (PrPC) into fibrillar assemblies of misfolded PrP. While this process has been extensively studied in vivo and in vitro, non-physiological reaction conditions of fibril formation in vitro have precluded the identification and mechanistic analysis of cellular proteins, which may alter PrP self-assembly and prion replication. Here, we have developed a fibril formation assay for recombinant murine and human PrP (23-231) under near-native conditions (NAA) to study the effect of cellular proteins, which may be risk factors or potential therapeutic targets in prion disease. Genetic screening suggests that variants that increase syntaxin-6 expression in the brain (gene: STX6) are risk factors for sporadic Creutzfeldt-Jakob disease (CJD). Analysis of the protein in NAA revealed, counterintuitively, that syntaxin-6 is a potent inhibitor of PrP fibril formation. It significantly delayed the lag phase of fibril formation at highly sub-stoichiometric molar ratios. However, when assessing toxicity of different aggregation time points to primary neurons, syntaxin-6 prolonged the presence of neurotoxic PrP species. Electron microscopy and super-resolution fluorescence microscopy revealed that, instead of highly ordered fibrils, in the presence of syntaxin-6 PrP formed less-ordered aggregates containing syntaxin-6. These data strongly suggest that the protein can directly alter the initial phase of PrP self-assembly and, uniquely, can act as an 'anti-chaperone', which promotes toxic aggregation intermediates by inhibiting fibril formation.

HnRNPs in antiviral innate immunity.

During virus infection, many host proteins are redirected from their normal cellular roles to restrict and terminate infection. Heterogeneous nuclear ribonucleoproteins (hnRNPs) are cellular RNA-binding proteins critical to host nucleic acid homeostasis, but can also be involved in the viral infection process, affecting virus replication, assembly and propagation. It has become evident that hnRNPs play important roles in modulation of host innate immunity, which provides critical initial protection against infection. These novel findings can potentially lead to the leveraging of hnRNPs in antiviral therapies. We review hnRNP involvement in antiviral innate immunity, in humans, mice and other animals, and discuss hnRNP targeting as a potential novel antiviral therapeutic.

Graphene Oxide Nanosheets Toxicity in Mice Is Dependent on Protein Corona Composition and Host Immunity.

Two-dimension graphene oxide (GO) nanosheets with high and low serum protein binding profiles (high/low hard-bound protein corona/HChigh/low) are used in this study as model materials and screening tools to investigate the underlying roles of the protein corona on nanomaterial toxicities in vivo. We proposed that the in vivo biocompatibility/nanotoxicity of GO is protein corona-dependent and host immunity-dependent. The hypothesis was tested by injecting HChigh/low GO nanosheets in immunocompetent ICR/CD1 and immunodeficient NOD-scid II2rγnull mice and performed histopathological and hematological evaluation studies on days 1 and 14 post-injection. HClow GO induced more severe acute lung injury compared to HChigh GO in both immunocompetent and immunodeficient mice, with the effect being particularly pronounced in immunocompetent animals. Additionally, HClow GO caused more significant liver injury in both types of mice, with immunodeficient mice being more susceptible to its hepatotoxic effects. Moreover, administration of HClow GO resulted in increased hematological toxicity and elevated levels of serum pro-inflammatory cytokines in immunocompromised and immunocompetent mice, respectively. Correlation studies were conducted to explore the impact of distinct protein corona compositions on resulting toxicities in both immunocompetent and immunodeficient mice. This facilitated the identification of consistent patterns, aligning with those observed in vitro, thus indicating a robust in vitro-in vivo correlation. This research will advance our comprehension of how hard corona proteins interact with immune cells, leading to toxicity, and will facilitate the development of improved immune-modulating nanomaterials for therapeutic purposes.

Protein Intake and Mortality in Older Adults With Chronic Kidney Disease.

Avoiding high protein intake in older adults with chronic kidney disease (CKD) may reduce the risk of kidney function decline, but whether it can be suboptimal for survival is not well known. To estimate the associations of total, animal, and plant protein intake with all-cause mortality in older adults with mild or moderate CKD and compare the results to those of older persons without CKD. Data from 3 cohorts (Study on Cardiovascular Health, Nutrition and Frailty in Older Adults in Spain 1 and 2 and the Swedish National Study on Aging and Care in Kungsholmen [in Sweden]) composed of community-dwelling adults 60 years or older were used. Participants were recruited between March 2001 and June 2017 and followed up for mortality from December 2021 to January 2024. Those with no information on diet or mortality, with CKD stages 4 or 5, or undergoing kidney replacement therapy and kidney transplant recipients were excluded. Data were originally analyzed from June 2023 to February 2024 and reanalyzed in May 2024. Cumulative protein intake, estimated via validated dietary histories and food frequency questionnaires. The study outcome was 10-year all-cause mortality, ascertained with national death registers. Chronic kidney disease was ascertained according to estimated glomerular filtration rates, urine albumin excretion, and diagnoses from medical records. The study sample consisted of 8543 participants and 14 399 observations. Of the 4789 observations with CKD stages 1 to 3, 2726 (56.9%) corresponded to female sex, and mean (SD) age was 78.0 (7.2) years. During the follow-up period, 1468 deaths were recorded. Higher total protein intake was associated with lower mortality among participants with CKD; adjusted hazard ratio (HR) for 1.00 vs 0.80 g/kg/d was 0.88 (95% CI, 0.79-0.98); for 1.20 vs 0.80 g/kg/d, 0.79 (95% CI, 0.66-0.95); and for 1.40 vs 0.80 g/kg/d, 0.73 (95% CI, 0.57-0.92). Associations with mortality were comparable for plant and animal protein (HRs, 0.80 [95% CI, 0.65-0.98] and 0.88 [95% CI, 0.81-0.95] per 0.20-g/kg/d increment, respectively) and for total protein intake in participants younger than 75 years vs 75 years or older (HRs, 0.94 [95% CI, 0.85-1.04] and 0.91 [95% CI, 0.85-0.98] per 0.20-g/kg/d increment in total protein intake, respectively). However, the hazards were lower among participants without CKD than in those with CKD (HRs, 0.85 [95% CI, 0.79-0.92] and 0.92 [95% CI, 0.86-0.98] per 0.20-g/kg/d increment, respectively; P = .02 for interaction). In this multicohort study of older adults, higher intake of total, animal, and plant protein was associated with lower mortality in participants with CKD. Associations were stronger in those without CKD, suggesting that the benefits of proteins may outweigh the downsides in older adults with mild or moderate CKD.

Regulation of the p53/SLC7A11/GPX4 Pathway by Gentamicin Induces Ferroptosis in HEI-OC1 Cells.

Gentamicin is a commonly used aminoglycoside antibiotic, with ototoxicity as a significant side effect. Ferroptosis, an iron-dependent form of cell death, has been implicated in a variety of disorders. Whether ferroptosis impacts gentamicin ototoxicity is not yet known. The current work used an in-vitro model to examine the influence of gentamicin-induced ferroptosis on cochlear hair cell damage and probable molecular biological pathways. House Ear Institute-Organ of Corti 1 (HEI-OC1) cells were treated with different concentrations of gentamicin for 24 hours, with or without ferrostatin-1 pretreatment, to observe gentamicin-induced ferroptosis. The role of p53/solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) signaling in gentamicin-induced ferroptosis was explored by pretreating cells with the p53 inhibitor pifithrin-α (PFT-α). We investigated the effect of gentamicin on cells by assessing cell viability. Cellular proteins were isolated and Western blots were performed to detect changes in the expression of p53, SLC7A11, and GPX4. Fluorescence staining was used to assess levels of reactive oxygen species. An enzymatic detection kit was used to detect glutathione, Fe, and malondialdehyde markers. Gentamicin reduced cell viability, glutathione content, and SLC7A11 and GPX4 protein levels, and increased levels of p53 protein, reactive oxygen species, malondialdehyde, and Fe. These effects were largely blocked by pretreatment with ferrostatin-1. Pretreatment with the p53 inhibitor PFT-α prevented the gentamicin-induced reduction in SLC7A11 and GPX4, which alleviated several features of ferroptosis including glutathione depletion, iron overload, and lipid peroxidation build-up. Gentamicin induces ferroptosis in the HEI-OC1 cell line, and the mechanism may be related to the p53/SLC7A11/GPX4 signaling pathway.

Do we neglect nutrition in childhood interstitial lung disease?

Growth failure and inadequate weight gain are common problems in childhood interstitial lung diseases (chILD) and these children usually need high calories. It is important to manage both pulmonary functions and nutrition as part of their overall treatment plan and early interventions will help children to improve their quality of life and slow the progression of chronic lung disease. Nutritional evaluation on routine clinical follow-up is important, although there are not any specific guidelines for chILD. Nutritional education, high balanced energy, protein, and fat diet will assist to improve weight gain and maintenance of adequate nutrition status in children with ILD.

Modification of an AIE Fluorescent Probe for Monitoring the Polarity of Lipid Droplets Based on a series of Synthesized Aryl Naphthalizes.

Lipid droplets (LDs) are subcellular organelles that are dynamic and play a central role in energy homeostasis and lipid metabolism. They also contribute to the transport and maturation of cellular proteins and are closely associated with several diseases. The important role of the cellular microenvironment in maintaining cellular homeostasis. Changes in cell polarity, particularly in organelles, have been found to be strongly linked to inflammation, Alzheimer's disease, cancer, and other illnesses. It is essential to check the polarity of the LDs. A series of arylated naphthalimide derivatives were synthesized using the Suzuki reaction. Modification of synthesized aryl naphthalimides using oligomeric PEG based on intramolecular charge transfer (ICT) mechanism. A series of fluorescent probes were designed to target LDs and detect their polarity. Nap-TPA-PEG3 probe exhibited high sensitivity to polarity. The addition of oligomeric polyethylene glycol (PEG) to the probe not only significantly improved its solubility in water, but also effectively reduced its cytotoxicity. In addition, the probe exhibited excellent aggregation-induced luminescence (AIE) properties and solvent discolouration effects. Nap-TPA-PEG3 probe exhibited high Pearson correlation coefficient (0.957163) in lipid droplet co-localization in cells. Nap-TPA-PEG3 could be used as an effective hand tool to monitor cell polarity.

Helicobacter pylori upregulates PAD4 expression via stabilising HIF-1α to exacerbate rheumatoid arthritis

ObjectiveHelicobacter pylori infection has been reported to aggravate rheumatoid arthritis (RA), but the relevant mechanism remains unclear. This study aimed to investigate the underlying pathogenic mechanism of H. pylori infection...

A comparative analysis of crustacean exoskeletons: structural, microstructural, morphological, and UV absorption studies

Abstract This study aims to investigate the structural, thermal, and spectral characteristics along with ultra-violet (UV) absorption of various marine benthos exoskeletons, such as various species of crabs (Portunus sanguinolentus, Portunus pelagicus, Charybdis feriata) and mantis shrimp (Oratosquilla oratoria). Their unique properties and ability to survive in harsh oceanic environments make them interesting research subjects. This research utilized powder X-ray diffraction (XRD) analysis to determine the crystal structure of the benthic varieties. The sample surface was analysed using high-resolution micrographs obtained from field-emission scanning electron microscopy (FESEM) identified the presence of chitin and calcite in the marine benthos, which was further confirmed by differential scanning calorimetry (DSC), and Fourier transform infrared spectroscopy (FTIR). The optical characteristics were investigated using UV-visible spectroscopy. The proximate analysis revealed high protein content in the mantis shrimp exoskeleton compared to other crab species, revealing its excellent UV-absorption characteristics. Overall, this research has the potential to broaden our understanding of marine organisms, which can have potential applications in biotechnology and materials science to develop nature-inspired innovative materials sustainably.

Effects of dietary protein levels on growth performance, hemato–biochemical, and immune parameters in ship sturgeon (Acipenser nudiventris) juveniles

A feeding trial was conducted to detect the optimal dietary protein requirement for ship sturgeon (Acipenser nudiventris) juveniles. Four isoenergetic diets were formulated to contain different protein levels i.e. 40 % (P40), 45 % (P45), 50 % (P50), and 55 % (P55). A total of 180 ship sturgeon juveniles, with an average weight of 63.5 ± 1.3 g, were randomly distributed into triplicated groups (15 fish per tank) and fed on the experimental diets for 10 weeks. Significant (P < 0.05) increases in growth indices and feed intake were noted with increasing dietary protein levels up to 50 % after which (P55) fish growth was declined. The highest values of protein and lowest value of lipids and ash contents were noted in the whole-body of fish fed on the P50 diet. The levels of red blood cells, hemoglobin, and hematocrit were markedly influenced (P < 0.05) by varying dietary protein concentrations, and the most elevated values were identified in the P50 treatment. Linear increases were observed in alanine aminotransferase and alkaline phosphate levels; while quadratic increases were observed in aspartate aminotransferase activities due to the increase in dietary protein levels. Quadratic increases in total protein (TP), total immunoglobulin (total Ig), and lysozyme (LYZ) were observed due to feeding the fish on higher protein levels. The highest level of TP was significantly (P < 0.05) found in P50 and P55 with no significant (P > 0.05) difference between them. Levels of total Ig and LYZ were significantly (P > 0.05) highest in the P50 treatment. Based on the regression fitting curves of growth indices, the optimum dietary protein level for ship sturgeon (A. nutiventis) juveniles is found to be 46.5 % – 50 %. Data AvailabilityAll data of this study are included in this article.

PRNP is a pan-cancer prognostic and immunity-related to EMT in colorectal cancer

BackgroundPrion protein gene (PRNP) is widely expressed in a variety of tissues. Although the roles of PRNP in several cancers have been investigated, no pan-cancer analysis has revealed its relationship with tumorigenesis and immunity.MethodsComprehensive analyses were conducted on The Cancer Genome Atlas (TCGA) Pan-Cancer dataset from the University of California Santa Cruz (UCSC) database to determine the expression of PRNP and its potential prognostic implications. Immune infiltration and enrichment analysis methods were used to ascertain correlations between PRNP expression levels, tumor immunity, and immunotherapy. Additionally, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) methods were employed to examine possible signaling pathways involving PRNP. In vitro experiments using CCK-8 assay, Wound healing assay, and Transwell assay to detect the effect of Cellular prion protein (PrPC) on proliferation, migration, and invasion in colorectal cancer (CRC) cells. The expression levels of epithelial-mesenchymal transition (EMT)-related proteins (N-cadherin, E-cadherin, Vimentin and Snail) were detected by western blot.ResultsAmong most cancer types, PRNP is expressed at high levels, which is linked to the prognosis of patients. PRNP expression is strongly associated with immune infiltrating cells, immunosuppressive cell infiltration and immune checkpoint molecules. In addition to tumor mutation burden (TMB), substantial correlations are detected between PRNP expression and microsatellite instability (MSI) in several cancers. In vitro cell studies inferred that PrPC enhanced the proliferation, migration, invasion, and EMT of CRC cells.ConclusionPRNP serves as an immune-related prognostic marker that holds promise for predicting outcomes related to CRC immunotherapy while simultaneously promoting cell proliferation, migration, and invasion activities. Furthermore, it potentially plays a role in governing EMT regulation within CRC.

The Impact of Combined Application of Donepezil and Nimodipine on Patients with Comorbid Cerebral Small Vessel Disease and Cognitive Dysfunction: Efficacy and Influence on Nutritional Status

Background and objective: Cerebral small vessel disease (CSVD) often coexists with cognitive dysfunction in patients, leading to significant challenges in treatment and management. This study aimed to examine the efficacy of combined application of donepezil and nimodipine on patients with comorbid CSVD and cognitive dysfunction and the effects on patients' albumin and prealbumin levels. Methods: The records of 112 patients with comorbid CSVD and cognitive dysfunction treated at the People's Hospital of Suzhou New District from January 2019 to December 2022 were analysed retrospectively. A total of 50 patients receiving donepezil were allocated to the control group, and 62 patients receiving both nimodipine and donepezil to the study group. Outcomes compared between the two groups included serum homocysteine (Hcy), high sensitivity C-reactive protein (hs-CRP), albumin, and prealbumin before and after therapy, efficacy, and adverse reactions. Additionally, logistic regression was performed to analyze the risk factors impacting patient prognosis. Results: Prior to therapy, the two groups did not differ significantly in Hcy and hs-CRP levels (p &gt; 0.05), whereas after therapy, the levels in both groups dropped significantly (p &lt; 0.01), with more obvious lower levels in the study group (p &lt; 0.05). After treatment, the study group presented significantly higher albumin and prealbumin levels than the control group (p &lt; 0.001). An obvious higher overall response rate was observed in the study group compared to the control group (p = 0.012). No significant inter-group discrepancy was found regarding the total incidence of adverse reactions (p = 0.752). Univariate analysis identified age, course of disease, heart rate (HR), Montreal Cognitive Assessment (MoCA) score, diastolic blood pressure (DBP), systolic blood pressure (SBP), drinking history, as well as medication regimen as risk factors impacting patient prognosis. Multivariate logistic regression analysis identified SBP, DBP, and medication regimen as the independent risk factors. Conclusion: Combined application of donepezil and nimodipine can effectively treat patients with comorbid CSVD and cognitive dysfunction. It can significantly lower the Hcy and hs-CRP levels and improve the nutritional status without increasing the frequency of adverse reactions. In addition, for CSVD patients with cognitive dysfunction, age, course of disease, MoCA score, HR, SBP, DBP, drinking history, and medication regimen are risk factors impacting patient prognosis, while SBP, DBP, and medication regimen are independent risk factors.

Effect of an intensive nutrition intervention of a high protein and low glycemic load diet on weight of kidney transplant recipients: a randomized clinical trial.

The purpose of this study is to evaluate the effect of a high protein and low glycemic load diet in preventing weight gain after kidney transplantation. We designed a prospective, single-center, open-label, randomized controlled study to compare the efficacy of a high protein (1.3-1.4 g/kg/day) and low glycemic load diet versus a conventional diet (0.8-1.0 g/kg/day of protein and no recommendations on glycemic load) in preventing weight gain (ClinicalTrials.gov identifier: NCT02883777). A total of 120 patients were evaluated. Patients were followed for 12 months, and the primary outcome was weight maintenance or weight gain lower than 5%. There were no differences in total energy intake, carbohydrates, and total fats between groups. Intervention group (IG) increased protein intake to 1.38 ± 0.56 g/kg/day and decreased the glycemic load to 87.27 ± 4.54 g/day, while control group (CG) had a dietary protein intake of 1.19 ± 0.43 g/kg/day and a glycemic load of 115.60 ± 7.01 g/day. Total fiber intake was greater and trans-fat was lower in IG. Dietetic cholesterol increased in IG over time and was significantly different between groups. Overall, patients had an increase in body weight over time, with a mean increment of 4.1 ± 5.5 kg (5.75%). The percentage of patients who achieved the primary outcome was 50% of sample size, without differences between groups. The glomerular filtration rate improved over time in both groups. Considering 24-h proteinuria and albuminuria, a similar rise was observed in both groups. The present dietary intervention was safe, but had no effect on weight gain in kidney transplant subjects. Our findings suggest that other strategies, including alternative dietary and/or pharmacological and psychological interventions might be tested in randomized control trials in order to improve patients' body weight outcomes after transplant.

Optimization of malted cowpea compositions and extrusion parameters for best quality Ethiopian emmer wheat-based snack food

Childhood malnutrition is a major concern in developing countries, particularly in Ethiopia. This study aims to address this problem by assessing the nutritional properties of snacks made from locally available raw materials. The study used a twin-screw extruder machine to study the effects of different processing conditions (barrel temperatures, feed moisture content, and cowpea/emmer wheat blending ratio) on the nutritional properties of extruded products. The results showed a highly significant effect of these processing variables on the nutritional properties of snacks. Increasing cowpea in the blend boosted the protein and fiber content of the snack products. Increasing the barrel temperature from 80 to 120 °C notably decreased the protein and fat levels in the snack. The optimization of the combined interactive effects on an extruded snack made from 19.83% cowpea and 80.17% emmer wheat, cooked at 120 °C barrel temperature with 22.04% feed moisture content, produced acceptable extruded snack items. The products developed with optimized parameters contained 16.68%, 1.55%, 2.50%, 70.95%, and 364.43 kcal/g for protein, fat, fiber, carbohydrate, and gross energy, respectively. The study concludes that blending Emmer wheat and cowpea in extruded snacks can provide high protein and gross energy. This suggests a locally viable solution for addressing protein-energy malnutrition in developing regions, particularly Ethiopia.

Exploring Arthropods as Sustainable Protein Source and the Effect of Cockroach Meal Supplemented Diet on the Growth of Fish

This comprehensive study centres on exploring the arthropods as sustainable protein sources, addressing both human dietary needs and livestock feed requirements. A diverse set of twenty arthropod species were collected from different locations of Kanyakumari District and the species were identified based on the identification keys pertaining to morphological characters. The surveyed animals comprises of Insects: Dragon Fly (Pantala sp.), Western subterranean termite (Reticulitermes sp.), Longhorn beetle (Callipogon sp.), Asiatic rhinoceros beetle (Oryctes rhinoceros), Brown grasshopper (Melanoplus sp.), Bush cricket (Tettigonia sp.), Grasshopper (Diabolocatantops sp.), Housefly (Musca domestica), Cockroach (Periplaneta americana), Termite (Macrotermes sp.), Black carpenter ant (Camponotus sp.), Red carpenter ant (Camponotus sp.); Crabs: Mottled light footed crab (Grapsus albolineatus), Sartoriana sp., Asian paddle crab (Charybdis sp.,),Box crab (Calappa sp.,), Asian shore crab (Hemigrapsus sp.); Millipedes: Yellow spotted millipede (Harpaphe haydeniana), Pill millipede (Arthrosphaera disticta) and African giant millipede (Thyropygus descriptus) were analyzed for the protein content. The samples were tested for protein using Biuret’s method. Among the different arthropods tested insects- Callipogon sp. and Periplaneta americana, crabs - Calappa sp. and Charybdis sp. and Myriapod - Harpaphe haydeniana, exhibited the highest protein content. Crustaceans demonstrated the highest protein content when compared to insects and millipedes. However, since cockroaches are generally considered as a pest and repugnant in human dwelling, it would be ideal to convert cockroach as a sustainable protein source in aquaculture. Moreover. since cockroach showed a higher protein content of 6.146 2.024 mg/ml it was tried as a supplement feed for fish. Cockroach meal was prepared in two different proportions (2% and 3%) to evaluate its impact on the growth of Koi carp fish in comparison with cockroach free meal as control. The inclusion of 3% cockroach powder in the fish meal augmented the growth of Koi carp when compared to the control during a fifteen-day study period. Thus, this study has tapped a potential protein source from insects. Cockroach can be used as an ideal fish feed as it contributes to the pursuit of sustainable and environmentally friendly animal feed source.

Impact of Dietary Variations on Kuruma Shrimp (Penaeus japonicus) Assessed through Individual-Based Rearing and Insights into Individual Differences

This study developed an individual-rearing method to compare the effects of live feed (sandworms Perinereis aibuhitensis), formulated pellet diets, and a mixture of live feed and formula feed on the Kuruma shrimp Penaeus japonicus, aiming to minimize the influence of non-dietary factors on the growth of P. japonicus, like cannibalism. Results indicated that live feed, with its higher protein, essential amino acids, and fatty acid content, led to significantly better growth and feeding performance in P. japonicus (p &lt; 0.05) compared to pellet diets. A mixed diet resulted in a lower average daily protein intake yet maintained a growth and feeding performance comparable to live feed. The intestinal microbiota of shrimp, dominated by Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria, showed significant shifts with diet changes. Specifically, formulated feed increased the relative abundance of Vibrio and Photobacterium while decreasing Shimia and Rhodobacterales (p &lt; 0.05), and feeding live food resulted in a more complex and stable bacterial network. Notably, individual variances in growth and feeding were observed among shrimps, with some on formulated diets showing growth comparable to those on live feed. Each shrimp’s final weight, specific growth rate, protein efficiency rate, and average daily food intake positively correlated with its initial body weight (p &lt; 0.05), and daily intake varied cyclically with the molting cycle. These findings suggest that individual-rearing is an effective approach for detailed feed evaluation and monitoring in P. japonicus, contributing to improved feed selection, development, and feeding strategies.

Growth and biomass of Wolffia arrhiza with different sources of organic fertilizer

Wolffia arrhiza has a growth rate and biomass as well as a good nutritional content for the growth of cultivated fish, especially herbivorous fish. Due to the high protein content (up to 45% of the dry weight), Wolffia is an excellent plant for the production of various biologically active substances, especially proteins. These plants can double every 1-6 days, similar to algae or mushrooms, and the number of leaves doubles every 2-3 days. This study aims to determine the effect of different fertilizer sources on doubling time, relative growth rate, population density and biomass of Wolffia arrhiza and water quality. The method used in this research is experimental. The research design used a completely randomized design (CRD) consisting of 4 treatments and 3 replications. The treatments were as follows: treatment A: chicken manure, B: cow manure, C: buffalo manure and D: goat manure with each dose of 200 grams. Parameters observed included doubling time, relative growth rate, population density, biomass and water quality parameters. The results showed that treatment B (cow manure) gave the highest value for doubling time, 1.59 days, relative growth rate 0.392 g/day, population density 209.75 ind/cm2, biomass 94.97. Keyword: Biomass; Fertilizer; Growth; Wolffia

Genome-Wide Identification of the Brassinosteroid Signal Kinase Gene Family and Its Profiling under Salinity Stress

Alfalfa (Medicago L.) is a high-quality perennial leguminous forage with the advantages of salt tolerance, mowing tolerance, high protein content, and other economically valuable characteristics. As the sixth class of plant hormones, brassinosteroids (BRs) play indispensable roles in modulating a variety of plant growth, maturation, and environmental adaptation processes, thereby influencing vegetal expansion and development. Brassinosteroid signal kinases (BSKs) are key cytoplasmic receptor kinases downstream of the BR signaling transduction pathway, participating in plant growth, development, and stress regulation. However, the phylogenetic and expression pattern analyses of the BSK gene family among the five alfalfa species have rarely been reported; in this study, 52 BSK family members were found in the genomes of the five subspecies, and phylogenetic trees were constructed according to protein sequences, allowing us to categorize all BSKs into seven distinct groups. Domain, conserved motif, and exon–intron structural analyses showed that most BSK members were relatively conserved, except for MtBSK3-2, MtBSK7-1, and MtBSK7-2, which may be truncated members. Intra-species collinearity and Ka/Ks analyses showed that purifying selection influenced BSK genes during evolution; most of the cis-acting elements in the promoter region were associated with responses, such as light, defense, and stress, anaerobic induction, MeJA, and abscisic acid. Expression pattern analysis indicated that the majority of alfalfa genes exhibited downregulation after reaching a peak at 0.5 h after treatment with 250 mM NaCl, especially for MsBSK14, MsBSK15, MsBSK17, MsBSK19, and MsBSK21; meanwhile, MsBSK4, MsBSK7, and MsBSK9 increased and were highly expressed at 12 h, demonstrating significantly altered expression patterns under salt stress; furthermore, MsBSK4, MsBSK7, and MsBSK9 exhibited expression specifically in the leaves. qRT-PCR analysis confirmed the expression trends for MsBSK4, MsBSK7, MsBSK9, MsBSK14, MsBSK15, and MsBSK16 matched the transcriptome data. However, the trends for MsBSK17, MsBSK19, and MsBSK21 diverged from the transcriptome data. Our study may provide a foundation for further functional analyses of BSK genes in growth, development, and salt stress tolerance in alfalfa.