High Protein Research Articles

A metal-trap tests and refines blueprints to engineer cellular protein metalation with different elements

It has been challenging to test how proteins acquire specific metals in cells. The speciation of metalation is thought to depend on the preferences of proteins for different metals competing at intracellular metal-availabilities. This implies mis-metalation may occur if proteins become mis-matched to metal-availabilities in heterologous cells. Here we use a cyanobacterial MnII-cupin (MncA) as a metal trap, to test predictions of metalation. By re-folding MncA in buffered competing metals, metal-preferences are determined. Relating metal-preferences to metal-availabilities estimated using cellular metal sensors, predicts mis-metalation of MncA with FeII in E. coli. After expression in E. coli, predominantly FeII-bound MncA is isolated experimentally. It is predicted that in metal-supplemented viable cells metal-MncA speciation should switch. MnII-, CoII-, or NiII-MncA are recovered from the respective metal-supplemented cells. Differences between observed and predicted metal-MncA speciation are used to refine estimated metal availabilities. Values are provided as blueprints to guide engineering biological protein metalation.

Rapid Development of High Concentration Protein Formulation Driven by High-Throughput Technologies.

High concentration protein formulation (HCPF) development needs to balance protein stability attributes such as conformational/colloidal stability, chemical stability, and solution properties such as viscosity and osmolality. A three-phase design is established in this work. In Phase 1, conformational and colloidal stability are measured by 384-well-based high-throughput (HT) biophysical screening while viscosity reduction screening is performed with HT viscosity screening. Collectively, the biophysical and viscosity screening data are leveraged to design the phase 2 of short-term stability study, executed using 96-well plates under thermal and freeze/thaw stresses. In phase 2, samples are analyzed by stability-indicating assays and processed with pair-wise Student's t-test analyses to choose the final formulations. In phase 3, the final formulations are then confirmed through a one-month accelerated stability in glass vials. Using a model antibody A (mAb-A), the initial HT screening successfully established the 384-well based platform. A lead formulation was chosen from the second round based on statistical analyses and subsequently tested against the commercial formulation of mAb-A as a control. Compared to the control, the lead formulation reduced the viscosity of mAb-A by 30% and decreased subvisible particles after thermal stress by 80%. HT biophysical screening in 384-well plates was demonstrated to effectively guide the rational design of a high-throughput stability screening study using 96-well plates. This platform enables the identification of a high concentration formulation within seven weeks within the first two phases of study that strategically balance stability with solution properties, thus achieving a rapid development of HCPF.

Deltacoronavirus HKU11, HKU13, PDCoV (HKU15) and HKU17 spike pseudoviruses enter avian DF-1 cells via clathrin-mediated endocytosis in a Rab5-, Rab7- and pH-dependent manner

Porcine deltacoronavirus (PDCoV), also known as HKU15, is a swine enteropathogenic virus that is believed to have originated in birds. PDCoV belongs to the genus Deltacoronavirus (DCoV), the members of which have mostly been identified in diverse avian species. We recently reported that chicken or porcine aminopeptidase N (APN), the major cellular receptor for PDCoV, can mediate cellular entry via three pseudotyped retroviruses displaying spike proteins from three avian DCoVs (HKU11, HKU13, and HKU17). In the present work, to better understand how avian-origin CoVs may be transmitted to pigs, we investigated the unknown DCoV entry pathway in avian cells. We show that clathrin-mediated endocytosis is involved in the entry of these DCoV pseudoviruses into chicken-origin DF-1 cells. Pseudovirus entry was suppressed by means of pharmacological inhibitors, dominant-negative mutants, and siRNAs targeting various cellular proteins and signalling molecules, suggesting that PDCoV and avian DCoV pseudovirus entry into DF-1 cells depends on clathrin, dynamin-2, cathepsins and a low-pH environment but is independent of caveolae and macropinocytosis. Furthermore, we found that DCoV pseudovirus entry was linked to Rab5- and Rab7-dependent pathways. This is the first report demonstrating that these DCoVs utilize clathrin-mediated endocytosis pathways to enter avian-origin cells, providing new insights into interspecies transmission of DCoVs.

Nicotinamide Prevents The Plasticity Impairments And The Cognitive Dysfunction Caused By Bone Fracture In Older Mice.

Postoperative delirium (POD), an acute cognitive dysfunction linked to morbidity and mortality, is characterized by memory impairments and disturbances in consciousness, particularly in patients aged 65 and older. Neuroinflammation and NAD+ imbalance are key mechanisms behind POD, leading to synaptic and cognitive deterioration. However, how surgery contributes to POD and neuroinflammation remains unclear, and effective treatments are lacking. Here we used a rodent model of bone fracture to examine the impact of surgery on synaptic plasticity, inflammation, and cognition. Additionally, we explored whether treatment with Nicotinamide (NAM), a NAD+ precursor, reduced the neuroinflammation and metabolic imbalance caused by surgery. Female C57BL/6J mice aged 20-22 months underwent tibial fracture surgery and received pre- and post-surgery NAM treatment. Neuroinflammation, synaptic plasticity, and cognition were assessed 72 hours post-surgery via long-term potentiation (LTP) assays, dendritic spine counting, and behavioral tests (open field maze and Y-maze). Tibial fracture surgery decreased LTP, dendritic spine density, and hippocampal-dependent memory function, and increased hippocampal inflammatory markers (IL-1 beta mRNA, CD38, and SIRT1 protein content); NAM pretreatment prevented these changes. Given surgery adverse effects on LTP and dendritic spine density, we assessed cellular oxidative state and BDNF protein levels. We found that surgery increased the oxidation of ryanodine receptor calcium channels (cellular redox sensors), and decreased BDNF protein levels; NAM supplementation mitigated both effects and prevented the cognitive decline and synaptic plasticity deficits while reducing inflammation post-surgery by lowering IL-1 beta and CD38 protein levels. We propose that the CD38 signaling pathway mediates these NAM protective effects.

High Serum S100B Protein Levels as a Risk Factor for Impaired Cognitive Function in Acute Non- Hemorrhagic Stroke: A Literature Review

The incidence of acute non-hemorrhagic stroke disease has a high disability rate and mortality rate and is a serious threat to people’s health and life and can burden families and society. The development of stroke after the onset of acute stroke, especially acute non-hemorrhagic stroke is not uncommon and has become a popular research topic in recent years. Various mechanism can occur, one of which is neuroinflammation. Stroke often leads to lifelong disability, including motor impairment, cognitive impairment, language impairment, and psychological impairment. Serum S100B is a calcium-binding protein that can determine disease severity and predict clinical outcomes to monitor disease progression and response to treatment. It was reported that serum S100B protein levels were detected to be higher in patients with cognitive impairment compared to normal ones. Keywords: Serum S100B protein, cognitive impairment, non-hemorrhagic stroke

Clinical characteristics and predictive factors of thrombotic complications in children with acute mastoiditis: a single center retrospective study.

Among acute mastoiditis (AM) complications, cerebral venous sinus thrombosis (CVST) is particularly severe, leading to increased intracranial pressure and potential neurological sequelae. Predicting the development of such complications is challenging. The aims of the present study were to evaluate the incidence, clinical characteristics, and risk factors for the development of CVST in AM. A retrospective study was conducted on children hospitalized with AM at the Meyer Children's Hospital between 2016 and 2024. Patients were divided into two groups: those with CVST (group A) and those without (group B), comparing demographic, clinical, and laboratory data. CVST was diagnosed using computed tomography (CT) and cerebral magnetic resonance angiography (MRA). To identify predictors of CVST complications, univariate and bivariate binary regression models were used. Out of 100 patients with AM, 15 (15%) developed CVST. Patients with CVST more frequently presented with fever, neurological symptoms (headache, vomiting), elevated CRP, and white blood cell counts compared to those who did not (p < 0.001, p < 0.001, p < 0.001, and p = 0.001, respectively). Streptococcus pyogenes and Streptococcus pneumoniae were more commonly encountered in patients with thrombotic complications (p = 0.024 and p = 0.05). Multivariate regression identified white blood cell (WBC) count and elevated C-reactive protein (CRP) as independent predictors of CVST in AM patients (OR: 1.14, 95% CI: 1.03-1.26, and OR: 1.10, 95% CI 1.00-1.21, p = 0.010 and p = 0.049, respectively). Conclusions: CVST is a frequent and serious complication of AM. Neurological symptoms and systemic inflammation (WBC count and CRP) are predictive indicators of CVST. Further studies are needed to develop risk algorithms for early diagnosis and to reduce sequelae. What is known: • Cerebral venous sinus thrombosis (CVST) are severe complications of acute mastoiditis, potentially leading to neurologic sequelae What is new: • An elevated inflammatory burden, namely a high C-reactive protein level and white blood cell count predict thrombotic complications in children with acute mastoiditis.

Frequency of viral etiology in community-acquired pneumonia.

The identification of etiology is very important when managing patients with community-acquired pneumonia (CAP). In Pakistan, studies regarding the viral etiology in CAP are scarce. The main objective of this study was to evaluate the frequency of viral etiology in CAP patients and analyze the clinical features and their impact on prognosis. Medical records of CAP patients admitted to Aga Khan University Hospital (Karachi, Pakistan) from March 2022 to February 2023 were retrospectively reviewed, patients who had microbiological tests performed within 48 hours of the hospital admission were included, and the frequency of viral and bacterial etiology was calculated. Patients who were immunocompromised were excluded. Epidemiological and clinical characteristics were examined, and the impact on prognosis was explored. A total of 166 patients were included; 115 (69.3%) patients were identified as having pneumonia with known causative microorganisms. A total of 83 (72.1%) patients had a viral etiology alone, 18 (15.6%) had only bacterial infection, and 14 (12.2%) had a viral and bacterial co-infection. Influenza A was most frequently detected (n=46/97; 47.4%), followed by Rhinovirus/Enterovirus (n=19/97; 19.6%). Staphylococcus aureus accounted for the majority (n=18; 56.3%) of cases among bacteria. Bacterial and viral-bacterial co-infection was significantly higher among non-survivors (38.1% vs. 16.6%, p=0.034). Confusion-Urea-Respiratory Rate-Blood Pressure-Age of 65 scores of 3-5 [odds ratio (OR) 4.234; 95% confidence interval 1.156-15.501], leukocytosis (OR 0.137; 0.030-0.636), high C-reactive protein (>10mg/L) (OR 1.008; 1.001-1.014), high serum procalcitonin level (≥0.5 ng/mL) (OR 10.731; 3.018-38.153), and mechanical ventilation required (OR 47.104; 13.644-162.625) were associated with mortality. Mechanical ventilation requirement was independently associated with increased odds of mortality (OR 43.407; 8.083-233.085). Of 166 patients, 21 (12.7%) had died, with the highest percentage (28.6%) seen in the viral-bacterial coinfection group (p=0.046). To conclude, respiratory viruses are increasingly being recognized as an important etiology in CAP, with higher mortality seen in bacterial infection, whether alone or with viral co-infection.

Fish protein hydrolysates from rainbow trout processing in replacement of feed protein sources: Effects on growth performances, liver status and body composition of gilthead sea bream, Sparus aurata L., juveniles

Fish protein hydrolysates from rainbow trout processing in replacement of feed protein sources: Effects on growth performances, liver status and body composition of gilthead sea bream, <i>Sparus aurata</i> L., juveniles

Enhanced sampling of protein conformational changes via true reaction coordinates from energy relaxation

The bottleneck in enhanced sampling lies in finding collective variables that effectively accelerate protein conformational changes; true reaction coordinates that accurately predict the committor are the well-recognized optimal choice. However, identifying them requires unbiased natural reactive trajectories, which, paradoxically, require effective enhanced sampling. Using the generalized work functional method, we uncover that true reaction coordinates control both conformational changes and energy relaxation, enabling us to compute them from energy relaxation simulations. Biasing true reaction coordinates accelerates conformational changes and ligand dissociation in PDZ2 domain and HIV-1 protease by 105 to 1015-fold. The resulting trajectories follow natural transition pathways, enabling efficient generation of unbiased reactive trajectories. In contrast, biased trajectories from empirical collective variables display non-physical features. Furthermore, our method uses a single protein structure as input, enabling predictive sampling of conformational changes. These findings unlock access to a broader range of protein functional processes in molecular dynamics simulations.

Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese

Grana Padano (GP) is an Italian hard cooked cheese characterized by a long ripening process and high protein and Ca contents. After in vitro static simulated gastrointestinal digestion, GP digest contained caseinophosphopeptides that were 6 to 24 amino acids in length, including tri-phosphorylated species incorporating the pSer-pSer-pSer-Glu-Glu cluster. Using rat ileum tissue, the digest was used to assess Ca absorption ex vivo, which showed significantly better results for the GP digest in comparison to the CaCO3 aqueous solution. An in vitro intestinal model based on Caco-2/HT-29 cell co-culture was able to mimic Ca absorption from GP digest, with Ca-rich water as a control. The metabolite-containing medium was then used to treat osteoblast-like SaOS-2 cells. As a consequence, metabolized GP digest significantly increased the number of osteoblasts, whereas the metabolized water did not exert this effect. Finally, the mice were fed diets containing GP or CaCO3 and pea isolate and the in vivo outcomes were assessed through fluorescent probe and computed tomography. Mice fed a diet containing GP showed a higher increase in bone remodeling and volume in comparison to those fed a control diet containing CaCO3 and pea isolate. Overall, the ex vivo, in vitro and in vivo experiments highlighted the effectiveness of GP in improving Ca absorption, osteoblast proliferation and bone remodeling and volume.

Energy availability and macronutrient intake over a 7-day training period in adolescent rugby players.

Understanding the dietary intake of elite adolescent athletes and its adequacy with sport nutrition recommendation is a key issue for health and player development, as well as performance and recovery. Energy availability needs to be considered to ensure optimal health and performance in young athletes. The present study aimed to quantify energy availability, energy expenditure and macronutrient intake in young male rugby union players competing at national level. Twelve male adolescent players (15.6±0.6 years) completed a 7-days prospective observational study (5 days of training and 2 days of full recovery). Total energy expenditure was estimated using indirect calorimetry and heart rate measurement. Energy intake was assessed using weighed food by a dietitian in cafeteria (training days) and image-based dietary (recovery days). Energy availability was calculated using (energy intake-exercise energy expenditure)/fat-free mass. Mean energy availability was 38.5±7.5, 40.2±5.4 and 47.8±5.1 kcal/kgFFM/d on heavy training, moderate training and recovery days, respectively. Players consumed a low carbohydrate (~5.0 g/kg/d), high protein (~2.0 g/kg/d) and high fat (~1.8 g/kg/d) diet on training and recovery days in relation to current international nutritional recommendations for young athletes. Athletes showed sub-optimal energy availability on training days, high energy availability on recovery days and did not comply with carbohydrate intake recommendations on training nor recovery days. These results highlight the short-term inadequacy of energy availability as a result of low carbohydrate intake, warning about the possible adverse short-term metabolic effects on health and performance of young athletes.

Direct observation of small molecule activator binding to single PR65 protein

The reactivation of heterotrimeric protein phosphatase 2A (PP2A) through small molecule activators is of interest to therapeutic intervention due to its dysregulation, which is linked to chronic conditions. This study focuses on the PP2A scaffold subunit PR65 and a small molecule activator, ATUX-8385, designed to bind directly to this subunit. Using a label-free single-molecule approach with nanoaperture optical tweezers (NOT), we quantify its binding, obtaining a dissociation constant of 13.6 ± 2.5 μM, consistent with ensemble fluorescence anisotropy results but challenging to achieve with other methods due to low affinity. Single-molecule NOT measurements reveal that binding increases optical scattering, indicating PR65 elongation. This interpretation is supported by all-atom molecular dynamics simulations showing PR65 adopts more extended conformations upon binding. This work highlights NOT’s utility in quantifying binding kinetics and structural impact, offering insights valuable for drug discovery.

Comprehensive Characterization of Raw and Processed Quinoa from Conventional and Organic Farming by Label-Free Shotgun Proteomics.

Quinoa is widely recognized for its exceptional nutritional properties, particularly its complete protein content. This study, for the first time, investigates the effects of processing methods (boiling and extrusion) and farming conditions (conventional and organic) on the proteomic profile. Following a label-free shotgun proteomics approach, a total of 1796 proteins were identified and quantified across all quinoa samples. Regarding processing, both boiling and extrusion produced protein extracts with lower total protein content, with the number of identified proteins decreasing from 1695 in raw quinoa to 957 in processed quinoa. Boiling led to a reduction in protein diversity and expression, while extrusion, which involves high temperatures and pressures, specifically decreased the abundance of high molecular mass proteins. Concerning cultivation practices, organic farming was associated with a broader protein diversity, especially proteins related to translation (28 vs 5%), while conventional farming showed a higher abundance of catalytic and enzymatic proteins (67 vs 46%). These findings highlight the distinct proteomic changes induced by different processing methods and farming conditions, offering valuable insights to manage quinoa's nutritional, bioactive, and functional properties across various production practices.

Restoration of Genetic Code in Macular Mouse Fibroblasts via APOBEC1-Mediated RNA Editing

RNA editing is a significant mechanism underlying genetic variation and protein molecule alteration; C-to-U RNA editing, specifically, is important in the regulation of mammalian genetic diversity. The ability to define and limit accesses of enzymatic machinery to avoid the modification of unintended targets is key to the success of RNA editing. Identification of the core component of the apoB RNA editing holoenzyme, APOBEC, and investigation into new candidate genes encoding other elements of the complex could reveal further details regarding APOBEC-mediated mRNA editing. Menkes disease is a recessive X-chromosome-linked hereditary syndrome in humans, caused by defective copper metabolism due to mutations in the ATP7A gene, which encodes a copper transport protein. Here, we generated plasmids encoding the MS2 system and the APOBEC1 deaminase domain and used a guide RNA with flanking MS2 sites to restore mutated Atp7a in fibroblasts from a macular mouse model of Menkes disease withs T&gt;C mutation. Around 35% of the mutated C nucleotide (nt) was restored to U, demonstrating that our RNA editing system is reliable and has potential for therapeutic clinical application. RNA base editing via human RNA-guided cytidine deaminases is a potentially attractive approach for in vivo therapeutic application and provides opportunities for new developments in this field.

Determination of collagen types and mineral contents in fish skin and collagen-containing skin-derived protein hydrolysates before and after in vitro simulated digestion.

An in vitro digestion model was established to characterize the types of collagens in skin of cod, white fish, and salmon as well as their collagen-containing skin-derived protein hydrolysates (CSPH) before and after digestion. Moreover, the mineral content and their bioaccessibility were evaluated. Finally, the presence of heavy metals was evaluated to assess the safety of these products. The results showed that white fish protein exhibited a high digestibility, reaching up to 92%. Among the collagen products, salmon collagen had the highest digestibility (∼73%). Protein identification revealed that the emPAI of type I collagen in digested skin and CSPH was higher than that of undigested samples. In addition, raw skins had higher contents of P, K, Ca and Mg, and the mineral content of CSPH was lower than that of unprocessed skins. Among the minerals studied, Ca and Cu showed the highest bioaccessibility in raw skin cod, being 32% and 26%, respectively. The bioaccessibility of Cu in raw skin salmon was also higher (∼34%). Moreover, in CSPH, Mg, K and Cu can be easily digested and absorbed. Regarding heavy metals, As and Pb were below the respective safe limits in all raw skins and CSPH, while Hg and Cd were not detected in the fish CSPH. Fish-derived collagen has gained significant attention due to its numerous health benefits, high bioavailability, and superior sustainability compared to animal collagen. Moreover, different types of collagens offer distance roles and advantages in the body. However, there are limited reports on how collagen structure and type may change during the digestive process. This study seeks to deepen our understanding of the economic value of fish collagen, as well as the mechanisms of its absorption and digestion. By investigating processes, the research aims to provide a clearer insight into the physiological effects of fish-derived collagen, which can inform the development of tailored collagen supplementation programs based on specific health needs.

Perch Hydrolysates from Upcycling of Perch Side Streams Accelerate Wound Healing by Enhancing Fibroblasts to Secrete Procollagen I, Fibronectin, and Hyaluronan

Wound healing incurs various challenges, making it an important topic in medicine. Short-chain peptides from fish protein hydrolysates possess wound healing properties that may represent a solution. In this study, perch hydrolysates were produced from perch side steams using a designed commercial complex enzyme via a proprietary pressure extraction technique. The average molecular weight of the perch peptides was 1289 kDa, and 62.60% of the peptides had a low molecular weight (≤1 kDa). Similarly to the beneficial amino acid sequence FPSIVGRP, FPSLVRGP accounted for 6.21% abundance may have a potential antihypertensive effect. The concentrations of collagen composition and branched-chain amino acids were 1183 and 1122 mg/100 g, respectively. In a fibroblast model, active perch peptides accelerated wound healing mainly by increasing the secretion of procollagen I, fibronectin, and hyaluronan. In an SD rat model established to mimic human wounds, orally administered perch hydrolysates with a molecular weight below 2.3 kDa accelerated wound healing, which mainly resulted from collagen-forming amino acids, branched-chain amino acids, and matrikine. Collectively, the residue of perch extract can be upcycled via a hydrolysis technique to produce not only bioactive sequences but also short-chain peptides. Considering the therapeutic potential to promote wound healing, such by-products are of great value and may be developed as dietary nutraceuticals.

Chitin Analysis in Insect-Based Feed Ingredients and Mixed Feed: Development of a Cost-Effective and Practical Method.

Insects are used as an alternative sustainable, protein-rich ingredient in fish, pet, pig and poultry diets. The significant difference between insect meals and common protein sources is the content of chitin. The nitrogen contained in chitin, which makes up 6.89% of the chitin mass, is detected as crude protein in the analysis and, therefore, deludes the crude protein content in a higher range. In this work, we developed a chitin analysis method that does not require expensive and specialized equipment within insect production and processing industries. The method is based on classical chemical methods such as crude fibre and nitrogen content, making it easily implementable within existing feed analysis. In the process of method validation, a recovery rate of over 95% for chitin in the presence of protein and a standard deviation of < 5% at concentrations as low as 2% was determined. Furthermore, determining chitin at a higher standard deviation of > 10% at concentrations as low as 2% is possible. The method was used to determine the chitin content in various products derived from insect breeding and processing. The chitin content was determined in four insect species (Hermetia Illucens; Tenebrio molitor; Acheta domesticus; Bombyx mori) and different developmental stages of the yellow mealworm (T. molitor), including larvae, pupae and beetles, as well as in commercial pet food. These results also allow for an estimation of the insect protein content, provided that the raw material is known.

Protein Intake and Its Association With Meal Patterns and Dietary Patterns in a Swedish Population of Older Adults.

Meeting protein intake recommendations is relevant for maintaining muscle mass. This study aimed to describe protein intake and its association with meal patterns and dietary patterns. An in-house designed, web-based 4-day record was used in the national dietary survey (in 2010/2011). Participants 60 years and older were included in the analysis (n = 533). Protein intake was described by hour of consumption, self-indicated meals and food source. Eating and drinking occasion (EDO) and food groups were defined, from which meal patterns and three a posteriori dietary patterns (using principal component analysis) were assessed. We observed a mean protein intake of just over 1 g/kg body weight (bw) in both men and women. Over 50% of the protein intake was sourced from the food groups meat, fish and milk/yoghurt. A bolus intake of 30 g protein per meal was observed in a small proportion of participants at breakfast and lunch, but was most common at dinner (41% women and 56% men). No strong correlations were observed between protein intake and neither dietary patterns nor the number of EDOs. A 5 g higher protein intake at any meal, but not higher EDO frequency, was associated with higher odds of meeting a protein intake over 1.1 g/kg bw. Protein intake over 1.1 g/kg bw was met by 44% of the participants. Lunch and dinner were the highest contributors to protein intake. Dietary and meal patterns were weakly associated with protein intake. Only total daily protein intake was associated with reaching > 1.1 g/kg bw.

Venlafaxine induces Teratogenesis and alters SHH Gene Expression and Protein Biochemistry of Developing Gallus sp. Embryos

The use of antidepressant drugs during pregnancy is increasing globally. SNRIs and SSRIs are most widely used for treating panic disorders. Venlafaxine is an antidepressant that inhibits serotonin and norepinephrine reuptake. Given the increasing prevalence of antidepressant use during pregnancy, Venlafaxine exposure during the early stages of development could lead to changes in protein dynamics and disrupt essential gene pathways, raising concerns about the potential teratogenic effects of antidepressant exposure during pregnancy. This study explores the developmental impact of Venlafaxine, a commonly prescribed serotonin-norepinephrine reuptake inhibitor (SNRI), on protein biochemistry and gene expression in the developing Gallus gallus domesticus embryos. Significant changes in protein profiles were observed among control and Venlafaxine-treated groups. Sonic Hedgehog (SHH) gene, a key regulator of embryonic patterning and organogenesis, specifically neural tube formation, and limb development, is known for its crucial role in embryonic development. qRT-PCR analysis of Venlafaxine-treated embryos showed changes in SHH gene expression suggesting that Venlafaxine may target SHH gene expression, and potentially alter signalling pathways crucial for normal embryogenesis. However, detailed studies are needed to understand the long-term consequences of prenatal and foetal antidepressant exposure on embryonic growth and development. These changes may lead to abnormal growth patterns or congenital defects, emphasizing the need for caution when prescribing Venlafaxine during pregnancy. Keywords: Antidepressants, Venlafaxine, SNRI, Gallus gallus domesticus, Sonic Hedgehog (SHH), Protein Biochemistry, qRT-PCR

Continuous high-soy protein soymilk intake affects ordinary walking speed in the Japanese pre-frail and frail elderly: a randomized controlled trial.

To investigate whether continuous intervention using soymilk containing high soy protein improves physical frailty, a randomized controlled trial was conducted among the Japanese pre-frail and frail elderly. Japanese pre-frail and frail elderly participants (n = 73) were randomly assigned to the high-soy protein and control groups, who then ingested soymilk containing 14.5g/200ml and 3.2g/200ml of soy protein, respectively. Before and after the 12-week intervention, walking speed, skeletal muscle mass, grip strength, and the revised Japanese CHS questionnaire regarding fatigue and physical activity were examined to evaluate the impact of each soymilk on physical frailty and compare the variation between the two groups. Physical activity (monitored using a pedometer), dietary intake (determined by questionnaire), and estimated protein intake (determined by casual urine testing) were also recorded before and after the intervention. For the final analysis of the entire cohort (n = 70), there were no significant differences in the endpoints between the two groups. In the subgroup analysis, among participants with a walking speed of at least 1m/s (n = 35, P = 0.012) and at least 5,000 steps/day before intervention (n = 27, P = 0.0083), the variation in walking speed after the 12-week intervention was significantly higher in the high-soy protein group than in the control group. Estimated protein intake was also significantly higher in the high-soy protein group than in the control group after the intervention. Regarding physical activity and dietary intake, no significant differences were observed between the groups before or after the intervention. The continuous 12-week intervention of high soy protein increased the walking speed among the Japanese pre-frail and frail elderly participants who had an ordinarily high walking speed and high step counts. UMIN Clinical Trials Registry, UMIN000044999. Registered July 29, 2021; https://center6.umin.ac.jp/cgi-open-bin/ctr/ctr_view.cgi?recptno=R000051409 .