Total Protein Synthesis Research Articles

Predominant synthesis of giant myofibrillar proteins in striated muscles of the long-tailed ground squirrel Urocitellus undulatus during interbout arousal

Molecular mechanisms underlying muscle-mass retention during hibernation have been extensively discussed in recent years. This work tested the assumption that protein synthesis hyperactivation during interbout arousal of the long-tailed ground squirrel Urocitellus undulatus should be accompanied by increased calpain-1 activity in striated muscles. Calpain-1 is known to be autolysed and activated in parallel. Western blotting detected increased amounts of autolysed calpain-1 fragments in the heart (1.54-fold, p < 0.05) and m. longissimus dorsi (1.8-fold, p < 0.01) of ground squirrels during interbout arousal. The total protein synthesis rate determined by SUnSET declined 3.67-fold in the heart (p < 0.01) and 2.96-fold in m. longissimus dorsi (p < 0.01) during interbout arousal. The synthesis rates of calpain-1 substrates nebulin and titin in muscles did not differ during interbout arousal from those in active summer animals. A recovery of the volume of m. longissimus dorsi muscle fibres, a trend towards a heart-muscle mass increase and a restoration of the normal titin content (reduced in the muscles during hibernation) were observed. The results indicate that hyperactivation of calpain-1 in striated muscles of long-tailed ground squirrels during interbout arousal is accompanied by predominant synthesis of giant sarcomeric cytoskeleton proteins. These changes may contribute to muscle mass retention during hibernation.

Caffeic Acid Phenethyl Ester (CAPE) Induced Apoptosis in Serous Ovarian Cancer OV7 Cells by Deregulation of BCL2/BAX Genes.

Ovarian cancer has the worst prognosis among all gynecological cancers. Therefore, it seems reasonable to seek new drugs that may improve the effectiveness of treatment or mitigate the adverse effects of chemotherapy. Caffeic acid phenethyl ester (CAPE) has many beneficial biological properties. The aim of the study was to assess the anticancer properties of CAPE against serum ovarian carcinoma cells. The morphology of the cells was evaluated in H-E staining and in transmission electron microscopy. The cytotoxic and proapoptotic activity of CAPE was investigated by using the XTT-NR-SRB assay, qRT-PCR analysis of BAX/BCL2 expression, and by cytometric evaluation. CAPE causes constriction in OV7 cells, numerous granulomas were observed in the cytoplasm, the cell nuclei were pyknotic. Autophagosomal vacuoles could suggest the occurrence of aponecrosis. CAPE significantly decreased the lysosomal activity and the total synthesis of cellular proteins. CAPE exhibited, dose and time dependent, cytotoxic activity against OV7 serum ovarian cancer cells. In OV7 cells CAPE induced apoptosis via dysregulation of BAX/BCL2 balance, while activated proapoptotic BAX gene expression level was 10 times higher than BCL2.

Dietary Protein and Amino Acid Deficiency Inhibit Pancreatic Digestive Enzyme mRNA Translation by Multiple Mechanisms.

Background & AimsChronic amino acid (AA) deficiency, as in kwashiorkor, reduces the size of the pancreas through an effect on mammalian target of rapamycin complex 1 (mTORC1). Because of the physiological importance of AAs and their role as a substrate, a stimulant of mTORC1, and protein synthesis, we studied the effect of acute protein and AA deficiency on the response to feeding.MethodsICR/CD-1 mice were fasted overnight and refed for 2 hours with 4 different isocaloric diets: control (20% Prot); Protein-free (0% Prot); control (AA-based diet), and a leucine-free (No Leu). Protein synthesis, polysomal profiling, and the activation of several protein translation factors were analyzed in pancreas samples.ResultsAll diets stimulated the Protein Kinase-B (Akt)/mTORC1 pathway, increasing the phosphorylation of the kinase Akt, the ribosomal protein S6 (S6) and the formation of the eukaryotic initiation factor 4F (eIF4F) complex. Total protein synthesis and polysome formation were inhibited in the 0% Prot and No Leu groups to a similar extent, compared with the 20% Prot group. The 0% Prot diet partially reduced the Akt/mTORC1 pathway and the activity of the guanine nucleotide exchange factor eIF2B, without affecting eIF2α phosphorylation. The No Leu diet increased the phosphorylation of eIF2α and general control nonderepressible 2, and also inhibited eIF2B activity, without affecting mTORC1. Essential and nonessential AA levels in plasma and pancreas indicated a complex regulation of their cellular transport mechanisms and their specific effect on the synthesis of digestive enzymes.ConclusionsThese studies show that dietary AAs are important regulators of postprandial digestive enzyme synthesis, and their deficiency could induce pancreatic insufficiency and malnutrition.

INFLUENCE OF MOLDING HOMOGENATES ON ELEMENT AND METABOLIC STATUS OF DOGS

In order to study the effect of the drone homogenate on the elemental, metabolic, hematological, and immune statuses of dogs, we created control and experimental groups of Labrador retrievers. The dogs were kept in the conditions of apartments in the village of Kochenevo, Novosibirsk Region. The diet of animals consisted of meat, cereal porridge, cottage cheese and vegetables. Dogs received daily vitamin and mineral supplements and apples. Labradors – retrievers of the control group daily, for two months, drone homogenate was drunk once a day at the rate of 15 mg per kg of body weight. The results of the studies showed that in the presence of a drone homogenate, dogs increase thyroxin synthesis, activate erythro and leukopoiesis, increase phagocytic activity and phagocytic index, synthesis of total blood protein, in particular globulins, and triglyceride levels. An analysis of the chemical composition of blood serum carried out before the start of the experiment revealed in dogs of both groups more or less pronounced microelementoses in terms of cobalt, chromium, iron, potassium, manganese, sodium, phosphorus, lead, selenium and zinc. After completion of the course of taking the homogenate, a correction of the macro- and microelement composition of the blood occurred. The concentration of lithium, chromium, sodium, iron, phosphorus was statistically significantly reduced, and the concentration of boron, calcium, copper, iodine, nickel and tin increased.

TGF-β1-induced expression of collagen type II and ACAN is regulated by 4E-BP1, a repressor of translation.

Eukaryotic initiation factor 4E (eIF4E)-binding protein 1 (4E-BP1) binds eIF4E and represses protein translation by displacing it from the mRNA. In this study, we investigated the influence of 4E-BP1 translational apparatus on the regulation of transforming growth factor-beta 1 (TGF-β1)-induced anabolic signaling in chondrocytes. The level of 4E-BP1 expression was significantly higher in human OA cartilage than normal cartilage. TGF-β1 increased total protein synthesis, including aggrecan (ACAN) and collagen type II (Col II), together with activation of Akt/mTOR signaling pathway. mTOR silencing significantly suppressed ACAN and Col II expressions through decreasing TGF-β1-induced phosphorylation of 4E-BP1. On the contrary, 4E-BP1 knockdown promoted total protein synthesis but suppressed Col II and ACAN expressions with decreased expression of Smad2/3 and Smad4 and increased expression of inhibitory Smad6 and Smad7. TGF-β1 suppressed the interaction of 4E-BP1 and eIF4E and subsequently enhanced protein synthesis. Furthermore, 4E-BP1 regulated translation levels of inhibitory Smads, which decreased the accumulation of nuclear Smad2/3 complexes on the promoter of ACAN and Col II genes, subsequently affecting transcription of ACAN and Col II. These results demonstrated that TGF-β1-modulated phosphorylation of 4EBP1 plays a role in the expression of Col II and ACAN through differential alteration of Smad signaling pathway.

Physiological Studies of Some Insecticides on Spodoptera littoralis Larvae

The present study aims to focus on the effect of the sublethal rates of Lambda-cyhalothrin +Thiamethoxam and Lambda-cyhalothrin, which belonging to groups 3 and 4 on the 4th larval instar of Spodoptera littoralisfield strain under laboratory conditions, and their effect on some physiological aspects, post-treatment with rates 50 and 100 ml/200L for 48 hrs. The results cleared that highly physiological disturbance was recognized as marked stimulating in total protein, total lipids, trehalase, AchE, and acid phosphatase synthesis however, inhibit amylase, invertase and alkaline phosphatase for lambda-cyhalothrin + Thiamethoxam. The tested sublethal rate of lambda-cyhalothrin, also be highly affected by stimulating total protein, invertase, trehalase, AchE, acid and alkaline phosphatases. On the other hand, inhibits total lipids and amylase synthesis in insect cells.

Influence of Bisphosphonates on the Behavior of Osteoblasts Seeded Onto Titanium Discs.

Among other factors, types of bisphosphonates and treatment regimens seem to be strongly associated with the success or failure of installation of osseointegrated implants. This study investigated the influence of two bisphosphonates, sodium alendronate (SA) and zoledronic acid (ZA), on the metabolism of osteoblasts. Human osteoblasts (Saos-2) were seeded onto machined or acid-treated titanium discs previously placed on 24-well plates in complete culture medium. After 24 h, cells were exposed to bisphosphonates at 0.5, 1 or 5 µM for 24 h, 48 h or 7 days. The effects of SA and ZA on osteoblasts were assessed based on the adhesion of these cells to the titanium surfaces by direct fluorescence, cell viability, total protein and collagen synthesis. Alkaline phosphatase activity and mineral nodule deposition by these cells were also evaluated. Data were evaluated by ANOVA and Tukey tests (α=0.05). Decreased adhesion of cells to the titanium discs was observed when exposed to both bisphosphonates; however, this lack of cell adhesion was more evident for ZA-treated cells. In addition, the exposure of osteoblasts to ZA decreased the viability, ALP activity and mineral nodule deposition, which may be related to poor osseointegration after implant installation.

Determining Ideal Balance Among Branched-Chain Amino Acids (BCAA) as a Proof of Concept Study in Healthy Children

Abstract Objectives Medical foods for children with in-born errors of metabolism (propionic academia, PROP) are formulated with imbalanced mixture of the BCAA (high leucine, to minimal or no valine and isoleucine), and therefore their use is controversial. The objective of the current study was to determine an ideal BCAA ratio at which total body protein synthesis is optimized in healthy children using the indicator amino acid oxidation (IAAO) method. Methods The study design was based on the oxidation of the stable isotope L-[1–,13C] phenylalanine to ,13CO2 to compare protein synthesis among seven different BCAA ratios. Leucine intake was gradually reduced from current high doses in medical foods; isoleucine and valine were kept constant. This study was done as a proof of concept in healthy children, to allow characterization of the metabolic responses to wide range of leucine test intakes, to help design narrow BCAA ratio range to test in children with PROP. Results A total of 8 healthy children were studied, completing 42 study days. ANOVA showed significant differences in F13CO2 with different BCAA ratios; P value &amp;lt;0.001. A BCAA ratio of (leucine: Isoleucine: Valine = 1:0:0) was associated with the highest F13CO2 compared to other ratios (p value &amp;lt;0.001), indicating low total body protein synthesis. By reducing leucine intake, with BCAA ratio between (1: 0.26: 0.28 to 1:0.35:0.4) protein synthesis was optimized. Conclusions Using the IAAO method in healthy children, we tested for the first time the effect of different BCAA ratios on protein synthesis. Results from this study confirmed that the BCAA ratio of (leucine: Isoleucine: Valine = 1:0:0), which is similar to the ratio in medical foods, limited total body protein synthesis. Furthermore, a balanced BCAA ratio that optimized protein synthesis was found to be between (1: 0.26: 0.28) and (1: 0.35:0.4). Thus, we propose reformulating the BCAA mixture in medical foods, by reducing leucine content by 50%. Future research in children with PROP with our proposed BCAA ratio is necessary to confirm improved patient growth outcomes. Funding Sources Faculty of Applied Medical Sciences, Department of Clinical Nutrition, King Abdulaziz University and BC Children's Hospital Research Institute.

Catching up to nature's ribosomes

Flow chemistry enables total chemical synthesis of proteins without solution-phase ligation

Styrene Oxide Caused Cell Cycle Arrest and Abolished Myogenic Differentiation of C2C12 Myoblasts.

Contaminations of chemicals in foods and drinks are raising public concerns. Among these, styrene, a monomer for plastic production, receives increasing interest due to its ability to leach from the packaging and contaminate in foods and drinks causing many health problems. The present study was designed to investigate the effects of styrene monomer (STR) and its metabolite styrene oxide (STO) on C2C12 myoblast proliferation and differentiation. Based on an MTT assay, both STR and STO showed no cytotoxic effect at 10–100 μM. However, at 50–100 μM STO, but not STR, significantly inhibited cell proliferation. The STO-treated cells were accumulated in S-phase of cell cycles as revealed by flow cytometry. The antioxidant enzyme (catalase and superoxide dismutase) activities and the gene expressing these enzymes of the arrested cells were decreased and ultimately led to nuclear condensation and expression of apoptotic markers such as cleaved caspase-3 and-9, but not cleaved caspase-8. In addition, STO significantly suppressed myogenic differentiation by decreasing both the number and size of differentiated myotubes. Biochemical analysis showed attenuations of total protein synthesis and myosin heavy chain (MHC) protein expression. In conclusion, a metabolite of styrene, STO, leached from plastic packaging of foods and beverages suppressed both myoblast proliferation and differentiation, which would affect skeletal muscle development and regeneration.

Chitosan-Gentamicin Conjugate Hydrogel Promoting Skin Scald Repair.

Our earlier research indicated that chitosan-gentamicin conjugate (CS-GT) possesses superior antimicrobial activity and good water solubility. To develop CS-GT-based scald dressings, the antibacterial properties of CS-GT were further studied, and the biosafety of CS-GT and the healing mechanism of CS-GT hydrogel was systematically explored in this article. It was found that cell viability shows a declined inclination with the prolonged culture time and the increased concentration of CS-GT. After three day’s culture, the cell viability could still remain at 79.72% when CS-GT concentration was as high as 1000 μg/mL. On the other hand, the hemolysis rate of CS-GT was lower than 5% when its concentration is 800 μg/mL. Therefore CS-GT has good cytocompatibility and hemocompatibility. A wound-healing experiment has shown that the skin healing rate of CS-GT hydrogel was the highest at 99.61%, followed by the positive control (wet burn ointment) 94.98%, GT hydrogel 87.50%, and matrix 77.39%. The blank control group, however, possessed the lowest healing rate of 75.45%. Further analysis indicated that CS-GT hydrogel could promote the synthesis of total protein (TP) in skin granulation tissue, resulting in the enhanced hydroxyproline (HYP) content, which facilitated collagen fibrogenesis, reduced cytokine expression in an inflammatory response, and, ultimately, accelerated wound healing. To sum up, CS-GT hydrogel is a promising scald dressing.

Actions of Prostaglandins on Human Nucleus Pulposus Metabolism Inferred by Cyclooxygenase 2 Inhibition of Cytokine Activated Cells.

ObjectiveLow back pain is frequently treated with nonsteroidal anti-inflammatory drugs (NSAIDs), but little is known about intervertebral disc metabolism of the prostaglandins that are diminished by these drugs. Hence, this study aimed at delineating prostaglandin actions in cytokine activated disc cells by comparing the response of nucleus pulposus (NP) cells to the pro-inflammatory cytokine interleukin (IL)-1β with and without cyclooxygenase 2 (COX-2) inhibition. MethodsNP cells cultured in alginate beads were activated with IL-1β ± the COX-2 inhibitor Sc-58125. Media harvested from cultured cells were analyzed for prostaglandin E2 (PGE2), prostaglandin F2 alpha (PGF2α), IL-6, and matrix metalloproteinase (MMP)-3 by enzyme-linked immunosorbent assay and nitric oxide by Griess Reaction. Gene expression along with proteoglycan, collagen, and total protein synthesis were also measured. ResultsIL-1β increased culture media PGE2 and PGF2α, but decreased proteoglycan and collagen syntheses as well as mRNA expression of the matrix genes aggrecan, versican, collagen I, and collagen II. COX-2 inhibition partially rescued proteoglycan and collagen syntheses and collagen I mRNA, but decreased collagen II mRNA IL-1β activated NP cells. COX-2 inhibition initially enhanced and subsequently reduced IL-1β induced inducible nitric oxide synthase, without altering medium nitrite. IL-1β induction of MMP-3 mRNA was increased by COX-2 inhibition at 24 and 48 hours. ConclusionCOX-2 inhibition alters the response of NP cells to IL-1β, suggesting IL-1β action on disc cells is mediated at least in part through COX-2 and its prostaglandins. COX-2 inhibition produces minimal effects on several key catabolic mediators, with the exception of MMP-3. Blocking COX-2 might be beneficial for maintaining disc matrix since it provides an overall rescue of IL-1 induced loss of matrix protein synthesis.

Feeding diets varying in forage proportion and particle length to lactating dairy cows: I. Effects on ruminal pH and fermentation, microbial protein synthesis, digestibility, and milk production

Physically effective neutral detergent fiber (peNDF) content of dairy cow diets was modified by varying the theoretical chop length of alfalfa silage and forage:concentrate (F:C) ratio, and effects on nutrient intakes, ruminal fermentation, site and extent of digestion, microbial protein synthesis, and milk production were evaluated. Estimates of dietary peNDF contents were compared with recommendations, and predictions of ruminal pH from peNDF and the recently developed physically adjusted neutral detergent fiber (paNDF) system were compared with observed pH. The experiment was designed as a triple 4 × 4 Latin square using 12 mid-lactating dairy cows with 4 intact, 4 ruminally cannulated, and 4 ruminally and duodenally cannulated cows. Site and extent of digestion and microbial protein synthesis were measured in a single 4 × 4 Latin square. Treatments were a 2 × 2 factorial arrangement; 2 forage particle lengths (FPL) of alfalfa silage (short and long) were combined with low (35:65) and high (60:40) F:C ratios [dry matter (DM) basis]. The peNDF contents were determined by multiplying the proportion (DM basis) of total mixed ration retained on 2 (8 and 19 mm; peNDF8.0) or 3 (1.18, 8, and 19 mm; peNDF1.18) sieves of the Penn State Particle Separator by the neutral detergent fiber content of the diet. The dietary peNDF contents ranged from 10.7 to 17.5% for peNDF8.0 or from 23.1 to 28.2% for peNDF1.18. Interactions between F:C ratio and FPL content were few. Increasing peNDF content of diets by increasing F:C ratio decreased DM intake, milk yield, and milk protein yield, whereas apparent total-tract DM digestibility and milk efficiency improved. Increasing F:C ratio improved ruminal pH status but decreased total volatile fatty acid concentration and microbial protein synthesis. Increasing peNDF content of diets via dietary FPL increased mean ruminal pH, but did not affect DM intake, total-tract digestibility, or milk production. The results indicate that feeding dairy cows a low F:C diet helps increase DM intake, milk production, and microbial protein synthesis, but may adversely affect feed digestibility and milk efficiency due to increased risk of subacute ruminal acidosis. Increased FPL improved ruminal pH status, but had minimal effects on feed intake, ruminal fermentation, nutrient digestibility, and milk production. The results indicate a trade-off between reducing the risk of subacute ruminal acidosis and maximizing ruminal fermentation, feed digestibility, and milk production of dairy cows. The paNDF model showed improvement in the predictability of ruminal pH over the peNDF model, but the accuracy of predictions varied depending upon the diet and ruminal fermentation variables considered in the equations.

An activator of G protein-coupled receptor and MEK1/2-ERK1/2 signaling inhibits HIV-1 replication by altering viral RNA processing.

The ability of HIV-1 to evolve resistance to combined antiretroviral therapies (cARTs) has stimulated research into alternative means of controlling this infection. We assayed >60 modulators of RNA alternative splicing (AS) to identify new inhibitors of HIV-1 RNA processing-a segment of the viral lifecycle not targeted by current drugs-and discovered compound N-[4-chloro-3-(trifluoromethyl)phenyl]-7-nitro-2,1,3-benzoxadiazol-4-amine (5342191) as a potent inhibitor of both wild-type (Ba-L, NL4-3, LAI, IIIB, and N54) and drug-resistant strains of HIV-1 (IC50: ~700 nM) with no significant effect on cell viability at doses tested. 5342191 blocks expression of four essential HIV-1 structural and regulatory proteins (Gag, Env, Tat, and Rev) without affecting total protein synthesis of the cell. This response is associated with altered unspliced (US) and singly-spliced (SS) HIV-1 RNA accumulation (~60% reduction) and transport to the cytoplasm (loss of Rev) whereas parallel analysis of cellular RNAs revealed less than a 0.7% of host alternative splicing (AS) events (0.25-0.67% by ≥ 10-20%), gene expression (0.01-0.46% by ≥ 2-5 fold), and protein abundance (0.02-0.34% by ≥ 1.5-2 fold) being affected. Decreased expression of Tat, but not Gag/Env, upon 5342191 treatment was reversed by a proteasome inhibitor, suggesting that this compound alters the synthesis/degradation of this key viral factor. Consistent with an affect on HIV-1 RNA processing, 5342191 treatment of cells altered the abundance and phosphorylation of serine/arginine-rich splicing factor (SRSF) 1, 3, and 4. Despite the activation of several intracellular signaling pathways by 5342191 (Ras, MEK1/2-ERK1/2, and JNK1/2/3), inhibition of HIV-1 gene expression by this compound could be reversed by pre-treatment with either a G-protein α-subunit inhibitor or two different MEK1/2 inhibitors. These observations demonstrate enhanced sensitivity of HIV-1 gene expression to small changes in host RNA processing and highlights the potential of modulating host intracellular signaling as an alternative approach for controlling HIV-1 infection.

Intracellular vesicle clusters are organelles that synthesize extracellular vesicle–associated cargo proteins in yeast

Extracellular vesicles (EVs) play important roles in cell-cell communication. In budding yeast (Saccharomyces cerevisiae), EVs function as carriers to transport cargo proteins into the periplasm for storage during glucose starvation. However, intracellular organelles that synthesize these EV-associated cargo proteins have not been identified. Here, we investigated whether cytoplasmic organelles-called intracellular vesicle clusters (IVCs)-serve as sites for the synthesis of proteins targeted for secretion as EV-associated proteins. Using proteomics, we identified 377 IVC-associated proteins in yeast cells grown under steady-state low-glucose conditions, with the largest group being involved in protein translation. Isolated IVCs exhibited protein synthesis activities that required initiation and elongation factors. We have also identified 431 newly synthesized proteins on isolated IVCs. Expression of 103Q-GFP, a foreign protein with a long polyglutamine extension, resulted in distribution of this protein as large puncta that co-localized with IVC markers, including fructose-1,6-bisphosphatase (FBPase) and the vacuole import and degradation protein Vid24p. We did not observe this pattern in cycloheximide-treated cells or in cells lacking VID genes, required for IVC formation. The induction of 103Q-GFP on IVCs adversely affected total protein synthesis in intact cells and on isolated IVCs. This expression also decreased levels of EV-associated cargo proteins in the extracellular fraction without affecting the number of secreted EVs. Our results provide important insights into the functions of IVCs as sites for the synthesis of EV-associated proteins targeted for secretion to the periplasm.

Dietary supplemental plant oils reduce methanogenesis from anaerobic microbial fermentation in the rumen

Ruminants contribute to the emissions of greenhouse gases, in particular methane, due to the microbial anaerobic fermentation of feed in the rumen. The rumen simulation technique was used to investigate the effects of the addition of different supplemental plant oils to a high concentrate diet on ruminal fermentation and microbial community composition. The control (CTR) diet was a high-concentrate total mixed ration with no supplemental oil. The other experimental diets were supplemented with olive (OLV), sunflower (SFL) or linseed (LNS) oils at 6%. Rumen digesta was used to inoculate the fermenters, and four fermentation units were used per treatment. Fermentation end-products, extent of feed degradation and composition of the microbial community (qPCR) in digesta were determined. Compared with the CTR diet, the addition of plant oils had no significant (P > 0.05) effect on ruminal pH, substrate degradation, total volatile fatty acids or microbial protein synthesis. Gas production from the fermentation of starch or cellulose were decreased by oil supplementation. Methane production was reduced by 21–28% (P < 0.001), propionate production was increased (P < 0.01), and butyrate and ammonia outputs and the acetate to propionate ratio were decreased (P < 0.001) with oil-supplemented diets. Addition of 6% OLV and LNS reduced (P < 0.05) copy numbers of total bacteria relative to the control. In conclusion, the supplementation of ruminant diets with plant oils, in particular from sunflower or linseed, causes some favorable effects on the fermentation processes. The addition of vegetable oils to ruminant mixed rations will reduce methane production increasing the formation of propionic acid without affecting the digestion of feed in the rumen. Adding vegetable fats to ruminant diets seems to be a suitable approach to decrease methane emissions, a relevant cleaner effect that may contribute to alleviate the environmental impact of ruminant production.

The Effects of &amp;lt;i&amp;gt;Sida corymbosa&amp;lt;/i&amp;gt; Leaf Extract Against Abnormal Bilirubin and Total Protein in Albino Rats by Carbon Tetrachloride (CCl&amp;lt;sub&amp;gt;4&amp;lt;/sub&amp;gt;)

The Sida corymbosa plant has been used for centuries in traditional medicine in different countries for prevention and treatment of different diseases such as liver and cardiac diseases. This work aimed at investigating if <i>Sida corymbosa</i> ethanolic leaf extract has preventive and therapeutic effects against abnormal serum bilirubin and total protein in Albino rats by carbon tetrachloride (CCl₄). This study took place at Nnamdi Azikiwe University, Nnewi Campus, Anambra State. The rats were grouped into those given 5000, 3000 and 1000 mg/kgbw of the extract before and after CCl₄ administration from 7 d to 28 d, those injected CCl₄ only which stayed from 7 d to 28 d and those without CCl₄ and extract. Male Albino rats weighing 170-190 g were used. Injection of CCl₄ (0.4 ml/kgbw i.p) was done using olive as vector before and after oral administration of the <i>Sida corymbosa</i> extract. Serum bilirubin was estimated using Powel’s method while serum total protein was estimated using the Biuret method. Results were analyzed statistically using SPSS statistical software version 21. Analysis done so far revealed that serum bilirubin of rats injected CCl₄ without extract administration (negative control) was significantly higher (P<0.05) than other groups, those administered 5000, 3000 and 1000 mg/kgbw of extract before and after CCl₄ injection and those without CCl₄ nor extract (positive control). The reverse was the case for serum total protein, suggesting protective and therapeutic potentials of the extract. <i>Sida corymbosa</i> ethanolic leaf extract may, therefore, has some protective and therapeutic effects against abnormal serum bilirubin and total protein synthesis in Albino rats by CCl₄ thereby improving the liver function.

Peptide Hydrazides as Thioester Equivalents for the Chemical Synthesis of Proteins.

The chemical synthesis of proteins allows for the precise control of structural information at the atomic level, overcoming the limits of protein expression. Peptide hydrazides are widely used as thioester equivalents in the total chemical synthesis and semisynthesis of proteins as they can be easily prepared using standard solid phase peptide synthesis (SPPS) and recombinant peptide techniques. Via treatment with NaNO2 and subsequent thiolysis, peptide hydrazides can be rapidly converted to peptide thioesters, which then selectively react with recombinant protein containing an N-terminal cysteine (Cys) to form a native peptide bond, thereby linking the two peptide segments without isolating any intermediates.

Branched-chain volatile fatty acids and folic acid accelerated the growth of Holstein dairy calves by stimulating nutrient digestion and rumen metabolism

The combined addition of branched-chain volatile fatty acids (BCVFAs) and folic acid (FA) could improve growth performance and nutrient utilization by stimulating ruminal microbial growth and enzyme activity. This study was conducted to evaluate the effects of BCVFA and FA addition on growth performance, ruminal fermentation, nutrient digestibility, microbial enzyme activity, microflora and excretion of urinary purine derivatives (PDs) in calves. Thirty-six Chinese Holstein weaned calves (60 ± 5.4 days of age and 107 ± 4.7 kg of BW) were assigned to one of four groups in a randomized block design. Treatments were control (without additives), FA (with 10 mg FA/kg dietary DM), BCVFA (with 5 g BCVFA/kg dietary DM) and the combined addition of FA and BCVFA (10 mg/kg DM of FA and 5 g/kg DM of BCVFA). Supplements were hand-mixed into the top one-third of total mixed ration. Dietary concentrate to maize silage ratio was 50 : 50 on a DM basis. Dietary BCVFA or FA addition did not affect dry matter intake but increased average daily gain (ADG) and feed conversion efficiency. Ruminal pH and ammonia N were lower, and total volatile fatty acids (VFAs) concentration was higher for BCVFA or FA addition than for control. Dietary BCVFA or FA addition did not affect acetate proportion but decreased propionate proportion and increased acetate to propionate ratio. Total tract digestibility of DM, organic matter, CP and NDF was higher for BCVFA or FA addition than for control. Dietary BCVFA or FA addition increased activity of carboxymethyl cellulase and cellobiase, population of total bacteria, fungi, Ruminococcus albus, R. flavefaciens, Fibrobacter succinogenes and Prevotella ruminicola as well as total PD excretion. Ruminal xylanase, pectinase and protease activity and Butyrivibrio fibrisolvens population were increased by BCVFA addition, whereas population of protozoa and methanogens was increased by FA addition. The BCVFA × FA interaction was significant for acetate to propionate ratio, cellobiase activity and total PD excretion, and these variables increased more with FA addition in diet without BCVFA than in diet with BCVFA. The data indicated that supplementation with BCVFA or FA increased ADG, nutrient digestibility, ruminal total VFA concentration and microbial protein synthesis by stimulating ruminal microbial growth and enzyme activity in calves.

Antibacterial mechanism of the synergistic combination between streptomycin and alcohol extracts from the Chimonanthus salicifolius S. Y. Hu. leaves.

Chimonanthus salicifolius S. Y. Hu. Is a unique traditional medicinal plant in ancient China, and it can eliminate turbid pathogens with aromatics, clear heat, detoxify, prevent colds and influenza, Xinhua Compendium of Materia Medica records that. In previous study, we investigated the regulation of ethanol extracts (EEs) from C. salicifolius S. Y. Hu. leaves on three common antibiotics (chloramphenicol, streptomycin, imipenem) by the checkerboard method. The combination exhibited the best synergy among all combinations, which were composed of streptomycin and 50% EE (SE) from the C. salicifolius S. Y. Hu. leaves. The aim of this study was to investigate the antibacterial mechanism of the SE against Escherichia coli (E. coli, G-) and Staphylococcus aureus (S. aureus, G+). The antibacterial mechanism of the SE was explored by the time-kill test, the phosphorus metabolism, cell membrane integrity assays, the SDS-PAGE, the SEM and TEM observation. The time-kill test illustrated that the SE was bacteriostatic with a time-dependent relationship, not sterilization. The phosphorus metabolism indicated that the SE lowered phosphorus consumption. The cell membrane integrity assays demonstrated that the cell membrane was damaged, with the nucleic acid flowing out. The SDS-PAGE analysis found that the SE inhibited the synthesis of the total protein. The SEM and TEM results revealed that the surface and internal ultrastructure of bacteria were damaged. The surface of the bacteria was shriveled and deformed, and the internal structure of the cells was also mutilated. The SE damaged the cell membrane, with the cytoplasm flowing out, disturbed the synthesis of total protein and phosphorus metabolism, and ultimately killed the bacteria.