Levels Of Various Proteins Research Articles

Serum deprivation-response protein regulates aldehyde dehydrogenase 1 through integrin-linked kinase signaling in endometrioid carcinoma cells.

Endometrioid carcinoma (EC) is one of the most common malignancies of the female genital system. We reported previously that aldehyde dehydrogenase 1 (ALDH1), a predominant isoform of the ALDH family in mammals and a potential marker of normal and malignant stem cells, is related to the tumorigenic potential of EC. We compared the levels of various proteins in human EC cells with high and low ALDH1 expression using shotgun proteomics and found that serum deprivation‐response protein (SDPR) was preferentially expressed in cells with high ALDH1 expression. Also known as cavin‐2, SDPR is a member of the cavin protein family, which is required for the formation of caveolae. Using SDPR‐knockout EC cells generated using the CRISPR/Cas9 system, we revealed that SDPR was correlated with invasion, migration, epithelial‐mesenchymal transition, and colony formation, as well as the expression of ALDH1. RNA sequencing showed that integrin‐linked kinase (ILK) signaling is involved in the effect of SDPR on ALDH1. Immunohistochemical analysis revealed that the localization of ILK at the cell cortex was disrupted by SDPR knockout, potentially interfering with ILK signaling. Moreover, immunohistochemical analysis of clinical samples showed that SDPR is related to histological characteristics associated with invasiveness, such as poor differentiation, lymphatic invasion, and the microcystic, elongated, and fragmented histopathological pattern. This is, to our knowledge, the first report that SDPR is related to tumor progression.

Monitoring the Immunoproteasome in Live Cells Using an Activity-Based Peptide-Peptoid Hybrid Probe.

Activity-based probes have greatly improved our understanding of the intrinsic roles and expression levels of various proteins within cells. To be useful in live cells, probes must be cell permeable and provide a read-out that can be measured without disrupting the cells or the activity of the target. Unfortunately, probes for the various forms of the proteasome that can be utilized in intact cells are limited; commercially available probes are most effectively used with purified protein or cell lysate. The proteasome, both the 26S and various isoforms of the 20S CP, is an important target with reported roles in cancer, autoimmune disorders, and neurodegenerative diseases. Here, we present the development of a selective probe for the immunoproteasome, a specialized isoform of the 20S proteasome, that becomes expressed in cells that encounter an inflammatory signal. Using a one-bead, one-compound library of small peptides, we discovered a trimer sequence efficiently cleaved by the immunoproteasome with significant selectivity over the standard proteasome. Upon conjugating this sequence to rhodamine 110 and a peptoid, we generated a probe with a considerable improvement in sensitivity compared to that of current aminomethylcoumarin-based proteasome probes. Importantly, our probe was capable of labeling immunoproteasome-expressing cells while maintaining its selectivity over other cellular proteases in live cell cultures. We anticipate this probe to find wide utility for those that wish to study the immunoproteasome's activity in a variety of cell lines and to be used as a reporter to discover small molecules that can perturb the activity of this proteasome isoform.

Economic Assessment of Various Levels of Protein and Energy in Pigeon Squabs Diet

This study was conducted to evaluate the effect of nine diets through the application of 3×3 randomized design which included three levels of energy (2600, 2800 and 3000 Kcal/kg) with 3 supplemental protein levels (14, 16 and 18%) on growth performance, blood parameters, carcass yields and economic efficiency of Baladi pigeon squabs from 28 days old until 6 months of age (age of sexual maturity). A total of 135 pairs of squabs were randomly and independently allocated to the nine dietary treatments, each containing 15 pairs (3 replicate of 5 pairs). The diets were formulated in mash form fortified with vitamin and mineral premix. Moreover, feed and water were provided for ad-libitum consumption. The results showed that, squabs fed higher energy and crude protein diets (3000 ME+16% CP) had higher live body weight with a coincident significant decrease of the feed intake (P<0.0001). In addition, dressing (%), breast and thigh meat yield (%) increased significantly with increasing levels of both energy and protein, where they hit the peak values in squabs fed on 3000 ME+16% CP diet. Furthermore, increasing energy levels did not affect serum proteins (albumin and globulin), urea and uric acid. However, it had a significant positive effect on serum lipids (cholesterol, triglyceride, high density lipoprotein and low density lipoprotein). In terms of economic analysis, 3000 ME+16% CP diet revealed the lowest cost per Kg live weight of birds 81.87 L.E per pair, along with the highest economic return 110.3 L.E per pair and net profit 10.43 L.E per pair. In conclusion, squabs can efficiently utilize diet containing high energy (3000 Kcal/kg) and crude protein (16%) diets, consequently, it can be used to optimize their growth performance and maintain the maximum economic return

Proteins and trypsin inhibitors in seeds of various plants

The potential trypsin inhibitor sources were identified from seeds of ten plants, based on the reaction of trypsin with synthetic substrate Nα-Benzoyl-D, L-arginine 4-nitroanilide hydrochloride (BApNA). All the tested ones contained various levels of protein (20.87 to 155.68 mg g−1 of seed) and trypsin inhibitor units (TIU) (245.77 to 11235.82 TIU g−1 seed). Among them, the Red lucky seed (Adenanthera pavonina L.) was identified as the richest source of trypsin inhibitor with 11235.82 TIU and 96.06 mg protein/g of seed with specific activity of 116.97 TIU mg−1 protein. The next best ones the seeds of bitter gourd (Momordica charantia L.) and babul (Acacia nilotica L.) with the TIU g−1 seed as 9100.03 and 8690.72, protein content of 155.68 and 88.11 mg g−1 seed with the specific activity of 58.45 and 98.63 TIU mg−1 protein, respectively. This study is the first report of Zingiber officinale Rosc. as the source of trypsin inhibitor.

Metformin Induces Apoptosis and Inhibits Proliferation through the AMP-Activated Protein Kinase and Insulin-like Growth Factor 1 Receptor Pathways in the Bile Duct Cancer Cells.

Background/Aims: Metformin has been found to have antineoplastic activity in some cancer cells. This study was performed to determine whether metformin inhibits the proliferation of bile duct cancer cells by inducing apoptosis and its effects on the expression of gene-related proteins involved in cancer growth.Methods: Human extrahepatic bile duct cancer cells (SNU-245 and SNU-1196) were cultured. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays were performed to determine the effect of metformin on the cell proliferation. Apoptosis was measured by a cell death detection enzyme-linked immunosorbent assay and a caspase-3 activity assay. Expression levels of various proteins, with or without specific small interfering ribonucleic acid-induced gene disruption, were measured by Western blot analysis. The migratory activity of the cancer cells was evaluated by wound healing assay.Results: Metformin suppressed cell proliferation in bile duct cancer cells by inducing apoptosis. Metformin inhibited mammalian target of rapamycin (mTOR) by activation of tuberous sclerosis complex 2 (TSC-2) through phosphorylation of adenosine monophosphate-activated protein kinase at threonine-172 (AMPKThr172). Hyperglycemia impaired metformin-induced AMPKThr172 activation and enhanced phosphorylation of AMPK at serine-485 (AMPKSer485). Metformin blocked the inhibitory effect of insulin-like growth factor 1 receptor (IGF-1R)/insulin receptor substrate 1 (IRS-1) pathway on TSC-2, and hyperglycemia impaired metformin-induced inhibition of IGF-1R/IRS-1 pathway and modulated the invasiveness of bile duct cancer cells; however, this effect was impaired by hyperglycemia.Conclusions: Metformin has antineoplastic effects in bile duct cancer, and hyperglycemic environment interrupts the effect of metformin. In addition, AMPK and IGF-1R play a key role in the proliferation of bile duct cancer cells.

Food Consumption Utilization, and Life History Parameters of Helicoverpa armigera (Lepidoptera: Noctuidae) Reared on Diets of Varying Protein Level.

Helicoverpa armigera (Hübner) is an important pest of crops worldwide, and several studies have focused on the development of this species on different artificial diets. However, studies evaluating the insect’s food consumption and utilization using nutritionally different diets are scarce. The aim of this study was to evaluate the biology and to compare the consumption and use of food by H. armigera larvae on diets with different protein levels provided by several dietary ingredients used in the diets. The nutritional index, the relative consumption rate, the relative metabolic rate, the relative growth rate, and the apparent digestibility were higher in the diet with higher than the optimum level of protein. On the other hand, the conversion efficiency of digested food was lower, resulting in a higher metabolic cost. In terms of biological aspects, larval survival was higher for the diet with optimal protein content and lower for the diet with a higher protein level. The pupal period was longer for the diet with a higher protein content, while pupal survival was lower. Among the evaluated diets, the diet with an optimal protein containing white bean 75 g, wheat germ 60 g, soy bran 30 g, milk powder 30 g, brewer’s yeast 37.5 g as the protein sources resulted in a higher net reproductive rate, a shorter time for the population to double in number, and the highest rates of population growth. The results suggest that lower or higher protein contents in the diets of H. armigera negatively affect the biological aspects of this species.

Potent anti-cancer activity of Alnus nitida against lung cancer cells; in vitro and in vivo studies

Alnus nitida is used for multiple disorders in norther areas of Pakistan. In this study we have evaluated methanol extract of leaf (ANL) and stem bark (ANB) of A. nitida against two lung cancer cell lines; A-549 and H460 (Human non-small lung cancer cell lines) during in vitro assays for growth inhibition. Treatment with ANL and ANB markedly inhibited the growth of both cancer cell lines. Exposure of A-549 and H460 cancer cell lines to ANL and ANB inhibited cell survival, colony growth and migration of cells. Further, treatment of A-549 and H460 with ANL and ANB indicated alteration in actin fibers after staining with rhodamine-phalloidin. Both extracts cause shrinkage and cell cycle arrest during G1 phase. Treatment of A-549 and H460 cancer cells with ANL and ANB repressed the expression of anti-apoptotic proteins Bcl-2 and Bcl-xL along with downregulation of NFκB, cyclin D1 and PI3-K protein. In addition, intraperitoneal injection of ANL and ANB (10 mg/kg bw and 20 mg/kg bw) to C57BL/6 J mice implanted with B16F10 (Mouse melanoma cancer cell line) cells significantly (p < 0.01) decreased the number of nodules per lung and the level of various proteins reciprocating the in vitro studies. These results suggest that ANL and ANB be explored further for therapeutic use in lung cancer.

Potential protein markers for breast cancer recurrence: a retrospective cohort study.

We evaluated five key proteins involved in various cancer-related pathways and assessed their relation to breast cancer recurrence. We used the Kentucky Cancer Registry to retrospectively identify primary invasive breast cancer cases (n = 475) that were diagnosed and treated at University of Kentucky Medical Center between 2000 and 2007. Breast cancer recurrence was observed in 62 cases during the 5-year follow-up after diagnosis. Protein expression or activity level was analyzed from surgery tissue using immuno-histochemical assays. Compared to ER+/PR+/HER2- patients without recurrence, those with recurrence had higher TWIST expression (p = 0.049) but lower ABL1/ABL2 activity (p = 0.003) in primary tumors. We also found that triple-negative breast cancer patients with recurrence had higher SNAI1 expression compared to those without recurrence (p = 0.03). After adjusting for potential confounders, the higher ABL1/ABL2 activity in primary tumors was associated with a decreased risk of recurrence (OR 0.72, 95% CI 0.85-0.90) among ER+/PR+/HER2- patients. In addition, among patients with recurrence we observed that the activity level of ABL1/ABL2 was significantly increased in recurrent tumors compared to the matched primary tumors regardless of the subtype (p = 0.013). These findings provide evidence that the expression/activity level of various proteins may be differentially associated with risk of recurrence of breast tumor subtypes.

Influence of protein concentration and coagulation temperature on rennet-induced gelation characteristics and curd microstructure

This study characterized the coagulation properties and defined the cutting window (CW; time between storage modulus values of 35 and 70 Pa) using rheometry for milk standardized to 4, 5, or 6% protein and set at 28, 32, or 36°C. Milks were standardized to a protein-to-fat ratio of approximately 1 by blending ultrafiltration retentate, skim milk, and whole milk. The internal curd microstructure for selected curd samples was analyzed with transmission electron microscopy and scanning electron microscopy. Lowering the coagulation temperature caused longer rennet coagulation time and time to reach storage modulus of 35 Pa, translating into a wider CW. It also led to a lower maximum curd-firming rate (MCFR) with lower firmness at 40 min at a given protein level. Increasing protein levels resulted in the opposite effect, although without an effect on rennet coagulation time at a given temperature. On coagulation at 28°C, milk with 5% protein resulted in a similar MCFR (∼4 Pa/min) and CW (∼8.25 min) compared with milk with 4% protein at 32°C, which reflects more standard conditions, whereas increasing milk to 6% protein resulted in more than doubling of the curd-firming rate (MCFR = 9.20 Pa/min) and a shorter CW (4.60 min). Gels set at 28°C had lower levels of rearrangement of protein network after 40 min compared with those set at 36°C. Protein levels, on the other hand, had no influence on the levels of protein network rearrangement, as indicated by loss tangent values. The internal structure of curd particles, as investigated by both scanning electron microscopy and transmission electron microscopy, appeared to have less cross-linking and smaller casein aggregates when coagulated at 28°C compared with 36°C, whereas varying protein levels did not show a marked effect on aggregate formation. Overall, this study showed a marked interactive effect between coagulation temperature and protein standardization of milk on coagulation properties, which subsequently requires adjustment of the CW during cheesemaking. Lowering of the coagulation temperature greatly altered the curd microstructure, with a tendency for less syneresis during cutting. Further research is required to quantify the changes in syneresis and in fat and protein losses to whey due to changes in the microstructure of curd particles arising from the different coagulation conditions applied to the protein-fortified milk.

Novel Chemically Synthesized, Alpha-Mangostin-Loaded Nano-Particles, Enhanced Cell Death Through Multiple Pathways Against Malignant Glioma.

Mangostin is a hydrophobic agent with potential anticancer activity. Molecular dynamics computer simulation indicated methoxy poly(ethylene glycol)-poly(lactide) (MPEG-PLA) and α-mangostin (α-M) have good compatibility. The α-M-loaded nano-particles acting as an anticancer agents against growth of glioma cells were prepared by self-assembly methods. In this study, the effects of α-M-loaded nano-particles, α-M/MPEG-PLA with drug loading of 15% and a mean particle size of 32 nm, on the growth of glioma cells were examined both in vitro and in vivo. The occurrence of changes in the cell signaling molecules and expression levels of various proteins related to cell death and glioma xenograft models (i.e., zebra fish, subcutaneous-mouse and orthotopic-mouse) growth following the administration of α-M/MPEG-PLA were investigated. The novel α-M/MPEG-PLA inhibited the growth of malignant glioma cells, induced cells apoptosis with cleaved caspases expression and DNA fragmentation, along with down-regulation of anti-apoptotic molecules and up-regulation of apoptotic molecules. Furthermore, decreased proliferation as well as vascularization of the tumor in vivo models were significantly achieved. A dramatic induction of programmed cell death was found in malignant glioma cells after treatment with synthetic α-M/MPEG-PLA. These results suggest that synthetic α-M/MPEG-PLA could be a promising novel anticancer agent for performing chemotherapy against malignant glioma.

Effect and mechanisms of celastrol on the apoptosis of HOS osteosarcoma cells.

Osteosarcoma is the most common primary malignant tumor of the bone found predominantly in children and teenagers and results in early metastasis and poor prognosis. The present study primarily focused on the impact of celastrol on apoptosis and autophagy of osteosarcoma HOS cells, as well as the related mechanisms. Following the appropriate treatment, the human osteosarcoma cell line HOS was assessed for viability, Ca2+ in cells, apoptosis and changes in cell morphology using Cell Counting Kit-8, flow cytometry, inverted phase contrast microscope, Hoechst staining and transmission electron microscopy. The expression levels of various proteins, including endoplasmic reticulum stress (ERS)-related proteins (Bip, PERK, p-PERK, IRE1α, calnexin, PDI and Erol‑Lα), apoptosis-related proteins (CHOP, cleaved caspase‑12), mitochondrial apoptosis-related proteins (Bax, Bcl-2 and cytochromec), cleaved caspase-3, and autophagy-related proteins (LC3-Ⅰ, LC3-Ⅱ and P62) and β-actin, were assessed with western blotting. Celastrol significantly inhibited the viability of HOS cells in a dose-dependent manner and promoted the expression of ERS-related, apoptosis-related and mitochondrial apoptosis-related proteins. The ERS inhibitor tauroursodeoxycholate promoted celastrol-induced autophagy and apoptosis of HOS cells. Pretreatment with the PERK inhibitor GSK2656157 significantly promoted celastrol-induced death and attenuated HOS cell autophagy. Our results indicated that the ERS pathway and the mitochondrial pathway were involved in celastrol-induced apoptosis of HOS cells. The ERS/PERK pathway may protect HOS cells from apoptosis by celastrol and may play a complicated role in the process of autophagy.

Abstract 448: The novel role of ecdysoneless (ECD) in maintain ER Ca2+ homeostasis

Abstract Background and Significance: Calcium (Ca2+) is an essential signal transduction element involved in the regulation of several cellular activities such as gene transcription, proliferation and apoptosis. Endoplasmic Reticulum (ER) is the Ca2+ reservoir inside the cell that strictly orchestrate Ca2+ gradient across the Plasma Membrane(PM). Importantly, intracellular Ca2+ flux, is a key to orderly regulate cell proliferation, if altered can promote uncontrolled proliferation leading to cancer. Ca2+ entry through the [Ca2+]PM channels during [Ca2+]ER depletion is called the Store Operated Ca2+ Entry (SOCE). Impairment of endogenous SOCE leads to sustained [Ca2+]ER depletion and results in accumulation of unfolded proteins in the ER (known as ER stress). Understanding the basic molecular mechanism regulating PM to ER Ca2+ flow and developing novel strategies to target ER Ca2+ signaling could be beneficial from cancer therapeutic perspective. We have identified Ecdysoneless (ECD), an evolutionary conserved novel cell cycle regulator protein. ECD is overexpressed in several cancers, including breast and pancreatic cancers and its overexpression in breast cancer correlates with poor prognosis and short survival of patients. Recently our lab demonstrates that ECD is a negative regulator of ER stress; however, there is no mechanistic insights of how ECD senses and acts to maintain ER homeostasis. Experimental Design: We have utilized mouse embryonic fibroblasts (MEFs) from Tet(O)-FhECD-IRES-eGFP-rtTA and Ecdflox/flox mice transgenic mice to regulate ECD levels in cells to assess the role of ECD in Ca2+ signaling in a highly controlled system. Structured illumination microscopy(SIM) and biochemical fractionation techniques were performed to identify localization of ECD. Live cell imaging of cells stained with Ca2+ binding dyes like Fluo4-AM and Fluo5N-AM were used to access [Ca2+]Cyto and [Ca2+]ER levels respectively. Fluorescence resonance energy transfer (FRET) and immunoprecipitation methods were conducted to analyze SOCE pathway. Western blotting was used to assess the levels of various proteins involved in Ca2+ signaling. Results: We demonstrate several novel findings: (i) ECD is a smooth ER associated protein; (ii) ECD regulates assembly of ER-PM protein complex to drive Ca2+ entry through SOCE into the ER, maintaining [Ca2+]ER homeostasis; iii) ECD is overexpressed upon Ca2+ depletion and provides survival benefit to cells; v) ECD regulates Ca2+ mediated cell proliferation via CaMKII phosphorylation. Conclusion: Our results demonstrate ECD is a sensor of ER stress, induced by Ca2+deprivation, and overexpressed ECD promotes cell proliferation and survival under ER stress. These findings suggest ECD could be a potential target of Cancer therapy. Citation Format: Aniruddha Sarkar, Irfana Saleem, Sameer Mirza, Keshore Bidasee, Rajini Rao, Hamid Band, Vimla Band. The novel role of ecdysoneless (ECD) in maintain ER Ca2+ homeostasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 448.

Economic Assessment of Various Levels of Protein and Energy in Pigeon Squabs Diet

Economic Assessment of Various Levels of Protein and Energy in Pigeon Squabs Diet

Phytochemical Analysis by HPLC–HRESI-MS and Anti-Inflammatory Activity of Tabernaemontana catharinensis

Tabernaemontana catharinensis (Apocynaceae) has been popularly used by folk medicine because of its anti-inflammatory, analgesic, and antiophidic properties. This study aims to analyze the flavonoids composition of the hydroethanolic extract and of the ethyl acetate (EtOAc) and butanol (BuOH) fractions of T. catharinensis leaves, as well as to evaluate their anti-inflammatory activity using in vivo models. The phytochemical profile, determined by High-Performance Liquid Chromatography–High-Resolution Electrospray Ionization-Mass Spectrometry (HPLC–HRESI-MS), showed the presence of flavonoids mainly having an isorhamnetin nucleus. The anti-inflammatory activity was evaluated in carrageenan-induced paw edema (pre- and post-treatment) with oral administration of a T. catharinensis hydroethanolic extract (50, 100, and 150 mg/kg) and of organic fractions (50 mg/kg). The extract and fractions showed antiedematogenic activity by decreasing myeloperoxidase (MPO) production. In the zymosan-air-pouch model, the extract and fractions inhibited leukocyte migration and significantly decreased the levels of various proteins, such as MPO, interleukin (IL)-1β, and tumor necrosis factor (TNF)-α. The cytotoxicity was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, which revealed no cytotoxicity of the extract and the fractions. These results suggest that the hydroethanolic extract and organic fractions of T. catharinensis leaves have sufficient anti-inflammatory activity to support the popular use of this plant in the treatment of inflammatory disorders.

Differential research of inflammatory and related mediators in BPH, histological prostatitis and PCa.

Prostate cancer (PCa) is one of the most common male malignancies in the world. It was aimed to investigate differential expression of inflammatory and related factors in benign prostatic hyperplasia (BPH), prostate cancer (PCa), histological prostatitis (HP) and explore the role of Inducible nitric oxide synthase (iNOS), (VEGF) Vascular endothelial growth factor, androgen receptor (AR) and IL-2, IL-8 and TNF-α in the occurrence and development of prostate cancer. RT-PCR was used to detect the mRNA expression level of iNOS, VEGF, AR and IL-2, IL-8 and TNF-α in BPH, PCa and BPH+HP. Western blotting and immunohistochemical staining were used to detect the protein levels of various proteins in three diseases. The results showed the mRNA and protein levels of iNOS, VEGF and IL-2, IL-8 and TNF-α were significantly increased in PCa and BPH+HP groups compared with BPH group (p<.05), while the AR was significantly lower than those in PCa and BPH+HP groups (p<.05). There was no significant difference in the mRNA and protein levels of iNOS, VEGF, AR and IL-2, IL-8 and TNF-α between PCa and BPH+HP groups (p>.05). iNOS, VEGF, AR and IL-2, IL-8 and TNF-α are involved in the malignant transformation of prostate tissue and play an important role in the development and progression of Prostate cancer (PCa).

Histopathological and Behavioral Assessments of Aging Effects on Stem Cell Transplants in an Experimental Traumatic Brain Injury.

Traumatic brain injury (TBI) displays cognitive and motor symptoms following the initial injury which can be exacerbated by secondary cell death. Aging contributes significantly to the morbidity of TBI, with higher rates of negative neurological and behaviors outcomes. In the recent study, young and aged animals were injected intravenously with human adipose-derived mesenchymal stem cells (hADSCs) (Tx), conditioned media (CM), or vehicle (unconditioned media) following TBI. The beneficial effects of hADSCs were analyzed using various molecular and behavioral techniques. More specially, DiR-labeled hADSCs were used to observe the biodistribution of the transplanted cells. In addition, a battery of behavior tests was conducted to evaluate the neuromotor function for each treatment group and various regions of the brain were analyzed utilizing Nissl, hematoxylin and eosin (H&E), and human nuclei (HuNu) staining. Finally, flow cytometry was also performed to determine the levels of various proteins in the spleen. Here, we discuss the protocols for characterizing the histopathological and behavioral effects of transplanted stem cells in an animal model of TBI, with an emphasis on the role of aging in the therapeutic outcomes.

Effect of varying protein levels on growth and digestive enzyme activities of pengba Osteobrama belangeri (Valenciennes, 1844)

Pengba, Osteobrama belangeri (Valenciennes, 1844) is considered as one of the candidate species for diversification of aquaculture. However, only scanty information is available on its nutrient requirements. Hence, the present study was conducted on growth and digestive enzyme activities of pengba fry in response to different dietary protein levels. Six semi-purified isocaloric diets were formulated with different levels of protein at 5% increments (25, 30, 35, 40, 45 and 50%) and fed to different groups of pengba fry for a period of 60 days. Survival of fish was significantly influenced by different protein levels (p&lt;0.05). The highest weight gain% and specific growth rate (4.17% day-1) was obtained at 45% protein level which thereafter declined. The best feed conversion ratio (1.46) was observed at 45% protein level and the highest protein efficiency ratio (2.14) was obtained at 25% protein levelwith significant difference (p&lt;0.05). The crude protein content of fish was significantly affected by dietary protein levels (p&lt;0.05) and highest was recorded at 45% protein diet. Moisture, ether extract and ash content did not show significant (p&gt;0.05) variation among dietary treatments. The activity of proteolytic enzymes (proteases, typsin and chymotrypsin) and acid phosphatase were signicantly (p&lt;0.05) enhanced by dietary protein levels up to 45%, which were found to decline thereafter. Amylase activity was significantly decreased as protein levels increased in the diet. Lipase and alkaline phosphatase activities were not influenced by dietary protein levels. In the present study, penbga fry fed with 45% protein diet, showed the best growth performance and highest digestive enzyme activities. The activity of digestive enzymes in the gut clearly demonstrated that the proteolytic enzymes are activated in response to dietary protein utilisation and also revealed that pengba fry may require high protein (45%) diet for better growth and survival. Second order polynomial regression analysis revealed that the maximum protein requirement of pengba fry was 45.1% (r2 = 0.90). The findings of the study is expected to have significant implications for feed development for pengba aquaculture.

Herbal prescription, Danggui-Sayuk-Ga-Osuyu-Senggang-Tang, inhibits TNF-α-induced epithelial-mesenchymal transition in HCT116 colorectal cancer cells.

Tumor necrosis factor‑α‑mediated (TNF‑α) epithelial‑mesenchymal transition (EMT) is associated with distant metastasis in patients with colorectal cancer with poor prognosis. Although traditional herbal medicines have long been used to treat colorectal cancer, the incidence and mortality in patients with colorectal cancer has continued to increase. Danggui‑Sayuk‑Ga‑Osuyu‑Saenggang‑Tang (DSGOST) has long been used for treatment of chills, while few studies have reported its anticancer effect. This study aimed to demonstrate the inhibitory effect of DSGOST on TNF‑α‑mediated invasion and migration of colorectal cancer HCT116 cell lines. MTT was used to measure cell viability. Wound healing and Τranswell invasion assay were used to detect migration and invasion of cells, respectively. The intracellular localization of proteins of interest was assessed by immunocytochemistry. Western blotting was performed to determine the expression level of various proteins. A non‑toxic dose of DSGOST (50µg/ml) on HCT116 cells was determined by MTT assay. Furthermore, DSGOST prevented the TNF‑α‑induced invasive phenotype in HCT116 cells. DSGOST inhibition of the invasive phenotype was also associated with increased expression of EMT markers. Furthermore, DSGOST treatment blocked TNF‑α‑induced migration and invasion of HCT116 cells. In addition, DSGOST treatment inhibited TNF‑α‑mediated nuclear translocation of Snail. DSGOST treatment also downregulated TNF‑α‑induced phosphorylation of AKT and glycogen synthase kinase‑3β. Therefore, the findings of the current study suggest that DSGOST exhibits anti‑migration and anti‑invasion effects in TNF‑α‑treated HCT116 human colorectal cells.

Mycobacterium smegmatis PafBC is involved in regulation of DNA damage response

Two genes, pafB and pafC, are organized in an operon with the Pup-ligase gene pafA, which is part of the Pup-proteasome system (PPS) present in mycobacteria and other actinobacteria. The PPS is crucial for Mycobacterium tuberculosis resistance towards reactive nitrogen intermediates (RNI). However, pafB and pafC apparently play only a minor role in RNI resistance. To characterize their function, we generated a pafBC deletion in Mycobacterium smegmatis (Msm). Proteome analysis of the mutant strain revealed decreased cellular levels of various proteins involved in DNA damage repair, including recombinase A (RecA). In agreement with this finding, Msm ΔpafBC displayed increased sensitivity to DNA damaging agents. In mycobacteria two pathways regulate DNA repair genes: the LexA/RecA-dependent SOS response and a predominant pathway that controls gene expression via a LexA/RecA-independent promoter, termed P1. PafB and PafC feature winged helix-turn-helix DNA binding motifs and we demonstrate that together they form a stable heterodimer in vitro, implying a function as a heterodimeric transcriptional regulator. Indeed, P1-driven transcription of recA was decreased in Msm ΔpafBC under standard conditions and induction of recA expression upon DNA damage was strongly impaired. Taken together, our data indicate an important regulatory function of PafBC in the mycobacterial DNA damage response.

Galectin-3 rs4652 A>C polymorphism is associated with the risk of gastric carcinoma and P-glycoprotein expression level.

Galectin-3 serves an important function in cancer development and progression. The present study aimed to explore the association between single nucleotide polymorphisms in galectin-3, and the susceptibility to chemotherapy drug resistance of gastric carcinoma. The present study was a case-control study including 479 patients with gastric carcinoma and 458 cancer-free controls in a population from the Fujian province in Southeast China. Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry was used to determine the genotype of rs4644 and rs4652, and immunohistochemistry was used to identify the expression level of various proteins associated with chemotherapeutic drug resistance. The results revealed that individuals exhibiting the rs4652 CA/AA genotype had a significantly increased risk of developing gastric carcinoma compared with the rs4652 CC genotype (adjusted odds ratio, 1.51; 95% confidence interval, 1.05-2.18; adjusted P=0.03). In addition, it was demonstrated that there were significant differences in the P-glycoprotein expression level depending on rs4652 genotypic distributions (χ2=9.063; P=0.028). Therefore, the present study demonstrated that rs4652 single nucleotide polymorphisms of the galectin-3 gene contribute to the susceptibility to and chemotherapeutic drug resistance of gastric carcinoma.