Development and characterization of brain cell line from Trachinotus blochii and its application in virological and gene expression studies.

A permanent cell line, SPB (Snubnose pompano brain) was established from Trachinotus blochii by the explant culture method. It has been sub-cultured more than 75 passages and showed optimal growth at 28°C using L-15 medium supplemented with 15% to 20% FBS. The SPB cells were cryopreserved at different passage levels for various applications. SPB cells were composed of fibroblastic and epithelial-like cells. The SPB cells were tested for mycoplasma contamination which was found to be negative. The origin of the SPB cell line from T. blochii was confirmed by amplification of the mitochondrial cytochrome oxidase I (COI) gene. The transfection efficiency of SPB cell line is 15% assessed by expression of green fluorescent protein using pEGFP-N1 plasmid. In addition, two CMV promotor plasmids pFNCPE42-DNA and pcDNAVP28 were transfected to SPB cells and it shows high expression levels of FNCP of fish nodavirus and VP28 protein of white spot syndrome virus by immunostaining. The SPB cells showed susceptibility to SJNNV and the infection was confirmed by RT-PCR, Western blot, ELISA, TCID50 and RT-qPCR. Experimental infection was carried out in T. blochii using SJNNV propagated in SPB cell line and found 100% mortality with clinical signs. The infection was confirmed by RT-PCR. The SPB cell line can be used for propagation of fish viral pathogens and production of the recombinant proteins.

Starch bio-based composite active edible film functionalized with Carum carvi L. essential oil: antimicrobial, rheological, physic-mechanical and optical attributes.

In the present study, the antimicrobial, rheological, mechanical, barrier and optical properties of Carrageenan and Manihot esculenta (composite) starch biobased edible film incorporated with caraway (Carum carvi L.) essential oil (EO) were investigated. The Minimum Inhibitory Concentration (MIC) of caraway oil against B. cereus, E. coli, P. aeruginosa and S. aureus were found to be 0.6, 1.4, 1.4 and 0.8% respectively. The Gas Chromatography- Mass Spectroscopy (GC-MS) of caraway EO expressed a distinct chromatogram peak for phenolic compounds. Rheological results of Film-Forming Solution (FFS) revealed solid-like viscoelastic behavior. Incorporation of caraway EO in the film caused significant (P < 0.05) increase in moisture, moisture absorption, bio-degradability in terms of film solubility, L value, total color difference (ΔE), haziness and transparency value, however, significantly (P < 0.05) decreased tensile strength and whiteness index were observed. The zone of inhibition of caraway EO incorporated films against all test bacteria were highly significant (P < 0.01) than control whereas antibacterial activity was found more towards gram-positive bacteria than gram-negative bacteria. No significant (P>0.05) changes in thickness, density, water activity, swelling, elongation at break, water vapor transmission rate, a and b value were observed with increasing caraway EO concentration. These results with some good rheological, physic-mechanical, antimicrobial and optical characteristics suggest the application of such active film into a variety of foods with improved food safety and quality. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13197-021-05028-1.

Effect of amaranth and quinoa seed flour on rheological and physicochemical properties of goat meat nuggets.

An attempt was made through the present study to prepare gluten free goat meat nuggets by replacing refined wheat flour from product formulation with healthy, dietary fibre rich amaranth (A) and quinoa (Q) flour at different levels. A total of five different treatments viz., AI (1.5% amaranth), AII (3% amaranth), QI (1.5% quinoa) and QII (3% quinoa) were prepared. The physicochemical, colour, texture, sensory and rheological properties of these pseudocereal-added products were evaluated against control (3% refined wheat flour). Emulsion stability of meat batter was significantly affected (P < 0.05) due to addition of amaranth flour (1.5% and 3%) and quinoa (3%). Treatment groups AII and QI had significantly low (P < 0.05) moisture content with respect to control while the amount of fat content showed a reverse trend. Addition of amaranth and quinoa significantly increased (P < 0.05) the dietary fibre in meat products. Rheology of meat batter was affected by types and level of pseudocereal incorporation. Treatment AII had low (P < 0.05) hunter colour lightness value, whereas redness value was low for treatment QI. Effect of added amaranth and quinoa flour was observed on the textural parameters like adhesiveness, cohesiveness, gumminess and chewiness. High scores for almost all the sensory parameters were recorded in pseudocereal-added meat products. Though, all the products were very much acceptable, product with 1.5% quinoa flour was found to have high (P < 0.05) overall acceptability score.

A gene expression signature distinguishes innate response and resistance to proteasome inhibitors in multiple myeloma.

Extensive interindividual variation in response to chemotherapy is a major stumbling block in achieving desirable efficacy in the treatment of cancers, including multiple myeloma (MM). In this study, our goal was to develop a gene expression signature that predicts response specific to proteasome inhibitor (PI) treatment in MM. Using a well-characterized panel of human myeloma cell lines (HMCLs) representing the biological and genetic heterogeneity of MM, we created an in vitro chemosensitivity profile in response to treatment with the four PIs bortezomib, carfilzomib, ixazomib and oprozomib as single agents. Gene expression profiling was performed using next-generation high-throughput RNA-sequencing. Applying machine learning-based computational approaches including the supervised ensemble learning methods Random forest and Random survival forest, we identified a 42-gene expression signature that could not only distinguish good and poor PI response in the HMCL panel, but could also be successfully applied to four different clinical data sets on MM patients undergoing PI-based chemotherapy to distinguish between extraordinary (good and poor) outcomes. Our results demonstrate the use of in vitro modeling and machine learning-based approaches to establish predictive biomarkers of response and resistance to drugs that may serve to better direct myeloma patient treatment options.

Quality and Acceptability of Meat Nuggets with Fresh Aloe vera Gel.

Aloe vera has been used worldwide for pharmaceutical, food, and cosmetic industries due to its wide biological activities. However, quality improvement of low fat meat products and their acceptability with added Aloe vera gel (AVG) is scanty. The aim of this study was to explore the feasibility of using fresh AVG on physicochemical, textural, sensory and nutritive qualities of goat meat nuggets. The products were prepared with 0%, 2.5%, and 5% fresh AVG replacing goat meat and were analyzed for proximate composition, physicochemical and textural properties, fatty acid profile and sensory parameters. Changes in lipid oxidation and microbial growth of nuggets were also evaluated over 9 days of refrigerated storage. The results showed that AVG significantly (p<0.05) decreased the pH value and protein content of meat emulsion and nuggets. Product yield was affected at 5% level of gel. Addition of AVG in the formulation significantly affected the values of texture profile analysis. The AVG reduced the lipid oxidation and microbial growth in nuggets during storage. Sensory panelists preferred nuggets with 2.5% AVG over nuggets with 5% AVG. Therefore, AVG up to 2.5% level could be used for quality improvement in goat meat nuggets without affecting its sensorial, textural and nutritive values.

Rapid Identification of Novel Inhibitors of the Human Aquaporin-1 Water Channel.

Aquaporins (AQPs) are a family of membrane proteins that function as channels facilitating water transport in response to osmotic gradients. These play critical roles in several normal physiological and pathological states and are targets for drug discovery. Selective inhibition of the AQP1 water channel may provide a new approach for the treatment of several disorders including ocular hypertension/glaucoma, congestive heart failure, brain swelling associated with a stroke, corneal and macular edema, pulmonary edema, and otic disorders such as hearing loss and vertigo. We developed a high-throughput assay to screen a library of compounds as potential AQP1 modulators by monitoring the fluorescence dequenching of entrapped calcein in a confluent layer of AQP1-overexpressing CHO cells that were exposed to a hypotonic shock. Promising candidates were tested in a Xenopus oocyte-swelling assay, which confirmed the identification of two lead classes of compounds belonging to aromatic sulfonamides and dihydrobenzofurans with IC50 s in the low micromolar range. These selected compounds directly inhibited water transport in AQP1-enriched stripped erythrocyte ghosts and in proteoliposomes reconstituted with purified AQP1. Validation of these lead compounds, by the three independent assays, establishes a set of attractive AQP1 blockers for developing novel, small-molecule functional modulators of human AQP1.

Single-cell analysis of targeted transcriptome predicts drug sensitivity of single cells within human myeloma tumors.

Multiple myeloma (MM) is characterized by significant genetic diversity at subclonal levels that have a defining role in the heterogeneity of tumor progression, clinical aggressiveness and drug sensitivity. Although genome profiling studies have demonstrated heterogeneity in subclonal architecture that may ultimately lead to relapse, a gene expression-based prediction program that can identify, distinguish and quantify drug response in sub-populations within a bulk population of myeloma cells is lacking. In this study, we performed targeted transcriptome analysis on 528 pre-treatment single cells from 11 myeloma cell lines and 418 single cells from 8 drug-naïve MM patients, followed by intensive bioinformatics and statistical analysis for prediction of proteasome inhibitor sensitivity in individual cells. Using our previously reported drug response gene expression profile signature at the single-cell level, we developed an R Statistical analysis package available at https://github.com/bvnlabSCATTome, SCATTome (single-cell analysis of targeted transcriptome), that restructures the data obtained from Fluidigm single-cell quantitative real-time-PCR analysis run, filters missing data, performs scaling of filtered data, builds classification models and predicts drug response of individual cells based on targeted transcriptome using an assortment of machine learning methods. Application of SCATT should contribute to clinically relevant analysis of intratumor heterogeneity, and better inform drug choices based on subclonal cellular responses.

Management of relapsed multiple myeloma: recommendations of the International Myeloma Working Group.

The prognosis for patients multiple myeloma (MM) has improved substantially over the past decade with the development of new, more effective chemotherapeutic agents and regimens that possess a high level of anti-tumor activity. In spite of this important progress, however, nearly all MM patients ultimately relapse, even those who experience a complete response to initial therapy. Management of relapsed MM thus represents a vital aspect of the overall care for patients with MM and a critical area of ongoing scientific and clinical research. This comprehensive manuscript from the International Myeloma Working Group provides detailed recommendations on management of relapsed disease, with sections dedicated to diagnostic evaluation, determinants of therapy, and general approach to patients with specific disease characteristics. In addition, the manuscript provides a summary of evidence from clinical trials that have significantly impacted the field, including those evaluating conventional dose therapies, as well as both autologous and allogeneic stem cell transplantation. Specific recommendations are offered for management of first and second relapse, relapsed and refractory disease, and both autologous and allogeneic transplant. Finally, perspective is provided regarding new agents and promising directions in management of relapsed MM.

Risk factors for and outcomes of patients with POEMS syndrome who experience progression after first-line treatment.

Although clinical improvement is almost universal with therapy in patients with POEMS (an acronym for polyneuropathy, organomegaly, endocrinopathies, monoclonal protein and a variety of skin changes) syndrome, outcomes and management of patients who relapse or progress (R/P) after first-line treatment have not been described. We retrospectively identified 262 patients with POEMS syndrome treated at the Mayo Clinic from 1974 to 2014 and who had follow-up information. The 5-year progression-free survival (PFS) and overall survival (OS) was 58% and 78%, respectively. Median time to R/P was 42 months. Seventy-nine patients (30%) had an R/P, with 52 (19%) experiencing a symptomatic R/P. Eighteen patients relapsed with symptoms or signs that were not documented at diagnosis. Median times to vascular endothelial growth factor, hematologic, radiographic and clinical R/P were 35 months (range, 4-327 months), 72 months (range, 4-327 months), 51 months (range, 4-327 months) and 48 months (range, 6-311 months), respectively. On multivariate analyses, low albumin at diagnosis and failure to achieve a complete hematologic response to first-line therapy were independent risk factors for PFS. Thirty patients had documentation of a second R/P at a median of 26 months from diagnosis of the first R/P. An early R/P was a risk factor for death, but most patients with an R/P had salvageable disease. A majority of patients are still without R/P at 5 years from diagnosis.

Conversion from tacrolimus to cyclosporine in patients with new-onset diabetes after renal transplant: an open-label randomized prospective pilot study.

BACKGROUND: New-onset diabetes after transplant (NODAT) is associated with serious morbidity and mortality. The incidence of NODAT is higher with tacrolimus (Tac) compared with cyclosporine (CsA); however, the effects of switching from Tac to CsA in NODAT have not been studied well. MATERIALS AND METHODS: This was a single-center, open-label, prospective, randomized study, including renal transplant recipients who were on Tac-based immunosuppression and developed NODAT. Those with pretransplant diabetes, hypersensitivity to CsA or Tac, severe infections, and denying consent were excluded. Subjects were randomized to either switch to CsA or to continue on Tac. Fasting and postprandial plasma glucose, fasting insulin and C-peptide levels, insulin and oral hypoglycaemic agents (OHA) use were monitored monthly for 3 months, whereas glycosylated haemoglobin (HbA1c) was checked at baseline and 3 months. RESULTS: Sixty-seven subjects were randomized to switch to CsA (n = 32) or continuation of Tac (n = 35). Both groups had similar baseline characteristics. After randomization, there was significant improvement in fasting plasma glucose, fasting insulin levels, C-peptide levels, and insulin requirement in both groups, whereas HbA1c improved significantly only in the CsA group. The decline in fasting plasma glucose and insulin requirement was more significant in subjects on CsA. An equal number of subjects in each group (59.4% in CsA group and 40% in Tac group, P = ns) had resolution of NODAT. Weight gain was more significant in the CsA group; however, there was no difference in other side effects or rejection episodes. CONCLUSIONS: A switch from tacrolimus to cyclosporine is a safe and effective strategy in patients with NODAT.

Texture analysis of (125)I-A5B7 anti-CEA antibody SPECT differentiates metastatic colorectal cancer model phenotypes and anti-vascular therapy response.

BACKGROUND: We aimed to test the ability of texture analysis to differentiate the spatial heterogeneity of (125)I-A5B7 anti-carcinoembryonic antigen antibody distribution by nano-single photon emission computed tomography (SPECT) in well-differentiated (SW1222) and poorly differentiated (LS174T) hepatic metastatic colorectal cancer models before and after combretastatin A1 di-phosphate anti-vascular therapy. METHODS: Nano-SPECT imaging was performed following tail vein injection of 20 MBq (125)I-A5B7 in control CD1 nude mice (LS174T, n=3 and SW1222, n=4), and CA1P-treated mice (LS174T, n=3; SW1222, n=4) with liver metastases. Grey-level co-occurrence matrix textural features (uniformity, homogeneity, entropy and contrast) were calculated in up to three liver metastases in 14 mice from control and treatment groups. RESULTS: Before treatment, the LS174T metastases (n=7) were more heterogeneous than SW1222 metastases (n=12) (uniformity, P=0.028; homogeneity, P=0.01; contrast, P=0.045). Following CA1P, LS174T metastases (n=8) showed less heterogeneity than untreated LS174T controls (uniformity, P=0.021; entropy, P=0.006). Combretastatin A1 di-phosphate-treated SW1222 metastases (n=11) showed no difference in texture features compared with controls (all P>0.05). CONCLUSIONS: Supporting the potential for novel imaging biomarkers, texture analysis of (125)I-A5B7 SPECT shows differences in spatial heterogeneity of antibody distribution between well-differentiated (SW1222) and poorly differentiated (LS174T) liver metastases before treatment. Following anti-vascular treatment, LS174T metastases, but not SW1222 metastases, were less heterogeneous.

Microbial quality, instrumental texture, and color profile evaluation of edible by-products obtained from Barbari goats.

AIM: The study was conducted to estimate the contribution of edible byproducts of Barbari kids to their live and carcass weight as well as to assess textural and color characteristics and microbiological status of these byproducts. MATERIALS AND METHODS: Percent live weight, Percent carcass weight, Texture, color, and microbiological analysis was done for edible byproducts viz. liver, heart, kidney, spleen, brain and testicle and longissimus dorsi muscle was taken as a reference. RESULTS: The edible byproducts of Barbari kids constitute about 3% of the live weight of an animal of which liver contributed maximum (1.47%) followed by testicles (0.69%) and heart (0.41%). While the same constituted 3.57, 1.70, and 0.99%, respectively on carcass weight. There was significant (p<0.05) difference among all organs regarding textural properties. Liver required the maximum shear force and work of shear (121.48N and 32.19 kg-sec) followed by spleen and heart. All organs revealed characteristics color values (L*, a*, b*, chroma, and hue) which were significantly different (p<0.05) from muscle values. The total viable count, coliform count showed slight differences for all organs studied. The staphylococcus counts were low with little differences among organs. CONCLUSION: Edible byproducts have a significant contribution to carcass weight which could enhance total edible portion of the carcass. Efficient utilization of these by-products returns good source of revenue to the meat industries. Textural and color analysis give information for their incorporation in comminuted meat products, and microbial study tells about the storage study. However, study was in the preliminary and basic step forward toward better utilization of 3% of live animal which could increase the saleable cost of animal by 6.94%.

Effect of almond on technological, nutritional, textural and sensory characteristics of goat meat nuggets.

An attempt was made through the present study to develop meat based functional food by incorporating almond nut at two different 2.5 and 5 % (Formulation 1 and 2) levels and observe its impact on the different quality attributes against control goat meat nuggets. Almond improved (P < 0.05) the emulsion stability of formulation 1 and 2. Product with 2.5 % almond had higher (P < 0.05) cooking yield than other two products, whereas expressible water was lower. Emulsion and products pH values increased with the addition of almond. Emulsion and products with almond had higher (P < 0.05) proximate values except moisture content. Textural properties of all the products did not differ significantly. Percent SFAs decreased (P < 0.05) and MUFAs were increased with the addition of almond. The fatty acids C16:0, C18:1, C18:2 were significantly higher in formulation 1 and 2 products. Almond incorporation can be a very good approach to enhance nutritional profile of the meat products without affecting acceptability.

Effect of different fat level on microwave cooking properties of goat meat patties.

The study was carried out to evaluate the effect of various fat levels on the cooking and sensory properties of goat meat patties cooked by microwave energy. Goat meat patties were prepared with refined vegetable oil to get fat level of 5, 10, 15 and 20%. Each patty was cooked in a microwave oven with full power (700 W) operating at 2450 MHz to an internal temperature of 75-80 °C. pH value of raw patties with 5% fat level were lower compared to patties with 10, 15 and 20% fat level. Fat level did not affect emulsion stability of batter but it decreased as fat level increased. Microwave cooking time decreased as fat levels increased. With an increase in fat contents, protein and moisture in raw patties decreased and in cooked meat patties with 5% fat had higher protein and moisture content than those with more fat. Patties with 5% level showed lower cooking loss than other fat level. Water activity of patties was affected by fat level and patties with 15 and 20% fat had lower water activity than patties with 5 and 10% fat. As fat level increased, shear force value decreased indicating soft texture. Subjective colour evaluation indicated that 5% patties were darker and redder than patties with more fat. Sensory analysis revealed that goat meat patties with 5 and 10% fat had less flavour and juicer than patties with 15 and 20% fat. Goat meat patties with 20% fat were the juiciest. Tenderness and oiliness increased significantly with an increase in fat level. Patties with 15% fat were rated higher overall palatability than others.

Guava (Psidium guajava L.) Powder as an Antioxidant Dietary Fibre in Sheep Meat Nuggets.

This study was conducted to explore the antioxidant potential and functional value of guava (Psidium guajava L.) powder in muscle foods. Guava powder was used as a source of antioxidant dietary fibre in sheep meat nuggets at two different levels i.e., 0.5% (Treatment I) and 1.0% (Treatment II) and its effect was evaluated against control. Guava powder is rich in dietary fibre (43.21%), phenolics (44.04 mg GAE/g) and possesses good radical scavenging activity as well as reducing power. Incorporation of guava powder resulted in significant decrease (p<0.05) in pH of emulsion and nuggets, emulsion stability, cooking yield and moisture content of nuggets while ash and moisture content of emulsion were increased. Total phenolics, total dietary fibre (TDF) and ash content significantly increased (p<0.05) in nuggets with added guava powder. Product redness value was significantly improved (p<0.05) due to guava powder. Textural properties did not differ significantly except, springiness and shear force values. Guava powder was found to retard lipid peroxidation of cooked sheep meat nuggets as measured by TBARS number during refrigerated storage. Guava powder did not affect sensory characteristics of the products and can be used as source of antioxidant dietary fibre in meat foods.

Quantitative assessment of the relative antineoplastic potential of the n-butanolic leaf extract of Annona muricata Linn. in normal and immortalized human cell lines.

Natural products have been the target for cancer therapy for several years but there is still a dearth of information on potent compounds that may protect normal cells and selectively destroy cancerous cells. The present study was aimed to evaluate the cytotoxic potential of n-butanolic leaf extract of Annona muricata L. on WRL-68 (normal human hepatic cells), MDA-MB-435S (human breast carcinoma cells) and HaCaT (human immortalized keratinocyte cells) lines by XTT assay. Prior to cytotoxicity testing, the extract was subjected to phytochemical screening for detecting the presence of compounds with therapeutic potential. Their relative antioxidant properties were evaluated using the reducing power and DPPH* radical scavenging assay. Since most of the observed chemo-preventive potential invariably correlated with the amount of total phenolics present in the extract, their levels were quantified and identified by HPLC analysis. Correlation studies indicated a strong and significant (P<0.05) positive correlation of phenolic compounds with free radical scavenging potential. The results revealed that the extract was moderately cytotoxic to normal cells with a mean IC50 value of 52.4 µg when compared with those obtained for cancerous cells (IC50 values of 29.2 µg for MDA-MB-435S and 30.1 µg for HaCaT respectively). The study confirms the presence of therapeutically active antineoplastic compounds in the n-butanolic leaf extract of Annona muricata. Isolation of the active metabolites from the extract is in prospect.

Antioxidant effects of broccoli powder extract in goat meat nuggets.

The antioxidant potential of broccoli powder extract (BPE) was determined and evaluated in goat meat nuggets at three different levels 1, 1.5 and 2%, compared with control and butylated hydroxyl toluene (100ppm BHT). Total phenolics in 5mg broccoli powder was higher (P<0.05) than 100ppm BHT. Free radical scavenging activity of 2.25mg and 3mg broccoli powder was found similar to 50 and 100ppm BHT. Reducing power of 10mg broccoli powder was comparable to the 100ppm BHT. Incorporation of 1.5 and 2% BPE decreased (P<0.05) the pH value of the products. Total phenolics in product with 2% BPE was similar to BHT nuggets. Chroma value of products with 1.5 and 2% BPE was lower (P<0.05) than control and BHT nuggets. Thiobarbituric acid reactive substances number of BPE nuggets was lower (P<0.05) than control throughout the storage. Thus 2% BPE can be used as natural antioxidant in goat meat nuggets without affecting product acceptability.

Apoptosis induction in MDA-MB-435S, Hep3B and PC-3 cell lines by Rheum emodi rhizome extracts.

The study was aimed at evaluating apoptosis induction potentials of methanolic and aqueous extracts of Rheum emodi Wall. ex Meissn. rhizome. The ability of the extracts to induce apoptosis in MDA-MB-435S (human breast carcinoma), Hep3B (human hepatocellular carcinoma) and PC-3 (human prostate cancer) cell lines were tested by ELISA to detect cellular DNA fragmentation. Results obtained from the present study confirm that the extracts target the cancerous cells towards apoptosis. The study concludes that R. emodi possess anticancer metabolites that can be isolated and used as precursors in development of anticancer drugs. Suppression of apoptosis might contribute to tumor development by means of accumulation of continuously proliferating cells. The strategy employed in this study, to induce apoptosis in the tumor cells, could be a potential target of therapeutic intervention of cancers.