Optimization of NPK Medium with the Addition of Mung Bean Sprout Extract for Biomass and Lipid Production of Stanieria HS-48 as a Potential Biofuel

Stanieria strain HS-48 from Ciater hot spring, Indonesia contains lipid content in the form of fatty acids that can be used as a raw material for biodiesel. Optimization of the growth medium was used to produce high contents of microalgal lipids. The growth medium in this study utilized NPK medium with the addition of variations in the concentration of mung bean sprout extract 2% and 4%. This study aimed to determine the best concentration of mung bean sprout extract in NPK medium for the highest biomass and lipid production of Stanieria HS-48. The experiments were conducted in airlift photobioreactors (APBR) for 19 days. The highest total cell density of Stanieria HS-48 in NPK medium with the addition of variations in the concentration of mung bean sprout extract 2% and 4% were 6.165 x 106 and 4.702 x 106 cell/ml. The lipid content of the biomass from Stanieria HS-48 on NPK medium with the addition of mung bean sprout extract 2% and 4% were 62.9% and 44.8%. The study showed that the NPK medium with the addition of the concentration of mung bean sprout extract 2% was a good growth medium for increasing biomass and lipid of Stanieria HS-48.

Peptide-Based Formulation from Lactic Acid Bacteria Impairs the Pathogen Growth in Ananas Comosus (Pineapple)

Worldwide, street vending commerce has grown exponentially, representing in some countries, including Ecuador, a significant proportion of food consumed by the urban population. Pineapple is one of the common fruits sold as ready-to-eat slices by ambulant vendors in the street or on public transport at risk of contamination by various microorganisms. Previously, we selected Lactobacillus plantarum UTNCys5-4 and Lactococcus lactis subsp. lactis Gt28 strains producing peptides with high capacity to inhibit pathogen growth in vitro. In this study, the effect of different edited formulations containing a mixture of Cys5-4/Gt28 peptides was evaluated in vitro and ex vitro against a pathogenic cocktail containing E. coli (2), Salmonella (2) and Shigella (1). The growth of bacterial cocktail co-inoculated with cell-free supernatant containing peptides (formulation T1) and precipitated peptides (formulation T6), in a ratio of Cys5-4/Gt28:1:1 (v/v), results in a decrease of total cell viability with 1.85 and 1.2 log CFU/mL orders of magnitude at 6 h of incubation. About the same decrease (1.9 log CFU/g) was observed when pineapple slices artificially inoculated with the pathogenic cocktail were coated with T1 formulation, indicating the capacity to diminish simultaneous pathogens in situ, thus demonstrating its great biological control and protection. However, the E. coli cell counts reduced by 2.08 log CFU/g while Salmonella and Shigella cell counts reduced by 1.43 and 1.91 log CFU/g, respectively, at 5 days of refrigeration. In the untreated pineapple slices, the total cell density was maintained during storage, suggesting the adaptation of the pathogens to the fruit matrix. The peptide-based formulation exerted a bacteriolytic mode of action inducing pathogenic cell death. The results indicate that coating pineapple slices with peptide-based formulation is a promising approach to protect them from further contamination by microbial spoilage as well as an alternative to increase the food safety.

Low Hodgkin/Reed-Sternberg Cell Density Assessed with Automated Image Analysis Is a Poor Prognostic Factor in Classic Hodgkin Lymphoma Patients

Background: Classical Hodgkin lymphoma (cHL) is the most common lymphoid malignancy under the age of 30. Despite the relatively high curability, about 20-30% of patients relapse after front-line therapy. Since the International Prognostic Factors Project score became obsolete in the era of dose-intensive regimens (BEACOPP), a clinically applicable tool for outcome prediction is lacking. Recent years have brought much novel information about close Hodgkin/Reed-Sternberg (HRS) microenvironment cross-talk promoting tumor growth. Many studies have tested the predictive value of microenvironment cells using immunohistochemistry but none have been concerned with HRS cell density. Aim: To assess the prognostic role of HRS cell density in lymph node biopsies using a novel automated system for scanning large tumor sample areas in cHL patients. Methods: Thirty-eight high-quality tissue samples obtained at time of cHL diagnosis were analyzed. The median age at diagnosis was 35 (17.5-93) years; the male-to-female ratio was 0.66:1. The lymphoma subtypes were nodular sclerosis (NS) in 25 (66%) and mixed cellularity (MC) in 13 (34%) patients. At the time of analysis, detailed baseline clinical information was available in 35 (92%) patients. Ann Arbor stages I through IV were observed in 2, 18, 9 and 6 patients, respectively. Systemic symptoms were present in 26 (75%) cases and large lymphoma mass (≥5 cm) was detected in 19 (56%) patients. German Hodgkin Study Group stages were: limited in 4 (12%), intermediate in 8 (23%) and advanced in 23 (66%) patients. Chemotherapy was given to 35 patients (92%): BEACOPP in 20 (57.1%), ABVD in 6 (17.1%), Stanford V in 4 (11.4%) and other (COPP, COPP/ABV) in 5 (14.3%) cases. Involved-field radiotherapy was applied in 11 (29%) cases. Tissue array analyses were performed using the TissueFAXS (TissueGnostics, Austria) system combining detailed morphologic information offered by microscopy with the scientific accuracy of multi-channel flow cytometry. Data were analyzed with TissueQuest software. Paraffin-embedded biopsies were prepared from pre-selected diagnostic samples stained with hematoxylin-eosin for morphology analyses and using CD30 (HRS) for cell density analyses. Each sample was previewed by the scanning software and only significant tumor tissue was manually gated by an experienced pathologist for further analyses. Fibrotic (≥10%), necrotic or residual lymphatic structures were excluded. Results: After treatment, 31 (82%) patients achieved complete remission, three partial remission, one stable disease and three patients progressed. After a median follow-up of 64.3 months, 9 (24%) patients relapsed or progressed and 7 (18%) of them died. Five-year overall survival (5-y OS) reached 81.0% (95%CI 0.68-0.94); 5-year progression survival (PFS) was 68.3% (95%CI 0.53-0.83). Tissue data: The mean scanned area covered 27.2±13.5 mm2 with the mean (median) total number of cells 461,504±286,491 (466,138). The mean (median) total cell density and CD30+ HRS cell density were 17,886±7,884 (16,557) and 279±232 (181) cells per mm2, respectively. Total cell density was 1.28-fold higher (p=0.14) and HRS cell density 1.44-fold (p=0.19) higher in MC compared to NS. HRS cell density did not correlate with sex (p=0.46), systemic symptoms (p=0.38), tumor bulk (p=0.34) or Ann Arbor stage (p=0.59). There was a trend for older age among low HRS cell density (below the median) patients (p=0.19). Low HRS cell density was associated with inferior 5-y PFS (44.4% vs 90.0%, p=0.0024) and OS (57.5% vs 100%, p=0.002). Multivariate Cox regression identified low HRS cell density as an unfavorable prognostic factor for PFS (p=0.037, HR=5.50) independent of age and lymphoma subtype. Conclusions: This is the first evidence about the possible predictive role of HRS cell density in cHL patients. Low density at diagnosis was associated with 5.5-fold higher risk of lymphoma progression or death. Automated image analysis is a new tool overcoming technical limitations caused by small (microarray) samples in lymphomas with high intra-tumor heterogeneity. Further analyses will define the role of HRS cell analysis in cHL in the context of risk-adapted and CD30-targeted therapies. Acknowledgment - Supported by: Palacky University Olomouc (IGA-LF-2015-001), Takeda Restricted Educational Grant and J. W. Fulbright Commission. Figure 1. Figure 1. Disclosures Prochazka: Takeda: Research Funding.

Studies on Species Composition of Phytoplankton in Fuxian Lake of Yunnan, China

It was investigated and analyzed about the species composition of phytoplankton, and their relations to environmental factors sampled in spring, summer and autumn in 2001-2011 in Fuxian Lake (24o 21'~24o 38' N, 102o 49'12 "~102o 57'26"E). The phytoplankton from Fuxian lake included 78 species belong to 41 genera 25 family, 7 phyla. Chlorophyta was predominant, with consisted of 43.5% of total species number. Species number of phytoplankton appeared visible monthly variation. The main predominant phytoplankton species was Mougeotia bloodlei, which accounted for 42.94%~84.94% of total cell density of phytoplankton. The community structures of phytoplankton were consist of Mougeotia bloodlei, Chroococcus spp., Cyclotella spp., Ceratium hirundinella and Dinobryon sertularia.

Polychlorinated Biphenyls Exert Selective Effects on Cellular Composition of White Matter in a Manner Inconsistent with Thyroid Hormone Insufficiency

Developmental exposure to polychlorinated biphenyls (PCBs) is associated with a variety of cognitive deficits in humans, and recent evidence implicates white matter development as a potential target of PCBs. Because PCBs are suspected of interfering with thyroid hormone (TH) signaling in the developing brain, and because TH is important in oligodendrocyte development, we tested the hypothesis that PCB exposure affects the development of white matter tracts by disrupting TH signaling. Pregnant Sprague Dawley rats were exposed to the PCB mixture Aroclor 1254 (5 mg/kg), with or without cotreatment of goitrogens from gestational d 7 until postnatal d 15. Treatment effects on white matter development were determined by separately measuring the cellular density and proportion of myelin-associated glycoprotein (MAG)-positive, O4-positive, and glial fibrillary acidic protein (GFAP)-positive cells in the genu of the corpus callosum (CC) and in the anterior commissure (AC). Hypothyroidism decreased the total cell density of the CC and AC as measured by 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) staining and produced a disproportionate decrease in MAG-positive oligodendrocyte density with a simultaneous increase in GFAP-positive astrocyte density. These data indicate that hypothyroidism reduces cellular density of CC and AC and fosters astrocyte development at the expense of oligodendrocyte density. In contrast, PCB exposure significantly reduced total cell density but did not disproportionately alter MAG-positive oligodendrocyte density or change the ratio of MAG-positive oligodendrocytes to GFAP-positive astrocytes. Thus, PCB exposure mimicked some, but not all, of the effects of hypothyroidism on white matter composition.

Pattern formation of vascular smooth muscle cells subject to nonuniform fluid shear stress: mediation by gradient of cell density

Smooth muscle cells (SMCs) are organized in various patterns in blood vessels. Whereas straight blood vessels mainly contain circumferentially aligned SMCs, curved blood vessels are composed of axially aligned SMCs in regions with vortex blood flow. The vortex flow-dependent feature of SMC alignment suggests a role for nonuniform fluid shear stress in regulating the pattern formation of SMCs. Here, we demonstrate that, in experimental models with vascular polymer implants designed for the observation of neointima formation and SMC migration under defined fluid shear stress, nonuniform shear stress possibly plays a role in regulating the direction of SMC migration and alignment in the neointima of the vascular implant. It was found that fluid shear stress inhibited cell growth, and the presence of nonuniform shear stress influenced the distribution of total cell density and induced the formation of cell density gradients, which in turn directed SMC migration and alignment. In contrast, uniform fluid shear stress in a control model influenced neither the distribution of total cell density nor the direction of SMC migration and alignment. In both the uniform and nonuniform shear models, the gradient of total cell density was consistent with the alignment of SMCs. These observations suggest that nonuniform shear stress may regulate the pattern formation of SMCs, possibly via mediating the gradient of cell density in the neointima of vascular polymer implants.