Studying Microflora of Semi-hard Cheese, and Sporulation within Contact Surface Biofilms

The first study was conducted on commercial Italian semi-hard cheese samples that were analyzed microbiologically to understand the effect of fast and slow rate of cooling on cheese microflora during ripening process at different temperatures. A cheese plant noticed an active growth of non-starter lactic acid bacteria, especially the heterofermentative bacteria, in ripened cheese blocks, which weren’t fully cooled. For that, three cheese sample sets, each having 8 samples, were received from the cheese plant for microbiological analysis. The first set included a group of fast cooled cheese samples to 38°F, and the other group was slow cooled to 50°F. The second set was cooled as the first set but further ripened for 2 months at either 38°F or 50°F, while the third set was ripened for 6 months after cooling. We were targeting the desired and undesired non-starter lactic acid bacteria (NSLAB), the total aerobic viable count for mesophiles / thermophiles, the thermoduric mesophiles / thermophiles, gas producers, Leuconostoc spp., Lactobacillus wasatchensis, and Clostridium spp. Non-selective and different types of selective media were used in order to enumerate and isolate each type of microorganism. The media were incubated aerobically or anaerobically at different temperatures depending on the targeted isolate’s optimum growth conditions. Random colonies on the selective media were picked and sent to another analytical laboratory for further analysis using Matrix Assisted Laser Desorption Ionization mass spectrometry - Time of flight technique MALDI-TOF. The total aerobic counts showed, in general, a declining trend during 6 months of ripening. Initial NSLAB counts in the 12 days old samples cooled to 38°F started at 5.87°0.13 logs CFU/g, and 6.19°0.13 logs CFU/g in the samples cooled to 50°F, and the two were significantly different. The fast-cooled samples thus showed lower counts than the slow cooled samples. According to MALDITOF identification, the added adjunct cultures were dominating in the whole study, which were Pediococcus acidilactici, Lactobacillus paracasei, and Lactobacillus rhamnosus. None of the samples had any of the Clostridium spp., undesired Lactobacillus spp. or Leuconostoc spp. Our results suggest that if accelerated ripening at 50°F is the goal, the rate of cooling to 38°F or 50°F will not make a difference from the bacterial count aspect in the presence of Pediococcus acidilactici as adjunct. However, bacterial distribution in cheese due to different rate of cooling might matter the most. The second study was aimed to understand the sporulation within biofilm matrix that formed on stainless steel surface by common sporeformers in the dairy industry. Undisturbed biofilm works as a reservoir for spore forming contaminants that might produce spores within the biofilm matrix itself. The results of this study showed that the duration of the biofilm formation and sporulation is variable among the different species and their strains. A strain of B. licheniformis ATCC⌐6634™ formed biofilm within 4 hours, but formed no spores even after an extended duration of incubation up to 21 days. However, the second strain of G. stearothermophilus ATCC⌐ 15952™ not only formed biofilms in 4 hours, but also sporulated within the biofilm matrix in 48 hours. On the other hand, B. licheniformis ATCC®14580™ took 7 days to produce spores within the NFDM and the biofilm. Keeping in mind the application of our research for future projects, the first study was conducted with cheese samples that contained high levels of Pediococcus acidilactici as a part of the adjunct cultures, which did not allow true predominance to emerge. It may be more useful to conduct this study without added Pediococcus, so that actual microbial interactions in cheese matrix could show up. The second study provided a proof of concept on the sporulation behavior of sporeformers within biofilm matrix. This would be useful to design cleaning strategies for resilient biofilms formed on plate heat exchangers and other equipment in dairy plants

The crustal uplift determined at the Jakobshavn glacier (West Greenland) using ATM and GPS data

The Greenland ice sheet has experienced record melting in recent years. In order to estimate the ice loss we can make use of the earth's natural elasticity to weigh the ice. Ice bends down the bedrock so when the ice melts away, the bedrock rises measurably in response. Throughout this abstract we present both a predicted and observed crustal uplift for the Jakobshavn glacier using ATM data (Airborne Topographic Mapper) from NASA ATM flights during 1997, 2005 and 2010 supplemented with data provided from continuous Global Positioning System (GPS), measurements made on bedrock between 2005-2010. In order to compute the crustal uplift in response to the ice mass loss of the Jakobshavn area from the GPS stations, the convolution of the gridded thinning rates has been computed with the vertical-displacement Green's function as described in [1]. Several manipulations of data were required in order to achieve a good prediction of the crustal uplift. In this sense the programs Matlab and Geogrid-Gravsoft were used along with some Fortran executable files. Furthermore, the GPS data which presents the difference in uplift is provided processed as a difference of data from the permanent GPS stations KAGA, ILUL and QEQE relative to the AASI station (Figure 1). Also, in order to compare the predicted uplift from ATM data with the observed uplift from GPS data the post-glacial rebound (PGR) rates have been subtracted. The results obtained for the predicted crustal uplift for KAAS is 11.62 mm/yr while the observed value was 16.321 mm/yr, for ILAS 1.74 mm/yr and 1.53 mm/yr, for QEAS -0.189 mm/yr and 1.15 mm/yr. That being a difference of 4.701 mm/yr is found for KAAS, 0.21 mm/yr for ILAS and 1.339 mm/yr for QEAS. The uncertainties associated both with the ATM and GPS results are 0.8 mm/yr for ATM and 0.5 mm/yr for GPS. The total ice mass loss in km3 of water predicted from the ATM data concerning the Jakobshavn area is -88.815 between 2010 and 2005 and -83.599 between 2005 and 1997. It seems fair to state that this differences, between the predicted and observed rates, may also be due to the fact that not all the errors have been taken into account when computing the observed results and also due to the fact that, perhaps, ice is melting in Greenland much faster than predicted. REFERENCES Farrell, W. E. (1972), Deformation of the Earth by surface loads, Rev. Geophys., 10(3), 761– 797, doi:10.1029/RG010i003p0076

Are all fibrinogen concentrates the same? The effects of two fibrinogen therapies in an afibrinogenemic patient and in a fibrinogen deficient plasma model. A clinical and laboratory case report

The choice of treatments for inherited, or acquired, fibrinogen deficient states is expanding and there are now several fibrinogen concentrate therapies commercially available. Patients with the rare inherited bleeding disorder, afibrinogenemia, commonly require life-long replacement therapy with fibrinogen concentrate to prevent hemorrhagic complications. Recent reports in the setting of acquired bleeding, namely trauma hemorrhage, have highlighted the potential importance of the different compositions of fibrinogen supplements, including cryoprecipitate and the various plasma- derived concentrates. Clot strength and the subsequent susceptibility of a clot to lysis is highly dependent on the amount of fibrinogen as well as its structural composition, the concentration of pro- and anti-coagulant factors, as well as fibrinolytic regulators, such as factor XIII (FXIII). This report details the effects of two commercially available fibrinogen concentrates (Riastap®, CSL Behring and Fibryga®, Octapharma) on important functional measures of clot formation and lysis in a patient with afibrinogenemia. Our report offers insights into the differential effects of these concentrates, at the clot level, according to the variable constituents of each product, thereby emphasizing that the choice of fibrinogen concentrate can influence the stability of a clot in vivo. Whether this alters clinical efficacy is yet to be understood

A Cloud-Based Global Flood Disaster Community Cyber-Infrastructure: Development and Demonstration

Flood disasters have significant impacts on the development of communities globally. This study describes a public cloud-based flood cyber-infrastructure (CyberFlood) that collects, organizes, visualizes, and manages several global flood databases for authorities and the public in real-time, providing location-based eventful visualization as well as statistical analysis and graphing capabilities. In order to expand and update the existing flood inventory, a crowdsourcing data collection methodology is employed for the public with smartphones or Internet to report new flood events, which is also intended to engage citizen-scientists so that they may become motivated and educated about the latest developments in satellite remote sensing and hydrologic modeling technologies. Our shared vision is to better serve the global water community with comprehensive flood information, aided by the state-of-the- art cloud computing and crowdsourcing technology. The CyberFlood presents an opportunity to eventually modernize the existing paradigm used to collect, manage, analyze, and visualize water-related disasters

A cloud-based global flood disaster community cyber-infrastructure: Development and demonstration

Flood disasters have significant impacts on the development of communities globally. This study describes a public cloud-based flood cyber-infrastructure (CyberFlood) that collects, organizes, visualizes, and manages several global flood databases for authorities and the public in real-time, providing location-based eventful visualization as well as statistical analysis and graphing capabilities. In order to expand and update the existing flood inventory, a crowdsourcing data collection methodology is employed for the public with smartphones or Internet to report new flood events, which is also intended to engage citizen-scientists so that they may become motivated and educated about the latest developments in satellite remote sensing and hydrologic modeling technologies. Our shared vision is to better serve the global water community with comprehensive flood information, aided by the state-ofthe- art cloud computing and crowd-sourcing technology. The CyberFlood presents an opportunity to eventually modernize the existing paradigm used to collect, manage, analyze, and visualize water-related disasters

Targeting the Microtubule-Network Rescues CTL Killing Efficiency in Dense 3D Matrices

Efficacy of cytotoxic T lymphocyte (CTL)-based immunotherapy is still unsatisfactory against solid tumors, which are frequently characterized by condensed extracellular matrix. Here, using a unique 3D killing assay, we identify that the killing efficiency of primary human CTLs is substantially impaired in dense collagen matrices. Although the expression of cytotoxic proteins in CTLs remained intact in dense collagen, CTL motility was largely compromised. Using light-sheet microscopy, we found that persistence and velocity of CTL migration was influenced by the stiffness and porosity of the 3D matrix. Notably, 3D CTL velocity was strongly correlated with their nuclear deformability, which was enhanced by disruption of the microtubule network especially in dense matrices. Concomitantly, CTL migration, search efficiency, and killing efficiency in dense collagen were significantly increased in microtubule-perturbed CTLs. In addition, the chemotherapeutically used microtubule inhibitor vinblastine drastically enhanced CTL killing efficiency in dense collagen. Together, our findings suggest targeting the microtubule network as a promising strategy to enhance efficacy of CTL-based immunotherapy against solid tumors, especially stiff solid tumors

Multiple myeloma and its treatment contribute to increased platelet reactivity

Multiple myeloma (MM) and its precursor states, smoldering myeloma (SM) and monoclonal gammopathy of undetermined significance (MGUS) are associated with increased incidence of thrombosis, however the cause of this is unknown. Lenalidomide treatment of MM substantially improves patient survival, although significantly increases thrombotic risk by an unknown mechanism. This pilot study aimed to establish the impact of MM and its treatment with Lenalidomide on platelet function. We analyzed platelet function in MGUS, SM and MM compared to healthy controls. We report an increase in platelet reactivity in MGUS, SM, and MM where increases in fibrinogen binding, P-selectin exposure, altered receptor expression, elevated levels of aggregation and enhanced sensitivity to agonist stimulation were observed. We also demonstrate an increase in patient platelet reactivity post Lenalidomide treatment compared to pre-treatment. We show Lenalidomide treatment of platelets ex vivo increased reactivity that was associated with formation of larger thrombi at arterial shear rates but not venous shear rates. This study demonstrates a clear increase in platelet reactivity and prothrombotic potential in patients with MGUS, SM and MM which is elevated further upon treatment with Lenalidomide. Our observations suggest that more detailed studies are warranted to determine mechanisms of thrombotic complications to enable the development of new preventative strategies that specifically target platelets

Molecular Cloning, Characterization and Predicted Structure of a Putative Copper-Zinc SOD from the Camel, Camelus dromedarius

Superoxide dismutase (SOD) is the first line of defense against oxidative stress induced by endogenous and/or exogenous factors and thus helps in maintaining the cellular integrity. Its activity is related to many diseases; so, it is of importance to study the structure and expression of SOD gene in an animal naturally exposed most of its life to the direct sunlight as a cause of oxidative stress. Arabian camel (one humped camel, Camelus dromedarius) is adapted to the widely varying desert climatic conditions that extremely changes during daily life in the Arabian Gulf. Studying the cSOD1 in C. dromedarius could help understand the impact of exposure to direct sunlight and desert life on the health status of such mammal. The full coding region of a putative CuZnSOD gene of C. dromedarius (cSOD1) was amplified by reverse transcription PCR and cloned for the first time (gene bank accession number for nucleotides and amino acids are JF758876 and AEF32527, respectively). The cDNA sequencing revealed an open reading frame of 459 nucleotides encoding a protein of 153 amino acids which is equal to the coding region of SOD1 gene and protein from many organisms. The calculated molecular weight and isoelectric point of cSOD1 was 15.7 kDa and 6.2, respectively. The level of expression of cSOD1 in different camel tissues (liver, kidney, spleen, lung and testis) was examined using Real Time-PCR. The highest level of cSOD1 transcript was found in the camel liver (represented as 100%) followed by testis (45%), kidney (13%), lung (11%) and spleen (10%), using 18S ribosomal subunit as endogenous control. The deduced amino acid sequence exhibited high similarity with Cebus apella (90%), Sus scrofa (88%), Cavia porcellus (88%), Mus musculus (88%), Macaca mulatta (87%), Pan troglodytes (87%), Homo sapiens (87%), Canis familiaris (86%), Bos taurus (86%), Pongo abelii (85%) and Equus caballus (82%). Phylogenetic analysis revealed that cSOD1 is grouped together with S. scrofa. The predicted 3D structure of cSOD1 showed high similarity with the human and bovine CuZnSOD homologues. The Root-mean-square deviation (rmsd) between cSOD1/hSOD1 and cSOD1/bSOD1 superimposed structure pairs were 0.557 and 0.425 A. The Q-score of cSOD1-hSOD1 and cSOD1-bSOD1 were 0.948 and 0.961, respectively

Horizontal DNA transfer mechanisms of bacteria as weapons of intragenomic conflict

Horizontal DNA transfer (HDT) is a pervasive mechanism of diversification in many microbial species, but its primary evolutionary role remains controversial. Much recent research has emphasised the adaptive benefit of acquiring novel DNA, but here we argue instead that intragenomic conflict provides a coherent framework for understanding the evolutionary origins of HDT. To test this hypothesis, we developed a mathematical model of a clonally descended bacterial population undergoing HDT through transmission of mobile genetic elements (MGEs) and genetic transformation. Including the known bias of transformation toward the acquisition of shorter alleles into the model suggested it could be an effective means of counteracting the spread of MGEs. Both constitutive and transient competence for transformation were found to provide an effective defence against parasitic MGEs; transient competence could also be effective at permitting the selective spread of MGEs conferring a benefit on their host bacterium. The coordination of transient competence with cell-cell killing, observed in multiple species, was found to result in synergistic blocking of MGE transmission through releasing genomic DNA for homologous recombination while simultaneously reducing horizontal MGE spread by lowering the local cell density. To evaluate the feasibility of the functions suggested by the modelling analysis, we analysed genomic data from longitudinal sampling of individuals carrying Streptococcus pneumoniae. This revealed the frequent within-host coexistence of clonally descended cells that differed in their MGE infection status, a necessary condition for the proposed mechanism to operate. Additionally, we found multiple examples of MGEs inhibiting transformation through integrative disruption of genes encoding the competence machinery across many species, providing evidence of an ongoing "arms race." Reduced rates of transformation have also been observed in cells infected by MGEs that reduce the concentration of extracellular DNA through secretion of DNases. Simulations predicted that either mechanism of limiting transformation would benefit individual MGEs, but also that this tactic's effectiveness was limited by competition with other MGEs coinfecting the same cell. A further observed behaviour we hypothesised to reduce elimination by transformation was MGE activation when cells become competent. Our model predicted that this response was effective at counteracting transformation independently of competing MGEs. Therefore, this framework is able to explain both common properties of MGEs, and the seemingly paradoxical bacterial behaviours of transformation and cell-cell killing within clonally related populations, as the consequences of intragenomic conflict between self-replicating chromosomes and parasitic MGEs. The antagonistic nature of the different mechanisms of HDT over short timescales means their contribution to bacterial evolution is likely to be substantially greater than previously appreciated

Perceived risk of infection and death from COVID-19 among community members of low- and middle-income countries: A cross-sectional study [version 1; peer review: awaiting peer review]

Background: Risk perceptions of coronavirus disease 2019 (COVID-19) are considered important as they impact community health behaviors. The aim of this study was to determine the perceived risk of infection and death due to COVID-19 and to assess the factors associated with such risk perceptions among community members in low- and middle-income countries (LMICs) in Africa, Asia, and South America. Methods: An online cross-sectional study was conducted in 10 LMICs in Africa, Asia, and South America from February to May 2021. A questionnaire was utilized to assess the perceived risk of infection and death from COVID-19 and its plausible determinants. A logistic regression model was used to identify the factors associated with such risk perceptions. Results: A total of 1,646 responses were included in the analysis of the perceived risk of becoming infected and dying from COVID-19. Our data suggested that 36.4% of participants had a high perceived risk of COVID-19 infection, while only 22.4% had a perceived risk of dying from COVID-19. Being a woman, working in healthcare-related sectors, contracting pulmonary disease, knowing people in the immediate social environment who are or have been infected with COVID-19, as well as seeing or reading about individuals infected with COVID-19 on social media or TV were all associated with a higher perceived risk of becoming infected with COVID-19. In addition, being a woman, elderly, having heart disease and pulmonary disease, knowing people in the immediate social environment who are or have been infected with COVID-19, and seeing or reading about individuals infected with COVID-19 on social media or TV had a higher perceived risk of dying from COVID-19. Conclusions: The perceived risk of infection and death due to COVID-19 are relatively low among respondents; this suggests the need to conduct health campaigns to disseminate knowledge and information on the ongoing pandemic