Semicontinuous system for the production of recombinant mCherry protein in Chlamydomonas reinhardtii

Biotechnology advances have allowed bacteria, yeasts, plants, mammalian and insect cells to function as heterologous protein expression systems. Recently, microalgae have gained attention as an innovative platform for recombinant protein production, due to low culture media cost, compared to traditional systems, as well as the fact that microalgae such as Chlamydomonas reinhardtii are considered safe (GRAS) by the Food and Drug Administration (FDA). Previous studies showed that recombinant protein production in traditional platforms by semicontinuous process increased biomass and bio product productivity, when compared to batch process. As there is a lack of studies on semicontinuous process for recombinant protein production in microalgae, the production of recombinant mCherry fluorescent protein was evaluated by semicontinuous cultivation of Chlamydomonas reinhardtii in bubble column photobioreactor. This semicontinuous cultivation process was evaluated in the following conditions: 20%, 40%, and 60% culture portion withdrawal. The highest culture withdrawal percentage (60%) provided the best results, as an up to 161% increase in mCherry productivity (454.5 RFU h⁻¹ – Relative Fluorescence Unit h⁻¹), in comparison to batch cultivation (174.0 RFU h⁻¹) of the same strain. All cultivations were carried out for 13 days, at pH 7, temperature 25°C and, by semicontinuous process, two culture withdrawals were taken during the cultivations. Throughout the production cycles, it was possible to obtain biomass concentration up to 1.36 g L⁻¹.     

Assessing environmental requirements effects on forest fragmentation sensitivity in two arboreal rodents

The study of the effect of habitat fragmentation on species that inhabit residual patches requires the investigation of the relationship existing between species distribution and landscape components. To understand which components of landscape mosaics are more influential for species’ persistence, we compared the distribution of two arboreal rodents proved to be sensitive to habitat fragmentation, the hazel dormouse Muscardinus avellanarius and the red squirrel Sciurus vulgaris. Their occurrence in residual oak woods in central Italy was studied with nest-boxes and hair-tubes, developing a new method for hair analysis. Their distributions were analysed considering patch, matrix composition and configuration, and landscape vegetation variables. The two species showed a different degree of plasticity, with the squirrel being significantly more specialised at the landscape scale. The comparison of the two distribution patterns highlighted the influence of different ecological constraints and the existence of different strategies to cope with fragmentation. Patch size and patch attributes were generally weaker determinants of occurrence, compared to landscape metrics. The squirrel presence was significantly influenced by the presence of shared perimeter between hedgerows and woods and by the lack of isolation of the residual patches, suggesting the use of several fragments to compensate the low habitat quality. Conversely the hazel dormouse seemed to be more affected by the internal management of the woods, and in particular by the mean DBH. Our results highlight how the recognition of the extrinsic constraints and the influence of multi-scale habitat selection may help guiding land use management, to ensure species conservation in profoundly exploited landscapes.     

Effects of different light wavelengths at equal irradiance on testis weight, protein content, and K-paranitrophenylphosphatase activity in the Syrian hamster

The effects of different light wavelengths at equal irradiance on testis weight, testis protein content, and testis K-paranitrophenylphosphatase (K-pNPPase) activity were studied in the Syrian hamster. One group (long photoperiod) was maintained on a light:dark cycle of 14:10, and another group (short photoperiod) on a cycle of 10:14. Five other groups were maintained on a cycle of 10:14 but with a one hour pulse of equally intense illumination in the middle of the dark period with UV, blue, green, yellow or red light. Animals exposed to a long photoperiod or UV, blue or green light pulses had significantly greater testis weights--up to eightfold greater than those in the yellow or red or short photoperiod groups. Organ protein content closely paralleled organ weight, but the protein/wet weight ratio was consistently higher in the large organ groups. K-pNPPase and Mg-pNPPase activities were significantly higher in the large organ groups, even when expressed per mg protein. Therefore, at a balanced irradiance of 0.2uW/cm2, light wavelength exerts a profound effect on testicular weight, protein content, and K-pNPPase and Mg-pNPPase activities. Testicular involution is a process that is selective with regard to protein biosynthesis.     

Low level lead inhibits the human brain cation pump.

The impact of low level lead exposure on human central nervous system function is a major public health concern. This study addresses the inhibition of the cation pump enzyme Na, K-ATPase by low level lead. Human brain tissue was obtained at autopsy and frozen until use. Brain homogenates were preincubated with PbCl2 for 20 min at 0 degrees C. Inhibition of K-paranitrophenylphosphatase (pNPPase), a measure of the dephosphorylation step of Na,K-ATPase, reached steady state within 10 min. K-pNPPase activity, expressed (mean +/- SEM) as a percentage of control (45.2 +/- 2.7 nmol/mg/min), fell to 96.3 +/- 0.9% at 0.25 uM [PbCl2] to 82.0 +/- 1.6% at 2.5 uM [PbCl2] in homogenates prepared from normal brain. Similar results were obtained with homogenates prepared from brains of patients with a history of alcohol abuse and of those with other miscellaneous conditions. Since the mean blood level of lead in the United States has ranged recently from 9.2 to 16.0 ug/dl (0.44 to 0.77 uM), these results indicate that current in vivo levels of lead exposure may impair important human brain function.     

Different effects of aging on the opioid mechanisms controlling gonadotropin and cortisol secretion in man

The present study was undertaken in order to assess the influence of aging on the endogenous opioid control of gonadotropin and adrenocorticotropin/cortisol secretion in man. For this purpose, the capability of the opioid antagonist naloxone to increase circulating levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and cortisol was tested in male subjects of different ages. Thirty normal men were randomly chosen and divided into 3 groups by age: group I = 22-40 years (n = 10); group II = 41-59 years (n = 10); group III = 62-80 years (n = 10). Since the men of group III showed higher basal serum gonadotropin concentrations than the subjects of group I and group II, we selected from a large population a fourth group of elderly men with normal basal LH and FSH levels: group IV = 61-82 years (n = 7). All subjects were tested for 120 min during the intravenous administration of naloxone (4 mg given in an intravenous bolus at time 0, plus 10 mg infused for 2 h). Control tests with normal saline instead of naloxone were performed in all groups. All subjects had similar blood testosterone and cortisol levels, whereas LH and FSH concentrations were significantly higher in group III than in groups I, II and IV. Naloxone increased plasma cortisol concentrations by 50% in all groups. The cortisol secretory response followed a similar pattern regardless of age.(ABSTRACT TRUNCATED AT 250 WORDS)     

Nutrient content and seasonal fluctuations in the leaf component of coark-oak (Quercus suber L.) litterfall

Nutrient content and seasonality of the leaf component in cork-oak litterfal were studied over a two year period in two cork-oak forest sites differing in biomass and edaphic condition in the north-eastern Iberian peninsula. Fallen senescent leaves compared to young leaves showed higher non-mobile nutrient concentrations and lower mobile nutrient concentrations, specially P, N, K, and Mg. At both sites, seasonal fluctuations affected both leaf production and leaf mineral content. The maximum leaf fall period correspond to the start of the vegetative growth and to the lowest N and P concentration in the falling leaves. The opposite was true for the winter, when litterfal was minimal and N and P content in falling leaves was at a maximum.The comparison between falling leaves and canopy leaves suggests that the analysis of fallen leaves can be a useful measure of N and P nutrition in cork-oak. We found site dependent differences in nutrient content and nutrient remobilization. In comparison with Q. ilex, although litter production was in the same range, nutrient retranslocation was greater for Q. suber.