Customized media based on miniaturized screening improve growth rate and cell yield of methane-oxidizing bacteria of the genus Methylomonas

The growth of twelve methanotrophic strains within the genus Methylomonas, including the type strains of Methylomonas methanica and Methylomonas koyamae, was evaluated with 40 different variations of standard diluted nitrate mineral salts medium in 96-well microtiter plates. Unique profiles of growth preference were observed for each strain, showing a strong strain dependency for optimal growth conditions, especially with regards to the preferred concentration and nature of the nitrogen source. Based on the miniaturized screening results, a customized medium was designed for each strain, allowing the improvement of the growth of several strains in a batch setup, either by a reduction of the lag phase or by faster biomass accumulation. As such, the maintenance of fastidious strains could be facilitated while the growth of fast-growing Methylomonas strains could be further improved. Methylomonas sp. R-45378 displayed a 50 % increase in cell dry weight when grown in its customized medium and showed the lowest observed nitrogen and oxygen requirement of all tested strains. We demonstrate that the presented miniaturized approach for medium optimization is a simple tool allowing the quick generation of strain-specific growth preference data that can be applied downstream of an isolation campaign. This approach can also be applied as a first step in the search for strains with biotechnological potential, to facilitate cultivation of fastidious strains or to steer future isolation campaigns.     

Temperature effect on acetate and propionate consumption by sulfate-reducing bacteria in saline wastewater

Seawater toilet flushing, seawater intrusion in the sewerage, and discharge of sulfate-rich industrial effluents elevates sulfate content in wastewater. The application of sulfate-reducing bacteria (SRB) in wastewater treatment is very beneficial; as for example, it improves the pathogen removal and reduces the volume of waste sludge, energy requirement and costs. This paper evaluates the potential to apply biological sulfate reduction using acetate and propionate to saline sewage treatment in moderate climates. Long-term biological sulfate reduction experiments at 10 and 20 °C were conducted in a sequencing batch reactor with synthetic saline domestic wastewater. Subsequently, acetate and propionate (soluble organic carbon) conversion rate were determined in both reactors, in the presence of either or both fatty acids. Both acetate and propionate consumption rates by SRB were 1.9 times lower at 10 °C than at 20 °C. At 10 °C, propionate was incompletely oxidized to acetate. At 10 °C, complete removal of soluble organic carbon requires a significantly increased hydraulic retention time as compared to 20 °C. The results of the study showed that biological sulfate reduction can be a feasible and promising process for saline wastewater treatment in moderate climate.     

IL-4 Haplotype -590T, -34T and Intron-3 VNTR R2 Is Associated with Reduced Malaria Risk among Ancestral Indian Tribal Populations

Background

Interleukin 4 (IL-4) is an anti-inflammatory cytokine, which regulates balance between T_H1 and T_H2 immune response, immunoglobulin class switching and humoral immunity. Polymorphisms in this gene have been reported to affect the risk of infectious and autoimmune diseases.

Methods

We have analyzed three regulatory IL-4 polymorphisms; -590C>T, -34C>T and 70 bp intron-3 VNTR, in 4216 individuals; including: (1) 430 ethnically matched case-control groups (173 severe malaria, 101 mild malaria and 156 asymptomatic); (2) 3452 individuals from 76 linguistically and geographically distinct endogamous populations of India, and (3) 334 individuals with different ancestry from outside India (84 Brazilian, 104 Syrian, and 146 Vietnamese).

Results

The -590T, -34T and intron-3 VNTR R2 alleles were found to be associated with reduced malaria risk (P<0.001 for -590C>T and -34C>T, and P = 0.003 for VNTR). These three alleles were in strong LD (r²>0.75) and the TTR2 (-590T, -34T and intron-3 VNTR R2) haplotype appeared to be a susceptibility factor for malaria (P = 0.009, OR = 0.552, 95% CI = 0.356 –0.854). Allele and genotype frequencies differ significantly between caste, nomadic, tribe and ancestral tribal populations (ATP). The distribution of protective haplotype TTR2 was found to be significant (χ² ₃ = 182.95, p-value <0.001), which is highest in ATP (40.5%); intermediate in tribes (33%); and lowest in caste (17.8%) and nomadic (21.6%).

Conclusions

Our study suggests that the IL-4 polymorphisms regulate host susceptibility to malaria and disease progression. TTR2 haplotype, which gives protection against malaria, is high among ATPs. Since they inhabited in isolation and mainly practice hunter-gatherer lifestyles and exposed to various parasites, IL-4 TTR2 haplotype might be under positive selection.     

Selection of trichloroethene (TCE) degrading bacteria that resist inactivation by TCE

Two isoprene (2-methyl-1,3-butadiene) utilizing bacteria, Alcaligenes denitrificans ssp. xylosoxidans JE 75 and Rhodococcus erythropolis JE 77, were identified as highly efficient cooxidizers of TCE, cis- and transdichloroethene, 1,1-dichloroethene and vinylchloride. Isoprene grown cells eliminate chloride from TCE in stoichiometric amounts and tolerate high concentrations of TCE.     

The isolation and characterization of gibberellin-deficient mutants in tomato

Summary In tomato, nine independent EMS-induced mutants representing recessive mutations at three different loci (gib-1, gib-2, and gib-3) were isolated. Six of these have an almost absolute gibberellin requirement for seed germination and elongation growth. In addition, the leaves are darker green, smaller, and changed in structure as compared to wild type. The three other mutants, which germinate without GA, are allelic to specific, nongerminating mutants and have less severe mutant characteristics. The respective loci are situated on three different chromosomes. The genes identified by these mutants control steps in gibberellin biosynthesis, as endogenous gibberellins are strongly reduced.     

Protein kinase C inhibition by calmodulin and its fragments

Inhibition of protein kinase C (PKC) by calmodulin is investigated and we describe the localization of inhibitory sequences within the calmodulin molecule. We present evidence that calmodulin inhibits PKC through an inhibition of the activation of PKC associated with lipid membranes: Binding of PKC to lipid vesicles is not affected, but activation is abolished. The potent calmodulin antagonist R24571 (calmidazol) did not affect the inhibition of PKC by calmodulin at concentrations up to 10^–5 M. Two tryptic fragments of calmodulin were isolated which inhibited PKC. They were only slightly less potent than intact calmodulin with an IC₅₀ of 6 µ M compared to 1 µ M of intact calmodulin. They were identified as Ser³⁸-Arg⁷⁴ and His¹⁰⁷-Lys¹⁴⁸. Each of the inhibiting fragments contains an intact Ca²⁺-binding domain with complete helix-loop-helix structure (EF hand). Other calmodulin peptides showed only weak inhibitory activity. Both fragments did not stimulate cAMP phosphodiesterase even at concentrations 100-fold higher than the calmodulin concentration needed for maximal stimulation. None of the fragments acted as a calmodulin antagonist.     

Readiness to lay in the blowfly Lucilia cuprina is unaffected by oviposition site-deprivation or egg-load

Oviposition by Lucilia cuprina Wiedemann (Diptera, Calliphoridae) was examined in relation to period of oviposition site-deprivation and egg-load. Effects of oviposition site-deprivation were examined by comparing oviposition performance of individual females that had matured their batch of oocytes within the previous 24 h with that of females which had reached ovarian maturity 8 days previously. Egg-load was manipulated by causing females of this anautogenous species to consume different amounts of protein-rich material. In no-choice experiments, individual females of the different categories were given access for 4 h to oviposition substrate, soaked with (i) liver exudate, (ii) the exudate diluted 16-fold or (iii) the undilated exudate containing the oviposition deterrent sodium chloride at a concentration of 2 M. These solutions elicited oviposition from different proportions of females, but neither these proportions, nor the interval between introduction of the oviposition site and the initiation of oviposition, was significantly affected by the period of oviposition site-deprivation or the number of eggs matured by the females.

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Résumé L'effet de la privation de lieu de ponte a été étudié en comparant les pontes de femelles isolées ayant formé leurs ufs mûrs dans les 24 heures précédentes, à celles de femelles ayant atteint leur maturité sexuelle 8 jours avant. La rétention ovocytaire est provoquée en faisant consommer aux femelles de cette espèce anautogène différentes quantités d'aliments riches en protéines. La ponte de femelles dont le contingent total de leurs ovocytes s'est développé, — c'est-à-dire 260 —, après consommation ad libitum de foie de mouton pendant 48 heures, a été comparée à celle de femelles ayant formé 190 ovocytes mûrs après ingestion d'une quantité limitée de jus de foie.Dans des expériences sans choix, les femelles isolées de différences catégories ont eu accès pendant 4 heures au substrat de ponte trempé: 1) dans du jus de foie, 2) dans du jus dilué 16 fois, 3) dans du jus de foie non dilué mais contenant NaCl (inhibiteur de la ponte) à la concentration de 2 M. Le jus non dilué a provoqué une forte stimulation, induisant la ponte de 80% des femelles. Le jus dilué et celui contenant NaCl n'ont induit la ponte que de 40% des femelles avec des niveaux de stimulation bien plus faibles. La date d'introduction du lieu de ponte et le taux de rétention des ovocytes mûrs n'ont eu auçun effet sur la proportion de femelles réagissant à ces 3 types de stimulation.

     

Subpicosecond resonance Raman spectra of the early intermediates in the photocycle of bacteriorhodopsin

The resonance Raman spectra are presented for the species formed during the photocycle of bacteriorhodopsin (bR) on a timescale of 800-900 fs. In the ethylenic stretch region two intermediates were found with frequencies of 1,510 and 1,518 cm^-1, corresponding to species with optical absorption maxima at 660 and 625 nm, respectively. This leads to the assignment of the 1,518 cm^-1 band to the J₆₂₅ intermediate. In the fingerprint region, the appearance of a vibration at 1,195 cm^-1 strongly suggests that the isomerization indeed has taken place in a time less than the pulsewidth of our laser. This supports the previous proposals made on the basis of the optical spectra. The spectra are compared with those observed in tens of picoseconds up to nanoseconds.     

Coupling of insulin-responsive glucose transport to receptors for insulin-like growth factor 1 in primary human fibroblasts

We have recently described an insulin-resistant patient with leprechaunism (leprechaun G.) having a homozygous leucine----proline mutation at amino acid position 233 in the alpha-chain of the insulin receptor. The mutation results in a loss of insulin binding to cultured fibroblasts. Fibroblasts from the patient and control individuals were used to quantify the stimulation of 2-deoxyglucose uptake by insulin and insulin-like growth factor 1 (IGF-1). Insulin hardly stimulates basal 2-deoxyglucose uptake in the patient's fibroblasts whereas in control fibroblasts the uptake of 2-deoxyglucose is stimulated by insulin approximately 1.7 times. In contrast, IGF-1 stimulates hexose uptake in the patient's fibroblasts 1.8 times, a similar value to that obtained by stimulation of control fibroblasts with insulin or IGF-1. With both types of fibroblasts, maximal IGF-1 response is reached at about 10 nM IGF-1, the ED50 being approximately 4 nM. The results indicate that the insulin responsive glucose transport in primary fibroblasts is functionally linked to the receptor for IGF-1. Insulin binds with an approximately 200-fold lower affinity to IGF-1 receptors, compared to homologous IGF-1 binding. As an insulin concentration of 10 microM is unable to give maximal stimulation of glucose uptake in the patient's fibroblasts, which is already seen with 10 nM IGF-1, it seems that occupation of IGF-1 receptors by insulin on the patient's cells is less efficient at stimulating hexose uptake compared to homologous activation.