Dipeptidyl aminopeptidase yspI mutants of Schizosaccharomyces pombe: Genetic mapping of dpa1+ on chromosome III

Abstract A mutant strain of Schizosaccharomyces pombe lacking dipeptidyl aminopeptidase yspI was isolated from a strain already defective in aminopeptidase activity by means of a staining technique with the chromogenic substrate ala-pro-4-methoxy-β-naphthylamide to screen colonies for the absence of the enzyme. The defect segregated 2⁺ :2⁻ in meiotic tetrads, indicating a single chromosomal gene mutation, which was shown to be recessive. Gene dosage experiments indicated that the mutation resides in the structural gene of dipeptidyl aminopeptidase yspI, dpa 1⁺. The dpa 1⁺ gene was located on chromosome III by using l m- fluorophen-ylalanine-induced haploidization and mitotic analysis. dpa1 mutants did not show any obvious phenotype under a variety of conditions tested.     

Using FTA® cards to store avian blood samples for genetic studies. Their application in sex determination

FTA® cards were used for long‐term storage of avian blood samples. Blood DNA was extracted by a simple method and used in PCR for sex identification of adult and nestling Great Grey Shrikes Lanius excubitor.     

Can silicon partially alleviate micronutrient deficiency in plants? a review

Silicon protects plants against various biotic and abiotic stresses, including metal toxicity. Under a high metal concentration, Si can externally decrease metal availability to the plant by its precipitation in the growth media, and Si also affects the metal distribution inside the plant, diminishing the damage. Could Si also protect plants against metal deficiency stress? Recently, the physiological role of Si in relation to micronutrients deficiency symptoms has been assessed in several plant species in hydroponics. In cucumber, Si supply mitigated the symptoms of Fe deficiency, but this effect was not clear under Zn- or Mn-deficiency conditions. The main factor controlling this beneficial effect seems to be the Si contribution to the formation of metal deposits in the root and/or leaves apoplast and its role in their following remobilization when required. The enhancement of the content of long-distance transport molecules (such as citrate) due to Si addition should also contribute to the metal transport from root to shoot, which will diminish deficiency symptoms.     

Mutation analysis of phenylketonuria in Spain: prevalence of two Mediterranean mutations

Summary We have screened seven Spanish phenylketonuric (PKU) families for the most prevalent Mediterranean and Caucasian mutations, and have subsequently found mutations P281L and IVS10. We have analyzed these two mutations in 23 of our patients. The frequencies found correspond to those of Mediterranean countries, such as Italy. This is the first report of a mutation analysis in the Spanish PKU population.     

Cytosolic Free Calcium and Gene Expression During Chemical Hypoxia

Understanding the cellular response to hypoxia may help elucidate the role of altered oxidation in neuronal death or abnormal cell function. In PC12 cells, 30 min of chemical hypoxia (i.e., KCN) reduced ATP concentrations by 92%, but diminished viability by only 10%. Ten minutes of hypoxia increased cytosolic free calcium ([Ca2+]i) 2.5-fold above control, but after 30 min of hypoxia, [Ca2+]i was slightly below that of nonhypoxic cells. Short periods of hypoxia also exaggerated the K(+)-induced elevation of [Ca2+]i, but by 30 min these ATP-depleted cells reestablished a calcium gradient that was equal to nonhypoxic, K(+)-depolarized cells. Thus, 30 min of severe ATP depletion left [Ca2+]i and viability relatively unaffected. Nerve growth factor caused slight, but significant, improvements in ATP and viability of hypoxic cells, but had no effect on [Ca2+]i. Although [Ca2+]i was equivalent in control and hypoxic cells after 30 or 60 min, hypoxia abolished the K(+)-stimulated elevation of [Ca2+]i. The nerve growth factor induction of c-fos, an indicator of the genomic response, was diminished by approximately 80%. Thus, hypoxic PC12 cells with greatly reduced ATP stores maintained normal [Ca2+]i, but their ability to respond to external stimulation was impaired. Further, the reduced oxidation that occurs in the brain in a variety of pathological conditions may interfere with the cellular response to stimulation and growth factors.     

Engineering defensin α‐helix to produce high‐affinity SARS‐CoV‐2 spike protein binding ligands

The binding of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) spike protein to the angiotensin‐converting enzyme 2 (ACE2) receptor expressed on the host cells is a critical initial step for viral infection. This interaction is blocked through competitive inhibition by soluble ACE2 protein. Therefore, developing high‐affinity and cost‐effective ACE2 mimetic ligands that disrupt this protein–protein interaction is a promising strategy for viral diagnostics and therapy. We employed human and plant defensins, a class of small (2–5 kDa) and highly stable proteins containing solvent‐exposed alpha‐helix, conformationally constrained by two disulfide bonds. Therefore, we engineered the amino acid residues on the constrained alpha‐helix of defensins to mimic the critical residues on the ACE2 helix 1 that interact with the SARS‐CoV‐2 spike protein. The engineered proteins (h‐deface2, p‐deface2, and p‐deface2‐MUT) were soluble and purified to homogeneity with a high yield from a bacterial expression system. The proteins demonstrated exceptional thermostability (Tm 70.7°C), high‐affinity binding to the spike protein with apparent K _d values of 54.4 ± 11.3, 33.5 ± 8.2, and 14.4 ± 3.5 nM for h‐deface2, p‐deface2, and p‐deface2‐MUT, respectively, and were used in a diagnostic assay that detected SARS‐CoV‐2 neutralizing antibodies. This work addresses the challenge of developing helical ACE2 mimetics by demonstrating that defensins provide promising scaffolds to engineer alpha‐helices in a constrained form for designing of high‐affinity ligands.     

Seasonal changes in condition, nutrition, gonad maturation and energy content in barbel, Barbus sclateri, inhabiting a fluctuating river

The objective of this study was to investigate the seasonal cycle of condition, nutrition and gonad development, as well as the magnitude of seasonal variations in energy content of somatic and gonad tissues in juveniles, males and females of Sclater's barbel, Barbus sclateri, from the upper Guadalete River (south Spain). The influence of reproductive cycle on somatic changes was also investigated and discussed. Measurements of condition factor (K), somatic index (SI), gonadosomatic index (GSI) and somatic and gonad energy content (J g^-1 dw) were made in individual specimens taken from the Guadalete River monthly for 12 months. This freshwater ecosystem is characterised by strong seasonal fluctuations in water and flow level, temperature and food supply. It was found that in general juveniles, males and females of barbel exhibited a similar condition, nutrition and somatic energy cycle throughout the year, with the highest values in spring and the lowest in summer. Depletion of K, SI and somatic energy storage from spring until mid-summer seems to be associated with high metabolic demands during this period, and in adult fish also with spawning-related activity. The gonad index and energy content of the gonad were the highest in April and the lowest in summer for both sexes. Spawning took place during late spring – early summer, with fish quiescent by mid summer. The energy required for ovarian development (3970 J g^-1 dw) was greater than the one for testes development (2763 J g^-1 dw). Data on gonad energy content indicated a period (March to April) of intense energy accumulation (64% males and 37% females) which was related to the decline in the average somatic energy content in males and females. The somatic energy content was linearly related to K and SI. In the same way, GSI correlated positively with gonad energy. Linear trends were found between somatic parameters (K, SI and energy content) and gonad parameters (GSI and energy content). This revised version was published online in July 2006 with corrections to the Cover Date.     

Extensive gaps and biases in our knowledge of a well‐known fauna: implications for integrating biological traits into macroecology

Aim Ecologists seeking to describe patterns at ever larger scales require compilations of data on the global abundance and distribution of species. Comparable compilations of biological data are needed to elucidate the mechanisms behind these patterns, but have received far less attention. We assess the availability of biological data across an entire assemblage: the well‐documented demersal marine fauna of the United Kingdom. We also test whether data availability for a species depends on its taxonomic group, maximum body size, the number of times it has been recorded in a global biogeographic database, or its commercial and conservation importance. Location Seas of the United Kingdom. Methods We defined a demersal marine fauna of 973 species from 15 phyla and 40 classes using five extensive surveys around the British Isles. We then quantified the availability of data on eight key biological traits (termed biological knowledge) for each species from online databases. Relationships between biological knowledge and our predictors were tested with generalized linear models. Results Full data on eight fundamental biological traits exist for only 9% (n= 88) of the UK demersal marine fauna, and 20% of species completely lack data. Clear trends in our knowledge exist: fish (median biological knowledge score = six traits) are much better known than invertebrates (one trait). Biological knowledge increases with biogeographic knowledge and (to a lesser extent) with body size, and is greater in species that are commercially exploited or of conservation concern. Main conclusions Our analysis reveals deep ignorance of the basic biology of a well‐studied fauna, highlighting the need for far greater efforts to compile biological trait data. Clear biases in our knowledge, relating to how well sampled or ‘important’ species are suggests that caution is required in extrapolating small subsets of biologically well‐known species to ecosystem‐level studies.     

Late Holocene ecological history of Pinus pinaster forests in the Sierra de Gredos of central Spain

This article describes the patterns and processes of vegetation change and fire history in the Late Holocene (c. 2400 calendar year BP) palaeoecological sequence of Lanzahíta, Sierra de Gredos in central Spain, and provides the first Iberian pollen sequence undertaken within a monospecific Pinus pinaster woodland. These new data reassess not only the autochthonous nature of this pine species in the region and the Iberian Peninsula, but also the naturalness of well-developed cluster pine forests. Conflicts of palaeoecological evidence with phytosociological models of vegetation dynamics in the study region, and the relationships of P. pinaster with fire occurrence in Mediterranean-type ecosystems, are discussed.