Mitochondria mediate septin cage assembly to promote autophagy of Shigella

Septins, cytoskeletal proteins with well‐characterised roles in cytokinesis, form cage‐like structures around cytosolic Shigella flexneri and promote their targeting to autophagosomes. However, the processes underlying septin cage assembly, and whether they influence S. flexneri proliferation, remain to be established. Using single‐cell analysis, we show that the septin cages inhibit S. flexneri proliferation. To study mechanisms of septin cage assembly, we used proteomics and found mitochondrial proteins associate with septins in S. flexneri‐infected cells. Strikingly, mitochondria associated with S. flexneri promote septin assembly into cages that entrap bacteria for autophagy. We demonstrate that the cytosolic GTPase dynamin‐related protein 1 (Drp1) interacts with septins to enhance mitochondrial fission. To avoid autophagy, actin‐polymerising Shigella fragment mitochondria to escape from septin caging. Our results demonstrate a role for mitochondria in anti‐Shigella autophagy and uncover a fundamental link between septin assembly and mitochondria.     

Anatomical changes in stem and root of soybean plants submitted to salt stress

• The soybean is a legume that is widely cultivated in many countries due to the high levels of protein and oil contained in its seed, and is used for human and animal nutrition. However, salinity affects more than 800 million hectares worldwide, limiting global agricultural production.
• The aim of this research was to evaluate the structural behaviour of the roots and stems under progressive salt stress, detailing the possible anatomical modifications to these organs in soybean plants during this stress. The plants were randomized into five treatments (0, 50, 100, 150 and 200 mm NaCl).
• All the root regions studied and exposed to 100 mm Na⁺ exhibited increases in the epidermis and endodermis and formation of lysogenic aerenchyma with increasing salinity, revealing the protective roles of these structures in reducing Na⁺ influx. In the stem, increases in the cortex and pith in the first internode subject to 100 mm Na⁺ suggest anatomical responses that aim to minimize oxidative stress.
• Soybean plants subjected to progressive salt stress (>50 mm Na⁺) avoided cavitation and loss of function linked to vessel elements, reducing the metaxylem in all the root and stem regions analysed. Finally, our results confirm anatomical changes to the roots and stems.

     

Maternal clutch reduction in the pied flycatcher Ficedula hypoleuca: an undescribed clutch size adjustment mechanism

During eight years of study of a population of pied flycatchers Ficedula hypoleuca breeding in central Spain, we observed throughout incubation 22 cases of intact eggs being placed on the nest rim and clearly outside of the nest bowl and remaining there, with consequent embryo mortality. We assume that the removal of eggs had been performed by the incubating female. There is no evidence that pied flycatcher parents can detect embryo mortality and eject unhatchable eggs. Maternal egg ejection constitutes a direct mechanism of clutch size adjustment during incubation and before hatching of any young of the brood. This adjustment of clutch size happened when the mean and minimum temperature of the day before ejection were especially low. Also, clutch-reducing females were significantly more prone to show moult-breeding overlap than other females, suggesting a lower disposition to invest in reproduction. Clutch-reducing females were also frequently older than four years and had indications of a lower immunocompetence than females not ejecting eggs. Our results support the hypothesis that adverse conditions can elicit parental family size adjustment at every stage of reproduction without the need to depend on mechanisms of sibling competition.     

Study of the acclimation stage and of the effect of the biodegradability on the performance of a microbial fuel cell

In this work, it has been studied the production of electricity and the oxidation of the pollutants contained in a synthetic wastewater fed with glucose and peptone of soybean as carbon sources, using a mediator-less microbial fuel cell (MFC). Special attention has been paid to the acclimation stage, in which it was found that with high hydraulic and solid retention times it is possible to obtain a very efficient process with a 90% COD removal and practically total conversion of COD into electricity (considering the typical stoichiometric yield of heterotrophic biomass). The influence of concentration sludge was studied working with three different amounts of suspended solids, from 120 to 14000 mg. The maximum power density increased exponentially with the concentration sludge from 2.1 mW m⁻² to 11 mW m⁻² at the highest concentration sludge. More over, the percentage of the influent COD used to produce electricity was higher than 100% when the highest sludge concentration was used. This was explained taking into account the endogenous metabolism of micro-organisms presented in the system.     

Mycobacterium leprae induces insulin-like growth factor and promotes survival of Schwann cells upon serum withdrawal

Peripheral nerve lesions are considered the most relevant symptoms of leprosy, a chronic infectious disease caused by Mycobacterium leprae . The strategies employed by M. leprae to infect and multiply inside Schwann cells (SCs), however, remain poorly understood. In this study, it is shown that treatment of SCs with M. leprae significantly decreased cell death induced by serum deprivation. Not displayed by Mycobacterium smegmatis or Mycobacterium bovis BCG , the M. leprae survival effect was both dose dependent and specific . The conditioned medium (CM) of M. leprae -treated cultures was seen to mimic the protective effect of the bacteria, suggesting that soluble factors secreted by SCs in response to M. leprae were involved in cell survival. Indeed, by quantitative RT-PCR and dot blot/ELISA, it was demonstrated that M. leprae induced the expression and secretion of the SC survival factor insulin-like growth factor-I. Finally, the involvement of this hormone in M. leprae -induced SC survival was confirmed in experiments with neutralizing antibodies. Taken together, the results of this study delineate an important strategy for the successful colonization of M. leprae in the nerve based on the survival maintenance of the host cell through induction of IGF-I production.     

Ex vivo expansion of human mesenchymal stem cells: A more effective cell proliferation kinetics and metabolism under hypoxia

The low bone marrow (BM) MSC titers demand a fast ex vivo expansion process to meet the clinically relevant cell dosage. Attending to the low oxygen tension of BM in vivo, we studied the influence of hypoxia on human BM MSC proliferation kinetics and metabolism. Human BM MSC cultured under 2% (hypoxia) and 20% O₂ (normoxia) were characterized in terms of proliferation, cell division kinetics and metabolic patterns. BM MSC cultures under hypoxia displayed an early start of the exponential growth phase, and cell numbers obtained at each time point throughout culture were consistently higher under low O₂, resulting in a higher fold increase after 12 days under hypoxia (40 ± 10 vs. 30 ± 6). Cell labeling with PKH26 allowed us to determine that after 2 days of culture, a significant higher cell number was already actively dividing under 2% compared to 20% O₂ and BM MSC expanded under low oxygen tension displayed consistently higher percentages of cells in the latest generations (generations 4–6) until the 5th day of culture. Cells under low O₂ presented higher specific consumption of nutrients, especially early in culture, but with lower specific production of inhibitory metabolites. Moreover, 2% O₂ favored CFU‐F expansion, while maintaining BM MSC characteristic immunophenotype and differentiative potential. Our results demonstrated a more efficient BM MSC expansion at 2% O₂, compared to normoxic conditions, associated to an earlier start of cellular division and supported by an increase in cellular metabolism efficiency towards the maximization of cell yield for application in clinical settings. J. Cell. Physiol. 223: 27–35, 2010. © 2009 Wiley‐Liss, Inc.     

Transcriptional response to alcohol exposure in Drosophila melanogaster

Background  

Alcoholism presents widespread social and human health problems. Alcohol sensitivity, the development of tolerance to alcohol and susceptibility to addiction vary in the population. Genetic factors that predispose to alcoholism remain largely unknown due to extensive genetic and environmental variation in human populations. Drosophila, however, allows studies on genetically identical individuals in controlled environments. Although addiction to alcohol has not been demonstrated in Drosophila, flies show responses to alcohol exposure that resemble human intoxication, including hyperactivity, loss of postural control, sedation, and exposure-dependent development of tolerance.     

De novo production of diverse intracellular antibody libraries

Many therapeutic targets are intracellular proteins and molecules designed to interact with them must effectively bind to their target inside the cell. Intracellular antibodies (intrabodies) recognise and bind to proteins in cells and various methods have been developed to produce such molecules. Intracellular antibody capture (IAC) is based on a genetic screening approach and is a facile methodology with which effective intracellular antibodies can be obtained. During the development of the IAC technology, consensus immunoglobulin variable frameworks were identified which can form the basis of intrabody libraries for direct screening. In this paper, we describe the de novo synthesis of intrabody libraries based on the IAC consensus sequence. The procedure comprises in vitro production of a single antibody gene fragment from oligonucleotides and diversification of CDRs of the immunoglobulin variable domain by mutagenic PCR. Completely de novo intrabody libraries can be rapidly generated in vitro by these approaches. As an example, a single immunoglobulin VH domain intrabody library was screened directly in yeast with an oncogenic BCR-ABL antigen bait and distinct antigen binders were isolated illustrating the functional utility of the library. This second generation IAC approach (IAC²) has many practical advantages, in particular the ability to isolate intrabodies by direct genetic selection, which obviates the need for in vitro production of antigen for pre-selection of antibody fragments.     

A bird's eye survey of Central American planorbid molluscs

In the course of two trips to Central America (June 1967 and JulyAugust 1976) I had the opportunity of collecting topotypic specimens of Planorbis nicaraguanus Morelet, 1849, anatomically defined in this paper, and of P. yzabalensis Crosse & Fischer, 1879, the identity of the latter with Drepanotrema anatinum (Orbigny, 1835) is confirmed. The following planorbid species were also found: Helisoma trivolvis (Say, 1817) in Nicaragua, Guatemala, Costa Rica and Belize; H. duryi (Wetherby, 1879) in Costa Rica; Biomphalaria helophila (Orbigny, 1835) in Guatemala, Belize, Nicaragua, Costa Rica and El Salvador; B. kuhniana (Clessin, 1883) in Panama; B. obstructa (Morelet,1849) in Guatemala, Belize and El Salvador; B. straminea (Dunker, 1848) in Costa Rica; B. subprona (Martens, 1899) in Guatemala; D. anatinum (Orbigny,1835) in Belize, Guatemala, Nicaragua and Costa Rica; D. depressissimum (Moricand,1839) in Nicaragua, Costa Rica and Panama; D. lucidum (Pfeiffer, 1839) in Guatemala, Belize and Nicaragua; D. surinamense (Clessin, 1884) in Costa Rica and Panama; and Gyraulus percarinatus sp. n. in Panama. The occurrence of B. kuhniana and D. surinamense is first recorded in Central America, and Gyraulus percarinatus is the first representative of the genus provenly occurring in the American continent south of the United States. The following synonymy is proposed: Planorbis declivis Tate, 1870 = Biomphalaria helophila (Orbigny, 1835); Planorbis isthmicus Pilsbry, 1920 = Biomphalaria kuhniana (Clessin, 1883); Planorbis cannarum Morelet, 1849 and Segmentina donbilli Tristram, 1861 = Biomphalaria obstructa (Morelet, 1849); and Planorbis yzabalensis Crosse & Fischer, 1879 = Drepanotrema anatinum (Orbigny, 1835), confirming Aguayo (1933).