Serratia marcescens Suppresses Host Cellular Immunity via the Production of an Adhesion-inhibitory Factor against Immunosurveillance Cells

Injection of a culture supernatant of Serratia marcescens into the bloodstream of the silkworm Bombyx mori increased the number of freely circulating immunosurveillance cells (hemocytes). Using a bioassay with live silkworms, serralysin metalloprotease was purified from the culture supernatant and identified as the factor responsible for this activity. Serralysin inhibited the in vitro attachment of both silkworm hemocytes and murine peritoneal macrophages. Incubation of silkworm hemocytes or murine macrophages with serralysin resulted in degradation of the cellular immune factor BmSPH-1 or calreticulin, respectively. Furthermore, serralysin suppressed in vitro phagocytosis of bacteria by hemocytes and in vivo bacterial clearance in silkworms. Disruption of the ser gene in S. marcescens attenuated its host killing ability in silkworms and mice. These findings suggest that serralysin metalloprotease secreted by S. marcescens suppresses cellular immunity by decreasing the adhesive properties of immunosurveillance cells, thereby contributing to bacterial pathogenesis.     

Sialyltransferase ST3Gal IV deletion protects against temporal lobe epilepsy

Temporal lobe epilepsy (TLE) often becomes refractory, and patients with TLE show a high incidence of psychiatric symptoms, including anxiety and depression. Therefore, it is necessary to identify molecules that were previously unknown to contribute to epilepsy and its associated disorders. We previously found that the sialyltransferase ST3Gal IV is up‐regulated within the neural circuits through which amygdala‐kindling stimulation propagates epileptic seizures. In contrast, this study demonstrated that kindling stimulation failed to evoke epileptic seizures in ST3Gal IV‐deficient mice. Furthermore, approximately 80% of these mice failed to show tonic–clonic seizures with stimulation, whereas all littermate wild‐type mice showed tonic–clonic seizures. This indicates that the loss of ST3Gal IV does not cause TLE in mice. Meanwhile, ST3Gal IV‐deficient mice exhibited decreased acclimation in the open field test, increased immobility in the forced swim test, enhanced freezing during delay auditory fear conditioning, and sleep disturbances. Thus, the loss of ST3Gal IV modulates anxiety‐related behaviors. These findings indicate that ST3Gal IV is a key molecule in the mechanisms underlying anxiety – a side effect of TLE – and may therefore also be an effective target for treating epilepsy, acting through the same circuits.

     

Reduction of Gibberellin by Low Temperature Disrupts Pollen Development in Rice

Microsporogenesis in rice (Oryza sativa) plants is susceptible to moderate low temperature (LT; approximately 19°C) that disrupts pollen development and causes severe reductions in grain yields. Although considerable research has been invested in the study of cool-temperature injury, a full understanding of the molecular mechanism has not been achieved. Here, we show that endogenous levels of the bioactive gibberellins (GAs) GA₄ and GA₇, and expression levels of the GA biosynthesis genes GA20ox3 and GA3ox1, decrease in the developing anthers by exposure to LT. By contrast, the levels of precursor GA₁₂ were higher in response to LT. In addition, the expression of the dehydration-responsive element-binding protein DREB2B and SLENDER RICE1 (SLR1)/DELLA was up-regulated in response to LT. Mutants involved in GA biosynthetic and response pathways were hypersensitive to LT stress, including the semidwarf mutants sd1 and d35, the gain-of-function mutant slr1-d, and gibberellin insensitive dwarf1. The reduction in the number of sporogenous cells and the abnormal enlargement of tapetal cells occurred most severely in the GA-insensitive mutant. Application of exogenous GA significantly reversed the male sterility caused by LT, and simultaneous application of exogenous GA with sucrose substantially improved the extent of normal pollen development. Modern rice varieties carrying the sd1 mutation are widely cultivated, and the sd1 mutation is considered one of the greatest achievements of the Green Revolution. The protective strategy achieved by our work may help sustain steady yields of rice under global climate change.Grain yields in rice plants (Oryza sativa) are often reduced by exposure to moderate low temperature (LT), which is termed cool-temperature damage. It is estimated that the net effect of cool-temperature damage is an annual loss of at least three to five million tons of rice in East Asia (Li and Guo, 1993). Unexpected climate change, such as abnormally hot or cool summer temperatures, has occurred repeatedly during recent years due to the El Niño/La Niña-Southern Oscillation (Intergovernmental Panel on Climate Change, 2007). Shifts in population demographics result in the abandonment of agricultural fields in rapidly industrialized areas and the establishment of new fields in mountainous areas. Studies on cool-temperature damage in rice have a long history and have identified physiological responses to LT, including abnormal enlargement of anther wall cells and tapetal cells, reduction in the numbers of mature pollen, and increased male sterility (Sakai, 1943; Nishiyama, 1976, 1982). These studies show that microsporogenesis is the most susceptible stage to LT during pollen development in rice. However, the mechanisms underlying these physiological changes have not been completely elucidated.Research on plants during the last decade has identified numerous cellular pathways that respond to abiotic environmental stresses. Several phytohormones are involved in the regulation of homeostasis, stress responses, and cross talk in different signaling pathways (Qin et al., 2011). Abscisic acid is a typical phytohormone that responds to abiotic stress (Raghavendra et al., 2010; Qin et al., 2011). GA is generally regarded as a growth-promoting phytohormone that positively regulates processes such as seed germination, vegetative growth, flowering, and fruit development (Olszewski et al., 2002; Sun and Gubler, 2004). GA functions to mediate both tolerance and intolerance pathways involved in the responses to different abiotic stresses. Application of exogenous GA to plant seeds reverses the salt stress- and heat stress-induced inhibition of germination and seedling establishment (Kabar and Baltepe, 1990; Kaur et al., 1998; Nasri et al., 2011). By contrast, salt stress in Arabidopsis (Arabidopsis thaliana) seedlings leads to a decrease of GA levels, the accumulation of DELLA proteins, and the suppression of plant growth, which ultimately confers tolerance to stress (Achard et al., 2006). A quadruple DELLA mutant lacking Gibberellic Acid Insensitive (GAI), Repressor of GA (RGA), RGA-Like1 (RGL1), and RGL2 is more sensitive to salt stress than the wild-type plant. In barley (Hordeum vulgare) seedlings, treatment with GA inhibitors leads to higher tolerance to heat and oxidative stresses (Sarkar et al., 2004). Barley seedlings, which have the largest concentrations of endogenous GAs, are most susceptible to these abiotic stresses. Although stress tolerance response pathways mediated by GA are complex, it is well known that GA-deficient and GA-insensitive mutants in several plant species display abnormal anther development (Nester and Zeevaart, 1988; Jacobsen and Olszewski, 1991; Cheng et al., 2004; Aya et al., 2009). The typical defect of abnormal tapetal cell enlargement observed in rice GA mutants (Aya et al., 2009) is quite similar to the observed effects induced by LT injury in wild-type cultivated rice plants (Nishiyama, 1976; Oda et al., 2010).In this paper, we studied the relationship between GA and LT damage of anther development in rice. The endogenous GA levels and expression of genes involved in GA biosynthesis were measured in developing anthers with or without exposure to LT. The sensitivity of the GA biosynthesis mutant semidwarf1 (sd1; Asano et al., 2011) to LT damage was monitored. The sd1 mutant is considered one of the greatest achievements of the Green Revolution (Monna et al., 2002; Sasaki et al., 2002). In addition, we studied the response to LT in other GA biosynthesis mutants, including dwarf Tan-Ginbozu (d35; Suge, 1975; Itoh et al., 2004), the gain-of-function slender rice1 (slr1-d) mutant (Asano et al., 2009), and the GA-insensitive gibberellin insensitive dwarf1 (gid1) mutant (Ueguchi-Tanaka et al., 2007). We also explored potential remedial strategies for LT damage by examining the effects of GA application on pollen development under cool-temperature conditions.     

Adsorbents for Apheresis Prepared from Polysaccharides of Algae that Threaten Ecosystem Services

In this work, we investigated whether materials isolated from algae that threaten ecosystems can be used for human benefit. We converted acidic polysaccharides (ulvan) from the alga Ulva pertusa into soft hydrogel materials. In addition to ulvan, the hydrogels also contained alginate in a polyion complex with chitosan. Cross‐linking the hydrogel with glutaraldehyde reduced polysaccharide elution from the polyion complex gel. We also found that both ulvan? chitosan and alginate? chitosan gels were able to remove urea and heavy metals from aqueous solution. This is clinically significant, since during apheresis, toxic compounds such as urea have to be removed from the bloodstream of patients. Importantly, albumin was not removed by the hydrogels, implying that this vital protein can be returned to the bloodstream following dialysis.     

Increased superoxide radicals generation from alveolar macrophages in immature guinea-pigs

To study the effect of maturation on abilities of superoxide radicals (O-2) generation in the airways, we compared stimuli-induced O-2 generation by alveolar macrophages in immature (aged 10+/-2 days) and adult (aged 90+/-2 days) guinea-pigs. The production of O-2 was assayed by chemiluminescence method, using a Cypridina luciferin analog as a highly sensitive and specific probe for O-2. Whereas no significant difference in cell components of bronchoalveolar lavage fluid was observed between immature and adult animals, O-2 generation induced by stimulation of alveolar macrophages was greater in immature than in adult animals, with significant differences observed after platelet-activating factor (100 nM) or phorbol myristate acetate (0.5 micro g/ml). The results suggest that alveolar macrophages from immature animals are far more potent O-2 generators than the same cells of adult animals.     

ESR dosimetry study on population of settlements nearby Ust-Kamenogorsk city, Kazakhstan

The method of electron spin resonance (ESR) dosimetry has been applied to human tooth enamel, to obtain individual absorbed doses of residents of settlements in vicinity of Ust-Kamenogorsk city, Kazakhstan (located about 400 km to the east from the epicenter of explosion at the Semipalatinsk Nuclear Test Site, SNTS). This region developed as a major mining and metallurgical center during the Soviet period (uranium production). Most of the investigated settlements (Ust-Kamenogorsk city, Glubokoe, Tavriya, Gagarino) are located near the central axis of the radioactive fallout trace that originated from the surface nuclear test on 24 August 1956, while the Kokpekty settlement (located 400 km to the Southeast from SNTS) was chosen as a control because it was not subjected to any radioactive contamination. In total, 44 samples were measured. It was found that the excess doses obtained after subtraction of natural background radiation ranged up to about 114 mGy for residents of Ust-Kamenogorsk city, whose tooth enamel was formed before 1956. For residents of Gagarino, excess doses did not exceed 47 mGy for all ages. For residents of Tavriya, the maximum excess dose was 54 mGy, while for residents of Glubokoe it was about 58 mGy. For the population of the Shemonaikha settlements located at a distance of about 70 km from the central axis of the radioactive fallout trace, highest excess doses were 110 mGy. These high doses may be due to the influence of uranium enterprises located in that region, but probably not due to dental X-ray irradiation. For a final conclusion on the radiological situation in this region, the number of samples was too small and, therefore, more work is required to obtain representative results.     

Systematics of <Emphasis Type="Italic">Mycosphaerella</Emphasis> species associated with the invasive weed <Emphasis Type="Italic">Fallopia japonica</Emphasis>, including the potential biological control agent <Emphasis Type="Italic">M. polygoni-cuspidati</Emphasis>

Native to Japan, Fallopia japonica, most frequently referred to as Japanese knotweed, is a highly problematic invasive weed, particularly in the UK and North America. During surveys for natural enemies of this plant in Japan, two species of Mycosphaerella were collected. One of these was identified as M. polygoni-cuspidati, and is redescribed and neotypified. Causing a damaging leaf spot disease of F. japonica throughout its natural range in Japan, it is absent from the host’s exotic range. The restriction of M. polygoni-cuspidati to F. japonica in its center of origin, together with its severe impact on host fitness, indicates that this is a coevolved natural enemy with high potential as a classical biological control agent for the long-term management of this ecologically and economically important weed. In the field, the fungus has a reduced life cycle, with only spermogonia and pseudothecia (ascomata) being formed. Ascospores are the primary source of infection, and studies show that the mycelium from in vitro cultures is also infective and hyphae penetrate mainly via the stomata. A further, undescribed species of Mycosphaerella co-occurs with M. polygoni-cuspidati, here proposed as the new species M. shimabarensis. Both species have been studied using cultural, morphological and molecular phylogenetic methods.     

Activation regimens for full‐term development of rabbit oocytes injected with round spermatids

The present study was designed to investigate the effect of activation regimens on full‐term development of rabbit oocytes after round spermatid injection (ROSI). In the first series, rabbit oocytes were treated with 5 µM ionomycin before ROSI, after ROSI, or before and after ROSI. In addition, non‐treated oocytes were subjected to intracytoplasmic sperm injection (ICSI) using ejaculated spermatozoa. Cleavage rate of ROSI oocytes activated before and after ROSI (55%) was comparable with that of ICSI oocytes (60%), and significantly higher than those of ROSI oocytes activated either before or after ROSI (29–39%; P < 0.05). No offspring were produced by transfer of the cleaving ROSI oocytes, while 8% of the cleaving ICSI oocytes transferred gave birth to offspring. In the second series, oocytes were exposed to 5, 10, or 20 µM ionomycin, followed by ROSI, 5 µM ionomycin treatment, and incubation with 5 µg/ml cycloheximide (CHX) + 2 mM 6‐dimethylaminopurine (DMAP). Significantly higher cleavage rates were derived from oocytes activated with 10 and 20 µM ionomycin before ROSI (91% and 82%, respectively; P < 0.05) compared to those activated with 5 µM ionomycin before ROSI (53%). Live offspring were obtained when the cleaving ROSI oocytes with the initial ionomycin treatment at 5 and 10 µM were transferred (offspring rate 2% and 4%, respectively). These activation regimens, however, were not valid for the ROSI using cryopreserved round spermatids. In conclusion, rabbit ROSI oocytes were capable of developing into full‐term when the oocytes were activated with a combined treatment of ionomycin and CHX/DMAP. Mol. Reprod. Dev. 76: 573–579, 2009. © 2008 Wiley‐Liss, Inc.     

Suppression of dynamin GTPase activity by sertraline leads to inhibition of dynamin-dependent endocytosis

Dynamin (Dyn) 1 plays a role in recycling of synaptic vesicles, and thus in nervous system function. We previously showed that sertraline, a selective serotonin reuptake inhibitor (SSRI), is a mixed-type inhibitor of Dyn 1 with respect to both GTP and l-α-phosphatidyl-l-serine (PS) in vitro, and we suggested that it may regulate the neurotransmitter transport by modulating synaptic vesicle endocytosis via inhibition of Dyn 1 GTPase. Here, we investigated the effect of sertraline on endocytosis of marker proteins in human neuroblastoma SH-Sy5Y cells and HeLa cells. Sertraline inhibited endocytosis in both cell lines. Western blotting showed that SH-Sy5Y expresses Dyn 1 and Dyn 2, while HeLa expresses only Dyn 2. GTPase assay showed that sertraline inhibited Dyn 2 as well as Dyn 1. Therefore, the effect of sertraline on endocytosis was mediated by Dyn 2, at least in HeLa cells, as well as by Dyn 1 in cell lines that express it. Moreover, the inhibition mechanism of transferrin (Tf) uptake by sertraline differed from that in cells expressing Dyn 1 K44A, a GTP binding-defective variant, and sertraline did not interfere with the interaction between Dyn 1 and PS-liposomes.