Impact of Placental Plasmodium falciparum Malaria on the Profile of Some Oxidative Stress Biomarkers in Women Living in Yaoundé, Cameroon

Background

Impact of the pathophysiology of Plasmodium falciparum placental malaria (PM) on the profile of some oxidative stress biomarkers and their relationship with poor pregnancy outcomes in women remain unknown.

Methods

Between 2013 and 2014, peripheral blood and placenta tissue from 120 Cameroonian women at delivery were assessed for maternal haemoglobin and, parasitaemia respectively. Parasite accumulation in the placenta was investigated histologically. The levels of oxidative stress biomarkers Malondialdehyde (MDA), Nitric Oxide (NO), Superoxide dismutase (SOD), Catalase (CAT) and Gluthatione (GSH) in the supernatant of teased placenta tissues were determined by Colorimetric enzymatic assays.

Results

Parasitaemia was inversely related to haemoglobin levels and birth weight (P <0.001 and 0.012, respectively). The level of lipid peroxide product (MDA) was significantly higher in the malaria infected (P = 0.0047) and anaemic (P = 0.024) women compared to their non-infected and non-anaemic counterparts, respectively. A similar trend was observed with SOD levels, though not significant. The levels of MDA also correlated positively with parasitaemia (P = 0.0024) but negatively with haemoglobin levels (P = 0.002). There was no association between parasitaemia, haemoglobin level and the other oxidative stress biomarkers. From histological studies, levels of MDA associated positively and significantly with placenta malaria infection and the presence of malaria pigments. The levels of SOD, NO and CAT increased with decreasing leukocyte accumulation in the intervillous space. Baby birth weight increased significantly with SOD and CAT levels, but decreased with levels of GSH.

Conclusions

Placental P. falciparum infection may cause oxidative stress of the placenta tissue with MDA as a potential biomarker of PM, which alongside GSH could lead to poor pregnancy outcomes (anaemia and low birth weight). This finding contributes to the understanding of the pathophysiology of P. falciparum placental malaria in women.     

Hypoglycin A Content in Blood and Urine Discriminates Horses with Atypical Myopathy from Clinically Normal Horses Grazing on the Same Pasture

Hypoglycin A (HGA) in seeds of Acer spp. is suspected to cause seasonal pasture myopathy in North America and equine atypical myopathy (AM) in Europe, fatal diseases in horses on pasture. In previous studies, this suspicion was substantiated by the correlation of seed HGA content with the concentrations of toxic metabolites in urine and serum (MCPA-conjugates) of affected horses. However, seed sampling was conducted after rather than during an outbreak of the disease. The aim of this study was to further confirm the causality between HGA occurrence and disease outbreak by seed sampling during an outbreak and the determination of i) HGA in seeds and of ii) HGA and MCPA-conjugates in urine and serum of diseased horses. Furthermore, cograzing healthy horses, which were present on AM affected pastures, were also investigated. AM-pastures in Germany were visited to identify seeds of Acer pseudoplatanus and serum (n = 8) as well as urine (n = 6) from a total of 16 diseased horses were analyzed for amino acid composition by LC-ESI-MS/MS, with a special focus on the content of HGA. Additionally, the content of its toxic metabolite was measured in its conjugated form in body fluids (UPLC-MS/MS). The seeds contained 1.7–319.8 μg HGA/g seed. The content of HGA in serum of affected horses ranged from 387.8–8493.8 μg/L (controls < 10 μg/L), and in urine from 143.8–926.4 μg/L (controls < 10 μg/L), respectively. Healthy cograzing horses on AM-pastures showed higher serum (108.8 ± 83.76 μg/L) and urine concentrations (26.9 ± 7.39 μg/L) compared to control horses, but lower concentrations compared to diseased horses. The range of MCPA-carnitine and creatinine concentrations found in diseased horses in serum and urine were 0.17–0.65 mmol/L (controls < 0.01), and 0.34–2.05 μmol/mmoL (controls < 0.001), respectively. MCPA-glycine levels in urine of cograzing horses were higher compared to controls. Thus, the causal link between HGA intoxication and disease outbreak could be further substantiated, and the early detection of HGA in cograzing horses, which are clinically normal, might be a promising step in prophylaxis.     

Lower foraging efficiency of offspring constrains use of optimal habitat in birds with extended parental care

After reproducing successfully, birds with extended parental care form family groups. Despite being the dominant social unit, such family groups have been reported to switch to alternative habitat earlier than adults without offspring, with potential negative carry‐over effects for the next breeding season. Here we test a proposed mechanism for this earlier habitat switch, namely a low foraging efficiency in juveniles. Such a test is best performed under controlled conditions because in the field families may occupy food patches of a different quality than singles or pairs without young. We studied this mechanism in Bewick's Swans Cygnus columbianus bewickii, which trample (or ‘treadle’) for food buried in the sediment. The gross intake rate of juveniles was as low as 60% of that of adults, depending on the burial depth of the food. Trampling effort did not differ between age classes, but differences in intake rate were related to body size, suggesting that larger or heavier birds were trampling more efficiently. Corresponding giving‐up densities in the field were calculated to be c. 60% higher for juveniles than for adults. Our findings are consistent with the hypothesis that the lower foraging efficiency of juveniles may be responsible for the segregation of family groups from adults without offspring.     

Seroepidemiological Survey of Paracoccidioidomycosis Infection Among Urban and Rural Dogs from Uberaba,Minas Gerais,Brazil

There is some evidence that dogs can be naturally infected by Paracoccidioides brasiliensis in endemic areas of paracoccidioidomycosis. In order to evaluate canine infection with this fungus, a survey with 149 urban and 126 rural dogs was carried out using ELISA and intradermal tests with the gp43 antigen of P. brasiliensis in Uberaba, Minas Gerais state of Brazil. Forty-one out of 149 urban dogs were euthanatized and had their lungs, liver and spleen removed. One slice from each viscera was processed for histopathological examination and the remaining was homogenized and then cultivated on mycobiotic agar at room temperature and Fava-Netto medium at 35°C and observed for 12 weeks. Of urban dogs, 75 (50.3%) were small adult females, 56 (36%) were strays, while 93 (64%) had been donated to the municipal zoonosis control center. Nine (6.2%) had a positive intradermal test without statistical differences regarding gender, race, nutritional status or origin. No colonies with microscopic or morphology appearances resembling P. brasiliensis were isolated, nor granulomatous process or fungal structures were observed from histopathological examination. Eighty (53.6%) of the urban dogs presented seroreactivity, without statistical differences regarding gender, race, nutritional state, origin, or positive intradermal test. Of 126 rural dogs, 102 (80.5%) presented antibodies against gp43 antigen, and this was statistically significant in relation to the reactivity detected in urban dogs (P = 0.0001). Thus, dogs are commonly infected with P. brasiliensis, but they probably present natural resistance to develop paracoccidioidomycosis.     

A multilocus approach to harvestman (Arachnida: Opiliones) phylogeny with emphasis on biogeography and the systematics of Laniatores

The internal phylogeny of the arachnid order Opiliones is investigated by including molecular data from five molecular markers for ca. 140 species totalling 43 families of Opiliones. The phylogenetic analyses consisted of a direct optimization (DO) approach using POY v. 4 and sophisticated tree search algorithms as well as a static alignment analysed under maximum likelihood. The four Opiliones suborders were well‐supported clades, but subordinal relationships did not receive support in the DO analysis, with the exception of the monophyly of Palpatores (=Eupnoi + Dyspnoi). Maximum‐likelihood analysis strongly supported the traditional relationship of Phalangida and Palpatores: (Cyphophthalmi ((Eupnoi + Dyspnoi) Laniatores)). Relationships within each suborder are well resolved and largely congruent between direct optimization and maximum‐likelihood approaches. Age estimates for the main Opiliones lineages suggest a Carboniferous diversification of Cyphophthalmi, while its sister group, Phalangida, diversified in the Early Devonian. Diversification of all suborders predates the Triassic, and most major lineages predate the Cretaceous. The following taxonomic changes are proposed. Dyspnoi: Hesperonemastoma is transferred to Sabaconidae. Insidiatores: Sclerobunidae stat. nov. is erected as a family for Zuma acuta. © The Willi Hennig Society 2009.     

Utilization of early soybeans for food and reproduction by the tarnished plant bug (Hemiptera: Miridae) in the delta of Mississippi

Commercially produced maturity group (MG) IV soybeans, Glycine max L., were sampled during bloom for tarnished plant bugs, Lygus lineolaris (Palisot de Beauvois), during May and June 1999 (3 fields) and 2001 (18 fields). The adults and nymphs were found primarily in single population peaks in both years, indicating a single new generation was produced during each year. The peak mean numbers of nymphs were 0.61 and 0.84 per drop cloth sample in 1999 and 2001, respectively. Adults peaked at 3.96 (1999) and 3.76 (2001) per sweep net sample (25 sweeps). Tests using laboratory-reared and field-collected tarnished plant bugs resulted in very poor survival of nymphs on 16 different soybean varieties (MG III, one; IV, four; V, nine; VI, two). A large cage (0.06 ha) field test found that the number of nymphs produced on eight soybean varieties after mated adults were released into the cages was lower than could be expected on a suitable host. These results indicated that soybean was a marginal host for tarnished plant bugs. However, the numbers of adults and nymphs found in the commercially produced fields sampled in the study may have been high enough to cause feeding damage to the flowering soybeans. The nature of the damage and its possible economic importance were not determined. Reproduction of tarnished plant bugs in the commercially produced early soybean fields showed that the early soybeans provided tarnished plant bugs with a very abundant host at a time when only wild hosts were previously available.     

Nitrogen metabolism and seed composition as influenced by foliar boron application in soybean

The physiological effects of foliar boron application (FB) on nitrogen metabolism and seed composition have not been well established in soybean [(Glycine max(L.)Merr.)]. Therefore, the effect of FB on nitrogen metabolism and seed composition was investigated. Nitrate assimilation was evaluated by measuring nitrate reductase activity (NRA) and nitrogen fixation was evaluated by measuring nitrogenase activity and natural abundance of ¹⁵N/¹⁴N. NRA were significantly (P?≤?0.05) higher in plants that received FB than the control plants. Higher rate of FB (One application of four times of commercial rate) inhibited nitrogen fixation as measured by natural abundance of ¹⁵N/¹⁴N ratio, but increased NRA. The higher activities of NR and nitrogenase by FB were accompanied with a higher B concentration in leaves. The significant (P?<?0.0001) enrichment of ¹⁵N/¹⁴N, accompanied with a higher rate of FB, suggested a possible mechanism where nitrate assimilation may compensate for the decrease in nitrogen fixation. FB increased seed protein by 13.7% and oleic acid by 30.9% compared to the control plants. This alteration was accompanied by a higher B concentration in leaves and seed. The results suggest that FB affects nitrogen metabolism and alters seed compositions, especially protein and unsaturated fatty acids.     

MODIFIED VACUOLE PHENOTYPE1 Is an Arabidopsis Myrosinase-Associated Protein Involved in Endomembrane Protein Trafficking

We identified an Arabidopsis (Arabidopsis thaliana) ethyl methanesulfonate mutant, modified vacuole phenotype1-1 (mvp1-1), in a fluorescent confocal microscopy screen for plants with mislocalization of a green fluorescent protein-δ tonoplast intrinsic protein fusion. The mvp1-1 mutant displayed static perinuclear aggregates of the reporter protein. mvp1 mutants also exhibited a number of vacuole-related phenotypes, as demonstrated by defects in growth, utilization of stored carbon, gravitropic response, salt sensitivity, and specific susceptibility to the fungal necrotroph Alternaria brassicicola. Similarly, crosses with other endomembrane marker fusions identified mislocalization to aggregate structures, indicating a general defect in protein trafficking. Map-based cloning showed that the mvp1-1 mutation altered a gene encoding a putative myrosinase-associated protein, and glutathione S-transferase pull-down assays demonstrated that MVP1 interacted specifically with the Arabidopsis myrosinase protein, THIOGLUCOSIDE GLUCOHYDROLASE2 (TGG2), but not TGG1. Moreover, the mvp1-1 mutant showed increased nitrile production during glucosinolate hydrolysis, suggesting that MVP1 may play a role in modulation of myrosinase activity. We propose that MVP1 is a myrosinase-associated protein that functions, in part, to correctly localize the myrosinase TGG2 and prevent inappropriate glucosinolate hydrolysis that could generate cytotoxic molecules.The plant endomembrane system is a complex network of subcellular compartments that includes the endoplasmic reticulum (ER), Golgi apparatus, vacuole, plasma membrane, secretory vesicles, and numerous intermediary compartments. Protein trafficking through the endomembrane system requires specific cargo recognition and delivery mechanisms that are mediated by a series of highly specific targeting signals (Surpin and Raikhel, 2004), whose proper recognition is critical for the function of numerous downstream processes, such as floral development (Sohn et al., 2007), gravitropism (Kato et al., 2002; Surpin et al., 2003; Yano et al., 2003), abiotic stress tolerance (Zhu et al., 2002), autophagy (Surpin et al., 2003; Bassham., 2007), pathogen defense (Robatzek, 2007), and turgor pressure and growth (De, 2000).The importance of protein trafficking for plant survival was demonstrated by the identification of the essential Arabidopsis (Arabidopsis thaliana) gene VACUOLELESS1 (VCL1; Rojo et al., 2001). VCL1 was identified as a homolog of Saccharomyces cerevisiae VPS16, which is critical for yeast vacuole biogenesis. Knockouts of yeast VPS16 lack discernible vacuoles but survive despite their severe phenotype. The absence of vacuoles in Arabidopsis vcl1-1 mutants results in embryo lethality (Rojo et al., 2001). The essential nature of trafficking in plants was also demonstrated by insertional mutagenesis of syntaxin genes, where lethality was observed after disruption of single genes in families with highly homologous members (Lukowitz et al., 1996; Sanderfoot et al., 2001). Thus, despite large families of endomembrane components with many homologous genes, many are not redundant in Arabidopsis.Although embryo-lethal mutations provide critical data, it is difficult to obtain additional information. Less severe mutations have proven successful for functional genetics studies of endomembrane trafficking proteins. For example, point mutations in the KATAMARI1/MURUS3 (KAM1/MUR3; Tamura et al., 2005) and KATAMARI2/GRAVITROPISM DEFECTIVE2 (KAM2/GRV2; Tamura et al., 2007; Silady et al., 2008) genes lead to disruption of endomembranes, resulting in the formation of perinuclear aggregates containing organelles. Nonlethal trafficking disruptions have also been generated using chemical genomics, where small molecules were used to perturb trafficking of a soluble cargo protein (Zouhar et al., 2004) and localization of endomembrane markers (Surpin et al., 2005; Robert et al., 2008). Such studies have provided valuable clues about these essential cellular processes.In order to obtain less severe, viable mutants with defects in endomembrane protein trafficking, we previously identified point mutants with defects in localization of a tonoplast reporter protein, GFP:δ-TIP (Avila et al., 2003). Two hundred one putative mutants were grouped into four categories based on the nature of their defects. One unique mutant, cell shape phenotype1, was recently characterized as a trehalose-6-phosphate synthase with roles in regulation of plant architecture, epidermal pavement cell shape, and trichome branching (Chary et al., 2008).Here, we describe an endomembrane trafficking mutant categorized by perinuclear aggregates of GFP:δ-TIP fluorescence (Avila et al., 2003). We refer to this mutant as modified vacuole phenotype1-1 (mvp1-1). At least five endomembrane fusion proteins are partially relocalized to these structures. Positional cloning identified MVP1 as a myrosinase-associated protein (MyAP) localized previously to the tonoplast by proteomics (Carter et al., 2004). mvp1-1 mutants showed reduced endomembrane system functionality, as demonstrated by defects in growth, utilization of stored carbon, gravitropic responsiveness, salt sensitivity, and increased susceptibility to a fungal necrotroph. MVP1 interacted specifically with THIOGLUCOSIDE GLUCOHYDROLASE2 (TGG2), a known myrosinase protein in Arabidopsis, and the mvp1-1 mutation had a significant effect on nitrile production during glucosinolate hydrolysis, suggesting a role in myrosinase function. Furthermore, MVP1 may function in quality control of glucosinolate hydrolysis by contributing to the proper tonoplast localization of TGG2.