Plant peroxiredoxins: catalytic mechanisms, functional significance and future perspectives

Peroxiredoxins (Prx) are a family of thiol dependent peroxidases found in almost all kingdoms. In plants, five major classes of Prx are known. They are known to catalyze the decomposition of peroxides and as they lack a prosthetic group, the catalytic cycle results in the generation of an inactive form of Prx. In order to regain the active form, Prx rely on external electron donors such as thioredoxins, glutaredoxins, cyclophilins, NADPH-dependent thioredoxin reductase C (NTRC) etc. In addition to their well established role in antioxidative defense, Prx are also reported to play an important role in growth and development, dessication tolerance in dormant seeds, protection of photosynthesis, defense against pathogens and redox signaling. Prx are also known to establish an alternate water–water cycle for the detoxification of H₂O₂, parallel to ascorbate-dependent H₂O₂ detoxification. But the relative contribution of Prx in detoxifying H₂O₂ compared to ascorbate peroxidase is not known so far due to experimental limitations. In view of the above, the present review focuses on the recent developments on Prxs.     

NAD(P)H: quinone oxidoreductase 1 deficiency conjoint with marginal vitamin C deficiency causes cigarette smoke induced myelodysplastic syndromes

Background

The etiology of myelodysplastic syndromes (MDS) is largely unknown. Exposure to cigarette smoke (CS) is reported to be associated with MDS risk. There is inconsistent evidence that deficiency of NAD(P)H-quinone: oxidoreductase 1 (NQO1) increases the risk of MDS. Earlier we had shown that CS induces toxicity only in marginal vitamin C-deficient guinea pigs but not in vitamin C-sufficient ones. We therefore considered that NQO1 deficiency along with marginal vitamin C deficiency might produce MDS in CS-exposed guinea pigs.

Methodology and Principal Findings

Here we show that CS exposure for 21 days produces MDS in guinea pigs having deficiency of NQO1 (fed 3 mg dicoumarol/day) conjoint with marginal vitamin C deficiency (fed 0.5 mg vitamin C/day). As evidenced by morphology, histology and cytogenetics, MDS produced in the guinea pigs falls in the category of refractory cytopenia with unilineage dysplasia (RCUD): refractory anemia; refractory thrombocytopenia that is associated with ring sideroblasts, micromegakaryocytes, myeloid hyperplasia and aneuploidy. MDS is accompanied by increased CD34(+) cells and oxidative stress as shown by the formation of protein carbonyls and 8-oxodeoxyguanosine. Apoptosis precedes MDS but disappears later with marked decrease in the p53 protein. MDS produced in the guinea pigs are irreversible. MDS and all the aforesaid pathophysiological events do not occur in vitamin C-sufficient guinea pigs. However, after the onset of MDS vitamin C becomes ineffective.

Conclusions and Significance

CS exposure causes MDS in guinea pigs having deficiency of NQO1 conjoint with marginal vitamin C deficiency. The syndromes are not produced in singular deficiency of NQO1 or marginal vitamin C deficiency. Our results suggest that human smokers having NQO1 deficiency combined with marginal vitamin C deficiency are likely to be at high risk for developing MDS and that intake of a moderately large dose of vitamin C would prevent MDS.     

Manipulating cellulose biosynthesis by expression of mutant Arabidopsis proM24::CESA3ixr1‐2 gene in transgenic tobacco

Manipulation of the cellulose biosynthetic machinery in plants has the potential to provide insight into plant growth, morphogenesis and to create modified cellulose for anthropogenic use. Evidence exists that cellulose microfibril structure and its recalcitrance to enzymatic digestion can ameliorated via mis‐sense mutation in the primary cell wall–specific gene AtCELLULOSE SYNTHASE (CESA)3. This mis‐sense mutation has been identified based on conferring drug resistance to the cellulose inhibitory herbicide isoxaben. To examine whether it would be possible to introduce mutant CESA alleles via a transgenic approach, we overexpressed a modified version of CESA3, AtCESA3^ixr1‐2 derived from Arabidopsis thaliana L. Heynh into a different plant family, the Solanceae dicotyledon tobacco (Nicotiana tabacum L. variety Samsun NN). Specifically, a chimeric gene construct of CESA3^ixr1‐2, codon optimized for tobacco, was placed between the heterologous M24 promoter and the rbcSE9 gene terminator. The results demonstrated that the tobacco plants expressing M24‐CESA3^ixr1‐2 displayed isoxaben resistance, consistent with functionality of the mutated AtCESA3^ixr1‐2 in tobacco. Secondly, during enzymatic saccharification, transgenic leaf‐ and stem‐derived cellulose is 54%–66% and 40%–51% more efficient, respectively, compared to the wild type, illustrating translational potential of modified CESA loci. Moreover, the introduction of M24‐AtCESA3^ixr1‐2 caused aberrant spatial distribution of lignified secondary cell wall tissue and a reduction in the zone occupied by parenchyma cells.     

Aberrant Expression of Dynein light chain 1 (DYNLT1) is Associated with Human Male Factor Infertility*

DYNLT1 is a member of a gene family identified within the t-complex of the mouse, which has been linked with male germ cell development and function in the mouse and the fly. Though defects in the expression of this gene are associated with male sterility in both these models, there has been no study examining its association with spermatogenic defects in human males. In this study, we evaluated the levels of DYNLT1 and its expression product in the germ cells of fertile human males and males suffering from spermatogenic defects. We screened fertile (n = 14), asthenozoospermic (n = 15), oligozoospermic (n = 20) and teratozoospermic (n = 23) males using PCR and Western blot analysis. Semiquantitative PCR indicated either undetectable or significantly lower levels of expression of DYNLT1 in the germ cells from several patients from across the three infertility syndrome groups, when compared with that of fertile controls. DYNLT1 was localized on head, mid-piece, and tail segments of spermatozoa from fertile males. Spermatozoa from infertile males presented either a total absence of DYNLT1 or its absence in the tail region. Majority of the infertile individuals showed negligible levels of localization of DYNLT1 on the spermatozoa. Overexpression of DYNLT1 in GC1-spg cell line resulted in the up-regulation of several cytoskeletal proteins and molecular chaperones involved in cell cycle regulation. Defective expression of DYNLT1 was associated with male factor infertility syndromes in our study population. Proteome level changes in GC1-spg cells overexpressing DYNLT1 were suggestive of its possible function in germ cell development. We have discussed the implications of these observations in the light of the known functions of DYNLT1, which included protein trafficking, membrane vesiculation, cell cycle regulation, and stem cell differentiation.The t-complex of the mouse occupies the proximal half of chromosome 17 and contains genes which have profound effects on spermatogenesis. Multiple mutations in several loci in the t-complex appear to interact to cause complete male sterility (1, 2). Tctex-1 (t-complex testis expressed-1), lately renamed as dynein light chain 1 (Dynlt1)¹, is identified as a candidate gene involved in male sterility in mice (1) and maps to the t-complex in mice (3). Dynlt1 is a member of a multigene family which is virtually germ cell-specific and is eightfold over expressed in t-homozygotes and 200-fold higher in testis than in other adult tissues (1). The human homologue of the mouse Dynlt1 is located on chromosome 6q25.2–25.3. The amino acid sequence shows a high degree of similarity to the predicted product of the Dynlt1 gene of the mouse t complex (4).DYNLT1 gene encodes a 14 kDa protein constituting the inner arm L1 of cytoplasmic and flagellar dynein complexes (5, 6). DYNLT1 is localized to Golgi complexes as well (7). DYNLT1 protein is present in sperm tails and oocytes (8, 9). A wide range of cellular events are brought about by cytoplasmic dynein and its association with the accessory intermediate, light intermediate, and light chain subunits. These subunits define the interaction of cytoplasmic dynein motor complex with other molecules (10). DYNLT1 is involved in cargo binding (11), lymphocyte division (8), vesicle transport (12–14), and human embryo implantation (15). DYNLT1 is known to undergo phosphorylation during apical delivery of rhodopsin (16) and during its interaction with the bone morphogenetic receptor type II (BMPRII) (17). DYNLT1 can function in dynein-independent fashion as a cell fate regulator by its interaction with G-protein β γ subunit regulating initial neurite sprouting (18), axonal specification, and elongation of hippocampal neurons in culture (11, 19). GEF-H1 is bound to microtubules by DYNLT1 and its release without microtubule depolymerization is mediated through the interaction of DYNLT1 with G proteins (20). DYNLT1 is a novel marker for neural progenitors in adult brain (21). DYNLT1 regulatory element was identified which selectively marked nestin⁺/GFAP⁺/Sox2⁺ neural stem-like cells in developing and adult brain (22). The genetic knockdown of DYNLT1 in radial precursors promoted neurogenesis (23). The use of GFP placed under the control of DYNLT1 promoter to mark adult neural stem cells and thus allowing the insertion of any nucleotide sequence selectively into neural progenitors has been patented (24).DYNLT1 is reported to have functional roles in non-murine germ cells as well. DYNLT1 was found to be essential during spermatid differentiation in Drosophila (10) and a mouse DYNLT1 homolog was identified in the dynein light chain of sea urchin sperm flagella (25, 26). However, the expression of DYNLT1 in human testicular germ cells and its association, if any, with human male factor subfertility are not yet evaluated. This study evaluates the association between DYNLT1expression and spermatogenesis in infertile human males and the possible function of DYNLT1 in spermatogonial cell division and differentiation.     

Effect of Antenatal Parasitic Infections on Anti-vaccine IgG Levels in Children: A Prospective Birth Cohort Study in Kenya

Background

Parasitic infections are prevalent among pregnant women in sub-Saharan Africa. We investigated whether prenatal exposure to malaria and/or helminths affects the pattern of infant immune responses to standard vaccinations against Haemophilus influenzae (Hib), diphtheria (DT), hepatitis B (Hep B) and tetanus toxoid (TT).

Methods and Findings

450 Kenyan women were tested for malaria, schistosomiasis, lymphatic filariasis (LF), and intestinal helminths during pregnancy. After three standard vaccinations at 6, 10 and 14 weeks, their newborns were followed biannually to age 36 months and tested for absolute levels of IgG against Hib, DT, Hep B, and TT at each time point. Newborns’ cord blood (CB) lymphocyte responses to malaria blood-stage antigens, soluble Schistosoma haematobium worm antigen (SWAP), and filaria antigen (BMA) were also assessed. Three immunophenotype categories were compared: i) tolerant (those having Plasmodium-, Schistosoma-, or Wuchereria-infected mothers but lacking respective Th1/Th2-type recall responses at birth to malaria antigens, SWAP, or BMA); ii) sensitized (those with infected/uninfected mothers and detectable Th1/Th2-type CB recall response to respective parasite antigen); or iii) unexposed (no evidence of maternal infection or CB recall response).Overall, 78.9% of mothers were infected with LF (44.7%), schistosomiasis (32.4%), malaria (27.6%) or hookworm (33.8%). Antenatal maternal malaria, LF, and hookworm were independently associated with significantly lower Hib-specific IgG. Presence of multiple maternal infections was associated with lower infant IgG levels against Hib and DT antigens post-vaccination. Post-vaccination IgG levels were also significantly associated with immunophenotype: malaria-tolerized infants had reduced response to DT, whereas filaria-tolerized infants showed reduced response to Hib.

Conclusions

There is an impaired ability to develop IgG antibody responses to key protective antigens of Hib and diphtheria in infants of mothers infected with malaria and/or helminths during pregnancy. These findings highlight the importance of control and prevention of parasitic infections among pregnant women.     

Mesenchymal stem cell-conditioned media: A novel alternative of stem cell therapy for quality wound healing

Mesenchymal stem cells-conditioned media (MSCs-CM) contains several growth factors and cytokines, thus may be used as a better alternative to stem cell therapy, which needs to be elucidated. The present study was conducted to evaluate the therapeutic potential of caprine, canine, and guinea pig bone marrow-derived MSCs-CM in excision wound healing in a guinea pig model. MSCs were obtained from bone marrow, expanded ex vivo and characterized as per ISCT criteria. CM was collected assayed by western blot to ascertain the presence of important secretory biomolecules. Quantitative estimation by enzyme-linked immunosorbent assay was done for a vascular epidermal growth factor (VEGF) and interleukin-6 (IL-6) in caprine MSCs-CM and optimum time for collection of CM was decided as 72 hr. CM from all the species was lyophilized by freeze-drying method. Full-thickness (2 × 2 cm²) excision skin wounds were created in guinea pigs (six animals in each group) and respective lyophilized CM mixed with laminin gel was applied topically at weekly interval. On Day 28, histopathological examinations of healed skin were done by hemotoxylin and eosin staining. MSCs were found to secrete important growth factors and cytokines (i.e., VEGF, transforming growth factor-β1, fibroblast growth factor-2, insulin-like growth factor-1, stem cell factor, and IL-6) as demonstrated by immunohistochemistry and western blot assay. It was found that allogenic and xenogenic application of CM significantly improved quality wound healing with minimal scar formation. Thus, MSCs-CM can be used allogenically as well as xenogenically for quality wound healing.     

Colour removal of anaerobically treated pulp and paper mill effluent by microorganisms in two steps bioreactor

In the present study sequential anaerobic and aerobic treatment in two steps bioreactor was performed for removal of colour in the pulp and paper mill effluent. In anaerobic treatment, colour (70%), lignin (25%), COD (42%), AOX (15%) and phenol (39%) were reduced in 15 days. The anaerobically treated effluent was separately applied in bioreactor in presence of fungal strain, Paecilomyces sp., and bacterial strain, Microbrevis luteum. Data of study indicated reduction in colour (95%), AOX (67%), lignin (86%), COD (88%) and phenol (63%) by Paecilomyces sp. where as M. luteum showed removal in colour (76%), lignin (69%), COD (75%) AOX (82%) and phenol (93%) by day third when 7 days anaerobically treated effluent was further treated by aerobic microorganisms. Change in pH of the effluent, and increase in biomass of microorganisms substantiated results of the study, which was concomitant to the treatment method.     

Multiresolution wavelet analysis of time-dependent physiological responses in syncopal youths

Our prior studies indicated that postural fainting relates to thoracic hypovolemia. A supranormal increase in initial vascular resistance was sustained by increased peripheral resistance until late during head-up tilt (HUT), whereas splanchnic resistance, cardiac output, and blood pressure (BP) decreased throughout HUT. Our aim in the present study was to investigate the alterations of baroreflex activity that occur in synchrony with the beat-to-beat time-dependent changes in heart rate (HR), BP, and total peripheral resistance (TPR). We proposed that changes of low-frequency Mayer waves reflect sympathetic baroreflex. We used DWT multiresolution analyses to measure their time dependence. We studied 22 patients, 13 to 21 yr old, 14 who fainted within 10 min of upright tilt (fainters) and 8 healthy control subjects. Multiresolution analysis was obtained of continuous BP, HR, and respirations as a function of time during 70 degrees upright tilt at different scales corresponding to frequency bands. Wavelet power was concentrated in scales corresponding to 0.125 and 0.25 Hz. A major difference from control subjects was observed in fainters at the 0.125 Hz AP scale, which progressively decreased from early HUT. The alpha index at 0.125 Hz was increased in fainters. RR interval 0.25 Hz power decreased in fainters and controls but was markedly increased in fainters with syncope and thereafter corresponding to increased vagal tone compared with control subjects at those times only. The data imply a rapid reduction in time-dependent sympathetic baroreflex activity in fainters but not control subjects during HUT.