Comparison of ELISA with shell vial cell culture method for the detection of human rotavirus in fecal specimens

The aim of the study was to compare an enzyme immunoassay method with shell vial cell culture method for detection of rotavirus in fecal specimens. In addition, the correlation between laboratory results and clinical scores of patients with gastroenteritis was evaluated. A total of 219 fecal specimens from children (ages 3 weeks to 5 years) with acute gastroenteritis submitted to pediatric emergency room were evaluated by both ELISA and shell vial cell culture. A Vesikari score was used for assessing the severity of the illness. Among 219 stool samples tested, 107 (48.9%) were determined to be positive. Two specimens were positive by shell vial cell culture method while they were ELISA negative. According to these results the calculated sensitivity, specificity, PPV, and NPV of ELISA were 98.1%, 100%, 100%, and 98.2%, respectively. The mean severity score for the 107 episodes of rotavirus diarrhoea was 11.0 +/- 3.6 compared to 4.5 +/- 1.9 for the 112 episodes of non-rotavirus diarrhea in the same population. Our study indicates that ELISA, which is easier to perform, faster and cheaper than cell culture methods may be suitable for routine diagnosis of rotavirus infections. The severity of rotavirus positive gastroenteritis was significantly higher than that of rotavirus negative patients.     

Comamonas testosteroni bacteremia in a patient with perforated acute appendicitis. Short communication

Comamonas testosteroni is an uncommon isolate in the clinical laboratory as a human pathogen. C. testosteroni most commonly emerges in abdominal pathologies especially in perforated appendicitis. In Turkey we report first time a case of bacteremia due to this organism, in a 22-year-old man with perforated acute appendicitis. The organism was shown to be susceptible to routine antibiotics so it was easily eliminated even after having caused a bacteremia.     

PHOTOSYSTEM II PROTEIN33, a Protein Conserved in the Plastid Lineage,Is Associated with the Chloroplast Thylakoid Membrane and Provides Stability to Photosystem II Supercomplexes in Arabidopsis

Photosystem II (PSII) is a multiprotein complex that catalyzes the light-driven water-splitting reactions of oxygenic photosynthesis. Light absorption by PSII leads to the production of excited states and reactive oxygen species that can cause damage to this complex. Here, we describe Arabidopsis (Arabidopsis thaliana) At1g71500, which encodes a previously uncharacterized protein that is a PSII auxiliary core protein and hence is named PHOTOSYSTEM II PROTEIN33 (PSB33). We present evidence that PSB33 functions in the maintenance of PSII-light-harvesting complex II (LHCII) supercomplex organization. PSB33 encodes a protein with a chloroplast transit peptide and one transmembrane segment. In silico analysis of PSB33 revealed a light-harvesting complex-binding motif within the transmembrane segment and a large surface-exposed head domain. Biochemical analysis of PSII complexes further indicates that PSB33 is an integral membrane protein located in the vicinity of LHCII and the PSII CP43 reaction center protein. Phenotypic characterization of mutants lacking PSB33 revealed reduced amounts of PSII-LHCII supercomplexes, very low state transition, and a lower capacity for nonphotochemical quenching, leading to increased photosensitivity in the mutant plants under light stress. Taken together, these results suggest a role for PSB33 in regulating and optimizing photosynthesis in response to changing light levels.PSII is a multiprotein complex in plants with 31 identified polypeptides (Wegener et al., 2011; Pagliano et al., 2013). It is associated with an extrinsic trimeric light-harvesting complex (LHC), forming the PSII-LHCII supercomplex. The PSII complex performs a remarkable biochemical reaction, the oxidation of water using light energy from the sun, which profoundly contributes to the overall biomass accumulation in the biosphere (Barber et al., 2004). Consequently, the stability and functional integrity of the PSII-LHCII supercomplex is crucially important for photosynthetic function. The energy of a photon, either absorbed directly by PSII or indirectly via energy transfer from adjacent antenna chlorophyll (Chl) molecules, excites the PSII reaction center P680. The excited state, P680*, can transfer an electron to pheophytin, producing the most powerful oxidant known in biology, P680⁺, which can remove electrons from water. Excessive input of excitation energy into PSII saturates the electron transfer system and causes either acceptor or donor site limitation in the complex. This results in increased production of reactive oxygen species (ROS): singlet oxygen at the PSII donor side and superoxide at the acceptor side (Munné-Bosch et al., 2013). Several protective mechanisms have been documented that decrease the production of singlet oxygen at the PSII donor side in photosynthetic eukaryotes. Notably, reducing energy transfer from LHC to PSII via nonphotochemical quenching (NPQ) is a key avoidance mechanism (Ruban and Murchie, 2012).Despite years of intensive study of PSII structure and function, new proteins that are associated with the PSII complex continue to be discovered, including an increasing number involved in the stability and organization of PSII-LHCII supercomplexes (García-Cerdán et al., 2011; Lu et al., 2011a; Wegener et al., 2011). Two complementary approaches (Merchant et al., 2007; Lu et al., 2008, 2011b; Ajjawi et al., 2010) that utilize phylogenomics (GreenCut) and large-scale phenotypic mutant screening (Chloroplast 2010 Project; http://www.plastid.msu.edu/) were employed by our groups to discover novel plant proteins with roles in photosynthesis. GreenCut identifies proteins found only in photosynthetic organisms, and it is likely that many of them are involved in biochemical processes associated with the structure, assembly, or function of the photosynthetic apparatus and the chloroplast that houses it (Merchant et al., 2007; Karpowicz et al., 2011). The Chloroplast 2010 Project was a large-scale reverse-genetic mutant screen in which thousands of homozygous Arabidopsis (Arabidopsis thaliana) transfer DNA (T-DNA) insertion lines were analyzed for defects in the rise and decay kinetics of Chl fluorescence (Lu et al., 2008, 2011a, 2011b; Ajjawi et al., 2010).The GreenCut and Chloroplast 2010 approaches both identified the Arabidopsis At1g71500 locus as encoding a protein of unknown function with potential relevance to photosynthesis. In this work, we demonstrate that plant lines carrying three independent mutations at this locus display severe light-induced photoinhibition due to a less stable supramolecular organization of PSII. Biochemical analyses revealed that this protein is associated with PSII complexes, and since the last described PSII protein was called PHOTOSYSTEM II PROTEIN32 (PSB32), we named the gene PSB33. The nuclear genome-encoded PSB33 is predicted to have a chloroplast transit peptide and a transmembrane domain. The biochemical analyses presented below indicate that PSB33 is required for the proper interaction and stability of PSII-LHCII supercomplexes and, in turn, in regulating photosynthesis in response to fluctuating light levels.     

Light acclimation involves dynamic re‐organization of the pigment–protein megacomplexes in non‐appressed thylakoid domains

Thylakoid energy metabolism is crucial for plant growth, development and acclimation. Non‐appressed thylakoids harbor several high molecular mass pigment–protein megacomplexes that have flexible compositions depending upon the environmental cues. This composition is important for dynamic energy balancing in photosystems (PS) I and II. We analysed the megacomplexes of Arabidopsis wild type (WT) plants and of several thylakoid regulatory mutants. The stn7 mutant, which is defective in phosphorylation of the light‐harvesting complex (LHC) II, possessed a megacomplex composition that was strikingly different from that of the WT. Of the nine megacomplexes in total for the non‐appressed thylakoids, the largest megacomplex in particular was less abundant in the stn7 mutant under standard growth conditions. This megacomplex contains both PSI and PSII and was recently shown to allow energy spillover between PSII and PSI (Nat. Commun., 6, 2015, 6675). The dynamics of the megacomplex composition was addressed by exposing plants to different light conditions prior to thylakoid isolation. The megacomplex pattern in the WT was highly dynamic. Under darkness or far red light it showed low levels of LHCII phosphorylation and resembled the stn7 pattern; under low light, which triggers LHCII phosphorylation, it resembled that of the tap38/pph1 phosphatase mutant. In contrast, solubilization of the entire thylakoid network with dodecyl maltoside, which efficiently solubilizes pigment–protein complexes from all thylakoid compartments, revealed that the pigment–protein composition remained stable despite the changing light conditions or mutations that affected LHCII (de)phosphorylation. We conclude that the composition of pigment–protein megacomplexes specifically in non‐appressed thylakoids undergoes redox‐dependent changes, thus facilitating maintenance of the excitation balance between the two photosystems upon changes in light conditions.     

The role of curcumin on intestinal oxidative stress,cell proliferation and apoptosis after ischemia/reperfusion injury in rats

The aim of this study was to demonstrate the role of curcumin on oxidative stress, cell proliferation and apoptosis in the rat intestinal mucosa after ischemia/reperfusion (I/R). A total of 30 male Wistar albino rats were divided into three groups: sham, I/R and I/R+ curcumin; each group contain 10 animals. Sham group animals underwent laparotomy without I/R injury. After I/R groups animals underwent laparotomy, 1 h of superior mesenteric artery ligation were followed by 1 h of reperfusion. In the curcumin group, 3 days before I/R, curcumin (100 mg/kg) was administered by gastric gavage. All animals were sacrificed at the end of reperfusion and intestinal tissues samples were obtained for biochemical and histopathological investigation in all groups. Curcumin treatment significantly decreased the elevated tissue malondialdehyde levels and increased of reduced superoxide dismutase, and glutathione peroxidase enzyme activities in intestinal tissues samples. I/R caused severe histopathological injury including mucosal erosions and villous congestion and hemorrhage. Curcumin treatment significantly attenuated the severity of intestinal I/R injury, with inhibiting of I/R-induced apoptosis and cell proliferation. These results suggest that curcumin treatment has a protective effect against intestinal damage induced by intestinal I/R. This protective effect is possibly due to its ability to inhibit I/R-induced oxidative stress, apoptosis and cell proliferation.     

Optimization of α-amylase production by Bacillus subtilis RSKK96: using the Taguchi experimental design approach

In this study, the Taguchi experimental design was applied to optimize the conditions for α-amylase production by Bacillus subtilis RSKK96, which was purchased from Refik Saydam Hifzissihha Industry (RSHM). Four factors, namely, carbon source, nitrogen source, amino acid, and fermentation time, each at four levels, were selected, and an orthogonal array layout of L(16) (4(5)) was performed. The model equation obtained was validated experimentally at maximum casein (1%), corn meal (1%), and glutamic acid (0.01%) concentrations with incubation time to 72 h in the presence of 1% inoculum density. Point prediction of the design showed that maximum α-amylase production of 503.26 U/mg was achieved under optimal experimental conditions.     

Effects of exogenous metallothionein on acute cadmium toxicity in rats

The present study was carried out to evaluate the effect of exogenously administered metallothionein (MT) to rats exposed to high cadmium levels. A total of 72 rats were used in the study. The animals were divided into three groups: controls, Cd administered, and Cd+MT. Cadmium was administered by subcutaneous injection of cadmium(II) chloride at a dose of 3.5 mg/kg for 7 d. In addition to CdCl₂, 30 μmol/kg MT was administered to the second group of rats (group II). Control rats received 0.5 mL physiologic serum via subcutaneous injection. Eight rats from each group were sacrificed on the 1st, 3rd, 5th, and 7th day after administration of the compounds. Liver, kidney, and blood samples were harvested. Levels of malondialdehyde (MDA), glutathione peroxidase (GSH-Px), serum ALT, AST, BUN, ALP, creatinine, and urea were measured. MDA levels in group I were observed to increase starting from d 1 compared to group II (p<0.05). Although MDA levels in group II were higher than controls (p<0.05), they were lower, especially in liver and blood, compared to group II. Erythrocyte GSH-Px activity levels were determined to decrease starting from d 1 in both groups (p<0.05). Decreases in GSH-Px activity levels in group II were less than group I. Serum creatinine levels in both groups were increased significantly compared to controls (p<0.05); the increase in group I was higher than group II. Serum ALT, AST, and ALP levels in group I increased to very high levels compared to controls, whereas increases in group II were at moderate levels (p<0.05). Although serum BUN levels were determined to be reduced, there was no significant change among the groups. Serum urea levels in both groups were higher than controls. Based on our results, it is possible to postulate that exogenous MT can act as antioxidant against Cd toxicity and lipid peroxidation.     

Synthesis of Two New Acetanilide Derivatives and Their Effect on the Serum Antioxidant Vitamins (A,E, and C) and the MDA Level in Rats

Acetanilide derivatives, 2,2′-thiobis[N-(4-nitrophenyl)acetamide] and 2,2′-thiobis[N-(4-chlorophenyl)acetamide], were synthesized and characterized. They were shown to cause a considerable oxidative stress in rats.