Association mapping for protein, total soluble sugars, starch, amylose and chlorophyll content in rice.

BACKGROUND: Protein, starch, amylose and total soluble sugars are basic metabolites of seed that influence the eating, cooking and nutritional qualities of rice. Chlorophyll is responsible for the absorption and utilization of the light energy influencing photosynthetic efficiency in rice plant. Mapping of these traits are very important for detection of more number of robust markers for improvement of these traits through molecular breeding approaches. RESULTS: A representative panel population was developed by including 120 germplasm lines from the initial shortlisted 274 lines for mapping of the six biochemical traits using 136 microsatellite markers through association mapping. A wide genetic variation was detected for the traits, total protein, starch, amylose, total soluble sugars, chlorophyll a, and chlorophyll b content in the population. Specific allele frequency, gene diversity, informative markers and other diversity parameters obtained from the population indicated the effectiveness of utilization of the population and markers for mapping of these traits. The fixation indices values estimated from the population indicated the existence of linkage disequilibrium for the six traits. The population genetic structure at K = 3 showed correspondence with majority of the members in each group for the six traits. The reported QTL, qProt1, qPC6.2, and qPC8.2 for protein content; qTSS8.1 for total soluble sugar; qAC1.2 for amylose content; qCH2 and qSLCHH for chlorophyll a (Chl. a) while qChl5D for chlorophyll b (Chl. b) were validated in this population. The QTL controlling total protein content qPC1.2; qTSS7.1, qTSS8.2 and qTSS12.1 for total soluble sugars; qSC2.1, qSC2.2, qSC6.1 and qSC11.1 for starch content; qAC11.1, qAC11.2 and qAC11.3 for amylose content; qChla8.1 for Chl. a content and qChlb7.1 and qChlb8.1 for Chl. b identified by both Generalized Linear Model and Mixed Linear Model were detected as novel QTL. The chromosomal regions on chromosome 8 at 234 cM for grain protein content and total soluble sugars and at 363 cM for Chl. a and Chl. b along with the position at 48 cM on chromosome 11 for starch and amylose content are genetic hot spots for these traits. CONCLUSION: The validated, co-localized and the novel QTL detected in this study will be useful for improvement of protein, starch, amylose, total soluble sugars and chlorophyll content in rice.

Curcumin protects against testis-specific side effects of irinotecan.

OBJECTIVE: Irinotecan (IR/CPT-11) is a semisynthetic, water-soluble derivative of the alkaloid camptothecin. It is a topoisomerase I group antineoplastic drug commonly used for the treatment of many cancer types, although it has side effects in tissues such as the testis. Curcumin (CRC) is a polyphenol compound produced from the Indian saffron root; it is used as food colouring and food flavouring. This study examined the testis-specific side effects of IR and the ability of CRC to protect against these side effects. MATERIALS AND METHODS: Forty male Sprague-Dawley rats were used in our study (n = 10). The rats were randomly divided into the following four groups: control, IR, IR + CRC, and CRC. IR 10 mg/kg/day was administered intraperitoneally and CRC 100 mg/kg was administered orally. Blood and testicular samples were collected from rats in all four groups on day 30 after drug administration. Histological, biochemical, and spermatological analyses were conducted. RESULTS: Testis tissue and blood samples were collected from the four groups. Tissue samples from the control and CRC groups demonstrated normal histological appearance on light microscopy. The IR group exhibited the following findings: vascular congestion in the tunica albuginea layer; tubular degeneration and vascular congestion in the interstitial area; oedema, vacuolisation, and luminised cells in the seminiferous tubule; and cells that temporarily stopped dividing at any stage of division in the seminiferous tubule epithelium. In the IR+CRC group, histopathological damage was significantly reduced by CRC treatment. Biochemical analysis showed that the level of thiobarbituric acid reactive substance (TBARS) was significantly increased in the IR group, compared with the other groups. CRC treatment significantly decreased this IR-mediated increase in TBARS level, and the TBARS level in the IR + CRC group approached the level observed in the control group. IR treatment caused significant decreases in glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) levels. However, CRC administration tended to ameliorate the decreases in GSH, SOD, CAT, and GPx levels. CONCLUSIONS: In this study, IR had some toxic effects in rat testis tissue; these effects were ameliorated by CRC treatment. Further studies are warranted to confirm our results.

Comparison of immunological, histological and oxidative effects of felbamate and levetiracetam in traumatic brain injury.

OBJECTIVE: We aimed to compare immunological, histological and oxidative effects of antiepileptic agents; felbamate and levetiracetam on head trauma in rats. MATERIALS AND METHODS: In this study, 32 Sprague-Dawley genus male rats were used. A closed head trauma mechanism was constituted in order to perform head trauma in rats. Rats were divided into 4 groups, and each group had 8 rats. Following head trauma, Group 1 (Control); normal saline was administered, Group 2; levetiracetam 50 mg/kg was administered, Group 3; felbamate 100 mg/kg was administered, and Group 4; levetiracetam 50 mg/kg and felbamate 100 mg/kg were administered with a combination. Injections were administered intraperitoneally once a day for 20 days. The rats were decapitated at the end of the 20th day. Blood and tissue samples were collected and analyzed for biochemical, immunohistochemical and histological parameters. RESULTS: Serum cytokine levels in Group 2, 3 and 4 were lower when compared to the control group. In Group 4, in which combined therapy was performed, cytokine levels were found to be the lowest. In Groups 2 and 3, a significant decrease in vascular congestion, mononuclear cell infiltration, hemorrhage, and neural degeneration was noticed in the pia mater. In Group 2, a decrease in vascular congestion and Purkinje cell degeneration was obtained in the cerebellum. However, the best outcomes were determined in Group 4. CONCLUSIONS: We determined that levetiracetam and felbamate alone are useful with respect to immunological, oxidative and histological alterations. However, their utility is better when used in a combination.

Early initiated feeding versus early reached target enteral nutrition in critically ill children: An observational study in paediatric intensive care units in Turkey.

AIM: Although early enteral nutrition (EN) is strongly associated with lower mortality in critically ill children, there is no consensus on the definition of early EN. The aim of this study was to evaluate our current practice supplying EN and to identify factors that affect both the initiation of feeding within 24 h after paediatric intensive care unit (PICU) admission and the adequate supply of EN in the first 48 h after PICU admission in critically ill children. METHODS: We conducted a prospective, multicentre, observational study in nine PICUs in Turkey. Any kind of tube feeding commenced within 24 h of PICU admission was considered early initiated feeding (EIF). Patients who received more than 25% of the estimated energy requirement via enteral feeding within 48 h of PICU admission were considered to have early reached target EN (ERTEN). RESULTS: Feeding was initiated in 47.4% of patients within 24 h after PICU admission. In many patients, initiation of feeding seems to have been delayed without an evidence-based reason. ERTEN was achieved in 43 (45.3%) of 95 patients. Patients with EIF were significantly more likely to reach ERTEN. ERTEN was an independent significant predictor of mortality (P < 0.001), along with reached target enteral caloric intake on day 2 associated with decreased mortality. CONCLUSIONS: There is a substantial variability among clinicians' perceptions regarding indications for delay to initiate enteral feeding in critically ill children, especially after the first 6 h of PICU admission. ERTEN, but not EIF, is associated with a significantly lower mortality rate in critically ill children.

Beneficial effects of hesperidin following cis-diamminedichloroplatinum-induced damage in heart of rats.

BACKGROUND: Increased oxidative stress and histopathological damage have been implicated in the cardiotoxicity that limits the clinical therapy of cisplatin (CP) as an anti-cancer drug. OBJECTIVES: This study aimed to investigate the protective effect of hesperidin (HP) against CP-induced cardiotoxicity in rats. MATERIALS AND METHODS: Rats were divided into four groups (n = 7/group), and the first group served as the control group. Animals in Group CP and Group CP + HP received a single dose of CP (CP - 7 mg/kg); animals in Group HP and Group CP + HP received 50 mg/kg/day HP with gavage for 14 days. At the end of day 14, cardiac tissue samples were histologically and biochemically examined. RESULTS: In this experimental study, thiobarbituric acid reactive substances levels in the cardiac tissue were significantly higher in the CP group, whereas glutathione (GSH), superoxide dismutase (SOD), and CAT levels were significantly lower in this group. On the other hand, GSH and SOD levels in the CP + HP group were similar to the control group. There was no significant difference in cardiac CAT levels between Group CP and Group CP + HP. CONCLUSION: Hesperetin treatment leads to a decrease in oxidative stress, and associated histological damage. The findings of the current study suggest that HP has a protective effect against CP-induced cardiotoxicity.

Protective role of Diospyros lotus on cisplatin-induced changes in sperm characteristics, testicular damage and oxidative stress in rats.

The aim of this study was to investigate the protective effect of Diospyros lotus (DL) on cisplatin (CP)-induced testicular damage in male rats. Twenty-eight male rats were randomly divided into four groups: group 1 - control, given isotonic saline solution; group 2 - CP 7 mg kg(-1) given intraperitoneally as single dose; group 3 - DL 1000 mg kg(-1) per day given orally for 10 days; group 4 - CP and DL given together at the same doses. CP caused a significant increase in thiobarbituric acid-reactive substances (TBARS) level and a significant decrease in superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and glutathione (GSH) levels in rats testis tissues compared to the control group. CP caused a significant increase in lipid peroxidation in testis tissues compared to the control group, whereas DL led to a significant increase in SOD and GSH levels. However, there were no statistically significant changes in GPx and CAT levels. In addition, serum testosterone levels, sperm concentration and sperm motility were significantly decreased, but abnormal sperm rate and histological changes were increased with CP. However, these effects of CP on sperm parameters, histological changes and the tissue weights were eliminated by DL treatment. In conclusion, our study showed that the reproductive toxicity caused by CP may be prevented by DL treatment.

Silent brain infarcts in two patients with zeta chain-associated protein 70 kDa (ZAP70) deficiency.

Zeta-chain associated protein 70 kDa deficiency (ZAP70) is a form of severe combined immunodeficiency (SCID). It is caused by defects in the signaling pathways associated with T-lymphocyte activation. ZAP70 deficiency is characterized by a marked reduction in peripheral CD8+ T-cells. In this report, we described two patients with ZAP70 deficiency who presented with recurrent infections, lung tuberculosis (TBC), congenital nephrotic syndrome (CNS), and silent brain infarcts (SBIs) as a common feature. The first patient initially presented with recurrent infections and TBC as in a classic SCID patient. At the age of 4, he was interned with febrile seizure. Cranial magnetic resonance imaging (MRI) showed SBIs. The second patient, an 8-month-old boy, presented with congenital nephrotic syndrome caused by cytomegalovirus (CMV) and he had also SBIs.

Hesperidin protects testicular and spermatological damages induced by cisplatin in rats.

The clinic usage of cisplatin, an anticancer drug, is limited due to it has many side effects in many systems and organs. In this context, it was aimed to investigate the protective effect of hesperidin, a citrus flavonoid, on testicular and spermatological damages induced by cisplatin in rats. The rats were randomly divided into four groups. The first group was kept as a control. In the second groups, cisplatin was given at the single dose of 7 mg kg(-1) intraperitoneally. In the third group, hesperidin was orally administered at the dose of 50 mg/kg day(-1) for 14 days. In the fourth group, cisplatin and hesperidin were given together at the same doses. Cisplatin treatment caused significant reductions enzymatic (SOD, CAT and GPx) and nonenzymatic (GSH) antioxidants and significant induction level of TBARS. In addition, cisplatin treatment caused decreased sperm motility, epididymal sperm concentration, increased abnormal sperm rate and histopathological damage. In contrast, hesperidin treatment significantly attenuated the harmful effects. In conclusion, this study clearly demonstrated that hesperidin has protective effects on cisplatin-induced reproductive system toxicity depending on its antioxidant properties. Thus, it is thought that hesperidin may be useful against cisplatin toxicity in patients with cancer in terms of reproductive system.

X-linked severe combined immunodeficiency due to a novel mutation complicated with hemophagocytic lymphohistiocytosis and presented with invagination: A case report.

Severe combined immunodeficiency (SCID) is an inherited disease with profoundly defective T cells, B cells, and natural killer (NK) cells. X-linked SCID (X-SCID) is its most common form. In this report, we describe a 4-month-old male with X-SCID who presented invagination and also showed hemophagocytic lymphohistiocytosis (HLH). The patient was admitted to our hospital with fever, cough, vomiting, monoliasis, and hepatosplenomegaly in postoperative period at the age of 3 months. The laboratory finding revealed no detectable T cells and hypogammaglobulinemia despite normal B-cell counts. Diagnosis of X-SCID was established by DNA analysis of the interleukin (IL)-2 receptor gamma chain gene (IL2RG); namely, we detected the novel mutation in the splice-site of exon 5 (c.595-1G>T). The patient died due to infection at the age of 4 months. Also, this case is the first report that describes the patient with X-SCID with presented invagination.

Sensitization of Cervical Cancer Cells to Cisplatin by Genistein: The Role of NFκB and Akt/mTOR Signaling Pathways.

Cervical cancer is among the top causes of death from cancer in women. Cisplatin-based chemotherapy has been shown to improve survival; however, cisplatin treatment is associated with toxicity to healthy cells. Genistein has been used as an adjunct to chemotherapy to enhance the activity of chemotherapeutic agents without causing increased toxicity. The present study was designed to investigate the effect of genistein (25 µM) on antitumor activity of cisplatin (250 nM) on HeLa cervical cancer cells. We have examined the alterations in expression of NF-κB, p-mTOR, p-p70S6K1, p-4E-BP1, and p-Akt protein levels in response to treatment. The combination of 25 µM genistein with 250 nM cisplatin resulted in significantly greater growth inhibition (P < 0.01). Genistein enhanced the antitumor activity of cisplatin and reduced the expression of NF-κB, p-mTOR, p-p70S6K1, p-4E-BP1, and p-Akt. The results in the present study suggest that genistein could enhance the activity of cisplatin via inhibition of NF-κB and Akt/mTOR pathways. Genistein is a promising nontoxic nutritional agent that may enhance treatment outcome in cervical cancer patients when given concomitantly with cisplatin. Clinical trials of genistein and cisplatin combination are warranted to test this hypothesis.

Notch1 intracellular domain increases cytoplasmic EZH2 levels during early megakaryopoiesis.

Notch pathway is a well-known factor in the development of lymphoid lineage. However, its role in the myeloid lineage has remained ambiguous. We looked into the effect of Notch1 on the megakaryocytic lineage commitment and found an increase in megakaryocyte-specific lineage markers upon transfection with Notch1 intracellular domain (NICD). This effect was mediated by Akt whereby constitutive activation of Akt increased the megakaryocyte markers, whereas inhibition of Akt signalling reduced these marker levels. Along with the change in differentiation status, NICD-induced initiation of early megakaryopoiesis was accompanied by an increased cytoplasmic enhancer of zeste homolog-2 (EZH2) expression. This process was found to be Akt-dependent, and inhibition or overexpression of Akt lead to concurrent changes in EZH2 levels. To elucidate the function of EZH2 in the cytoplasm, novel cytoplasmic interactors of EZH2 were identified by co-immunoprecipitation followed by matrix-assisted laser desorption ionization MS/MS-based protein identification, and thus, PDIA1 and LIM domain kinase-1 (LIMK1) were identified. Interaction of EZH2 with LIMK1 changed the activity of cofilin (a downstream target of LIMK1) towards actin filaments, thereby leading to lower filamentous actin content within these cells. Thus, Notch1 not only induces early megakaryopoiesis but also prepares these cells for subsequent morphological changes.

Homozygous ß-Thalassemia (FCS8-AA) and Hereditary Spherocytosis in the Same Patient.

A three-year old Turkish girl having both homozygous ß-thalassemia and hereditary spherocytosis and her family have been studied. The molecular defect causing thalassemia in the family was of the frame shift codon 8 (-AA) mutation type. The diagnosis of hereditary spherocytosis is based on osmotic fragility test in the patient and the family. However, the examination of erythrocyte membrane proteins has not been possible. ßthalassemia is in the heterozygous form in the mother, the father, and in two sisters. The mother, the father, and one of the sisters also have hereditary spherocytosis in addition to thalassemia. All those family members are asymptomatic. However, the patient who has frame shift codon 8 homozygosity along with hereditary spherocytosis presented with a severe form of hemolytic anemia.

Recombinant erythropoietin trial in children with transfusion-dependent homozygous beta-thalassemia.

Augmentation of gamma-gene synthesis by using recombinant human erythropoietin (r-Hu-EPO) represents a new approach to the therapy of beta-thalassemia. A prospective study was conducted in 26 transfusion-dependent beta-thalassemia major patients. r-Hu-EPO (Eprex/Cilag, Switzerland) was given to the patients at an initial dose of 500 IU/kg s.c. 3 times a week for at least 2 months during which no transfusion was applied. A sustained hemoglobin (Hb) level greater than 8 g/dl was considered as a response to EPO treatment. In the patients whose Hb levels remained under 8 g/dl or did not increase in comparison to pretreatment levels within 4 weeks, the dose of r-Hu-EPO was increased to 1,000 IU/kg 3 times a week and applied for another 4 weeks. Only 16 cases also received oral iron supplementation. The whole blood and reticulocyte counts, the biochemical tests including BUN, creatinine, AST, ALT, alkaline phosphatase and ferritin were done and the percentages of HbF and F cells were analyzed regularly. At the end of the 2nd month, 6 cases qualified to continue with the trial. At the end of the 6th month, r-Hu-EPO therapy was ceased in 3 cases of the 6 since their Hb levels had decreased below 7 g/dl. Only 3 cases (11.5%) continued with the r-Hu-EPO therapy without transfusion for up to 12 months. In conclusion, r-Hu-EPO may be useful in some selected transfusion-dependent patients with beta-thalassemia major. Selection criteria should include a mild beta-genotype of coinheritance of alpha-thalassemia, splenectomy and pretreatment reticulocyte response of the patients as well as the patients' compliance.

Fetal DNA in uterine vein blood.

OBJECTIVE: To investigate whether the ratio of fetal cells in the maternal circulation differs before and after the blood passes through the maternal lung. METHODS: We performed polymerase chain reaction-based Y-sequence analysis of DNA derived from antecubital vein blood obtained before and 2 hours after cesarean delivery, and from uterine vein blood obtained during cesarean of 14 women carrying male fetuses. RESULTS: Fetal DNA was detected in 17 tested specimens and, as estimated by comparison with parallel dilution series, the fetal-maternal DNA ratio was 1:10(5) to 1:10(6). However, there was no significant difference in the amount of Y-chromosomal DNA detectable between uterine vein blood and peripheral blood after polymerase chain reaction and Southern hybridization. In DNA derived from peripheral blood after delivery, the intensity of Y-specific fetal DNA sequences was also not significantly increased. CONCLUSION: Our results argue against the often-stated hypothesis of different ratios of fetal to maternal DNA in the uterine vein versus peripheral blood, and indicate that even delivery does not seem to increase the flow of fetal cells into the maternal circulation.

Nucleotide sequence of Aspergillus nidulans mitochondrial genes coding for ATPase subunit 6, cytochrome oxidase subunit 3, seven unidentified proteins, four tRNAs and L-rRNA.

The complete nucleotide sequence of a 14 kb segment of A. nidulans mtDNA reveals a rather compact organization of genes transcribed from the same strand and coding for two functionally known proteins, seven unidentified polypeptides (URFs), 24 tRNAs and two rRNAs. One of the URFs is located in the intron of the L-rRNA gene and codes for a basic protein of 410 residues. The other URFs are in spacer regions and code for hydrophobic proteins. URFa is homologous to human URF4, and URFb produces a polypeptide of 48 residues resembling the human URF6L product (hydrophobic N-terminus, basic C-terminus). The ATPase subunit 6 genes from mitochondria and E. coli appear to share a common ancestor. The codon frequencies of identified genes and URFs are similar, and codons ending with G or C are rarely used. The structures of tRNAs specific for arginine, asparagine, tyrosine and histidine are deduced from gene sequences.