Evaluation of Liver Function Tests in ß-Thalassemia Major Children.

Thalassemia is the most prevalent single gene defect in human beings worldwide. Repeated blood transfusions along with chelation therapy are mainstay of treatment in thalassemia patients. However these recurrent blood transfusions result in iron overload which along with chelation therapy causes deterioration of liver function. Aim of the study was to evaluate the liver function tests in ß-thalassemia major patients. This cross sectional study was conducted in the Department of Biochemistry, Dhaka Medical College, Dhaka, Bangladesh from January 2017 to December 2017. In this study, 50 diagnosed patients of ß Thalassemia major (Group A) and 50 apparently healthy children (Group B) of both sexes were selected from the department of Paediatrics, Dhaka medical college. The study parameters were serum ferritin, bilirubin, AST, ALT, ALP. The results were compared statistically between groups. Serum ferritin level (mean±SD) in thalassemic major patients in Group A (890±446.38 microgram/L) which is significantly higher above normal level. Serum bilirubin in Group A (3.27±2.62 mg/dl) and in Group B (0.48±0.24 mg/dl), Serum ALT in Group A (53.06±34.0 U/L) and in Group B (16.70±4.81 U/L), AST in Group A (84.56±33.54 U/L) and in Group B (11.60±2.72 U/L) and ALP levels in Group A (422.42±226.99 IU/L) and in Group B (221.86±80.54 IU/L). All the values were significantly higher (p<0.001) in ß-thalassemia patient than that of normal children. This study concludes that liver function parameters are significantly higher in ß thalassemia major patients. So routine evaluation of liver function tests may be advocated for thalassemic patients to predict early onset of hepatic dysfunction.

Plant-mediated gene silencing restricts growth of the potato late blight pathogen Phytophthora infestans.

Phytophthora infestans is an oomycete that causes severe damage to potato, and is well known for its ability to evolve rapidly in order to overcome resistant potato varieties. An RNA silencing strategy was evaluated here to clarify if small interfering RNA homologous to selected genes in P. infestans could be targeted from the plant host to reduce the magnitude of the infection. As a proof-of-concept, a hairpin RNA (hp-RNA) construct using the GFP marker gene was designed and introduced in potato. At 72 hpi, a 55-fold reduction of the signal intensity of a corresponding GFP expressing P. infestans strain on leaf samples of transgenic plants, compared with wild-type potato, was detected. This suggests that an RNA interference construct in the potato host could be processed and target a transcript of the pathogen. Three genes important in the infection process of P. infestans, PiGPB1, PiCESA2, and PiPEC, together with PiGAPDH taking part in basic cell maintenance were subsequently tested using an analogous transgenic strategy. Out of these gene candidates, the hp-PiGPB1 targeting the G protein ß-subunit (PiGPB1) important for pathogenicity resulted in most restricted disease progress. Further, Illumina sequencing of inoculated transgenic potato leaves revealed sRNAs of 24/25 nt size homologous to the PiGPB1 gene in the transgenic plants indicating post-transcriptional silencing of the target gene. The work demonstrates that a host-induced gene-silencing approach is functional against P. infestans but is highly dependent on target gene for a successful outcome. This finding broadens the arsenal of control strategies to this important plant disease.

Fragmentation of tRNA in Phytophthora infestans asexual life cycle stages and during host plant infection.

BACKGROUND: The oomycete Phytophthora infestans possesses active RNA silencing pathways, which presumably enable this plant pathogen to control the large numbers of transposable elements present in its 240 Mb genome. Small RNAs (sRNAs), central molecules in RNA silencing, are known to also play key roles in this organism, notably in regulation of critical effector genes needed for infection of its potato host. RESULTS: To identify additional classes of sRNAs in oomycetes, we mapped deep sequencing reads to transfer RNAs (tRNAs) thereby revealing the presence of 19-40 nt tRNA-derived RNA fragments (tRFs). Northern blot analysis identified abundant tRFs corresponding to half tRNA molecules. Some tRFs accumulated differentially during infection, as seen by examining sRNAs sequenced from P. infestans-potato interaction libraries. The putative connection between tRF biogenesis and the canonical RNA silencing pathways was investigated by employing hairpin RNA-mediated RNAi to silence the genes encoding P. infestans Argonaute (PiAgo) and Dicer (PiDcl) endoribonucleases. By sRNA sequencing we show that tRF accumulation is PiDcl1-independent, while Northern hybridizations detected reduced levels of specific tRNA-derived species in the PiAgo1 knockdown line. CONCLUSIONS: Our findings extend the sRNA diversity in oomycetes to include fragments derived from non-protein-coding RNA transcripts and identify tRFs with elevated levels during infection of potato by P. infestans.

Evidence for small RNAs homologous to effector-encoding genes and transposable elements in the oomycete Phytophthora infestans.

Phytophthora infestans is the oomycete pathogen responsible for the devastating late blight disease on potato and tomato. There is presently an intense research focus on the role(s) of effectors in promoting late blight disease development. However, little is known about how they are regulated, or how diversity in their expression may be generated among different isolates. Here we present data from investigation of RNA silencing processes, characterized by non-coding small RNA molecules (sRNA) of 19-40 nt. From deep sequencing of sRNAs we have identified sRNAs matching numerous RxLR and Crinkler (CRN) effector protein genes in two isolates differing in pathogenicity. Effector gene-derived sRNAs were present in both isolates, but exhibited marked differences in abundance, especially for CRN effectors. Small RNAs in P. infestans grouped into three clear size classes of 21, 25/26 and 32 nt. Small RNAs from all size classes mapped to RxLR effector genes, but notably 21 nt sRNAs were the predominant size class mapping to CRN effector genes. Some effector genes, such as PiAvr3a, to which sRNAs were found, also exhibited differences in transcript accumulation between the two isolates. The P. infestans genome is rich in transposable elements, and the majority of sRNAs of all size classes mapped to these sequences, predominantly to long terminal repeat (LTR) retrotransposons. RNA silencing of Dicer and Argonaute genes provided evidence that generation of 21 nt sRNAs is Dicer-dependent, while accumulation of longer sRNAs was impacted by silencing of Argonaute genes. Additionally, we identified six microRNA (miRNA) candidates from our sequencing data, their precursor sequences from the genome sequence, and target mRNAs. These miRNA candidates have features characteristic of both plant and metazoan miRNAs.

Growth performance and nutrient composition of juvenile Nile tilapia (Oreochromis niloticus) fed Spirulina flakes, rice bran and mustard oil cake.

INTRODUCTION: Tilapia (Oreochromis niloticus) is an important cultured fish that is widely distributed in Bangladesh. This study was conducted to improve the growth performance and nutrient contents of the fish using five different types of feeds. METHODS: Tilapia fingerlings were fed two types of commercial fish feeds (Feed-1 and Feed-2), Spirulina flakes (Feed-3), Feed-2 mixed with Spirulina flakes (Feed-4) and manually mixed feed made from a mixture of mustard oil cake and rice bran (Feed-5). After 4 weeks of being fed with the diets, growth parameters and meat nutrient composition of the tilapia fingerlings were recorded. RESULTS: Significant growth in length and weight was observed in juvenile tilapia fish fed with commercial Feed-1 only, while growth performance varied significantly among fingerlings fed other types of feeds. Body tissue calcium (92.8 mg/100 g), iron (1.29 mg/100 g) was higher in fishes fed with dry Spirulina flakes (Feed 3), while the highest amount of zinc (2.09 mg/100 g) was recorded in fishes fed Feed-5. Protein (13.32%) content was highest in fish fed Feed-2 mixed with Spirulina flakes (Feed-4). CONCLUSION: Meat nutritional quality of tilapia can be improved by combining commercial feeds with Spirulina flakes, compared with feeding commercial feeds in isolation.

Role of vitamin E supplementation on serum levels of copper and zinc in hemolytic anemic patients with G6PD deficiency.

Vitamin E scavenges free radicals and may prevent destruction of RBC in Glucose6-phosphate dehydrogenase (G6PD) deficient hemolytic anemia, where changes in copper (Cu) and zinc (Zn) may act as additional contributory factors for hemolysis. In the present study changes in serum Cu and Zn and role of vitamin E supplementation on these changes were observed in hemolytic anemic patients with G6PD deficiency. This study was conducted in the Department of Physiology, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka during July 2005-June 2006. For this, 102 subjects with age ranged 5-40 years of both sexes were included in the study. Among them 68 were G6PD deficient patients, of whom 34 were in supplemented group and 34 were non-supplemented group. The supplemented group received vitamin E for 60 consecutive days at a dose of 800 IU/day for adult and 400 IU/day for children < or =12 years (4 times daily). Age and sex matched 34 apparently healthy subjects with normal G6PD level were taken to observe the base line data (healthy control) and also for comparison. All the G6PD deficient patients were selected from the Out Patient Department (OPD) of Hematology, BSMMU, Dhaka, and all the healthy subjects from personal contact. Blood G6PD level was done by spectrophotometric method and serum Cu, Zn levels by atomic absorption spectrophotometric method. To observe the availability of binding proteins serum total protein, albumin, globulin and A:G ratio were done by standard laboratory techniques. All parameters were measured on day 1 of their 1st visit and also on day 60 in deficient groups. Data analysis was done by appropriate statistical method. Serum Cu was significantly (p<0.001) higher but serum Zn, total protein, albumin, A/G ratio were significantly (p<0.001) lower in G6PD deficient groups in comparison to those of healthy control on day 1. After vitamin E supplementation, values of these parameters were comparable with those of healthy control in supplemented group in comparison to those of their pre-supplemented and non-supplemented groups both on day 1 and day 60. So, vitamin E supplementation has got its effective role in restoration of normal serum concentration of Cu and Zn in this group of patients.