Isolation and Sequence Analyses of Salinity Tolerance Genes from Bruguiera Cylindrica (L.) Blume

Salinity is a major abiotic stress limiting the productivity of crop plants globally. The discovery of novel genes in stress adaptation will provide effective genetic engineering strategies leading to g reater stress tolerance. The objectives of this research are to identify and isolate salinity tolerance genes from the mangrove plant, Bruguiera cylindrica (L.) Blume through suppression subtractive hybridization (SSH) and bacterial functional assay. B. cylindrica propagules were grown in fresh water and 20 ppt salinity water. Root morphology differences between B. cylindrica grown in fresh water and 20 ppt salinity water were largely due to the need of roots to obtain more water and nutrients during salinity stress. B. cylindrica plants grew better in the presence of salt as higher mean values were obtained for all morphological measurements compared to B. cylindrica plants grown in fresh water. Four RNA extraction methods were attempted to obtain high yield and high purity RNA. The cesium chloride method was chosen for RNA extraction as it gave the highest amount of pure RNA. Subtracted cDNAs were prepared from the roots of the B. cylindrica seedlings that were grown in fresh water and salt water, respectively. A total of 84 subtracted cDNAs were cloned into pCR-BLUNT II TOPO and sequenced. A total of 51 subtracted cDNAs with good sequencing quality were assembled into 7 contigs and 10 singletons. These non-redundant sequences were grouped into unknown protein (41.18%), novel (29.41%), protein destination and storage (11.76%), energy (5.88%), intracellular traffic (5.88%) and protein synthesis (5.88%). Some motifs of novel and unknown sequences may involve in the salinity tolerance of B. cylindrica such as Kv1.3 voltage-gated K+ ion(s) channel signature, calcium-activated BK potassium channel alpha subunit and Kir2.1 inward rectifier K+ ion(s) channel signature. Meanwhile, a cDNA library was also constructed from the roots of B. cylindrica that were grown in fresh water. Bacterial functional assay was performed to identify cDNAs that confer salt tolerance. A total of 85 cDNA clones that were able to grow on 2x YT containing 400 mM NaCI were sequenced and 73 cDNAs with good sequence quality were assembled into 9 contigs and 53 singletons. The non-redundant sequences were also categorised into unknown protein (58.06%), metabolism (9.68%), transporters (9.68%), transcription (6.45%), energy (4.84%), cell growth/division (4.84%), novel (3.23%), miscellaneous (1.61%) and disease/defense (1.61%) A motif search on novel and unknown cDNAsequences had revealed some possible motifs that may be involved in salinity tolerance of B. cylindrica e.g. C. elegans Srg family integral membrane protein signature and 2Fe-2S ferredoxins, iron-sulfur binding region signature. Sequence analysis of subtracted cDNAs and putative salt tolerant cDNAs isolated by bacterial functional assay showed some putative proteins that may be involved in the salinity tolerance of B. cylindrica such as putative potassium transporter HAK 1p (M33), putative zinc finger protein (M3), ubiquitin (BC27) and L-ascorbate peroxidase (A46)

Isolation of salinity tolerant genes from the mangrove plant, Bruguiera cylindrica by using suppression subtractive hybridization (SSH) and bacterial functional screening

In this study, we have identified and isolated 126 salinity tolerant cDNAs from the root of a mangrove plant, Bruguiera cylindrica (L.) Blume by using suppression subtractive hybridization (SSH) and bacterial functional screening. Sequencing of 51 subtracted cDNA clones that were differentially expressed in the root of B. cylindrica exposed to 20 parts per thousand (ppt) NaCl water revealed 10 tentative unique genes (TUGs) with putative functions in protein synthesis, storage and destination, metabolism, intracellular trafficking and other functions; and 9 unknown proteins. Meanwhile, the 75 cDNA sequences of B. cylindrica that conferred salinity tolerance to Escherichia coli consisted of 29 TUGs with putative functions in transportation, metabolism and other functions; and 33 with unknown functions. Both approaches yielded 42 unique sequencess that have not been reported else where to be stress related and might provide further understanding of adaptations of this plant to salinity stress

Global variation in postoperative mortality and complications after cancer surgery: a multicentre, prospective cohort study in 82 countries

© 2021 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY-NC-ND 4.0 licenseBackground: 80% of individuals with cancer will require a surgical procedure, yet little comparative data exist on early outcomes in low-income and middle-income countries (LMICs). We compared postoperative outcomes in breast, colorectal, and gastric cancer surgery in hospitals worldwide, focusing on the effect of disease stage and complications on postoperative mortality. Methods: This was a multicentre, international prospective cohort study of consecutive adult patients undergoing surgery for primary breast, colorectal, or gastric cancer requiring a skin incision done under general or neuraxial anaesthesia. The primary outcome was death or major complication within 30 days of surgery. Multilevel logistic regression determined relationships within three-level nested models of patients within hospitals and countries. Hospital-level infrastructure effects were explored with three-way mediation analyses. This study was registered with ClinicalTrials.gov, NCT03471494. Findings: Between April 1, 2018, and Jan 31, 2019, we enrolled 15 958 patients from 428 hospitals in 82 countries (high income 9106 patients, 31 countries; upper-middle income 2721 patients, 23 countries; or lower-middle income 4131 patients, 28 countries). Patients in LMICs presented with more advanced disease compared with patients in high-income countries. 30-day mortality was higher for gastric cancer in low-income or lower-middle-income countries (adjusted odds ratio 3·72, 95% CI 1·70–8·16) and for colorectal cancer in low-income or lower-middle-income countries (4·59, 2·39–8·80) and upper-middle-income countries (2·06, 1·11–3·83). No difference in 30-day mortality was seen in breast cancer. The proportion of patients who died after a major complication was greatest in low-income or lower-middle-income countries (6·15, 3·26–11·59) and upper-middle-income countries (3·89, 2·08–7·29). Postoperative death after complications was partly explained by patient factors (60%) and partly by hospital or country (40%). The absence of consistently available postoperative care facilities was associated with seven to 10 more deaths per 100 major complications in LMICs. Cancer stage alone explained little of the early variation in mortality or postoperative complications. Interpretation: Higher levels of mortality after cancer surgery in LMICs was not fully explained by later presentation of disease. The capacity to rescue patients from surgical complications is a tangible opportunity for meaningful intervention. Early death after cancer surgery might be reduced by policies focusing on strengthening perioperative care systems to detect and intervene in common complications. Funding: National Institute for Health Research Global Health Research Unit

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

© 2022 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 licenseBackground: Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide. Methods: A multimethods analysis was performed as part of the GlobalSurg 3 study—a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital. Findings: Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3·85 [95% CI 2·58–5·75]; p<0·0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63·0% vs 82·7%; OR 0·35 [0·23–0·53]; p<0·0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer. Interpretation: Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised. Funding: National Institute for Health and Care Research