In vitro propagation and withaferin A production in Withania ashwagandha, a rare medicinal plant of India

Withania ashwagandha, belonging to the family Solanaceae, is an important medicinal herb of India with restricted geographic distribution. It is a rich source of withaferin A (WA) and other bioactive withanolides. In the present study a rapid in vitro mass propagation protocol of W. ashwagandha was developed from nodal explants. Nodal explants were cultured on MS medium supplemented with various concentrations and combinations of plant growth regulators (PGRs). The highest number of regenerated shoots per ex-plant (33±2.7) and highest WA (13.4±1.15 mg/g of DW) production was obtained on MS medium supplemented with 5.0 μM 6-benzyladenine (BA) and 1.0 μM Kinetin (Kn). In vitro raised shoots were further rooted on half-strength MS medium containing 2.0 μM Indole-3-butyric acid (IBA) and analyzed forWA production. The rooted plantlets when transferred to poly bags in the greenhouse showed 90 % survival frequency. Levels of WA were higher in the in vitro and ex vitro derived shoot and root tissues as compared to field grown mother plants. In an attempt to further maximize WA production, shoot cultures were further grown in liquid MS medium supplemented with 5.0 μM 6-benzyladenine (BA) and 1.0 μM Kinetin (Kn). Root cultures were grown on half strength MS liquid medium fortified with 2.0 μMof IBA.WA production in the liquid cultures was significantly higher compared to the static composition of the same media. This protocol, first of its kind in this plant, can be successfully employed for conservation, proliferation and large-scale production of WA. The regenerated plants can also be used in traditional medicine as an alternative to naturally collected plants.University of Pretoria and National Research Foundation (NRF).South Africa Vice-Chancellor and Freestanding Postdoctoral Fellowship.http://link.springer.com/journal/12298hb201

Enhanced production of withaferin-A in shoot cultures of Withania somnifera (L) Dunal

Withania somnifera (L) Dunal, commonly known as ashwagandha or Indian ginseng, is the source of large number of pharmacologically active withanolides. Withaferin-A (WS-3), a major withanolide of W. somnifera, has been proven to be an effective anti-cancer molecule. In this study, a liquid culture system for shoot proliferation, biomass accumulation and withaferin-A production of an elite accession (AGB002) of W. somnifera was investigated. The nodal explants cultured on Murashige and Skoog (MS) semi-solid medium supplemented with various concentrations of 6-benzyl adenine (BA) and Kinetin (Kn) elicited varied responses. The highest number of regenerated shoots per ex-plant (35 ± 3.25) and the maximum average shoot length (5.0 ± 0.25 cm) were recorded on MS medium supplemented with BA (5.0 μM). The shoots were further proliferated in half and full strength MS liquid medium supplemented with the same concentration BA. It was interesting to note that shoots cultured on MS half strength liquid medium fortified with 4 gL-1 FW (fresh weight) shoot inoculum mass derived from 5 week old nodal explants of W. somnifera showed highest accumulation of biomass and withaferin A content in 5 weeks. Withaferin A was produced in relatively high amounts (1.30 % and 1.10 % DW) in shoots cultured in half and full strength MS liquid media respectively as compared to natural field grown plants (0.85 % DW). A considerable amount of the withaferin A was also excreted in the culture medium. Successful proliferation of shoots in liquid medium and the synthesis of withaferin A in vitro opens new avenues for bioreactor scale-up and the large-scale production of the compound

Reproductive biology of Withania ashwagandha sp. novo (Solanaceae)

Withania ashwagandha Kaul (Solanaceae) is an annual plant species of immense medicinal importance. It is a repository of a large number of pharmacologically active secondary metabolites known as withanolides. Evidence for the delimitation of the species from W. somnifera has been provided by our group using multidisciplinary approaches. Knowledge of reproductive biology of medicinal plants is crucial for improvement, effective conservation and management plans to evolve genetically superior varieties. The present paper reports our findings on the floral biology, pollination behavior and breeding system of W. ashwagandha in natural populations grown at our experimental field under near natural conditions. Flowering (peak) takes place during April–July and anthesis occurs between 08:00 and 11:00 h. The period of stigma receptivity coincides with anther dehiscence. Fruit set on pollination treatments ranged from 90.8% (passive autogamy), 72% (assisted autogamy), 30.30% (xenogamy), and 56.50% (geitonogamy) through 50.40% (open pollination). Xenogamy brings about very low fruit set, seed-set and seed germination percentages. It is inferred that W. ashwagandha is predominantly an autogamous and self-compatible species. Self-compatibility is mainly accomplished due to close proximity of stigma and anthers. This work is the first report on the reproductive biology of W. ashwagandha and will be useful for conservation and development of improved varieties of this multipurpose herb.The author acknowledge the University of Pretoria for providing Vice-Chancellors Post Doctoral Fellowship.http://www.elsevier.com/locate/indcrophb201

Cold stress affects antioxidative response and accumulation of medicinally important withanolides in Withania somnifera (L.) Dunal

Withania somnifera (L.) Dunal (Indian ginseng) is a high value medicinal plant. It synthesizes a large array of biologically active withanolides. In this study, two month old seedlings of AGB002 (wild genotype) and AGB025 (cultivated genotype) of W. somnifera were subjected to cold stress (4 ◦C) under controlled envi-ronment. Plants were analyzed for three medicinally important secondary metabolites (withanolide A, withanone and withaferin A), lipid peroxidation (MDA), cell injury, superoxide radical (O2 •−) accumula-tion and anti-oxidative enzymes activities such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR). Increases in the titers of superoxide anion and MDA were observed from day 1 to day 7 in both genotypes, although the increase on the first day of exposure was significantly higher. Enzymatic activities of SOD, CAT, APX and GR also showed an increasing trend in both genotypes and reached a maximum on day 7 of the cold temperature exposure; however, this increase was higher in AGB002 than AGB025. Withanolide A (WS-1) in the roots of both genotypes sig-nificantly decreased on the first day of cold exposure and then showed a recovery until day 7. WS-1 was not detected in the leaves of either genotype. Withanone (WS-2) content in the leaves also decreased towards the end of the cold period in both genotypes. Cold stress also elicited the accumulation of WS-2 in AGB025, but was not detectable in control seedlings. At maturity, WS-2 was also detected in control plants. Furthermore, a significant increase in the leaf withaferin A (WS-3) content was recorded from day 1 to day 7 of the cold exposure in both the genotypes, suggesting the possible involvement of with-anolides in cold-protection. AGB002 showed comparatively higher accumulation of antioxidant enzymes and selected marker withanolides than AGB025, indicating that AGB002 is better adapted to cold than AGB025. It could be inferred from these observations that cold stress induces bioactive withanolide accu-mulation in W. somnifera as a mechanism for scavenging reactive oxygen species (ROS). These studies also provide an impetus for enhancing the withanolide accumulation in W. somnifera using controlled environment technology.CSIR-Indian Institute of Integrative Medicine (CSIR-IIIM).http://www.elsevier.com/locate/indcrop2016-11-30hb2016BiochemistryChemistryGenetic

Clear and independent associations of several HLA-DRB1 alleles with differential antibody responses to hepatitis B vaccination in youth

To confirm and refine associations of human leukocyte antigen (HLA) genotypes with variable antibody (Ab) responses to hepatitis B vaccination, we have analyzed 255 HIV-1 seropositive (HIV+) youth and 80 HIV-1 seronegatives (HIV−) enrolled into prospective studies. In univariate analyses that focused on HLA-DRB1, -DQA1, and -DQB1 alleles and haplotypes, the DRB1*03 allele group and DRB1*0701 were negatively associated with the responder phenotype (serum Ab concentration ≥ 10 mIU/mL) (P = 0.026 and 0.043, respectively). Collectively, DRB1*03 and DRB1*0701 were found in 42 (53.8%) out of 78 non-responders (serum Ab <10 mIU/mL), 65 (40.6%) out of 160 medium responders (serum Ab 10–1,000 mIU/mL), and 27 (27.8%) out of 97 high responders (serum Ab >1,000 mIU/mL) (P < 0.001 for trend). Meanwhile, DRB1*08 was positively associated with the responder phenotype (P = 0.010), mostly due to DRB1*0804 (P = 0.008). These immunogenetic relationships were all independent of non-genetic factors, including HIV-1 infection status and immunodeficiency. Alternative analyses confined to HIV+ youth or Hispanic youth led to similar findings. In contrast, analyses of more than 80 non-coding, single nucleotide polymorphisms within and beyond the three HLA class II genes revealed no clear associations. Overall, several HLA-DRB1 alleles were major predictors of differential Ab responses to hepatitis B vaccination in youth, suggesting that T-helper cell-dependent pathways mediated through HLA class II antigen presentation are critical to effective immune response to recombinant vaccines

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

Raspberry Pi Based Vision System for Foreign Object Debris (FOD) Detection

Background: The main purpose of this research is to design and develop a cost-effective system for detection of Foreign Object Debris (FOD), dedicated to airports. FOD detection has been a significant problem at airports as it can cause damage to aircraft. Developing such a device to detect FOD may require complicated hardware and software structures. The proposed solution is based on a computer vision system, which comprises of flexible off the shelf components such as a Raspberry Pi and Camera Module, allowing the simplistic and efficient way to detect FOD. Methods: The solution to this research is achieved through User-centered design, which implies to design a system solution suitably and efficiently. The system solution specifications, objectives and limitations are derived from this User-centered design. The possible technologies are concluded from the required functionalities and constraints to obtain a real-time efficient FOD detection system. Results: The results are obtained using background subtraction for FOD detection and implementation of SSD (single-shot multi-box detector) model for FOD classification. The performance evaluation of the system is analysed by testing the system to detect FOD of different size for different distances. The web design is also implemented to notify the user in real-time when there is an occurrence of FOD. Conclusions: We concluded that the background subtraction and SSD model are the most suitable algorithms for the solution design with Raspberry Pi to detect FOD in a real-time system. The system performs in real-time, giving the efficiency of 84% for detecting medium-sized FOD such as persons at a distance of 75 meters and 72% efficiency for detecting large-sized FOD such as cars at a distance of 125 meters, and the average frame per second (fps) that is the system ’s performance in recording and processing frames of the area required to detect FOD is 0.95

Effective Distribution of Roles and Responsibilities in Global Software Development Teams

Context. Industry is moving from co-located form of development to a distributed development in order to achieve different benefits such as cost reduction, access to skillful labor and around the clock working etc. This transfer requires industry to face different challenges such as communication, coordination and monitoring problems. Risk of project failure can be increased, if industry does not address these problems. This thesis is about providing the solutions of these problems in term of effective roles and responsibilities that may have positive impact on GSD team. Objectives. In this study we have developed framework for suggesting roles and responsibilities for GSD team. This framework consists of problems and casual dependencies between them which are related to team’s ineffectiveness, then suggestions in terms of roles and responsibilities have been presented in order to have an effective team in GSD. This framework, further, has been validated in industry through a survey that determines which are the effective roles and responsibilities in GSD. Methods. We have two research methods in this study 1) systematic literature review and 2) survey. Complete protocol for planning, conducting and reporting the review as well as survey has been described in their respective sections in this thesis. A systematic review is used to develop the framework whereas survey is used for framework validation. We have done static validation of framework. Results. Through SLR, we have identified 30 problems, 33 chains of problems. We have identified 4 different roles and 40 different responsibilities to address these chains of problems. During the validation of the framework, we have validated the links between suggested roles and responsibilities and chains of problems. Addition to this, through survey, we have identified 20 suggestions that represents strong positive impact on chains of problems in GSD in relation to team’s effectiveness. Conclusions. We conclude that implementation of effective roles and responsibilities in GSD team to avoid different problems require considerable attention from researchers and practitioners which can guarantee team’s effectiveness. Implementation of proper roles and responsibilities has been mentioned as one of the successful strategies for increasing team’s effectiveness in the literature, but which particular roles and responsibilities should be implemented still need to be addressed. We also conclude that there must be basic responsibilities associated with any particular role. Moreover, we conclude that there is a need for further development and empirical validation of different frameworks for suggesting roles and responsibilities in full scale industry trials

Effective Distribution of Roles and Responsibilities in Global Software Development Teams

Context. Industry is moving from co-located form of development to a distributed development in order to achieve different benefits such as cost reduction, access to skillful labor and around the clock working etc. This transfer requires industry to face different challenges such as communication, coordination and monitoring problems. Risk of project failure can be increased, if industry does not address these problems. This thesis is about providing the solutions of these problems in term of effective roles and responsibilities that may have positive impact on GSD team. Objectives. In this study we have developed framework for suggesting roles and responsibilities for GSD team. This framework consists of problems and casual dependencies between them which are related to team’s ineffectiveness, then suggestions in terms of roles and responsibilities have been presented in order to have an effective team in GSD. This framework, further, has been validated in industry through a survey that determines which are the effective roles and responsibilities in GSD. Methods. We have two research methods in this study 1) systematic literature review and 2) survey. Complete protocol for planning, conducting and reporting the review as well as survey has been described in their respective sections in this thesis. A systematic review is used to develop the framework whereas survey is used for framework validation. We have done static validation of framework. Results. Through SLR, we have identified 30 problems, 33 chains of problems. We have identified 4 different roles and 40 different responsibilities to address these chains of problems. During the validation of the framework, we have validated the links between suggested roles and responsibilities and chains of problems. Addition to this, through survey, we have identified 20 suggestions that represents strong positive impact on chains of problems in GSD in relation to team’s effectiveness. Conclusions. We conclude that implementation of effective roles and responsibilities in GSD team to avoid different problems require considerable attention from researchers and practitioners which can guarantee team’s effectiveness. Implementation of proper roles and responsibilities has been mentioned as one of the successful strategies for increasing team’s effectiveness in the literature, but which particular roles and responsibilities should be implemented still need to be addressed. We also conclude that there must be basic responsibilities associated with any particular role. Moreover, we conclude that there is a need for further development and empirical validation of different frameworks for suggesting roles and responsibilities in full scale industry trials