In vitro regeneration, detection of somaclonal variation and screening for mosaic virus in sugarcane (Saccharum spp.) somaclones

Three sugarcane accessions susceptible to sugarcane mosaic virus; HSF-240, S-2000-US-359, and S-2003-US-704 were evaluated for callogenesis and regeneration ability. For callogenesis, five different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) was used. The best callogenesis was obtained when Murashige and Skoog (MS) was portified with 3 mg/L 2,4-D and the highest regeneration was obtained on media containing MS + kinetin 0.5 + 0.5 mg/L naphthalene acetic acid (NAA). After succesful regeneration and rooting on half strength MS medium, with 1.5 mg/L indole-3-butyric acid supplementation, plantlets were shifted to green house. Enzyme linked immunosorbent assay (ELISA) test was performed to detect the presence of sugarcane mosaic virus (SCMV) in the regenerated plantlets and simple sequence repeat (SSR) markers were used to evaluate the genetic variation at DNA level between the parent’s plants and regenerated somaclones of the accession HSF-240. A total of 26 parent plants and 64 somaclones, among the regenerated plants were selected for the screening of virus through double antibody sandwich (DAS-ELISA) test. Four (4) parent plants out of the 26, showed negative reaction to the virus test. Ten (10) somaclones showed positive reaction to the disease, 9 somaclones showed mild reaction to virus and 45 somaclones showed negative reaction. For the detection of somaclonal variation, 38 primers pair were used and 15 simple sequence repeats (SSR) primer pairs were found to be polymorphic with 51.61% polymorphism. The study demonstrates that SSR genetic markers are the best tool for the investigation of genetic variation in sugarcane.Keywords: Callogenesis, somaclones, simple sequence repeats (SSR), genetic marker

Electrochemical investigations of unexplored anthraquinones and their DNA binding

The redox behaviour of two potential anticancer anthraquinones, 9,10-anthraquinone and 2-chloromethyl-9,10-anthraquinone was investigated in a wide pH range. Cyclic voltammetry based assay was developed for the assessment of the effect of medium, substituents, potential scan rate and number of scans on the voltammetric response of anthraquinones. The electrode reaction mechanism was suggested on the basis of cyclic and differential pulse voltammetric results. The effect of DNA on anthraquinones was also probed at physiological pH which could lead to further investigation of possible citotoxic activity in vitro. The results revealed that anthraquinones interact with DNA more strongly than the clinically used anticancer drug, epirubicin

Incidence of \u3ci\u3eSpodoptera litura\u3c/i\u3e (Lepidoptera: Noctuidae) and its feeding potential on various citrus (Sapindales: Rutaceae) cultivars in the Sargodha Region of Pakistan

Spodoptera litura F. (Lepidoptera: Noctuidae) adversely affects important crops such as cotton, okra, tomato, potato, and pumpkin. For the first time, this species was observed in a citrus nursery in the Sargodha Region of Pakistan. The principal objectives of this study were to determine the effects of several citrus (Sapindales: Rutaceae) cultivars, including Citrus reticulata Blanco (Feutrell’s Early, Seedless Kinnow, and Fairchild mandarin orange) and Citrus paradisi Macfad. (grapefruit), on feeding by S. litura, and the effects of these plants on the growth of the insect. Based on performance of 3rd instars, Feutrell’s Early and Seedless Kinnow supported maximum relative growth rates, high efficiencies of conversion of ingested food, and high levels of leaf consumption and larval weight gain, relative to the other citrus cultivars. Values of these parameters were lowest on grapefruit. Thus, the mandarin oranges Feutrell’s Early and Seedless Kinnow were more susceptible to damage than grapefruit. Spodoptera litura F. (Lepidoptera: Noctuidae) es una plaga muy importante que ataca adversamente a muchos cultivos importantes tales como algodón, okra, jitomate, papa, y calabaza. Se reporta por primera vez a S. litura atacando en un vivero de cítricos (Sapindales: Rutaceae) en la región de Sargodha, Pakistán. Los objetvos principales de este estudio fueron determinar los efectos del potencial alimenticio de S. litura sobre diferentes variedades de cítricos incluyendo a Citrus reticulata Blanco (Feutrell’s Early, Seedless Kinnow, Fairchild) and Citrus paradisi Macfad. (toronja). El potencial alimenticio de larvas de tercer estadio sugieren que Feutrell’s Early y Seedless Kinnow soportan mayor tasa de crecimiento relativo (TCR) así como los valores más altos de eficiencia de conversión de alimento ingerido (ECI), y consumo de área foliar entre las variedades de cítricos. Los valores más bajos de estos parámetros fueron observados en la variedad de toronja. Los resultados indicaron que Feutrell’s Early y Seedless Kinnow fueron las variedades más nutritivas y susceptibles entre las variedades cítricos ya que mostraron los valores más grandes de TCR, ECI, ganancia de peso larvario, consume de área foliar y pérdida de peso de follaje; por el contrario, en la variedad de toronja se corroboró como la menos nutritiva para S. litura, ya que produjo los valores más bajos de TCR and peso de excretas (PE)

Incidence of \u3ci\u3eSpodoptera litura\u3c/i\u3e (Lepidoptera: Noctuidae) and its feeding potential on various citrus (Sapindales: Rutaceae) cultivars in the Sargodha Region of Pakistan

Spodoptera litura F. (Lepidoptera: Noctuidae) adversely affects important crops such as cotton, okra, tomato, potato, and pumpkin. For the first time, this species was observed in a citrus nursery in the Sargodha Region of Pakistan. The principal objectives of this study were to determine the effects of several citrus (Sapindales: Rutaceae) cultivars, including Citrus reticulata Blanco (Feutrell’s Early, Seedless Kinnow, and Fairchild mandarin orange) and Citrus paradisi Macfad. (grapefruit), on feeding by S. litura, and the effects of these plants on the growth of the insect. Based on performance of 3rd instars, Feutrell’s Early and Seedless Kinnow supported maximum relative growth rates, high efficiencies of conversion of ingested food, and high levels of leaf consumption and larval weight gain, relative to the other citrus cultivars. Values of these parameters were lowest on grapefruit. Thus, the mandarin oranges Feutrell’s Early and Seedless Kinnow were more susceptible to damage than grapefruit. Spodoptera litura F. (Lepidoptera: Noctuidae) es una plaga muy importante que ataca adversamente a muchos cultivos importantes tales como algodón, okra, jitomate, papa, y calabaza. Se reporta por primera vez a S. litura atacando en un vivero de cítricos (Sapindales: Rutaceae) en la región de Sargodha, Pakistán. Los objetvos principales de este estudio fueron determinar los efectos del potencial alimenticio de S. litura sobre diferentes variedades de cítricos incluyendo a Citrus reticulata Blanco (Feutrell’s Early, Seedless Kinnow, Fairchild) and Citrus paradisi Macfad. (toronja). El potencial alimenticio de larvas de tercer estadio sugieren que Feutrell’s Early y Seedless Kinnow soportan mayor tasa de crecimiento relativo (TCR) así como los valores más altos de eficiencia de conversión de alimento ingerido (ECI), y consumo de área foliar entre las variedades de cítricos. Los valores más bajos de estos parámetros fueron observados en la variedad de toronja. Los resultados indicaron que Feutrell’s Early y Seedless Kinnow fueron las variedades más nutritivas y susceptibles entre las variedades cítricos ya que mostraron los valores más grandes de TCR, ECI, ganancia de peso larvario, consume de área foliar y pérdida de peso de follaje; por el contrario, en la variedad de toronja se corroboró como la menos nutritiva para S. litura, ya que produjo los valores más bajos de TCR and peso de excretas (PE)

Maksimalna proizvodnja intracelularne lipaze s pomoću mutanta Rhizopus oligosporus DGM 31: kinetička studija

Regulation and maximization of lipase production in a mutant derivative of R. oligosporus has been investigated using different substrates, inoculum sizes, pH of the medium, temperature, and nitrogen sources in shake flask experiments and batch fermentation in a fermentor. The production of intracellular lipase was improved 3 times following medium optimization involving one-at-a-time approach and aeration in the fermentor. Interestingly, intracellular lipase was poorly induced by oils, instead its production was induced by sugars, mainly starch, lactose, sucrose, xylose, glucose and glycerol. Dependent variables studied were cell mass, lipase activity, lipase yield, lipase specific and volumetric rate of formation. It was confirmed that lipase production in the derepressed mutant is sufficiently uncoupled from catabolite repression. The results of average specific productivities at various temperatures worked out according to the Arrhenius equation revealed that mutation decreased the magnitude of enthalpy and entropy demand in the inactivation equilibrium during product formation, suggesting that mutation made the metabolic network of the organism thermally more stable. The highest magnitudes of volumetric productivity (QP=490 IU/(L·h)) and other product attributes of lipase formation occurring on optimized medium in the fermentor are greater than the values reported by other workers. The purified enzyme is monomeric in nature and exhibits stability up to 80 °C and pH=6.0–8.0. Activation energy, enthalpy and entropy of catalysis at 50 °C, and magnitudes of Gibbs free energy for substrate binding, transition state stabilization and melting point indicated that this lipase is highly thermostable.Primjenom različitih supstrata, veličina inokuluma, pH-vrijednosti podloge, temperature i izvora dušika regulirana je i maksimizirana proizvodnja lipaze s pomoću mutanta Rhizopus oligosporus DGM 31 u pokusima na tresilici i šaržnim procesom u reaktoru. Proizvodnja intracelularne lipaze povećala se 3 puta optimiranjem podloge i prozračivanjem reaktora. Dodatak ulja malo je poboljšao proizvodnju lipaze, a znatno je poboljšana dodatkom šećera, osobito škroba, laktoze, saharoze, ksiloze, glukoze i glicerola. Ispitane su ove zavisne varijable: masa stanica, aktivnost i prinos lipaze, te specifična i volumetrijska brzina nastajanja enzima. Dokazano je da proizvodnja lipaze s pomoću mutanta ne ovisi o represiji s katabolitima. Vrijednosti prosječne specifične produktivnosti pri različitim temperaturama, izračunate prema Arrheniusovoj jednadžbi, pokazuju da mutacija smanjuje količinu entalpije i entropije potrebne za postizanje ravnotežne inaktivacije tijekom formiranja produkta, te da je omogućila odvijanje toplinski puno stabilnijeg metaboličkog procesa u organizmu. Korištenjem optimirane podloge u reaktoru postignuta je dosad najveća volumetrijska produktivnost lipaze (QP=7,3 IU/g škroba/h). Pročišćeni je enzim monomeran i stabilan čak i do 80 °C i pri pH-vrijednosti od 6,0 do 8,0. Veliku termostabilnost lipaze dokazuju: energija aktivacije, entalpija i entropija potrebne za provedbu katalize pri 50 °C, Gibbsova slobodna energija utrošena za vezanje supstrata, stabilnost prijelaznog stanja i točka taljenja

Diagnostic Accuracy of Barium Enema for the Diagnosis of Hirsch-sprung Disease in Children

Objective: To determine the diagnostic accuracy of Barium contrast enema for diagnosis of Hirsch-sprung disease, taking histopathology as the gold standard. Study Design: Cross-sectional validation study. Place and Duration of Study: Department of Paediatric Surgery, Pak Emirates Military Hospital, Rawalpindi Pakistan, from Nov 2019 to May 2021. Methodology: A total of 251 patients with symptoms of Hirsch-sprung disease were included. All patients were assessed with a Barium enema and full-thickness rectal biopsy. Barium enema was reported for the presence of contrast in the rectum after 24 hours, the presence or absence of transitional zone and a Recto Sigmoid Index less than 1. Full-thickness rectal biopsy was taken 2 cm above the dentate line, and histopathology was performed to see the ganglion cells in Meissner`s and Myenteric plexus. Results: Histo-pathologically confirmed cases were 120(true positive), where 12(false positive) had no Hirsch-sprung disease on histopathology. In Barium enema negative patients, 113 were true negatives while 06 were false negatives. Overall sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of Barium contrast enema for diagnosis of Hirsch-sprung disease were 95.24%, 90.40%, 90.91%, 94.96% and 92.83%, respectively. Conclusion: We concluded that the diagnostic accuracy of Barium contrast enema for diagnosis of Hirsch-sprung disease is quite high, taking histopathology as the gold standard

Development of a highly sensitive electrochemical sensing platform for the trace level detection of lead ions.

Herein we report for the first time a highly sensitive electrochemical platform for the trace level detection of Pb (ӏӏ) using glassy carbon electrode modifiedwith 1-dodecanoyl-3-phenylthiourea (DPT). The performance of the designed sensor was tested by electrochemical impedance spectroscopy, chronocoulometry, cyclic voltammetry and Square Wave Anodic Stripping Voltammetry (SWASV). The DPT was found to play an efficient role in enhancing the sensing response of the electrode for the detection of lead ions in aqueous samples. A number of experimental conditions such as deposition potential, accumulation time, surfactant concentration, pH, number of scans and supporting electrolytes were examined to optimize conditions for getting intense signal of the target analyte. Linear calibration curve was obtained using SWAS voltammetric data obtained under optimized conditions. The limit of detection with a value of 0.695 μg/L suggests that the designed sensor can sense lead ions even below the permissible concentration level (10 μg/L) recommended by the World Health Organization and Environmental Protection Agency of USA. The designed sensor demonstrated sensitivity, selectivity and stability for the targeted analyte. Percentage recoveries from real water samples with standard deviations of less than 2% suggested precision of the proposed method. Moreover, computational findings supported the experimental outcomes

Laparoscopy in management of appendicitis in high-, middle-, and low-income countries: a multicenter, prospective, cohort study.

BACKGROUND: Appendicitis is the most common abdominal surgical emergency worldwide. Differences between high- and low-income settings in the availability of laparoscopic appendectomy, alternative management choices, and outcomes are poorly described. The aim was to identify variation in surgical management and outcomes of appendicitis within low-, middle-, and high-Human Development Index (HDI) countries worldwide. METHODS: This is a multicenter, international prospective cohort study. Consecutive sampling of patients undergoing emergency appendectomy over 6 months was conducted. Follow-up lasted 30 days. RESULTS: 4546 patients from 52 countries underwent appendectomy (2499 high-, 1540 middle-, and 507 low-HDI groups). Surgical site infection (SSI) rates were higher in low-HDI (OR 2.57, 95% CI 1.33-4.99, p = 0.005) but not middle-HDI countries (OR 1.38, 95% CI 0.76-2.52, p = 0.291), compared with high-HDI countries after adjustment. A laparoscopic approach was common in high-HDI countries (1693/2499, 67.7%), but infrequent in low-HDI (41/507, 8.1%) and middle-HDI (132/1540, 8.6%) groups. After accounting for case-mix, laparoscopy was still associated with fewer overall complications (OR 0.55, 95% CI 0.42-0.71, p < 0.001) and SSIs (OR 0.22, 95% CI 0.14-0.33, p < 0.001). In propensity-score matched groups within low-/middle-HDI countries, laparoscopy was still associated with fewer overall complications (OR 0.23 95% CI 0.11-0.44) and SSI (OR 0.21 95% CI 0.09-0.45). CONCLUSION: A laparoscopic approach is associated with better outcomes and availability appears to differ by country HDI. Despite the profound clinical, operational, and financial barriers to its widespread introduction, laparoscopy could significantly improve outcomes for patients in low-resource environments. TRIAL REGISTRATION: NCT02179112

Postoperative outcomes in oesophagectomy with trainee involvement

BACKGROUND: The complexity of oesophageal surgery and the significant risk of morbidity necessitates that oesophagectomy is predominantly performed by a consultant surgeon, or a senior trainee under their supervision. The aim of this study was to determine the impact of trainee involvement in oesophagectomy on postoperative outcomes in an international multicentre setting. METHODS: Data from the multicentre Oesophago-Gastric Anastomosis Study Group (OGAA) cohort study were analysed, which comprised prospectively collected data from patients undergoing oesophagectomy for oesophageal cancer between April 2018 and December 2018. Procedures were grouped by the level of trainee involvement, and univariable and multivariable analyses were performed to compare patient outcomes across groups. RESULTS: Of 2232 oesophagectomies from 137 centres in 41 countries, trainees were involved in 29.1 per cent of them (n = 650), performing only the abdominal phase in 230, only the chest and/or neck phases in 130, and all phases in 315 procedures. For procedures with a chest anastomosis, those with trainee involvement had similar 90-day mortality, complication and reoperation rates to consultant-performed oesophagectomies (P = 0.451, P = 0.318, and P = 0.382, respectively), while anastomotic leak rates were significantly lower in the trainee groups (P = 0.030). Procedures with a neck anastomosis had equivalent complication, anastomotic leak, and reoperation rates (P = 0.150, P = 0.430, and P = 0.632, respectively) in trainee-involved versus consultant-performed oesophagectomies, with significantly lower 90-day mortality in the trainee groups (P = 0.005). CONCLUSION: Trainee involvement was not found to be associated with significantly inferior postoperative outcomes for selected patients undergoing oesophagectomy. The results support continued supervised trainee involvement in oesophageal cancer surgery