Characterization of sugarcane mutants developed through gamma irradiations for their lignin content and caffeic acid-O-methyl transferase (COMT) gene mutations.

PURPOSE: Bagasse, the residue left after extracting juice from sugarcane stalks, is rich in lignocellulosic biomass. The lignin present in this plant biomass is the key factor that hinders the efficient extraction of ethanol from the bagasse. In the current study, γ-irradiated sugarcane mutants were evaluated for variation in lignin content and its corresponding caffeic acid-O-methyl transferase (COMT) gene. MATERIALS AND METHODS: The acetyl bromide method was used to estimate lignin content in sugarcane mutants. PCR-based cloning of the COMT gene was performed in low lignin mutants as well as control plants in E. coli (strain DH5α) to understand the mechanism of variation at the molecular level. The Sanger sequencing for cloned gene was performed to check variation in gene sequence. RESULTS: In comparison to the control (21.5%), the mutant plants' lignin content ranged from 13 to 28%. The Sanger sequencing revealed approximately the same length of the gene from mutants as well as a control plant. In comparison to the reference gene, the mutated gene showed SNPs and indels in different regions, which may have an impact on lignin content. CONCLUSIONS: Therefore, γ-irradiated mutagenesis is an acceptable approach to develop novel mutants of sugarcane with low lignin content to enhance bioethanol production from waste material using bioprocess technology.

Defensive capabilities of contrasting sorghum genotypes against Atherigona soccata (Rondani) infestation.

Plants are equipped with a wide range of defensive mechanisms such as morphophysiological, biochemical, molecular, and hormonal signaling for protecting against insect-pest infestation. The infestation of a devastating pest shoot fly [Atherigona soccata (Rodani)] at seedling stage causes huge loss of sorghum crop productivity. In morphophysiological screening ICSV700, ICSV705, and IS18551 have been categorized as resistant, PSC-4 moderately resistant, SL-44 and SWARNA as susceptible. The present study focused on the role of defensive gene expression and its products viz: superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), polyphenol oxidase (PPO), phenyl alanine ammonia lyase (PAL), responsive enzymes, and metabolites restoring redox status in sorghum plants against shoot fly infestation. In both leaf and stem tissue of sorghum genotypes, shoot fly infestation induced SOD, APX, DHAR, GR, PAL, and PPO activities while CAT activity was significantly declined at 15 and 21 days after emergence (DAE). IS18551 with resistant behavior showed upregulation of SOD, GR, APX, and DHAR along with accumulation of ascorbate, glutathione enhancing redox status of the plant during shoot fly infestation at later stage of infestation. While SWARNA with susceptible response exhibited enhanced activity of phenylpropanoid pathway enzymes PAL and PPO which in turn increased the levels of secondary metabolites like o-dihydroxyphenol and other phenols deterring the insect to attack the plant. The qRT-PCR data predicted that stress-responsive genes were initially unregulated in SWARNA; however, at 21 DAE, multifold higher expression of SOD, CAT, APX, and PPO (24.8-, 37.2-, 21.7-, and 17.9-fold respectively) in 1S18551 indicates the resistance behavior of this genotype against insect infestation owing to sustainable development capability.

Genu valgum deformity - correction by a wedgeless implantless femoral "V" osteotomy.

Coronal malalignment of the knee joint is very common in developing countries especially because of nutritional rickets. Significant valgus deformity needs to be treated surgically to improve appearance, gait and function of the patient. The purpose of this prospective study was to evaluate the results of supracondylar "V" osteotomy as a surgical technique for correction of the valgus knee deformity. This study was conducted in a tertiary level teaching hospital and 30 cases were included in the study. For all the patients deformity was assessed using ana- tomical tibiofemoral angle, mechanical axis deviation and intermalleolar distance preoperatively and post- operatively. The average age of our patients was 13.7 years and the average follow up was3.29 years (1.39-14.22 yrs). Clinically the average value of intermalleolar distance preoperatively was 16cm and 3.2 cm postperatively. Average pre-operative tibiofemoral angle was 23° and the average postoperative angle was 6 0 which was found to be statistically significant using the Paired t test (p<0.005). The average value of preoperative mechanical axis deviation was 3.1 cm which decreased to an average value of 1.1 cm postoperatively. The results with this technique have been encouraging. The advantages of this technique are low morbidity, good stability allowing early ambulation, ability to adjust alignment postoperatively by casting and no need for internal fixation. Few studies have been conducted on osteotomies that do not require internal fixation and are inherently stable. This technique has the advantage of practically no occurrence of any infection or a second surgery to remove hardware in children and adolescents. Since no specialized instrumentation, image intensifier and implants are required, it is cost effective and can be used in any primary care or district level surgical setup in a developing country like ours.

Biodegradable composite films/coatings of modified corn starch/gelatin for shelf life improvement of cucumber.

Composite films comprising modified corn starch and gelatin were developed using a solvent casting technique. The effect of varied content of citric acid and gelatin on the functional properties of the corn starch and composite films, respectively, was investigated. Modified corn starch films enriched with 5% citric acid presented excellent film properties like solubility, swelling index, water vapor permeability, and mechanical property as compared to other films. Similarly, composite films enriched with a ratio (4:1) also exhibited excellent film properties like mechanical and opacity than other composite films. FTIR results confirmed the cross linking among the modified corn starch chains, which is responsible for the enhancement in the film properties. From SEM results, it was noticed that the modified corn starch films exhibited slightly rougher, less shiny than the composite films. Efficacy of the composite solution as a coating formulation in terms of the quality and shelf life of the cucumber investigated. From the results, it was noted that the composite film forming solution as a coating formulation has the potential to extend the shelf-life of the cucumbers additionally up to 16 days.

Morphophysiological and biochemical attributes influence intra-genotypic preference of shoot fly [Atherigona soccata (Rondani)] among sorghum genotypes.

Shoot fly [Atherigona soccata (Rondani)] is a destructive pest of sorghum at the seedling stage and causes huge losses to grain yield and green fodder. The host-plant resistance mechanism is the best approach to reduce the attack of insects in plants. The damage parameters, morphophysiological traits, and biochemical metabolites had been investigated in the leaves and stem of contrasting sorghum genotypes, viz., resistant (IS18551, ICSV705, ICSV700), moderately resistant (PSC-4), and susceptible (SWARNA and SL-44) at 15 and 21 days after emergence (DAE) against shoot fly infestation. The resistant genotypes recorded lowest shoot fly oviposition and incidence (0.3-0.7 eggs plant-1 and 10-15%) than the susceptible genotypes (2.4-3.0 eggs plant-1 and 70-80%), respectively. The susceptible genotype SWARNA recorded 50% and 80% higher deadheart formation than the resistant genotype IS18551 at 15 and 21 DAE, respectively. Resistant genotypes exhibited higher trichome density at adaxial and abaxial part of leaf (118-145 and 106-131) with pink colored leaf sheath (scale 1.50-3.25), glossy leaves (scale1.00-1.25), and lower leaf surface wetness (scale1.25-2.00) compared with susceptible genotype with 49.3-73.3 and 25.3-64.0, scale 2.50-4.00, scale 2.75-3.50, and scale 3.25-4.25 for the respective parameters. Another defense response of sorghum toward the insect attack was modulation of plant metabolism. The infested genotypes responded to insect attack by upregulation of total soluble sugar, total phenol, prussic acid, and chlorophyll content by 1.2-2.1-fold, 1.5-2.0-fold, 1.2-1.3-fold, and 1.2-3.9-fold with more induction in susceptible genotypes at 21 DAE. On the whole, the present study indicates that morphophysiological and biochemical attributes contribute toward the resistance mechanism in sorghum against shoot fly infestation.

Metabolic adjustments in forage oat (Avena sativa L.) genotypes under different sowing windows.

The current investigation was aimed to study the influence of planting dates (9 Oct, 24 Oct, 13 Nov, 26 Nov, 11 Dec, and 26 Dec) and maturity stages (40, 50, and 60 DAS) on nitrogen metabolic enzymes and biochemical constituents. The experiment was laid out in randomized block design during the winter season of 2019 by using two oat genotypes (OL-10 and OL-11) with variable growing ability. The activity pattern of studied enzymes revealed upregulated nitrate assimilation during optimum (Oct 24) and sub-optimum (Oct 9 and Nov 13) planting dates but a reverse trend was observed during late planting dates; LPD (Nov 26, Dec 11, and Dec 26). The environmental constraints during LPD resulted in the accrual of nitrate above toxic levels (> 2000 ppm). The regression analysis depicted a significant relationship of nitrate assimilating enzymes and nitrate-N with temperature and sunshine hour. Examination of ammonia assimilation and transaminases suggested that the enzyme activities got uplifted during late planting dates but were stable or slightly low at optimum and sub-optimum ones. Additionally, OL-10 proved to be an N-efficient genotype in comparison with the OL-11 genotype because of its high N assimilation potential. Therefore, gaining a better understanding of planting time is crucial for sustainable livestock production.

Moth bean starch (Vigna aconitifolia): isolation, characterization, and development of edible/biodegradable films.

In the current work, moth bean starch was separated from the moth bean seeds which were cultivated in the semi arid regions of Haryana and Rajasthan, India using alkali treatment technique and characterized. Isolated and purified moth bean starch comprised (12.89-20.37%) amylose, 0.8% protein, 0.4% ash, swelling index and solubility were 10.8-14.7% and 6.4-9.8% respectively. For wrapping application, starch was modified using citric acid as cross linking agent (1-7% w/w of total starch) and film was made using casting method, and investigated the influence of citric acid on the functional properties of the films such as moisture content, solubility, swelling index, water vapor permeability and mechanical property. Significant difference in the functional properties among native and modified moth bean starch films was noticed. Interactions among starch chains due to cross linking in the modified starch films were reported using FTIR spectra. Surface micrographs of both purfied starch granules and films were studied using scanning electron microscopy. From the outcomes, it was exhibited that obtained starch granules has range large to small size with distorted cylinder and oval shapes. While cross linked starch films showed compact, slightly rough and homogeneous surface. Tested modified moth bean starch films containing citric acid (CA) and sorbitol were utilized as wrapping films to improve the shelf life of fresh lemon. Outcomes showed that the film contains 5% CA could be most promising wrapping materials for lemon that enhanced the shelf life of lemon additionally up to 12 days.

Transverse Vaginal Septum With Secondary Infertility: A Rare Case.

STUDY OBJECTIVE: To demonstrate the technique of laparoscopic vaginal reconstruction in a rare case of mid-vaginal septum with secondary infertility. DESIGN: A step-by-step explanation of the technique using videos and pictures (Canadian Task Force classification IV). SETTING: Transverse vaginal septum is a rare condition, with an incidence of only 1 in 30,000 women. It is usually a congenital mullerian fusion defect; few cases of acquired septum have been reported. Roughly 40% of cases occur in the mid-vagina. Transverse vaginal septum typically presents with primary amenorrhea and hematocolpos. The goal of surgery is to create a patent vagina with restoration of fertility. The laparoscopic approach has proven superiority over the open technique. INTERVENTIONS: Laproscopic vaginal reconstruction was performed in a 24 year female with transverse vaginal septum. Dissection was done laproscopically up to mid vagina. Incision was given on vagina excising the septum followed by reconstruction. CONCLUSION: The laparoscopic approach to vaginal reconstruction avoids the abdominal incision, with its associated pain and possible adhesion formation. It also provides a better view for dissection. In this patient, a patent vagina was created in a single operation, with no postoperative dyspareunia, and fertility was restored.

Removal of mercury from water by fixed bed activated carbon columns.

The breakthrough curves for Hg(II) ions on a sample of granulated activated carbon (GAC) and a sample of activated carbon cloth (ACC) are generally S-shaped. The breakthrough time increases with increase in the bed depth but decreases on increasing the hydraulic loading rate (HLR) and the feed concentration. The adsorption of Hg(II) ions increases with HLR and attains a maximum value at HLR around 7m(3)/h/m(2). At low HLR, laminar flow conditions prevail so that the mass transfer takes place across a nearly stationary film of the liquid covering the carbon particles. This high resistance leads to low mass transfer and results in smaller adsorption. On increasing HLR, the interface resistance decreases resulting in an increase in adsorption. Beyond a certain HLR, the rate of adsorption decreases due to decrease in the residence time of the solution within the carbon bed and a lower time available for mass transfer. The adsorption zone parameters of the carbon column have been determined using the carbon bed column data and invoking the mathematical treatment suggested by Michaels. Bed Depth Service Time (BDST) theoretical model has been used to calculate the critical bed depth and the depth of the mass transfer zone. These have been found to be in agreement with the experimental values.

Xylanase production with xylan rich lignocellulosic wastes by a local soil isolate of Trichoderma viride / Produção de xilanase com resíduos lignocelulósicos ricos em xilana por uma cepa local de Trichoderma viride isolada de solo

In the present study, cultural and nutritional conditions for enhanced production of xylanase by a local soil isolate of Trichoderma viride, using various lignocellulosic substrates in submerged culture fermentation have been optimized. Of the lignocellulosics used, maize straw was the best inducer followed by jowar straw for xylanase production. The highest activity achieved was between 14 to 17 days of fermentation. A continuous increase in xylanase production was observed with increasing level of lignocellulosics in the medium and highest activity was observed with maize straw at 5 percent level. Xylanase production with higher levels of lignocellulosics (3 to 5 percent) of maize, jowar and barseem was found to be higher as compared to that with commercial xylan as carbon source. Sodium nitrate was the best nitrogen source among the six sources used. Maximum xylanase production was achieved with initial medium pH of 3.5-4.0 and incubation temperature of 25ºC.The enzyme preparation was effective in bringing about saccharification of different lignocellulosics. The xylanase production could be further improved by using alkali treated straw as carbon source.

Neste estudo, otimizou-se as condições culturais e nutricionais para produção aumentada de xilanase por uma cepa local de Trichoderma viride isolada de solo, empregando-se vários substratos lignocelulósicos, em fermentação submersa. Entre os substratos utilizados, o melhor indutor de produção de xilanase foi palha de milho, seguido de palha de sorgo. A atividade mais alta foi obtida entre 14 e 17 dias de fermentação. Com palha de milho observou-se um aumento contínuo na produção de xilanase com o aumento da concentração dos substratos lignocelulósicos no meio, sendo que a melhor atividade foi obtida com 5 por cento de palha de milho. A produção de xilanase com níveis mais altos de (3 a 5 por cento) de milho, sorgo e forragem verde (barseem) foi mais levada do que com xilana comercial como fonte de carbono. Entre as fontes de nitrogênio testadas, a melhor foi nitrato de sódio. Produção máxima de xilanase foi obtida quando o pH inicial do meio foi 3,5 4,0 e a temperatura de incubação 25ºC. A enzima foi eficiente na sacarificação de diferentes substratos lignocelulósicos. A produção de xilanase poderia ser aumentada empregando-se álcali ao invés de palha tratada como fonte de carbono.

Xylanase production with xylan rich lignocellulosic wastes by a local soil isolate of Trichoderma viride.

In the present study, cultural and nutritional conditions for enhanced production of xylanase by a local soil isolate of Trichoderma viride, using various lignocellulosic substrates in submerged culture fermentation have been optimized. Of the lignocellulosics used, maize straw was the best inducer followed by jowar straw for xylanase production. The highest activity achieved was between 14 to 17 days of fermentation. A continuous increase in xylanase production was observed with increasing level of lignocellulosics in the medium and highest activity was observed with maize straw at 5% level. Xylanase production with higher levels of lignocellulosics (3 to 5%) of maize, jowar and barseem was found to be higher as compared to that with commercial xylan as carbon source. Sodium nitrate was the best nitrogen source among the six sources used. Maximum xylanase production was achieved with initial medium pH of 3.5-4.0 and incubation temperature of 25ºC.The enzyme preparation was effective in bringing about saccharification of different lignocellulosics. The xylanase production could be further improved by using alkali treated straw as carbon source.

Use of phospholipid transfer protein as a probe to study the lipid dynamics and alkaline phosphatase activity in the brush border membrane of human term placenta.

Incubation of placental brush border membrane (BBM) along with sonicated vesicles of exogenous lipids (egg yolk PC) in the presence of phospholipid-transfer protein (PL-TP) showed a decrease in the alkaline phosphatase activity due to the change in the membrane micro-environment, such as fluidity. Effect of substrate concentration was tested by Lineweaver-Burk plot, which showed decreased V(max) and K(M). The effect of temperature was probed by the Arrhenius plot, which showed no change in transition temperature, but a decline in the energy of activation both below and above the transition temperature. The protein-catalyzed transfer of phospholipid from the donor unilamellar vesicles resulted in a substantial increase in the BBM phospholipid and a net decrease in cholesterol/phospholipid molar ratio. The change in membrane fluidity was assessed by translational as well as rotational diffusion of membrane extrinsic fluorescent probes, pyrene and diphenyl-hexatriene. An increased lateral mobility was recorded by the increased pyrene excimer formation. A decrease in fluorescent polarization of diphenyl-hexatriene was observed, which led to the decrease in fluorescence anisotropy and order parameter, and therefore, an increase in membrane fluidity (rotational diffusion). Mean anisotropy parameter was also decreased in the presence of PL-TP. Thus, the placental BBM alkaline phosphatase activity showed a distinct lipid dependence which may have important physiological consequences.

Is Chlamydia pneumoniae infection associated with stroke in children with sickle cell disease?

BACKGROUND: Stroke is often a devastating complication of sickle cell disease (SCD). Most children with SCD-related stroke have stenotic and occlusive disease of cerebral blood vessels due to intimal hyperplasia. This hyperplasia is hypothesized to result from an inflammatory response similar to that in atherosclerosis and has been attributed to infection by Chlamydia pneumoniae. OBJECTIVE: To determine whether C pneumoniae infection is associated with stroke and cerebrovascular disease, including transient ischemic attacks and abnormal transcranial Doppler examinations, in children with SCD. METHODS: Children with SCD on chronic transfusion due to a history of stroke, transient ischemic attack, or abnormal transcranial Doppler; children with SCD without stroke; healthy controls; and children being transfused for other reasons were enrolled. Peripheral blood and nasopharyngeal (NP) swab specimens were collected from all patients. In patients on transfusion, pretransfusion specimens and samples from the unit of packed red blood cells being transfused were obtained. Peripheral blood monocytic cells (PBMCs) and NP swab specimens were cultured for C pneumoniae in HEp-2 cells. C pneumoniae polymerase chain reaction was performed on PBMCs with a nested touch-down method with primers from the omp-1gene (in duplicate) and a second real-time polymerase chain reaction by using 16S ribosomal RNA primers. RESULTS: C pneumoniae DNA was detected in the PBMCs of 1 of 14 (7.1%) children with SCD on chronic transfusion, 1 of 10 (10%) sickle cell controls, 1 of 10 (10%) healthy controls, and none of the 5 children receiving chronic transfusion for other reasons. It was not detected in specimens from transfusion units. One child with SCD and stroke, 1 sickle cell control, and 1 transfusion control had positive NP cultures for C pneumoniae. C pneumoniae DNA was not detected in their PBMCs, and all 3 children were asymptomatic. C pneumoniae was not detected by culture of PBMCs from any of the patients after 7 passages. CONCLUSION: Stroke in children with SCD does not seem to be associated with C pneumoniae infection in our population.