Evaluation of free and immobilized cellulase on chitosan-modified magnetic nanoparticles for saccharification of sorghum residue.

Enzymatic hydrolysis plays a pivotal role in transforming lignocellulosic biomass. Addressing alternate techniques to optimize the utilization of cellulolytic enzymes is one strategy to improve its efficiency and lower process costs. Cellulases are highly specific and environmentally benign biocatalysts that break down intricate polysaccharides into simple forms of sugars. In contrast to the most difficult and time-consuming enzyme immobilization processes, in this research, we studied simple, mild, and successful techniques for immobilization of pure cellulase on magnetic nanocomposites using glutaraldehyde as a linker and used in the application of sorghum residue biomass. Fe3O4 nanoparticles were coated with chitosan from the co-precipitation method, which served as an enzyme carrier. The nanoparticles were observed under XRD, Zeta Potential, FESEM, VSM, and FTIR. The size morphology results presented that the Cs@Fe3O4 have 42.2 nm, while bare nanoparticles (Fe3O4) have 31.2 nm in size. The pure cellulase reaches to 98.07% of loading efficiency and 71.67% of recovery activity at optimal conditions. Moreover, immobilized enzyme's pH stability, thermostability, and temperature tolerance were investigated at suitable conditions. The kinetic parameters of free and immobilized enzyme were estimated as Vmax; 29 ± 1.51 and 27.03 ± 2.02 µmol min-1 mg-1, Km; 4.7 ± 0.49 mM and 2.569 ± 0.522 mM and Kcat; 0.13 s-1, and 0.89 s-1. Sorghum residue was subjected to 2% NaOH pre-treatment at 50 â„ƒ. Pre-treated biomass contains cellulose of 64.8%, used as a raw material to evaluate the efficiency of reducing sugar during hydrolysis and saccharification of free and immobilized cellulase, which found maximum concentration of glucose 5.42 g/L and 5.12 g/L on 72 h. Thus, our study verifies the use of immobilized pure cellulase to successfully hydrolyze raw material, which is a significant advancement in lignocellulosic biorefineries and the reusability of enzymes.

Cannabidiol-Based Prodrugs: Synthesis and Bioevaluation.

Cannabidiol (CBD 1) is a nonpsychotic cannabinoid-based drug approved by the U.S. FDA for treating refractory epilepsy, namely, Lennox-Gastaut and Dravet syndrome. However, its low aqueous solubility and oral bioavailability are compensated by administering high doses, and there is an increased demand for conjugates with improved properties. In this direction, the present work is focused on synthesizing CBD-based prodrugs to address the issue of poor solubility and oral bioavailability. Several CBD-based prodrugs were synthesized and studied in a battery of assays: viz, release kinetic (ex vivo), solubility (in vitro), chemical stability (in vitro), plasma stability (ex vivo), pharmacokinetics (in vivo), and efficacy studies (in vivo). Among the synthesized prodrugs, the morpholinyl CBD-based prodrugs 3a and 3aa showed good release behavior, stability, better solubility, and a plasma profile. Moreover, prodrug candidate 3aa showed better therapeutic efficacy. The present study identifies CBD-based prodrugs with improved physiochemical properties and oral exposure.

Effect of dextransucrase antibodies on biofilm formation and certain cariogenic activities in Streptococcus mutans.

Introduction. Dextransucrase produced by Streptococcus mutans plays a vital role in the formation of dental caries by synthesizing exopolysaccharides from sucrose, which helps in the attachment of microbes to the tooth surface, causing caries. Exploring antibody production against S. mutans antigens could be an effective method to protect against dental caries.Hypothesis. Dextransucrase antibodies may help in the prevention of caries formation by inhibiting essential cariogenic factors.Aims. The aim of this study was to investigate the effects of dextransucrase antibodies on biofilm formation and certain associated cariogenic factors of S. mutans.Methodology. Dextransucrase was purified from culture of S. mutans. The antisera against the enzyme were raised in rabbits. The effect of dextransucrase antibodies on biofilm formation was studied using scanning electron microscopy, fluorescence microscopy and quantitative real-time polymerase chain reaction. The effects of the antibodies on associated cariogenic factors were examined using established methods. The cross-reactivity of antibodies with human lung, liver, heart, thyroid and kidney tissues was evaluated by immunohistochemistry.Results. Our findings showed impaired biofilm formation in S. mutans in the presence of dextransucrase antibodies. Genes associated with biofilm formation such as gtfB, gtfC, brpA, relA, Smu.630 and vicK were downregulated (50-97 %) by dextransucrase antibodies in S. mutans. The adherence of S. mutans to glass surface was reduced by 58 % and hydrophobicity was reduced by 55.2 % in the presence of the antibodies compared to the controls. Immunohistochemistry studies revealed no cross-reactivity of human tissues with dextransucrase antibodies.Conclusions. These findings suggest that antibodies raised against dextransucrase exhibit a profound inhibitory effect on biofilm formation and vital cariogenic factors of S. mutans, which supports the contention that dextransucrase could be a promising antigen to study for its anticariogenic potential.

Impact of buffy coat reduction on the severity of febrile nonhemolytic transfusion reactions with red cell components.

BACKGROUND: Febrile nonhemolytic transfusion reactions (FNHTRs) are the most common adverse reaction reported under the Haemovigilance Programme of India, and the use of leukodepleted blood products is recommended. The severity of the reaction may affect the morbidity associated with the reaction. This study aims to calculate the incidence of various transfusion reactions in our blood center and to evaluate the impact of buffy coat reduction on the severity of febrile reaction and other hospital resource-consuming activities. MATERIALS AND METHODS: It was an observational retrospective study in which all reported FNHTRs were evaluated during the period July 1, 2018-July 31, 2019. Patient demographic details, component transfused, and clinical presentation were analyzed to identify factors affecting the severity of FNHTRs. RESULTS: The incidence of transfusion reaction in our study period was 0.11%. Out of total 76 reactions reported, 34 (44.7%) were febrile reactions. Other reactions included allergic reactions (36.8%), pulmonary reactions (9.2%), transfusion-associated hypotension (3.9%), and others (2.7%). The incidence of FNHTR in buffy coat-depleted packed red blood cells (PRBCs) and PRBCs is 0.03% and 0.05%, respectively. FNHTRs are seen more in females with prior history of transfusion (87.5%) as compared to males (66.67%) (P = 0.046). We also found that FNHTRs are less severe with buffy coat-depleted PRBC transfusion than PRBC transfusion as mean ± standard deviation temperature rise was less in buffy coat-depleted PRBC (1.3 ± 0.8) than PRBC (1.74 ± 1.129). The febrile response to buffy coat-depleted PRBC transfusion occurred at higher volume (145 ml) transfusion than PRBC transfusion (87.2 ml), and it was statistically significant (P = 0.047). CONCLUSION AND SUMMARY: Leukoreduction remains the main modality to prevent FNHTR, but in developing countries like India, the use of buffy coat-depleted PRBC over PRBC can reduce the incidence and severity of FNHTR.

Massive transfusion protocol: Need of the hour - A tertiary care centre experience.

Background and Aims: Massive transfusion (MT) in critically ill patients during major volume losses can lead to serious adverse outcomes. Studies have reported that rampant red cell infusion for maintaining perfusion support has had detrimental effects on patients' short- and long-term survival rates. Evidence-based studies quote the importance of maintaining blood product ratio during massive hemorrhage and ensuring good outcomes with the least morbidity and mortality. Material and Methods: It is an observational study to compare the ratio of usage of blood products and their role in the outcome of MT cases. Results: A total of 70 patients (29 females and 41 males) who received MT were included in the study. There was no fixed ratio of packed red blood cells (PRBC) to blood components for patients with massive hemorrhage. The average ratio of PRBC: fresh frozen plasma (FFP):platelet concentrate (PC) was 1:0.9:0.6. However, blood component therapy with PRBC: FFP ratio between 1 and 2 was associated with a significant rise in post-acute phase hemoglobin value (P value = 0.018). Conclusion: Appropriate blood component therapy during the acute bleeding phase in massively transfused patients can further decrease the transfusion demand and transfusion-related complications. There is a need to adhere to the MT protocol for the clinical areas requiring MT in the developing world too.

The spectrum of ABO haemolytic disease of the fetus and newborn in neonates born to group O mothers.

BACKGROUND AND OBJECTIVES: ABO haemolytic disease of the fetus and newborn (HDFN) is a lesser recognized entity; however, the severity may vary in neonates. This prospective observational study was performed to determine the severity and risk of ABO-HDFN in neonates born to O group mothers. MATERIALS AND METHODS: A total of 260 neonates born to non-alloimmunized blood group O mothers were recruited. Blood group O neonates were excluded from the study. Neonatal direct antiglobulin test (DAT) was performed using the column agglutination technique. They were monitored for clinical and laboratory parameters and followed up at 6-8 weeks. The maternal anti-A and anti-B titres (IgM and IgG) were also done. RESULTS: A total of 176 neonates with blood group A (77/260; 29.6%) and B (99/260; 38.1%) were finally included in the study, and 15 (8.5%) of them were DAT positive. Overall, 26.7% (47/176) neonates received phototherapy, 172 (97.7%) survived and none required readmission. The median (inter-quartile range [IQR]) maternal IgG anti-B titre (32 [32-64]) was significantly higher (p < 0.001) than the IgG anti-A titre (16 [8-64]). The maximum total serum bilirubin in neonates had a significant positive association with neonatal birth weight (p = 0.045), positive DAT (p = 0.006) and requirement of phototherapy (p < 0.001). The relative risk (95% CI) of a DAT-positive neonate requiring phototherapy was 4.55 (3.12-6.33). CONCLUSION: The frequency of ABO incompatibility in neonates born to group O mothers was 67.69% (176/260). The maternal IgG titre of ≥64 could be a good predictor for identifying the neonates at risk of developing hyperbilirubinaemia requiring phototherapy.

Exploration of COVID-19 related fears deterring from blood donation in India.

INTRODUCTION: The coronavirus pandemic (COVID-19) has impacted and pushed the healthcare settings to extremes across the globe. It was extremely challenging to sustain blood donation, and strategies could be formulated on knowing fears hindering blood donation. METHODS: A cross-sectional survey using Google Forms® through WhatsApp and email after obtaining the ethical clearance. The survey questionnaire was validated for content using the Delphi technique, and pilot tested for finalization. RESULTS: The survey was attempted by 1066 participants, and 749 participants who had not donated since pandemic were included in the study. A little more than half, 415 (55%) reported either one or more than one fear during the pandemic which hindered blood donation. They reported lack of confidence in the safety measures at the hospitals and fear of transmitting infection to family, in 415 (55%) of the participants each, respectively. The fear of COVID-19 hospital infection risk and hospital entry was statistically significant across the age groups that are eligible for blood donation. CONCLUSIONS: The clear and dedicated confidence building measures to sustain blood donation using all communication modalities clearly emerge as the most important strategies to augment blood donation in the COVID-19 pandemic. The measures should include information about implementation of safety measures to mitigate COVID-19 transmission at the blood centres and that the act of blood donation does not increase risk of COVID-19 and therefore the risk of transmission of infection to family.

A Study on the Effect of Pre-donation Salt Loading on Vasovagal Reactions in Young College Going Whole Blood Donors.

INTRODUCTION: The pathophysiology of vasovagal reactions (VVRs) involves both psychological and physiological components. Strategies which could allay physiological changes include interventions like pre-donation water intake and applied muscle tension have been published, however salt loading has not been tested. MATERIALS AND METHODS: Cross sectional study enrolling 1000 young college going whole blood donors with intervention 250 ml of salted loaded water or plain water as placebo. The immediate VVRs were recorded with respect to age, gender, donation status, blood volume, blood volume drawn and BMI. RESULTS: VVRs occurred in 25 out of 1000 (2.5%) young college going whole blood donors. Overall there were 18 VVRs in 526 (3.4%) donors in the placebo arm compared to 7 in 474 (1.5%) in salt loaded arm with odds of 2.36 (p = 0.049), however the difference in means of VVRs between the study arms could not achieve statistical significance on binary logistic regression. The independent risk factors including age, gender, blood volume, blood volume withdrawn and BMI or the donation status were not found to be effect modifiers on the occurrence of VVRs. CONCLUSION: Salt loading before blood donation in young college going whole blood donors does decrease the VVRs in the immediate post donation period; however the decrease was limited to a trend and could not attain statistical significance.

First report of the impact on voluntary blood donation by the blood mobile from India.

INTRODUCTION: The blood mobile is one of the modern methods of mobile blood collection facility funded through the third phase of National AIDS Control Programme by the National Blood Transfusion Council of India. MATERIAL AND METHODS: A retrospective analysis of data in relation to the blood mobile was carried out with respect to the number of blood donation camps, number of blood units collected, adverse donor reactions, and the expenditure that occurred during the blood collection in the blood mobile from 1(st) January 2012 to 30(th) June 2014. RESULTS: There were 64, 84 and 62 blood donation camps conducted in the blood mobile with collection of 3301, 5166 and 2842 blood units during 2012, 2013 and the first half of 2014. The percentage of voluntary blood collection in blood mobile was 8.5% in 2012, increased to 12.4% in 2013 and stands at 14.39% in the first half of 2014. The difference in the means of the adverse donor reactions in the blood mobile and the outdoor camps was not statistically significant. DISCUSSION AND CONCLUSION: The blood mobile is definitely an asset as far as augmentation of voluntary blood donation is concerned, ensures stable collection of blood for better provision of blood and blood components. However the facility requires a comprehensive annual maintenance with incorporation of onsite quick response team both from the manufacturer of the vehicle, and the blood collection equipments. Adequate provision of funding for operational expenditure would in turn facilitate optimum utilization of this facility.

Retrorectal cystic hamartoma (tailgut cyst): report of a case and review of literature.

Retrorectal cystic hamartoma is a rare type of congenital cystic lesion usually diagnosed in middle-aged women. Although mostly asymptomatic, patients may present with symptoms resulting from local mass effect or with a complication. The most important complications of these cysts are infection with secondary fistulization and malignant degeneration. Because of such non-specific nature of symptoms and rare incidence, it is not unusual to have a delay in clinical diagnosis. MRI has evolved to be the investigation of choice for the evaluation of presacral tumors as it can provide excellent anatomic detail and soft tissue contrast. Role of preoperative biopsy is controversial especially with improvements in the imaging techniques. When diagnosed these lesions should be excised because of the risk of malignant transformation.

Autoimmune hemolytic anemia due to auto anti-N in a patient with sepsis.

We describe a case of autoimmune hemolytic anemia (AIHA) due to anti-N in a young male patient with cellulitis. There have been several reports of anti-N in N positive individuals. But in all these reports, auto anti-N was mostly associated with underlying immunological conditions. We report here a case of auto anti-N in a patient of bacterial sepsis without any underlying immune disorder.

Application of response surface analysis for biodegradation of azo reactive textile dye using Aspergillus foetidus.

This paper reports the application of experimental design methodology for the optimization of decolourization of azo reactive textile dye Remazol Red RR and reduction of chemical oxygen demand (COD) using fungal isolate Aspergillus foetidus. Response surface methodology (RSM), involving central composite design matrix in three most important input variables; temperature, pH and initial dye concentration was employed. A total of 20 experiments were conducted in the study towards the construction of a quadratic model. This demonstrated the benefits of approach in achieving excellent predictions, while minimizing the number of experiments required. Very high regression coefficient between the variables and the responses indicated excellent evaluation of experimental data. Under optimized conditions fungal isolate was capable to decolourize Remazol Red RR up to 86.21% and COD reduction up to 55.43% was achieved during the experimental setup. Enzymatic activity indicated excellent outcome under the optimal process conditions. The experimental values agreed with the predicted ones, indicating suitability of the model and success of RSM approach in optimizing the process.

Optimization of process variables for decolorization of Disperse Yellow 211 by Bacillus subtilis using Box-Behnken design.

Decolorization of textile azo dye Disperse Yellow 211 (DY 211) was carried out from simulated aqueous solution by bacterial strain Bacillus subtilis. Response surface methodology (RSM), involving Box-Behnken design matrix in three most important operating variables; temperature, pH and initial dye concentration was successfully employed for the study and optimization of decolorization process. The total 17 experiments were conducted in the study towards the construction of a quadratic model. According to analysis of variance (ANOVA) results, the proposed model can be used to navigate the design space. Under optimized conditions the bacterial strain was able to decolorize DY 211 up to 80%. Model indicated that initial dye concentration of 100 mgl(-1), pH 7 and a temperature of 32.5 degrees C were found optimum for maximum % decolorization. Very high regression coefficient between the variables and the response (R(2)=0.9930) indicated excellent evaluation of experimental data by polynomial regression model. The combination of the three variables predicted through RSM was confirmed through confirmatory experiments, hence the bacterial strain holds a great potential for the treatment of colored textile effluents.

Response surface methodological approach for the decolorization of simulated dye effluent using Aspergillus fumigatus fresenius.

The aim of our research was to study, effect of temperature, pH and initial dye concentration on decolorization of diazo dye Acid Red 151 (AR 151) from simulated dye solution using a fungal isolate Aspergillus fumigatus fresenius have been investigated. The central composite design matrix and response surface methodology (RSM) have been applied to design the experiments to evaluate the interactive effects of three most important operating variables: temperature (25-35 degrees C), pH (4.0-7.0), and initial dye concentration (100-200 mg/L) on the biodegradation of AR 151. The total 20 experiments were conducted in the present study towards the construction of a quadratic model. Very high regression coefficient between the variables and the response (R(2)=0.9934) indicated excellent evaluation of experimental data by second-order polynomial regression model. The RSM indicated that initial dye concentration of 150 mg/L, pH 5.5 and a temperature of 30 degrees C were optimal for maximum % decolorization of AR 151 in simulated dye solution, and 84.8% decolorization of AR 151 was observed at optimum growth conditions.

Open port placement of the first laparoscopic port: a safe technique.

BACKGROUND: Blind insertion of the Veress needle and of the first trocar is a significant cause of complications in laparoscopic surgery. Despite this risk, the closed technique is still more popular than the open one. Our aim is to report the results of our experience with the routine use of the modified open technique in laparoscopic surgery and to describe the technical details of the creation of pnuemoperitoneum by the open technique that we used. METHODS: A prospective study was conducted in the department of surgery at Maulana Azad Medical College and associated Lok Nayak Hospital, New Delhi. A modified method of open laparoscopy was performed on 755 consecutive patients requiring laparoscopy or laparoscopic surgery over a 5-year period from August 1998 to February 2003 in 1 surgical unit. RESULTS: The mean time taken was 4 minutes (range, 2 to 10). No intraoperative complications occurred during trocar insertion. Forty-nine (6.49%) patients had minor umbilical sepsis, 22 (2.91%) had periumbilical hematoma, but none had umbilical hernia during 3 months of follow-up after surgery. CONCLUSION: Based on our own experience, we recommend open laparoscopy as a safe and easy approach for routine laparoscopic interventions.

Retroperitoneoscopic lumbar sympathectomy for buerger's disease: a novel technique.

BACKGROUND: The conventional lumbar sympathectomy procedure through the extraperitoneal route requires a muscle cutting-splitting incision, which leads to significant postoperative pain and prolonged convalescence. With increasing experience in retroperitoneoscopic procedures, we did a pilot study to explore the role of retroperitoneoscopy in lumbar sympathectomy. We describe herein our technique used for the surgery. METHODS: The patient was placed in a lateral position. A 15-mm incision was made just below the 12th rib, and retroperitoneal space was created using blunt finger dissection. A custom-made, large balloon was inserted and inflated with the equivalent of 750 mL to 1000 mL of saline. The second 10-mm port was placed in line with the first port above the iliac crest. The third and fourth 5-mm ports were placed anterior to the first 2 ports. Peritoneum was retracted anteriorly. The medial border of the psoas muscle was used as a landmark and a chain identified immediately medial to it. Lumbar vessels were ligated on the right side. The first to fourth lumbar sympathetic ganglia were removed with the intervening chain. The port sites were closed without a drain. RESULTS: We attempted and successfully completed this procedure in 8 patients; 6 on the left side and 2 on the right side. The average operating time was 38 minutes. The mean hospital stay was 1.5 days. All patients had symptomatic pain relief and clinical improvement. CONCLUSIONS: Retroperitoneoscopic lumbar sympathectomy is a safe and effective procedure. It has a short convalescent time and minimal morbidity; hence, it is a viable alternative for the open procedure.